Cytosolic phospholipase A2 Group IValpha but not secreted phospholipase A2 Group IIA, V, or X induces interleukin-8 and cyclooxygenase-2 gene and protein expression through peroxisome proliferator-activated receptors gamma 1 and 2 in human lung cells.
It has been reported that interleukin-8 (IL-8) and cyclooxygenase-2 (COX-2) expression is regulated by peroxisome proliferator-activated receptor (PPAR)-gamma synthetic ligands. We have shown previously that cytosolic phospholipase A2 (cPLA2) is able to activate gene expression through PPAR-gamma response elements (Pawliczak, R., Han, C., Huang, X. L., Demetris, A. J., Shelhamer, J. H., and Wu, T. (2002) J. Biol. Chem. 277, 33153-33163). In this study we investigated the influence of cPLA2 and secreted phospholipase A2 (sPLA2) Group IIA, Group V, and Group X on IL-8 and COX-2 expression in human lung epithelial cells (A549 cells). We also studied the results of cPLA2 activation by epidermal growth factor (EGF) and calcium ionophore (A23187) on IL-8 and COX-2 reporter gene activity, mRNA level, and protein synthesis. cPLA2 overexpression and activation increased both IL-8 and COX-2 reporter gene activity. Overexpression and activation of Group IIA, Group V, or Group X sPLA2s did not increase IL-8 and COX-2 reporter gene activity. Methyl arachidonyl fluorophosphate, a cPLA2 inhibitor, inhibited the effect of A23187 and of EGF on both IL-8 and COX-2 reporter gene activity, steady state levels of IL-8 and COX-2 mRNA, and IL-8 and COX-2 protein expression. Small inhibitory RNAs directed against PPAR-gamma1 and -gamma2 blunted the effect of A23187 and of EGF on IL-8 and COX-2 protein expression. Moreover small inhibitory RNAs directed against cPLA2 decreased the effect of A23187 and EGF on IL-8 and COX-2 protein expression. These results demonstrate that cPLA2 has an influence on IL-8 and COX 2 gene and protein expression at least in part through PPAR-gamma.


Generation of a CRISPR/Cas9-edited Plakoglobin (JUP) knock-out (JMUi001-A-4) iPSC line to model the cardiac phenotype of arrhythmogenic cardiomyopathy.
Arrhythmogenic cardiomyopathy (ACM) represents the cardiac phenotype of Naxos disease, an autosomal recessive disease with an additional cutaneous phenotype. ACM is mainly caused by mutated desmosomal proteins, which are part of cardiac adherens junctions and provide mechanical and electrical stability. Here, we generated a knock-out (KO) of the junctional protein Plakoglobin (JUP-KO; JMUi001-A-4) using the CRISPR/Cas9 system in healthy control induced pluripotent stem cells (iPSCs, (JMUi001-A). JUP-KO iPSCs maintained pluripotency, differentiation potential and genomic integrity and provide an in vitro system modelling ACM when differentiated into cardiomyocytes.


Development and performance evaluation of a streamlined method for nucleic acid purification, denaturation, and multiplex detection of Bluetongue virus and Epizootic hemorrhagic disease virus.
Bluetongue virus (BTV) and Epizootic hemorrhagic disease virus (EHDV) possess similar structural and molecular features, are transmitted by biting midges (genus Culicoides), and cause similar diseases in some susceptible ruminants. Generally, BTV causes subclinical disease in cattle, characterized by a prolonged viremia. EHDV-associated disease in cattle is less prominent; however, it has emerged as a major economic threat to the white-tailed deer (Odocoileus virginianus) industry in many areas of the United States. The recent emergence of multiple BTV and EHDV serotypes previously undetected in the United States demonstrates the need for robust detection of all known serotypes and differential diagnosis. For this purpose, a streamlined workflow consisting of an automated nucleic acid purification and denaturation method and a multiplex one-step reverse transcription quantitative polymerase chain reaction for the simultaneous detection of BTV serotypes 1-24 and EHDV serotypes 1-7 was developed using previously published BTV and EHDV assays. The denaturation of double-stranded (ds) BTV and EHDV RNA was incorporated into the automated nucleic acid purification process thus eliminating the commonly used separate step of dsRNA denaturation. The performance of this workflow was compared with the World Organization of Animal Health BTV reference laboratory (National Veterinary Services Laboratory, Ames, Iowa) workflow for BTV and EHDV detection, and high agreement was observed. Implementation of the workflow in routine diagnostic testing enables the detection of, and differentiation between, BTV and EHDV, and coinfections in bovine blood and cervine tissues, offering significant benefits in terms of differential disease diagnosis, herd health monitoring, and regulated testing.


Effects of aggregation and electron injection on photovoltaic performance of porphyrin-based solar cells with oligo(phenylethynyl) links inside TiO(2) and Al(2)O(3) nanotube arrays.
Porphyrins with phenylethynyl links of varied length (PE1-PE4) were sensitized on vertically oriented, anodic titanium-oxide (ATO) nanotube arrays for application as dye-sensitized solar cells (DSSC). The efficiency of power conversion decreased systematically from the dye with a short link to the dye with a long link. We measured the efficiency of conversion of incident photons to current (IPCE), the photocurrent decay of the devices, and steady-state and time-resolved fluorescence spectra of the thin-film samples to understand how the cell performance depends on the length of the link. Measurements of femtosecond fluorescence confirmed that the efficiency of electron injection depended on length because of dye aggregation that significantly increased the rate of aggregate-induced energy transfer for porphyrins with a long link. The rate of electron injection depended on the length of the link with an attenuation factor beta approximately 0.1 A(-1). Resonant energy transfer (RET) kinetics of porphyrins sensitized on anodic aluminium-oxide (AAO) nanotube arrays were performed with picosecond time-correlated single-photon counting and four molecular densities for each porphyrin. The kinetic data of PE1 and PE2 are described satisfactorily according to a Förster model, whereas those of PE3 and PE4 conform to a Dexter formula. A formation of clusters is proposed to rationalize the observed density-dependent kinetics for the RET of porphyrins on semiconductor films.


Mechanisms of inhibition of carcinogenesis by tea.
Tea (Camellia sinensis) preparations have been shown to inhibit tumorigenesis at the initiation, promotion, and progression stages in different animal models. The anti-proliferative effects of tea polyphenols may be a key mechanism, especially in the NNK-induced lung tumorigenesis model with mice. Studies with cell lines have demonstrated that tea polyphenols inhibit cell proliferation and induce apoptosis. The effective concentrations used in these studies (20-100 microM) are usually higher than those observed in blood and tissues of humans and animals, which are in the low micromolar range. Glucuronide and sulfate conjugated and methylated catechins as well as ring fission products (due to intestinal microflora) have been observed in human plasma and urine. Purified green and black tea polyphenols inhibited the H-ras induced milogen-activated protein kinases, AP-1 activities, and the growth of 30.7b Ras 12 and BES21 cells. Among the catechins, both the galloyl structure on the B ring and the gallate moiety are important for the inhibition. Both (-)-epigallocatechin-3-gallate and theaflavin-3,3'-digallate inhibited the phosphorylation of c-jun and p44/42 (ERK 1/2). More mechanistic and human studies in these areas will help us to understand the possible inhibitory action of tea against carcinogenesis in humans.


Sympathetic-parasympathetic activation during spontaneous attacks of cluster headache: evaluation by spectral analysis of heart-rate fluctuations.
Twenty-four hour ECG Holter and blood-pressure monitorings were performed in eight patients suffering from cluster headache. Spectral analysis of heart-rate fluctuation was used to assess the autonomic balance under basal conditions, after head-up tilt, and during a spontaneous attack. Normal autonomic balance was found at rest and during sympathetic activation obtained with head-up tilt in the interparoxysmal period. Before the onset of headache, an increase in the low-frequency (LF) component of the power spectrum was apparent in all patients. This sign of sympathetic activation was followed by an increase in the high-frequency (HF) component that developed about 2000 beats after the onset of headache and rapidly overcame the LF component until the end of pain. Significant differences were found when comparing the spectral parameters [total spectral values (TP), power of the LF and HF components and LF/HF ratio] obtained before, during and after headache. During the attack, blood pressure increased and heart rate decreased in all subjects. There appears to be a primary activation of both sympathetic and parasympathetic functions in cluster headache attacks. The sympathetic component seems to be involved mostly in the development of the attack, whereas the parasympathetic activation seems to occur, following the onset of the attack, independently of the pain.


[Algorithm of treatment postthrombotic disease of the lower extremities].
The results of application of pathogenetically substantiated diagnostic algorithm for determination of the treatment method in patients, suffering postthrombotic disease of the lower extremities, are adduced. Using algorithm proposed a clinical state was estimated, subfascial pressure on the shin was determined, ultrasound duplex scanning (USDS) of venous system was conducted, the stimulation electroneuromyography of the shins done, and a level of D-dimer (DD) with activity of antithrombin-III (AT--III) in general and regional blood flow with calculation of its ratio were established. In 33 (31.1%) patients a conservative therapy was conducted, in 34 (32.1%)--a postponed surgical intervention, in 39 (36.8%)--surgical correction of the venous blood flow, in 22 (20.8%)--preparation and closure of trophic ulcers in accordance to the clinic method. Determination of the DD level and the AT-III activity together with data of USDS have permitted to establish differentially the indications for performance of a vein-correcting operative interventions.


Polytraumatization in an adult national sample and its association with psychological distress and self-esteem.
The objective of this study was to examine the prevalence of self-reported experiences of potential childhood traumas and polytraumatization, and to find cut-off values for different kinds of potential traumatic events in a national representative sample of adults in Sweden. In addition, to analyse the association between polytraumatization and both psychological distress and global self-esteem. A web-based survey - containing SCL-25 and Rosenberg Self-Esteem Scale, and Linköping Difficult Life Events Scale - Adult - was sent out to a nationally reprative sample and 5062 people chose to participate in the study. Results showed that almost everyone (97%) has experienced at least one potential traumatic event and that polytraumatization (the 10% of the participants with most reported traumas) was significantly (Z = 12.57, P < 0.001, r = 0.18) associated with psychological distress and global self-esteem. Gender differences were significant (Z = 8.44, P < 0.001, r = 0.12), in that men experience more noninterpersonal traumas but women report more symptoms. The effect sizes regarding the impact of potential trauma on self-esteem were largest for women with experience of polytraumatization in the age group 18-25 (r = 0.48). There was almost linear increase in psychological distress and linear decrease in self-esteem with increasing number of traumatic events experienced. Experience of polytrauma can be considered an important factor to take into account in psychiatric settings as well.


Single-cell genomic profiling of acute myeloid leukemia for clinical use: A pilot study.
Although bulk high-throughput genomic profiling studies have led to a significant increase in the understanding of cancer biology, there is increasing awareness that bulk profiling approaches do not completely elucidate tumor heterogeneity. Single-cell genomic profiling enables the distinction of tumor heterogeneity, and may improve clinical diagnosis through the identification and characterization of putative subclonal populations. In the present study, the challenges associated with a single-cell genomics profiling workflow for clinical diagnostics were investigated. Single-cell RNA-sequencing (RNA-seq) was performed on 20 cells from an acute myeloid leukemia bone marrow sample. Putative blasts were identified based on their gene expression profiles and principal component analysis was performed to identify outlier cells. Variant calling was performed on the single-cell RNA-seq data. The present pilot study demonstrates a proof of concept for clinical single-cell genomic profiling. The recognized limitations include significant stochastic RNA loss and the relatively low throughput of the current proposed platform. Although the results of the present study are promising, further technological advances and protocol optimization are necessary for single-cell genomic profiling to be clinically viable.


Effect of acute and daily neurotensin and enkephalin treatments on extracellular dopamine in the nucleus accumbens.
The injection of neurotensin or the enkephalin analog Tyr-D-Ala-Gly-MePhe-Gly(ol) (DAMGO) into the A10 region of rats produces a motor stimulant effect that is associated with an increase in the postmortem levels of dopamine metabolites in the nucleus accumbens. These behavioral and neurochemical effects are augmented following daily administration. In vivo dialysis in the nucleus accumbens of conscious rats was used to determine if the acute and augmented behavioral responses following neurotensin or DAMGO administration are associated with an increase in extracellular dopamine concentrations. The acute injection of DAMGO produced a dose-dependent (0.03-3.3 nmol) elevation in extracellular dopamine and its metabolites in the nucleus accumbens. Neurotensin also produced a dose-related (0.1-3.3 nmol) increase in dopamine and its metabolites. The elevation in extracellular dopamine produced by DAMGO, but not by neurotensin, was positively correlated with the increase in motor activity. Following daily treatment of either neurotensin (1.0 nmol X 4 d) or DAMGO (0.03 nmol X 4 d), a significant elevation in extracellular dopamine levels occurred in the nucleus accumbens compared to an acute injection. The time course of the change in extracellular dopamine after daily injections was similar to the time course of the behavioral stimulation for both compounds. These data demonstrate that enhanced dopamine release in the nucleus accumbens mediates the acute behavioral effect of DAMGO but does not entirely explain the motor effects of neurotensin. However, enhanced dopamine release may mediate the behavioral sensitization produced by daily injection of both peptides into the A10 region.


Practical Application of DaTQUANT with Optimal Threshold for Diagnostic Accuracy of Dopamine Transporter SPECT.
Evaluation of Parkinsonian Syndromes (PS) with Ioflupane iodine-123 dopamine transporter single photon emission computed tomography (DaT-SPECT), in conjunction with history and clinical examination, aids in diagnosis. FDA-approved, semi-quantitative software, DaTQUANTTM (GE Healthcare, Chicago, IL, USA) is available to assist in interpretation. This study aims to evaluate the optimal variables and thresholds of DaTQUANT to yield the optimal diagnostic accuracy. It is a retrospective review with three different patient populations. DaT-SPECT images from all three study groups were evaluated using DaTQUANTTM software, and both single and multi-variable logistic regression were used to model PS status. The optimal models were chosen via accuracy, sensitivity, and specificity, then evaluated on the other study groups. Among single variable models, the posterior putamen yielded the highest accuracy (84% to 95%), while balancing sensitivity and specificity. Multi-variable models did not substantially improve the accuracy. When the optimal single variable models for each group were used to evaluate the remaining two groups, comparable results were achieved. In typical utilization of DaT-SPECT for differentiation between nigrostriatal degenerative disease (NSDD) and non-NSDD, the posterior putamen was the single variable that yielded the highest accuracy across three different patient populations. The posterior putamen's recommended thresholds for DaTQUANT are SBR ≤ 1.0, z-score of ≤-1.8 and percent deviation ≤ -0.34.


Clinically Approved Monoclonal Antibodies-Based Immunotherapy: Association With Glycemic Control and Impact Role of Clinical Pharmacist for Cancer Patient Care.
Compared with more conventional, nonspecific therapy options, such as radiotherapy and chemotherapy, monoclonal antibodies (mAbs) constitute a crucial approach of cancer treatment. Multiple autoimmune diseases have been observed during treatment with mAb medications, including autoimmune diabetes mellitus (DM). This study provides a narrative review of clinically approved mAbs in cancer treatment and focuses on the development of hyperglycemia and DM arising from using these therapies. Furthermore, it highlights the critical role of oncology clinical pharmacists in the management of autoimmune DM and patient care while using these medications in an oncology setting. An extensive literature search was conducted using various sources of electronic databases, such as Scopus, Embase, Web of Science, and PubMed, and search engines, such as Google Scholar, for studies on mAb classification, types, mechanisms of action, pharmacokinetic properties, current clinical applications, and the associated common adverse effects with significant recommendations for patient care in an oncology setting, along with focusing on the proposed mechanisms and clinical studies that reported the association of DM after the use of these therapies. There are 4 types (murine, chimeric, humanized, and human) and 3 classes (unconjugated, conjugated, and bispecific) of mAbs with several mechanisms of action that can destroy cancer cells, including preventing tumor cell survival cascades, inhibiting tumor growth by interfering with tumor angiogenesis, evading programmed cell death, and bypassing immune checkpoints. However, multiple endocrinopathies, autoimmune diseases, and complications were reported from the use of these medications, including the development of autoimmune DM and diabetic ketoacidosis. These autoimmune disorders were reported most with the use of immune checkpoint inhibitors, including inhibitors of the programmed cell death protein 1 (nivolumab and pembrolizumab), its ligand (atezolizumab, avelumab, and durvalumab), and cytotoxic T-lymphocyte-associated protein 4 (ipilimumab). mAbs are considered important approaches for the treatment of many cancer types. However, a high incidence of hyperglycemia, type 1 DM, and diabetic ketoacidosis is observed with the use of these medications, particularly immune checkpoint inhibitors. It is important for oncologic clinical pharmacists to be involved in addressing these autoimmune disorders from the use of these immunotherapies via the provision of patient education, medication adherence support, close monitoring, and follow-up, which will lead to better health-related outcomes and improved patient quality of life.


Mortality After Acute Coronary Syndrome in Human Immunodeficiency Virus Infection with Optimal Adherence: A Nationwide Study.
There have been few studies on the outcome of acute coronary syndrome (ACS) in human immunodeficiency virus (HIV) infection in the era when antiretroviral therapy (ART) is generalized and most of them have achieved viral suppression. Using claims data, we aimed to assess the mortality after atherosclerotic cardiovascular events in people with HIV (PWH) who maintain optimal adherence to ART. We used claims data from the National Health Insurance of the Korea to confirm newly diagnosed PWH from 2009 to 2019, and measured ART adherence. ACS and mortality were confirmed in PWH who showed optimal adherence to ART. Among 7,100 PWH with optimal adherence and during 27,387 person-year of follow-up duration, ACS was confirmed in 140 (2.0%) cases, which was 1.3 times greater than statistics of the Korean general population (511.0 vs. 383.1 per 100,000). Acquired immunodeficiency syndrome, hypertension, dyslipidemia, and diabetes mellitus were associated with the development of ACS in PWH with optimal adherence. Mortality was confirmed in 10 cases, which is 7.1% overall and 2.9% when limited to myocardial infarction. It was comparable with the mortality rate of the Korean general population after myocardial infarction (8.9%). ACS prevalence was higher in PWH even when optimal adherence was maintained. However, mortality after ACS was comparable to that in the HIV-negative population.


Adrenocortical carcinoma: nuclear deoxyribonucleic acid ploidy studied by flow cytometry.
Nuclear deoxyribonucleic acid (DNA) ploidy studies with use of paraffin-embedded specimens were performed by flow cytometry on 52 adrenocortical carcinomas. Specimens were prepared by the combined techniques of Hedley and Vindeløv. Clinical course was obtained by chart review and follow-up examination. Nine (17%) tumors had a normal (diploid) DNA pattern, 13 (25%) were DNA tetraploid, and 30 (58%) were DNA aneuploid. The DNA aneuploid group was subdivided: 18 tumors with one stemline and 12 tumors with two stemlines of abnormal DNA cells. For tumors that were resected for cure, the 5-year Kaplan-Meier disease-free survival rates of the five patients with DNA diploid tumors and of the six patients with DNA tetraploid tumors were 80% and 33%, respectively. For 21 patients of whom 12 had one-stemline and nine had two-stemline DNA aneuploid tumors, the survival was 67% and 0%, respectively. Following palliative resection, the 4-year survival rates of the four patients with DNA diploid, seven with DNA tetraploid, five (omitting one with short follow-up) with one-stemline DNA aneuploid, and three with two-stemline DNA aneuploid tumors were 0%, 0%, 0%, and 33%, respectively. Although adrenocortical carcinoma is in general markedly aggressive, the addition of nuclear DNA ploidy studies may help to identify certain groups of patients who have a relatively favorable prognosis.


Targeted agents in epithelial ovarian cancer: review on emerging therapies and future developments.
Epithelial ovarian cancer (EOC) remains a clinical challenge and there is a need to optimise the currently available treatment and to urgently develop new therapeutic strategies. Recently, there has been improved understanding of the molecular characteristics and tumour microenvironment of ovarian cancers. This has facilitated the development of various targeted agents used concurrently with chemotherapy or as maintenance. Most of the studies have explored the tumour angiogenesis pathways. In phase-III trials, bevacizumab showed a statistically significant improvement in progression-free survival, although there was no improvement in overall survival in selected high-risk cases. Although several multi-targeted tyrosine kinase inhibitors were found to be useful, the toxicity and survival benefit has to be weighed. Poly ADP ribose polymerase (PARP) inhibitors have been another marvellous molecule found to be effective in breast cancer 1, early onset (BRCA)-positive ovarian cancers. Several newer molecules targeting Her 2, Wee tyrsine kinases, PIP3/AKT/mTR-signalling pathways, folate receptors are under development and may provide additional opportunities in the future. This article focuses on the targeted agents that have successfully paved the way in the management of epithelial ovarian cancer and the newer molecules that may offer therapeutic opportunities in the future.


Amiodarone keratopathy: drug-induced lipid storage disease.
Therapeutic administration of amiodarone, an antiarrhythmic drug, to eight patients resulted in the formation of vortex-like figures within the anterior cornea. Clinical examination disclosed no visual loss or other ocular abnormality attributable to the drug. In one patient, light and transmission electron microscopy of corneal epithelium, bulbar conjunctiva, and cataractous lens revealed complex lipid deposits within lysosome-like intracytoplasmic inclusions in corneal, conjunctival, and lens epithelium, conjunctival fibrocytes, and conjunctival vascular endothelium. Amiodarone keratopathy is compared clinically and morphologically with the corneal alterations seen in Fabry's disease and in chloroquine use as an example of a drug-induced lipid storage disorder.


On the Influence of Heart Rate and Coupling Interval Prematurity on Heart Rate Turbulence.
Heart rate turbulence (HRT) has been successfully explored for cardiac risk stratification. While HRT is known to be influenced by the heart rate (HR) and the coupling interval (CI), nonconcordant results have been reported on how the CI influences HRT. The purpose of this study is to investigate HRT changes in terms of CI and HR by means of an especially designed protocol. A dataset was acquired from 11 patients with structurally normal hearts for which CI was altered by different pacing trains and HR by isoproterenol during electrophysiological study (EPS). The protocol was designed so that, first, the effect of HR changes on HRT and, second, the combined effect of HR and CI could be explored. As a complement to the EPS dataset, a database of 24-h Holters from 61 acute myocardial infarction (AMI) patients was studied for the purpose of assessing risk. Data analysis was performed by using different nonlinear ridge regression models, and the relevance of model variables was assessed using resampling methods. The EPS subjects, with and without isoproterenol, were analyzed separately. The proposed nonlinear regression models were found to account for the influence of HR and CI on HRT, both in patients undergoing EPS without isoproterenol and in low-risk AMI patients, whereas this influence was absent in high-risk AMI patients. Moreover, model coefficients related to CI were not statistically significant, p > 0.05, on EPS subjects with isoproterenol. The observed relationship between CI and HRT, being in agreement with the baroreflex hypothesis, was statistically significant ( ), when decoupling the effect of HR and normalizing the CI by the HR. The results of this study can help to provide new risk indicators that take into account physiological influence on HRT, as well as to model how this influence changes in different cardiac conditions.


Activation of the prolactin receptor but not the growth hormone receptor is important for induction of mammary tumors in transgenic mice.
Transgenic mice overexpressing the human growth hormone gene develop mammary carcinomas. Since human growth hormone gene can activate both the growth hormone receptor (GHR) and the prolactin (PRL) receptor (PRLR), it is not clear which receptor system is responsible for the malignant transformation. To clarify the receptor specificity, we created transgenic mice with two different genes: (a) transgenic mice overexpressing the bovine growth hormone (bGH) gene having high levels of bGH only activating the GHR and also high serum levels of IGF-I; and (b) transgenic mice overexpressing the rat PRL (rPRL) gene that have elevated levels of PRL (one line 150 ng/ml and one line 13 ng/ml) only binding to the PRLR and with normal IGF-I levels. When analyzed histologically, all of the PRL transgenic female mice developed mammary carcinomas at 11-15 mo of age. Only normal mammary tissue was observed among the bGH transgenic animals and the controls. Cell lines established from a tumor produced rPRL and expressed PRLR. In organ culture experiments, an auto/paracrine effect of rPRL was demonstrated. In conclusion, activation of the PRLR is sufficient for induction of mammary carcinomas in mice, while activation of the GHR is not sufficient for mammary tumor formation.


Asymmetric faces: Symbolic, spiritual, and representative.
In humans, physically attractive faces are measurably, though subtly, asymmetric. As asymmetry increases, it has a negative impact. Medically, asymmetry can be congenital or acquired. Symbolically, it has varied connotations from playfulness and complexity to despair and corruption. In Chinese opera, stylized make-up exaggerates the features, and aspects of the "mask" indicate qualities of the character. The asymmetric faces belong to characters who are corrupt, devious, or evil. In the Dan culture of western Africa, performance masks channel spirits in the community. A spirit with asymmetric facial mask exemplifies ugliness and moral failing. The Nasca culture of South America made generic figures of farmers, deities, and so on, but not of individuals. However, there is evidence of mutual influence between the Nasca and the Wari, with whom they traded. A clay figure apparently representing an individual, or at least a very specific recognized persona, is a ball player with facial asymmetry presumably due to injury. Here the message is one of fierceness and strength. The relative rarity of asymmetric facial depictions compared to symmetric ones is cross-cultural. This implies that asymmetry is special somehow, in all connotations of that term.


Cerebrospinal Fluid in a Small Cohort of Patients with Multiple Sclerosis Was Generally Free of Microbial DNA.
Multiple sclerosis (MS) is a common cause of non-traumatic neurologic disability with high incidence in many developed countries. Although the etiology of the disease remains elusive, it is thought to entail genetic and environmental causes, and microbial pathogens have also been envisioned as contributors to the phenotype. We conducted a metagenomic survey in cerebrospinal fluid (CSF) from 28 MS patients and 15 patients suffering other type of neurological conditions. We detected bacterial reads in eight out of the 15 non-MS patients and in a single MS patient, at an abundance >1% of total classified reads. Two patients were of special interest: one non-MS patient harbored ~73% bacterial reads, while an MS patient had ~83% bacterial reads. In the former case, Veillonella parvula, a bacterium occasionally found associated with meningitis was the predominant species, whilst Kocuria flava, apparently an environmental bacterium, predominated in the latter case. Thirty-four out of 43 samples contained <1% bacterial reads, which we regard as cross- or environmental contamination. A few viral reads corresponding to Epstein-Barr virus, cytomegalovirus, and parvovirus were also identified. Our results suggest that CSF of MS patients is often (but not always) free of microbial DNA.


Comparison of MicroSeal and System B/Obtura II obturation techniques.
To compare the proportion of gutta-percha, sealer and voids following the filling of simulated root canals in plastic blocks using two warm gutta-percha techniques. Ten simulated root canals in plastic blocks were prepared using System GT Rotary Files and a.02 taper size 40 Quantec rotary nickel-titanium instrument and then filled with MicroSeal and Kerr EWT sealer. A further 10 simulated canals were prepared solely with System GT Rotary Files and obturated with System B/Obtura II and Kerr EWT sealer. All blocks were sectioned at 1 mm intervals. The sectioned root fillings were photographed through a stereomicroscope and the resultant slides scanned. The images were analysed using adobe photoshop 6.0 to give the proportions of gutta-percha, sealer or void at each level. The data were analysed using unpaired Student's t-test and Mann-Whitney U-test. The MicroSeal technique produced a dense homogeneous gutta-percha fill at the apical 1 and 2 mm levels similar to the System B/Obtura II technique, but further coronally the sealer became mixed into the MicroFlow gutta-percha producing a heterogeneous mass with significantly less (P < 0.05) solid gutta-percha in the root filling compared to System B/Obtura II. The MicroSeal technique generally had more pooling of cement than the System B technique, however, this was only significantly different (P < 0.05) at the 4 mm level. The System B/Obtura II obturation gave an average of 99.27% +/- 1.09 gutta-percha fill at all levels. The MicroSeal technique produced a heterogeneous fill consisting of the MicroFlow Master Cone and gutta-percha from the MicroFlow Cartridge mixed with sealer. The System B/Obtura II technique produced a homogeneous fill at all levels.


Quantitative (31)P NMR spectroscopy and (1)H MRI measurements of bone mineral and matrix density differentiate metabolic bone diseases in rat models.
In this study, bone mineral density (BMD) of normal (CON), ovariectomized (OVX), and partially nephrectomized (NFR) rats was measured by (31)P NMR spectroscopy; bone matrix density was measured by (1)H water- and fat-suppressed projection imaging (WASPI); and the extent of bone mineralization (EBM) was obtained by the ratio of BMD/bone matrix density. The capability of these MR methods to distinguish the bone composition of the CON, OVX, and NFR groups was evaluated against chemical analysis (gravimetry). For cortical bone specimens, BMD of the CON and OVX groups was not significantly different; BMD of the NFR group was 22.1% (by (31)P NMR) and 17.5% (by gravimetry) lower than CON. For trabecular bone specimens, BMD of the OVX group was 40.5% (by (31)P NMR) and 24.6% (by gravimetry) lower than CON; BMD of the NFR group was 26.8% (by (31)P NMR) and 21.5% (by gravimetry) lower than CON. No significant change of cortical bone matrix density between CON and OVX was observed by WASPI or gravimetry; NFR cortical bone matrix density was 10.3% (by WASPI) and 13.9% (by gravimetry) lower than CON. OVX trabecular bone matrix density was 38.0% (by WASPI) and 30.8% (by gravimetry) lower than CON, while no significant change in NFR trabecular bone matrix density was observed by either method. The EBMs of OVX cortical and trabecular specimens were slightly higher than CON but not significantly different from CON. Importantly, EBMs of NFR cortical and trabecular specimens were 12.4% and 26.3% lower than CON by (31)P NMR/WASPI, respectively, and 4.0% and 11.9% lower by gravimetry. Histopathology showed evidence of osteoporosis in the OVX group and severe secondary hyperparathyroidism (renal osteodystrophy) in the NFR group. These results demonstrate that the combined (31)P NMR/WASPI method is capable of discerning the difference in EBM between animals with osteoporosis and those with impaired bone mineralization.


Evaluating the Impact of Data Representation on EHR-Based Analytic Tasks.
Different analytic techniques operate optimally with different types of data. As the use of EHR-based analytics expands to newer tasks, data will have to be transformed into different representations, so the tasks can be optimally solved. We classified representations into broad categories based on their characteristics, and proposed a new knowledge-driven representation for clinical data mining as well as trajectory mining, called Severity Encoding Variables (SEVs). Additionally, we studied which characteristics make representations most suitable for particular clinical analytics tasks including trajectory mining. Our evaluation shows that, for regression, most data representations performed similarly, with SEV achieving a slight (albeit statistically significant) advantage. For patients at high risk of diabetes, it outperformed the competing representation by (relative) 20%. For association mining, SEV achieved the highest performance. Its ability to constrain the search space of patterns through clinical knowledge was key to its success.


Zoledronic acid for the prevention of bone loss in postmenopausal women with low bone mass: a randomized controlled trial.
To evaluate the efficacy of zoledronic acid in the prevention of bone loss in postmenopausal women with low bone mass. In this 2-year, randomized, multicenter, double-blind, placebo-controlled study, postmenopausal women with low bone mass were selected randomly to receive either zoledronic acid 5 mg intravenously at randomization and at month 12 (zoledronic acid 2 x 5 mg), zoledronic acid 5 mg intravenously only at randomization and placebo at month 12 (zoledronic acid 1 x 5 mg), or placebo at randomization and at month 12 (placebo). The primary efficacy endpoint was the percentage change in lumbar spine bone mineral density (BMD) (lumbar spine BMD) at month 24 relative to baseline. Both zoledronic acid 2 x 5 mg and zoledronic acid 1 x 5 mg regimens significantly increased mean lumbar spine BMD compared with placebo at month 24 (5.18% and 4.42% compared with -1.32%, respectively, both P<.001). Similarly, significantly greater increases for both zoledronic acid regimens relative to placebo were observed for lumbar spine BMD at month 12 and for BMD at the proximal femur sites (total hip, femoral neck, trochanter) at month 12 and 24 (all P<.001). Both zoledronic acid regimens significantly reduced bone turnover markers over time relative to placebo (all P<.001), although changes with zoledronic acid 2 x 5 mg regimen were sustained greater during the second year relative to zoledronic acid 1 x 5 mg. The overall incidence of adverse events and serious adverse events were similar across all treatment groups. Both once-yearly dosing and a single dose of intravenous zoledronic acid 5 mg prevented bone loss for 2 years and were well-tolerated in postmenopausal women with low bone mass. ClinicalTrials.gov, www.clinicaltrials.gov, NCT00132808. I.


Aneurysmal bone cysts of the spine: treatment options and considerations.
Aneurysmal bone cysts (ABCs) are benign bone lesions with annual incidences ranging from 1.4 to 3.2 cases per million people. Approximately, 10-30% of ABCs are found in the spine. Such lesions are traditionally treated with curettage or other intralesional techniques. Because ABCs can be locally aggressive, intralesional resection can be incomplete and result in recurrence. This has led to increased use of novel techniques, including selective arterial embolization (SAE). This study aims to: (1) compare outcomes based on extent of surgical resection, and (2) compare the efficacy of SAE versus surgical resection. Clinical data pertaining to 71 cases of spinal ABCs were ambispectively collected from nine institutions in Europe, North America, and Australia. Twenty-two spinal ABCs were treated with surgery, 32 received preoperative embolization and surgery, and 17 were treated with SAE. Most tumors were classified as Enneking stage 2 (n = 29, 41%) and stage 3 (n = 29, 41%). Local recurrence and survival were investigated and a significant difference was not observed between treatment groups. However, all three local recurrences occurred following surgical resection. Surgical resection was further categorized based on Enneking appropriateness. Recurrences only occurred following intralesional Enneking inappropriate (EI) resections (P = 0.10), a classification that characterized 47% of all surgical resections. Furthermore, 56% of intralesional resections were EI, compared to only 10% of en bloc resections (P = 0.01). Although SAE treatment did not result in any local recurrences, 35% involved more than five embolization procedures. Spinal ABCs can be effectively treated with intralesional resection, en bloc resection, or SAE. Preoperative embolization should be considered before intralesional resection to limit intraoperative bleeding. Treatment plans must be guided by lesion characteristics and clinical presentation.


Genomic analysis of pterostilbene predicts its antiproliferative effects against pancreatic cancer in vitro and in vivo.
To investigate the inhibitory role of pterostilbene in pancreatic cancer, we conducted a genomic analysis of pterostilbene-treated pancreatic cancer cells. We also investigated the effect of pterostilbene upon the carcinogenic markers, manganese superoxide dismutase, cytochrome C, Smac/DIABLO, and STAT3 phosphorylation in vitro. The antiproliferative effects of pterostilbene were further evaluated in an in vivo model. Pancreatic cancer cells were treated with pterostilbene and evaluated with DNA microarray analysis. Pterostilbene-treated cells were analyzed for cytochrome C, Smac/DIABLO, manganese superoxide dismutase (MnSOD)/antioxidant activity, and STAT3 phosphorylation using ELISA. Data were statistically analyzed using ANOVA. Pterostilbene was then administered to nude mice for 8 weeks, and tumor growth rates were recorded and statistically analyzed. Microarray analysis of pterostilbene-treated cells revealed upregulation of pro-apoptosis genes. In vitro, pterostilbene treatment altered levels of phosphorylated STAT3, MnSOD/antioxidant activity, cytochrome C, and Smac/DIABLO. In nude mice, oral pterostilbene inhibited tumor growth rates. Pterostilbene alters gene expression in pancreatic cancer and increases the antiproliferative markers cytochrome C, Smac/DIABLO, and MnSOD/antioxidant activity. It was also shown to inhibit phosphorylated STAT3, a marker of accelerated tumorigenesis, and decrease pancreatic tumor growth in vivo. Further studies are warranted to elucidate the effects of pterostilbene in humans.


Clinical Integration of Genome Diagnostics for Congenital Anomalies of the Kidney and Urinary Tract.
Revolutions in genetics, epigenetics, and bioinformatics are currently changing the outline of diagnostics and clinical medicine. From a nephrologist's perspective, individuals with congenital anomalies of the kidney and urinary tract (CAKUT) are an important patient category: not only is CAKUT the predominant cause of kidney failure in children and young adults, but the strong phenotypic and genotypic heterogeneity of kidney and urinary tract malformations has hampered standardization of clinical decision making until now. However, patients with CAKUT may benefit from precision medicine, including an integrated diagnostics trajectory, genetic counseling, and personalized management to improve clinical outcomes of developmental kidney and urinary tract defects. In this review, we discuss the present understanding of the molecular etiology of CAKUT and the currently available genome diagnostic modalities in the clinical care of patients with CAKUT. Finally, we discuss how clinical integration of findings from large-scale genetic, epigenetic, and gene-environment interaction studies may improve the prognosis of all individuals with CAKUT.


Epithelial markers in pancreatic carcinoma: immunoperoxidase localisation of DD9, CEA, EMA and CAM 5.2.
Paraffin wax embedded, formalin fixed sections of 22 adenocarcinomas of the exocrine pancreas were stained with four mouse monoclonal antibodies: DD9-E7, an antibody raised against a human pancreatic tumour xenograft; carcino-embryonic antigen (CEA); epithelial membrane antigen (EMA); and cytokeratin (CAM 5.2). An indirect immunoperoxidase technique without enzyme pre-digestion and an affinity-purified sheep anti-mouse peroxidase conjugate were used. All of the tumours were positive for DD9-E7, EMA, and CAM 5.2. Twenty out of 22 were focally positive for CEA and the staining was often weak. As all of these adenocarcinomas were DD9-E7 positive, absence of staining for DD9-E7 in a tumour makes the diagnosis of adenocarcinoma of the exocrine pancreas very unlikely, and this is of value in distinction from endocrine carcinomas with a marked acinar pattern. The weak CEA staining distinguished pancreatic carcinomas from colorectal tumours. Because the distribution of staining for EMA and CAM 5.2 was no different from that previously seen in adenocarcinomas from other sites, these markers are likely to be of limited value in the differential diagnosis of abdominal adenocarcinomas of uncertain origin.


LINC00941: a novel player involved in the progression of human cancers.
LINC00941, also known as lncRNA-MUF, is an intergenic non-coding RNA located on chromosome 12p11.21. It actively participates in a complex competing endogenous RNA network, regulating the expression of microRNA and its downstream proteins. Through transcriptional and post-transcriptional regulation, LINC00941 plays a vital role in multiple signaling pathways, influencing cell behaviors such as tumor cell proliferation, epithelial-mesenchymal transition, migration, and invasion. Noteworthy is its consistently high expression in various tumor types, closely correlating with clinicopathological features and cancer prognoses. Elevated LINC00941 levels are associated with adverse clinical outcomes, including increased tumor size, extensive lymphatic metastasis, and distant metastasis, leading to poorer survival rates across different cancers. Additionally, LINC00941 and its associated genes are linked to various targeted drugs available in the market. In this comprehensive review, we systematically summarize existing studies, detailing LINC00941's differential expression, clinicopathological and prognostic implications, regulatory mechanisms, and associated therapeutic drugs. Our analysis includes relevant charts and incorporates bioinformatics analyses to verify LINC00941's differential expression in pan-cancer and explore potential transcriptional regulation patterns of downstream targets. This work not only establishes a robust data foundation but also guides future research directions. Given its potential as a significant cancer biomarker and therapeutic target, further investigation into LINC00941's differential expression and regulatory mechanisms is essential.


Mammary-derived growth inhibitor (MDGI) interacts with integrin α-subunits and suppresses integrin activity and invasion.
The majority of mortality associated with cancer is due to formation of metastases from the primary tumor. Adhesion mediated by different integrin heterodimers has an important role during cell migration and invasion. Protein interactions with the β1-integrin cytoplasmic tail are known to influence integrin affinity for extracellular ligands, but regulating binding partners for the α-subunit cytoplasmic tails have remained elusive. In this study, we show that mammary-derived growth inhibitor (MDGI) (also known as FABP-3 or H-FABP) binds directly to the cytoplasmic tail of integrin α-subunits and its expression inhibits integrin activity. In breast cancer cell lines, MDGI expression correlates with suppression of the active conformation of integrins. This results in reduced integrin adhesion to type I collagen and fibronectin and inhibition of cell migration and invasion. In tissue microarray of 1331 breast cancer patients, patients with MDGI-positive tumors had more favorable 10-year distant disease-free survival compared with patients with MDGI-negative tumors. Our data indicate that MDGI is a novel interacting partner for integrin α-subunits, and its expression modulates integrin activity and suppresses cell invasion in breast cancer patients. Retained MDGI expression is associated with favorable prognosis.


High-mobility group box-1 in ischemia-reperfusion injury of the heart.
High-mobility group box-1 (HMGB1) is a nuclear factor released by necrotic cells and by activated immune cells. HMGB1 signals via members of the toll-like receptor family and the receptor for advanced glycation end products (RAGE). Although HMGB1 has been implicated in ischemia/reperfusion (I/R) injury of the liver and lung, its role in I/R injury of the heart remains unclear. Here, we demonstrate that HMGB1 acts as an early mediator of inflammation and organ damage in I/R injury of the heart. HMGB1 levels were already elevated 30 minutes after hypoxia in vitro and in ischemic injury of the heart in vivo. Treatment of mice with recombinant HMGB1 worsened I/R injury, whereas treatment with HMGB1 box A significantly reduced infarct size and markers of tissue damage. In addition, HMGB1 inhibition with recombinant HMGB1 box A suggested an involvement of the mitogen-activated protein kinases jun N-terminal kinase and extracellular signal-regulated kinase 1/2, as well as the nuclear transcription factor nuclear factor-kappaB in I/R injury. Interestingly, infarct size and markers of tissue damage were not affected by administration of recombinant HMGB1 or HMGB1 antagonists in RAGE(-/-) mice, which demonstrated significantly reduced damage in reperfused hearts compared with wild-type mice. Coincubation studies using recombinant HMGB1 in vitro induced an inflammatory response in isolated macrophages from wild-type mice but not in macrophages from RAGE(-/-) mice. HMGB1 plays a major role in the early event of I/R injury by binding to RAGE, resulting in the activation of proinflammatory pathways and enhanced myocardial injury. Therefore, blockage of HMGB1 might represent a novel therapeutic strategy in I/R injury.


Newborn mice vaccination with BCG.HIVA²²² + MVA.HIVA enhances HIV-1-specific immune responses: influence of age and immunization routes.
We have evaluated the influence of age and immunization routes for induction of HIV-1- and M. tuberculosis-specific immune responses after neonatal (7 days old) and adult (7 weeks old) BALB/c mice immunization with BCG.HIVA(222) prime and MVA.HIVA boost. The specific HIV-1 cellular immune responses were analyzed in spleen cells. The body weight of the newborn mice was weekly recorded. The frequencies of HIV-specific CD8(+) T cells producing IFN-γ were higher in adult mice vaccinated intradermally and lower in adult and newborn mice vaccinated subcutaneously. In all cases the IFN-γ production was significantly higher when mice were primed with BCG.HIVA(222) compared with BCGwt. When the HIV-specific CTL activity was assessed, the frequencies of specific killing were higher in newborn mice than in adults. The prime-boost vaccination regimen which includes BCG.HIVA(222) and MVA.HIVA was safe when inoculated to newborn mice. The administration of BCG.HIVA(222) to newborn mice is safe and immunogenic and increased the HIV-specific responses induced by MVA.HIVA vaccine. It might be a good model for infant HIV and Tuberculosis bivalent vaccine.


Diagnosis and treatment of sinusitis.
Maxillary and ethmoid sinusitis may occur at any age. Frontal sinus infections first appear in adolescence. The bacteria that cause sinusitis are similar to those associated with otitis media. Streptococcus pneumoniae and Hemophilus influenzae are recovered from 50 to 70 percent of isolates. Anaerobes are also important pathogens in adults. Fungal sinusitis usually occurs only in diabetics or immunosuppressed individuals.


UV-B Induced Flavonoids Contribute to Reduced Biotrophic Disease Susceptibility in Lettuce Seedlings.
Biotrophic disease is one of the largest causes of decreased yield in agriculture. While exposure to ultraviolet B (UV-B) light (280-320 nm) has been previously observed to reduce plant susceptibility to disease, there is still a paucity of information regarding underlying biological mechanisms. In addition, recent advances in UV-LED technology raise the prospect of UV light treatments in agriculture which are practical and efficient. Here, we characterized the capability of UV-B LED pre-treatments to reduce susceptibility of a range of lettuce (Lactuca sativa) cultivars to downy mildew disease caused by the obligate biotroph Bremia lactucae. Innate cultivar susceptibility level did not seem to influence the benefit of a UV-B induced disease reduction with similar reductions as a percentage of the control observed (54-62% decrease in conidia count) across all susceptible cultivars. UV-B-induced reductions to conidia counts were sufficient to significantly reduce the infectivity of the diseased plant. Secondary infections caused by UV-B pre-treated plants exhibited yet further (67%) reduced disease severity. UV-B-induced flavonoids may in part mediate this reduced disease severity phenotype, as B. lactucae conidia counts of lettuce plants negatively correlated with flavonoid levels in a UV-B-dependent manner (r = -0.81). Liquid chromatography-mass spectrometry (LC-MS) was used to identify metabolic features which contribute to this correlation and, of these, quercetin 3-O-(6"-O-malonyl)-b-D-glucoside had the strongest negative correlation with B. lactucae conidia count (r = -0.68). When quercetin 3-O-(6"-O-malonyl)-b-D-glucoside was directly infiltrated into lettuce leaves, with those leaves subsequently infected, the B. lactucae conidia count was reduced (25-39%) in two susceptible lettuce cultivars. We conclude that UV-B induced phenolics, in particular quercetin flavonoids, may act as phytoanticipins to limit the establishment of biotrophic pathogens thus delaying or reducing their sporulation as measured by conidia count. These findings highlight the opportunity for UV-B morphogenesis to be exploited through the application of UV-LED technology, as part of the development of next-generation, sustainable disease control tools.


Localization of metal ions in biomolecules by means of pulsed dipolar EPR spectroscopy.
Metal ions are important for the folding, structure, and function of biomolecules. Thus, knowing where their binding sites are located in proteins or oligonucleotides is a critical objective. X-ray crystallography and nuclear magnetic resonance are powerful methods in this respect, but both have their limitations. Here, a complementary method is highlighted in which paramagnetic metal ions are localized by means of trilateration using a combination of site-directed spin labeling and pulsed dipolar electron paramagnetic resonance spectroscopy. The working principle, the requirements, and the limitations of the method are critically discussed. Several applications of the method are outlined and compared with each other.


Beta1-adrenoceptor-mediated relaxation with isoprenaline and the role of MaxiK channels in guinea-pig esophageal smooth muscle.
The possible functional coupling between beta1-adrenoceptor and MaxiK channels which results in smooth muscle relaxation was examined in the guinea-pig esophageal muscularis mucosae. Isoprenaline-elicited relaxation of esophageal smooth muscle was confirmed to be mediated through beta1-adrenoceptors as the response was competitively antagonized by a beta1-selective antagonist atenolol with a pA2 value of 7.01. Iberiotoxin (IbTx, 10(-7) M), a selective MaxiK channel inhibitor, substantially diminished the relaxant response to isoprenaline. The extent of the MaxiK channel contribution to the relaxant response was 15-40% of the control response when estimated as the E50%-Emax responses to isoprenaline. The relaxation to isoprenaline was also attenuated by high-KCl (80 mM) to the same degree as the relaxant response generated in the presence of IbTx, and thus the estimated extent of the K+ channel contribution was 10-40%. These findings indicate that beta1-adrenoceptors are substantially coupled with MaxiK channels to produce relaxation of esophageal smooth muscle in the guinea-pig. Although MaxiK channels account for the contribution of K+ channels to the beta1-adrenoceptor-mediated relaxation in this smooth muscle preparation, their contribution seems to be less when compared to the beta2-adrenoceptor-mediated relaxation of tracheal smooth muscle.


Comparison and Evaluation of Multiple Users' Usage of the Exposure and Risk Tool: Stoffenmanager 5.1.
Stoffenmanager is an exposure and risk assessment tool that has a control banding part, with risk bands as outcome and a quantitative exposure assessment part, with the 90th percentile of the predicted exposure as a default outcome. The main aim of the study was to investigate whether multiple users of Stoffenmanager came to the same result when modelling the same scenarios. Other aims were to investigate the differences between outcomes of the control banding part with the measured risk quota and to evaluate the conservatism of the tool by testing whether the 90th percentiles are above the measured median exposures. We investigated airborne exposures at companies in four different types of industry: wood, printing, metal foundry, and spray painting. Three scenarios were modelled and measured, when possible, at each company. When modelled, 13 users visited each company on the same occasion creating individual assessments. Consensus assessments were also modelled for each scenario by six occupational hygienists. The multiple users' outcomes were often spread over two risk bands in the control banding part, and the differences in the quantitative exposure outcomes for the highest and lowest assessments per scenario varied between a factor 2 and 100. Four parameters were difficult for the users to assess and had a large impact on the outcome: type of task, breathing zone, personal protection, and control measures. Only two scenarios had a higher measured risk quota than predicted by the control banding part, also two scenarios had slightly higher measured median exposure value than modelled consensus in the quantitative exposure assessment part. Hence, the variability between users was large but the model performed well.


The VA vascular injury study: A glimpse at quality of care in Veterans with traumatic vascular injury repair.
The high number of limb injuries among Post-9/11 Veterans and their long-term care pose significant challenges to clinicians. Current follow-up for extremity arterial vascular injury (EVI) is based on guideline-concordant care for treatment of peripheral vascular disease (GCC-PVD), including anticoagulant/antiplatelet or statin therapy and duplex ultrasound. No best practices exist for arterial EVI. Our goal was to determine correlates of GCC-PVD and other care among Post-9/11 Veterans with combat-related arterial EVI. We identified Post-9/11 Veterans with arterial EVI who underwent initial limb salvage repair or ligation (e.g., for single-vessel injury) attempt per DoD Trauma Registry validated by chart abstraction. Veterans Health Administration (VHA) data characterized the cohort in the first five years of VHA care. Models predicted (a) GCC-PVD, (b) pain clinic use, (c) mental/behavioral health care, (d) long-term opioid use, and (e) time to complication, controlling for injury severity and type, mental health parameters, and demographics. The 490-Veteran cohort with validated arterial injury was 77% White averaging 25.2 years at injury (range: 18-56). Mechanism of injury was primarily explosive (63%). Veterans had Injury Severity Scores classified as mild (60%), moderate (25%) and severe (15%). Approximately 25% received at least one component of VHA GCC-PVD including 8% arterial ultrasounds, 5% statins, and 11% anticoagulants/antiplatelets; 77% had mental/behavioral healthcare. GCC-PVD, as well as PTSD and substance use disorders, were associated with receipt of mental/behavioral health care. Complications affected 46% of the cohort and were more common among those prescribed 90+ days of opioids or receiving GCC-PVD. Despite injury severity (40% moderate/severe), only 25% of cohort patients received VHA GCC-PVD, and nearly half had complications from their arterial injury. Receiving GCC-PVD appeared to potentiate receiving care for mental and behavioral disorders. The treatment gap in Veterans with arterial EVI may be due to lack of appropriate guidelines, lack of vascular specialists in VHA or accessing care outside the VHA. Focused study of care options and their outcomes will help define optimal care processes for combat Veterans with arterial EVI.


Improved availability and quality of care with epilepsy nurse practitioners.
This study investigated the quality of care delivered by nurse practitioner (NP)-physician teams employed to expand clinic appointment availability for patients with epilepsy. We performed a retrospective observational cohort study of patients with epilepsy presenting to the Penn Epilepsy Center for a new patient appointment in 2014. During this time, patients were seen either by an NP-physician team care model or a more traditional physician-only care model. These care models were compared with regard to adherence to the 2014 American Academy of Neurology epilepsy quality measures at the initial visit. Clinical outcomes of seizure frequency, presentations to the Emergency Department, injury, and death were assessed over the subsequent year. A total of 169 patients were identified by our inclusion and exclusion criteria: 65 patients in the NP-physician team care model cohort and 104 patients in the physician-only care model cohort. The NP-physician team care model saw, on average, 3 more patients per clinic session. There were no meaningful differences between these cohorts in baseline characteristics. The NP-physician team care model showed equivalent adherence to the physician-only care model for the epilepsy quality measures, with superior adherence to the counseling measures of querying for side effects, provision of personalized epilepsy safety education, and screening for behavioral health disorders. The 2 care models performed similarly in all clinical outcomes. An NP-physician team care model employed to increase availability of care could also improve quality of care delivered.


Morphological and biochemical changes in supersensitive smooth muscle.
The effect of postganglionic denervation on the incidence of nexal contacts in the smooth muscle of the rat vas deferens was investigated. The chronically denervated tissue exhibited twice as many nexuses as control. This increase in the incidence of cell contacts may contribute to the supersensitivity and/or the increase in maximum response of the denervated vas deferens. The effects of denervation, decentralization, and pretreatment with reserpine on the concentration of adenosine triphosphate (ATP) in vasa deferentia of rats and guinea pigs were also investigated. One day after denervation there was a substantial decrease in the endogenous norepinephrine and ATP concentrations. The norepinephrine concentration remained low (less than 10% of control) throughout subsequent days (up to 14 days) whereas the ATP concentration, after the first postoperative day, rose significantly. The rise in ATP concentration was temporally correlated with the development of postjunctional supersensitivity. Decentralization and pretreatment with reserpine both resulted in a significant increase in ATP concentration which preceded by 2 to 3 days a significant increase in sensitivity of the vas deferens. It appears that a change in the tissue concentration of ATP may be one of the initial events that occurs following interruption of the neural contact to the smooth muscle of the vas deferens.


Seasonally and regionally determined indication potential of bioassays in contaminated river sediments.
River sediments are a dynamic system, especially in areas where floods occur frequently. In the present study, an integrative approach is used to investigate the seasonal and spatial dynamics of contamination of sediments from a regularly flooded industrial area in the Czech Republic, which presents a suitable model ecosystem for pollutant distribution research at a regional level. Surface sediments were sampled repeatedly to represent two different hydrological situations: spring (after the peak of high flow) and autumn (after longer period of low flow). Samples were characterized for abiotic parameters and concentrations of priority organic pollutants. Toxicity was assessed by Microtox test; genotoxicity by SOS-chromotest and green fluorescent protein (GFP)-yeast test; and the presence of compounds with specific mode of action by in vitro bioassays for dioxin-like activity, anti-/androgenicity, and anti-/estrogenicity. Distribution of organic contaminants varied among regions and seasonally. Although the results of Microtox and genotoxicity tests were relatively inconclusive, all other specific bioassays led to statistically significant regional and seasonal differences in profiles and allowed clear separation of upstream and downstream regions. The outcomes of these bioassays indicated an association with concentrations of polycyclic aromatic hydrocarbons (PAHs) and polychlorinated biphenyls (PCBs) as master variables. There were significant interrelations among dioxin-like activity, antiandrogenicity and content of organic carbon, clay, and concentration of PAHs and PCBs, which documents the significance of abiotic factors in accumulation of pollutants. The study demonstrates the strength of the specific bioassays in indicating the changes in contamination and emphasizes the crucial role of a well-designed sampling plan, in which both spatial and temporal dynamics should be taken into account, for the correct interpretations of information in risk assessments.


Comparing Commercial Metal-Coated AFM Tips and Home-Made Bulk Gold Tips for Tip-Enhanced Raman Spectroscopy of Polymer Functionalized Multiwalled Carbon Nanotubes.
Tip-enhanced Raman spectroscopy (TERS) combines the high specificity and sensitivity of plasmon-enhanced Raman spectroscopy with the high spatial resolution of scanning probe microscopy. TERS has gained a lot of attention from many nanoscience fields, since this technique can provide chemical and structural information of surfaces and interfaces with nanometric spatial resolution. Multiwalled carbon nanotubes (MWCNTs) are very versatile nanostructures that can be dispersed in organic solvents or polymeric matrices, giving rise to new nanocomposite materials, showing improved mechanical, electrical and thermal properties. Moreover, MWCNTs can be easily functionalized with polymers in order to be employed as specific chemical sensors. In this context, TERS is strategic, since it can provide useful information on the cooperation of the two components at the nanoscale for the optimization of the macroscopic properties of the hybrid material. Nevertheless, efficient TERS characterization relies on the geometrical features and material composition of the plasmonic tip used. In this work, after comparing the TERS performance of commercial Ag coated nanotips and home-made bulk Au tips on bare MWCNTs, we show how TERS can be exploited for characterizing MWCNTs mixed with conjugated fluorene copolymers, thus contributing to the understanding of the polymer/CNT interaction process at the local scale.


Soft matter from liquid crystals.
Liquid crystals (LCs) are fluids within which molecules exhibit long-range orientational order, leading to anisotropic properties such as optical birefringence and curvature elasticity. Because the ordering of molecules within LCs can be altered by weak external stimuli, LCs have been widely used to create soft matter systems that respond optically to electric fields (LC display), temperature (LC thermometer) or molecular adsorbates (LC chemical sensor). More recent studies, however, have moved beyond investigations of optical responses of LCs to explore the design of complex LC-based soft matter systems that offer the potential to realize more sophisticated functions (e.g., autonomous, self-regulating chemical responses to mechanical stimuli) by directing the interactions of small molecules, synthetic colloids and living cells dispersed within the bulk of LCs or at their interfaces. These studies are also increasingly focusing on LC systems driven beyond equilibrium states. This review presents one perspective on these advances, with an emphasis on the discovery of fundamental phenomena that may enable new technologies. Three areas of progress are highlighted; (i) directed assembly of amphiphilic molecules either within topological defects of LCs or at aqueous interfaces of LCs, (ii) templated polymerization in LCs via chemical vapor deposition, an approach that overcomes fundamental challenges related to control of LC phase behavior during polymerization, and (iii) studies of colloids in LCs, including chiral colloids, soft colloids that are strained by LCs, and active colloids that are driven into organized states by dissipation of energy (e.g. bacteria). These examples, and key unresolved issues discussed at the end of this perspective, serve to convey the message that soft matter systems that integrate ideas from LC, surfactant, polymer and colloid sciences define fertile territory for fundamental studies and creation of future transformative technologies.


When and How Should Clinicians View Discharge Planning as Part of a Patient's Care Continuum?
Inpatient admission of unhoused patients from an emergency department is becoming more frequent. Clinicians have ethical obligations to engage early in thorough discharge planning for these vulnerable patients, as discharge to the street or even to a shelter can produce poor health outcomes. This commentary on a case considers factors that influence safe discharge planning and execution, including linkage to follow-up, patient engagement, and multidisciplinary teamwork.


American College of Surgeons Oncology Group Z4099/Radiation Therapy Oncology Group 1021: a randomized study of sublobar resection compared with stereotactic body radiotherapy for high-risk stage I non-small cell lung cancer.
During the past decade, tremendous interest has arisen in the use of nonoperative therapies for patients with non-small cell lung cancer. Of these therapies, stereotactic body radiotherapy has become established as an effective modality for treating peripheral cancer in medically inoperable patients. Toxicity is low, and the treatment is effective, with excellent local control rates. Several investigators have suggested that stereotactic body radiotherapy could be effective for high-risk operable patients (usually treated with sublobar resection) and even perhaps for standard-risk operable patients (usually treated with lobectomy); however, this is less accepted. A direct comparison of stereotactic body radiotherapy and sublobar resection is difficult for a number of reasons. These include different definitions of recurrence, different populations of patients in these studies (with those undergoing stereotactic body radiotherapy tending to be the medically inoperable group), and different methods of classifying morbidity in the surgical and radiation oncology studies. Imaging follow-up has also not been standardized among the studies. Thus, a randomized study is necessary and timely. Investigators from the American College of Surgeons Oncology Group and the Radiation Therapy and Oncology Group have collaborated to develop a phase III randomized study comparing stereotactic body radiotherapy and sublobar resection (with or without brachytherapy) for high-risk operable patients with non-small cell lung cancer. This study (American College of Surgeons Oncology Group Z4099/Radiation Therapy Oncology Group 1021) has recently opened for accrual. It is hoped that this will help to better define the role of these therapies for patients with non-small cell lung cancer.


Selection bias in national surveys due to screening: implications from a county general population survey.
National alcohol surveys using face-to-face interviews, such as the US 1988 National Health Interview Survey Alcohol Supplement and the 1990 US National Alcohol Survey, for reasons of efficiency often use screener items to identify individuals who are likely to have experienced alcohol-related problems and only those individuals are chosen to respond to a list of alcohol-related problem questions. The consequence of screening is that only a subset of the current drinkers have complete data on such items. This paper examines the bias introduced by the exclusion of cases with incomplete information due to screening. Data from a regional general population survey were used to investigate possible bias due to screening because it included the screening questions used in NHIS and NAS but did not screen on those items. Risk curves and estimated probabilities from logistic regressions of three alcohol dependence symptom items and two problem indices were compared by gender across three subsamples: (1) all current drinkers (without screening); (2) those who passed the screener item for the NHIS and (3); the NAS, respectively. Results indicate that the effect of the screener items on the estimated prevalence of the measures concerned are minimal, supporting the practice, judiciously applied, when greater survey efficiency is required.


Essential Oil Microemulsions Inactivate Antibiotic-Resistant Bacteria on Iceberg Lettuce during 28-Day Storage at 4 °C.
The objective of this study was to investigate the antimicrobial activities of essential oil-based microemulsions in the wash water against Escherichia coli O157:H7 and Pseudomonas fluorescens on Iceberg lettuce. Evaluated wash microemulsions included oregano oil, lemongrass oil, and cinnamon oil, along with a plant-based emulsifier for improved solubility. Iceberg lettuce was inoculated for 2 min with E. coli O157:H7 (6.0 log CFU/g) or P. fluorescens (6.0 log CFU/g) and then dip-treated in a phosphate buffered saline (PBS) control, 50 ppm chlorine, 3% hydrogen peroxide treatment or a 0.1%, 0.3%, or 0.5% microemulsion solution. Treated leaves were stored at 4 °C, and analyzed for surviving bacteria on days 0, 3, 7, 10, 14, 21, and 28. Efficacies of the antimicrobials were concentration and storage-time dependent. There was a 1.26−4.86 log CFU/g reduction in E. coli O157:H7 and significant reductions (0.32−2.35 log CFU/g) in P. fluorescens during storage at days 0−28 (p < 0.05). The 0.1% oregano oil microemulsion resulted in the best visual appeal in Iceberg leaves inoculated with E. coli O157:H7 and showed better improvement in the quality of the Iceberg leaves inoculated with spoilage bacteria P. fluorescens. The results suggest that 0.5% cinnamon and 0.3% oregano oil treatments have the potential to provide natural, eco-friendly, and effective alternatives to chemicals for the decontamination of leafy greens, eliminating E. coli O157:H7 and P. fluorescens.


Spin rings in bistable planar semiconductor microcavities.
A remarkable feature of exciton-polaritons is the strongly spin-dependent polariton-polariton interaction, which has been predicted to result in the formation of spin rings in real space [Shelykh, Phys. Rev. Lett. 100, 116401 (2008)]. Here we experimentally demonstrate the spin bistability of exciton polaritons in an InGaAs-based semiconductor microcavity under resonant optical pumping. We observe the formation of spin rings whose size can be finely controlled in a spatial scale down to the micrometer range, much smaller than the spot size. Demonstration of optically controlled spin patterns in semiconductors opens way to the realization of spin logic devices and spin memories.


Teaching medical students social responsibility: the right thing to do.
As academic medicine has become more focused on the economic pressures of the marketplace, some educators have expressed concern about whether appropriate attention is being given to the character development and moral education of medical students. The authors conclude that medical schools do indeed have a duty to teach their medical students to be socially responsible. They define a socially responsible individual as a person who takes part in activities that contribute to the happiness, health, and prosperity of a community and its members. They suggest that medical students should participate in carefully designed, socially responsible activities in order to (1) practice and have reinforced such qualities as reliability, trustworthiness, dependability, altruism, and compassion; (2) partially reimburse society for the cost of their medical education; (3) increase their exposure to a population-based approach to health care; and (4) help medical schools fulfill their social contract with the public. The authors outline the process for developing a curriculum to teach social responsibility to medical students and list some of the key questions faculty and administrators must address in the processes of development and implementation. They conclude that while faculty responsible for implementing a curriculum in social responsibility must be highly committed and prepared to address numerous difficult questions concerning the curriculum's philosophy, structure, and function, the potential benefits of such a curriculum are well worth the effort.


T4 depresses olfactory responses to L-alanine and plasma T3 and T3 production in smoltifying Atlantic salmon.
During natural or induced smoltification, Atlantic salmon exhibit spring elevations in olfactory activity and plasma L-thyroxine (T4). To determine whether T4 influences olfactory activity, we administered T4 to late parr in early April and measured olfactory bulb electroencephalogram and olfactory epithelium electroolfactorogram responses to L-alanine nasal stimulation, plasma T4 and 3,5,3'-triiodo-L-thyronine (T3) levels, and hepatic and brain monodeiodination. T4 treatment raised plasma T4 to 15 ng/ml, simulating the smolt T4 surge, and depressed plasma T3 and electroencephalogram responses, without modifying electroolfactorogram responses. Decreased plasma T3 may be explained by inhibition of hepatic T4 outer-ring deiodination, generating T3, and stimulation of inner-ring deiodination, degrading T4 and T3. Brain T4 outer-ring deiodination was also strongly inhibited. In conclusion, creation of a high plasma T4 concentration simulating the natural smolt peak depressed olfactory bulb responses to L-alanine due to the high T4 concentration or the depressed T3 availability in brain induced by T4. T4 may terminate the period of heightened olfactory responsiveness during smoltification.


Induction of apoptosis by Concanavalin A and its molecular mechanisms in cancer cells.
Concanavalin A (Con A), a mannose/glucose-binding legume lectin, can induce cancer cell death through a mitochondria-mediated autophagic pathway; however, the precise mechanisms by which it mediates cell death are still only rudimentarily understood. In the present study, Con A possesses a remarkable antiproliferative effect on human melanoma A375 cells. Also, there is a link between the antiproliferative activity of Con A and its sugar-binding activity. Subsequently, Con A can induce human melanoma A375 cell apoptosis in a caspase-dependent manner. In addition, the treatment with Con A can cause mitochondrial transmembrane potential collapse, leading to cytochrome c release and caspase-9-caspase-3 activation. In conclusion, we demonstrate that there may be a close correlation between the antiproliferative activity of Con A and its sugar-binding activity. More importantly, we report for the first time that Con A can induce human melanoma A375 cell death in a caspase-dependent manner as well as via a mitochondrial apoptotic pathway.


The Challenge of Providing Renal Replacement Therapy in Developing Countries: The Latin American Perspective.
The costs of health care place developing countries under enormous economic pressure. Latin America is a region characterized by wide ethnic and per capita gross domestic product variations among different countries. Chronic kidney failure prevalence and incidence, as well as provision of renal replacement therapy (RRT), have increased in all Latin American countries over the last 20 years. From an ethical point of view, life-sustaining therapies such as RRT should be available to all patients with chronic kidney disease who might benefit. However, even among Latin American countries with similar per capita incomes and health care expenditures, only some have been able to achieve universal access to RRT. This indicates that it is not just a problem of wealth or distribution of scarce health care resources, but one of social justice. Strategies to increase the availability of RRT and renal palliative-supportive care, as well as implementation of interventions to prevent chronic kidney disease development and progression, are needed in Latin America and other developing countries.


Calibration of the channel that determines the omega-hydroxylation regiospecificity of cytochrome P4504A1: catalytic oxidation of 12-HALODOdecanoic acids.
The fatty acid omega-hydroxylation regiospecificity of CYP4 enzymes may result from presentation of the terminal carbon to the oxidizing species via a narrow channel that restricts access to the other carbon atoms. To test this hypothesis, the oxidation of 12-iodo-, 12-bromo-, and 12-chlorododecanoic acids by recombinant CYP4A1 has been examined. Although all three 12-halododecanoic acids bind to CYP4A1 with similar dissociation constants, the 12-chloro and 12-bromo fatty acids are oxidized to 12-hydroxydodecanoic acid and 12-oxododecanoic acid, whereas the 12-iodo analogue is very poorly oxidized. Incubations in H(2)(18)(2)O show that the 12-hydroxydodecanoic acid oxygen derives from water, whereas that in the aldehyde derives from O(2). The alcohol thus arises from oxidation of the halide to an oxohalonium species that is hydrolyzed by water, whereas the aldehyde arises by a conventional carbon hydroxylation-elimination mechanism. No irreversible inactivation of CYP4A1 is observed during 12-halododecanoic acid oxidation. Control experiments show that CYP2E1, which has an omega-1 regiospecificity, primarily oxidizes 12-halododecanoic acids to the omega-aldehyde rather than alcohol product. Incubation of CYP4A1 with 12,12-[(2)H](2)-12-chlorododecanoic acid causes a 2-3-fold increase in halogen versus carbon oxidation. The fact that the order of substrate oxidation (Br > Cl >> I) approximates the inverse of the intrinsic oxidizability of the halogen atoms is consistent with presentation of the halide terminus via a channel that accommodates the chloride and bromide but not iodide atoms, which implies an effective channel diameter greater than 3.90 Angstroms but smaller than 4.30 Angstroms.


Gold nanoparticles aggregation on graphene using Reactive force field: A molecular dynamic study.
We examine the aggregation behavior of AuNPs of different sizes on graphene as function of temperature using molecular dynamic simulations with Reax Force Field. In addition, the consequences of such aggregation on the morphology of AuNPs and the charge transfer behavior of AuNP-Graphene hybrid structure are analyzed. The aggregation of AuNPs on graphene is confirmed from the center of mass distance calculation. The simulation results indicate that the size of AuNPs and temperature significantly affect the aggregation behavior of AuNPs on graphene. The strain calculation showed that shape of AuNPs changes due to the aggregation and the smaller size AuNPs on graphene exhibit more shape changes than larger AuNPs at all the temperatures studies in this work. The charge transfer calculation reveals that, the magnitude of charge transfer is higher for larger AuNPs-graphene composite when compared with smaller AuNPs-graphene composite. The charge transfer trend and the trends seen in the number of Au atoms directly in touch with graphene are identical. Hence, our results conclude that, quantity of Au atoms directly in contact with graphene during aggregation is primarily facilitates charge transfer between AuNPs and graphene. Our results on the size dependent strain and charge transfer characteristics of AuNPs will aid in the development of AuNPs-graphene composites for sensor applications.


Vaccination with Vaxfectin(®) adjuvanted SIV DNA induces long-lasting humoral immune responses able to reduce SIVmac251 Viremia.
We evaluated the immunogenicity and efficacy of Vaxfectin(®) adjuvanted SIV DNA vaccines in mice and macaques. Vaccination of mice with Vaxfectin(®) adjuvanted SIV gag DNA induced higher humoral immune responses than administration of unadjuvanted DNA, whereas similar levels of cellular immunity were elicited. Vaxfectin(®) adjuvanted SIVmac251 gag and env DNA immunization of rhesus macaques was used to examine magnitude, durability, and efficacy of humoral immunity. Vaccinated macaques elicited potent neutralizing antibodies able to cross-neutralize the heterologous SIVsmE660 Env. We found remarkable durability of Gag and Env humoral responses, sustained during ~2 y of follow-up. The Env-specific antibody responses induced by Vaxfectin(®) adjuvanted env DNA vaccination disseminated into mucosal tissues, as demonstrated by their presence in saliva, including responses to the V1-V2 region, and rectal fluids. The efficacy of the immune responses was evaluated upon intrarectal challenge with low repeated dose SIVmac251. Although 2 of the 3 vaccinees became infected, these animals showed significantly lower peak virus loads and lower chronic viremia than non-immunized infected controls. Thus, Vaxfectin(®) adjuvanted DNA is a promising vaccine approach for inducing potent immune responses able to control the highly pathogenic SIVmac251.


Community proteogenomics highlights microbial strain-variant protein expression within activated sludge performing enhanced biological phosphorus removal.
Enhanced biological phosphorus removal (EBPR) selects for polyphosphate accumulating microorganisms to achieve phosphate removal from wastewater. We used high-resolution community proteomics to identify key metabolic pathways in 'Candidatus Accumulibacter phosphatis' (A. phosphatis)-mediated EBPR and to evaluate the contributions of co-existing strains within the dominant population. Overall, 702 proteins from the A. phosphatis population were identified. Results highlight the importance of denitrification, fatty acid cycling and the glyoxylate bypass in EBPR. Strong similarity in protein profiles under anaerobic and aerobic conditions was uncovered (only 3% of A. phosphatis-associated proteins exhibited statistically significant abundance differences). By comprehensive genome-wide alignment of 13,930 orthologous proteins, we uncovered substantial differences in protein abundance for enzyme variants involved in both core-metabolism and EBPR-specific pathways among the A. phosphatis population. These findings suggest an essential role for genetic diversity in maintaining the stable performance of EBPR systems and, hence, demonstrate the power of integrated cultivation-independent genomics and proteomics for the analysis of complex biotechnological systems.


Reduced ACh release at neuromuscular synapses of heterozygous leaner Ca(v)2.1-mutant mice.
Episodic ataxia type 2 (EA2) is an autosomal dominantly inherited neurological disorder. Patients have CACNA1A gene mutations resulting in truncation or single amino acid changes in the pore-forming subunit of Ca(v)2.1 (P/Q-type) Ca(2+) channels. These neuronal channels mediate synaptic neurotransmitter release. EA2 symptoms are thought to result from disturbed neurotransmission at cerebellar and neuromuscular synapses, caused by loss-of-function of Ca(v)2.1 channels. Heterozygous leaner (Ln/wt) mice, carrying a Cacna1a truncation mutation, as well as heterozygous Ca(v)2.1 null-mutant (KO/wt) mice may model synaptic aspects of EA2. We studied Ca(v)2.1-mediated acetylcholine (ACh) release at their neuromuscular junctions (NMJs) ex vivo. KO/wt mice did not show any ACh release abnormalities, not even at older age. However, Ln/wt mice had approximately 25% reduced spontaneous uniquantal ACh release and approximately 10% reduced nerve-stimulation evoked release, compared with wild-type. EA2 is treated with acetazolamide (AZA), but the pharmacotherapeutic mechanism is unknown. We tested the possibility of a direct influence on (mutant) presynaptic Ca(v)2.1 channel function by studying the acute effect of 50 muM AZA on ACh release at ex vivo NMJs of wild-type, KO/wt, and Ln/wt mice. No changes were found in any of the release parameters. Our results indicate that Ln-mutated Ca(v)2.1 channels at Ln/wt NMJs are either normally inserted in the presynaptic membrane but have reduced function, or that they inhibit wild-type channels by hampering their expression, trafficking, membrane insertion and/or function. In this respect Ln/wt NMJs may model EA2 synapses. Furthermore, AZA does not exert an acute, direct influence on the function of presynaptic (mutant) Ca(v)2.1 channels.


Brain natriuretic peptide and the risk of ventricular tachyarrhythmias in mildly symptomatic heart failure patients enrolled in MADIT-CRT.
There are limited data about the correlation between brain natriuretic peptide (BNP) levels and arrhythmic risk assessment in patients who receive device therapy for the treatment of heart failure (HF) or for the prevention of sudden cardiac death. We aimed to investigate the association between BNP levels and the risk of ventricular tachyarrhythmias among mildly symptomatic HF patients who receive an intracardiac defibrillator (ICD) with or without cardiac resynchronization therapy (respectively, CRT-D or CRT). The study population involved 1197 patients enrolled in MADIT-CRT. Plasma BNP was measured in a core laboratory at baseline and after 1-year follow-up. Ventricular tachycardia/fibrillation (VT/VF) events were identified from ICD/CRT-D interrogations. Multivariate Cox hazards regression modeling showed that elevated baseline (> median = 72 ng/L) and 1-year BNP were associated with a significant increase in the risk of VT/VF (HR = 1.36, P = .026; and HR = 1.79, P < .001, respectively); and VT/VF or death (HR = 1.37, P = .008; and HR = 1.84, P < .0001, respectively) during follow-up. At 1 year post device implantation, BNP levels were significantly lower among study patients treated with CRT-D as compared with those who received ICD only (P = .014). CRT-D patients who had greater than median reductions in BNP levels (greater than one-third reduction of initial value) experienced a significantly lower risk of subsequent VT/VF (HR = 0.61, P = .021) and VT/VF or death (HR = 0.45, P < .0001) as compared to patients without such reductions. In MADIT-CRT, elevated baseline and follow-up BNP levels were independent predictors of increased risk for subsequent ventricular tachyarrhythmias, whereas BNP reductions following CRT-D implantation identified patients with a lower incidence of VT/VF during follow-up.


Mortality after cancer diagnosis in HIV-infected individuals treated with antiretroviral therapy.
To evaluate survival and predictors of mortality after cancer diagnosis among HIV-infected persons receiving combination antiretroviral therapy (cART). Multisite cohort study. We examined all-cause mortality among HIV-infected patients treated with cART in routine care at eight US sites and diagnosed with cancer between 1996 and 2009, and predictors of mortality using Cox proportional hazards regression models. Non-AIDS-defining cancers (NADCs) were classified as related and unrelated to viral coinfections. Out of 20 677 persons in the Centers for AIDS Research Network of Integrated Clinical Systems cohort, 650 cART-treated individuals developed invasive cancer. Of these, 305 died during 1480 person-years of follow-up; crude mortality rate was 20.6 per 100 person-years [95% confidence interval (CI) 18.4, 23.1] and overall 2-year survival was 58% (95% CI 54, 62). Highest mortality was seen in primary central nervous system non-Hodgkin's lymphoma, liver, and lung cancer with rates of 90.6, 84.3, and 68.1 per 100 person-years, respectively. Adjusted hazard of death was higher among those who were older and had stage IV cancer. Adjusted hazard of death was lower among those with higher CD4 cell counts at cancer diagnosis, who achieved HIV-RNA suppression (≤400 copies/ml) on cART, received any cancer treatment, and had AIDS-defining cancer or infection-related NADCs compared to infection-unrelated NADCs. Independent predictors of mortality after cancer diagnosis among HIV-infected persons include poor immune status, failure to suppress HIV-RNA on cART, cancer stage, and lack of cancer treatment. Modification of these factors with improved strategies for the prevention and treatment of HIV and HIV-associated malignancies are needed.


Comparison of structural, thermodynamic, kinetic and mass transport properties of Mg(2+) ion models commonly used in biomolecular simulations.
The prevalence of Mg(2+) ions in biology and their essential role in nucleic acid structure and function has motivated the development of various Mg(2+) ion models for use in molecular simulations. Currently, the most widely used models in biomolecular simulations represent a nonbonded metal ion as an ion-centered point charge surrounded by a nonelectrostatic pairwise potential that takes into account dispersion interactions and exchange effects that give rise to the ion's excluded volume. One strategy toward developing improved models for biomolecular simulations is to first identify a Mg(2+) model that is consistent with the simulation force fields that closely reproduces a range of properties in aqueous solution, and then, in a second step, balance the ion-water and ion-solute interactions by tuning parameters in a pairwise fashion where necessary. The present work addresses the first step in which we compare 17 different nonbonded single-site Mg(2+) ion models with respect to their ability to simultaneously reproduce structural, thermodynamic, kinetic and mass transport properties in aqueous solution. None of the models based on a 12-6 nonelectrostatic nonbonded potential was able to reproduce the experimental radial distribution function, solvation free energy, exchange barrier and diffusion constant. The models based on a 12-6-4 potential offered improvement, and one model in particular, in conjunction with the SPC/E water model, performed exceptionally well for all properties. The results reported here establish useful benchmark calculations for Mg(2+) ion models that provide insight into the origin of the behavior in aqueous solution, and may aid in the development of next-generation models that target specific binding sites in biomolecules.


Random network behaviour of protein structures.
Geometric and structural constraints greatly restrict the selection of folds adapted by protein backbones, and yet, folded proteins show an astounding diversity in functionality. For structure to have any bearing on function, it is thus imperative that, apart from the protein backbone, other tunable degrees of freedom be accountable. Here, we focus on side-chain interactions, which non-covalently link amino acids in folded proteins to form a network structure. At a coarse-grained level, we show that the network conforms remarkably well to realizations of random graphs and displays associated percolation behavior. Thus, within the rigid framework of the protein backbone that restricts the structure space, the side-chain interactions exhibit an element of randomness, which account for the functional flexibility and diversity shown by proteins. However, at a finer level, the network exhibits deviations from these random graphs which, as we demonstrate for a few specific examples, reflect the intrinsic uniqueness in the structure and stability, and perhaps specificity in the functioning of biological proteins.


Increased KLRG1 and PD-1 expression on CD8 T lymphocytes in TB-IRIS.
Tuberculosis-associated immune reconstitution inflammatory syndrome (TB-IRIS) is an inflammatory complication in HIV-TB co-infected patients receiving antiretroviral therapy (ART). The exact contribution of T cells, natural killer (NK) cells, and monocytes to TB-IRIS development remains unclear. Here, we studied the expression of exhaustion markers on lymphocytes at different intervals during ART. We compared 13 HIV-TB patients who developed TB-IRIS with 13 patients who did not (HIV+TB+), 13 HIV-patients without TB (HIV+TB-) and 9 HIV/TB-negative controls (HIV-TB-). Patients did not differ in age, gender, or CD4-count prior to ART. Frozen peripheral blood mononuclear cells, collected before ART and during 3 months and 9 months of ART, were analysed using flow cytometry. We examined expression of KLRG1, PD-1 and IL-27R on CD4+ and CD8hi T cells, as well as CD3-negative CD8lo lymphocytes as an approximate subset of NK cells. In addition, expression of TLR2, TLR4, IL1RL1, and TRAILR on CD14+ monocytes were investigated. Prior to ART, TB-IRIS patients had higher percentages of CD8hi T cells that are KLRG1+PD-1+ compared to each control group (p≤0.034). Though PD-1 expression decreased during ART in all groups (p≤0.026), the percentage KLRG1+PD-1+CD8hi T cells remained higher in TB-IRIS patients after 3 months of ART (p≤0.013). Though these patterns were less pronounced in CD3-CD8lo lymphocytes, the percentage of KLRG1+ cells was higher in TB-IRIS patients prior to ART (p≤0.043). In contrast, no clear differences could be observed for CD4+ T cells or monocytes. TB-IRIS is preceded by a high level of exhausted (KLRG1+PD-1+) CD8hi T cells, which persists during 3 months of ART. This trait is potentially mirrored in a subpopulation of NK cells, but not CD4+ T cells. Since a dysfunctional CD8+ lymphocyte compartment could predispose patients to TB-IRIS, the functional role of these cells prior to TB-IRIS development should be further explored.


Immunoinformatic Risk Assessment of Host Cell Proteins During Process Development for Biologic Therapeutics.
The identification and removal of host cell proteins (HCPs) from biologic products is a critical step in drug development. Despite recent improvements to purification processes, biologics such as monoclonal antibodies, enzyme replacement therapies, and vaccines that are manufactured in a range of cell lines and purified using diverse processes may contain HCP impurities, making it necessary for developers to identify and quantify impurities during process development for each drug product. HCPs that contain sequences that are less conserved with human homologs may be more immunogenic than those that are more conserved. We have developed a computational tool, ISPRI-HCP, that estimates the immunogenic potential of HCP sequences by evaluating and quantifying T cell epitope density and relative conservation with similar T cell epitopes in the human proteome. Here we describe several case studies that support the use of this method for classifying candidate HCP impurities according to their immunogenicity risk.