CFTR: what's it like inside the pore?
The Cystic Fibrosis Conductance Regulator (CFTR) functions as a cAMP-activated, anion-selective channel, but the structural basis for anion permeation is not well understood. Here we summarize recent studies aimed at understanding how anions move through the CFTR channel, and the nature of the environment anions experience inside the pore. From these studies it is apparent that anion permeability selectivity and anion binding selectivity of the pore are consistent with a model based on a "dielectric tunnel." The selectivity pattern for halides and pseudohalides can be predicted if it is assumed that permeant anions partition between bulk water and a polarizable space that is characterized by an effective dielectric constant of about 19. Covalent labeling of engineered cysteines and pH titration of engineered cysteines and histidines lead to the conclusion that the CFTR anion conduction path includes a positively charged outer vestibule. A residue in transmembrane segment 6 (TM6) (R334) appears to reside in the outer vestibule of the CFTR pore where it creates a positive electrostatic potential that enhances anion conduction.


BERT-based Ranking for Biomedical Entity Normalization.
Developing high-performance entity normalization algorithms that can alleviate the term variation problem is of great interest to the biomedical community. Although deep learning-based methods have been successfully applied to biomedical entity normalization, they often depend on traditional context-independent word embeddings. Bidirectional Encoder Representations from Transformers (BERT), BERT for Biomedical Text Mining (BioBERT) and BERT for Clinical Text Mining (ClinicalBERT) were recently introduced to pre-train contextualized word representation models using bidirectional Transformers, advancing the state-of-the-art for many natural language processing tasks. In this study, we proposed an entity normalization architecture by fine-tuning the pre-trained BERT / BioBERT / ClinicalBERT models and conducted extensive experiments to evaluate the effectiveness of the pre-trained models for biomedical entity normalization using three different types of datasets. Our experimental results show that the best fine-tuned models consistently outperformed previous methods and advanced the state-of-the-art for biomedical entity normalization, with up to 1.17% increase in accuracy.


Endothelial cell adhesion molecules and cancer progression.
The role of cell adhesion molecules (CAMs), such as intercellular cell adhesion molecule-1 (ICAM-1), vascular endothelial cell adhesion molecule-1 (VCAM-1), E-selectin, and P-selectin, has been studied extensively in the process of inflammation. These molecules are responsible for recruiting leukocytes onto the vascular endothelium before extravasation to the injured tissues. Some circulating cancer cells have been shown to extravasate to a secondary site using a process similar to inflammatory cells. The most studied ligands for CAMs expressed on cancer cells, sialyl Lewis (a/x) antigens, are shown to be involved in adhesion to endothelial cells by binding to E-selectin. This process, shared by inflammatory cells and cancer cells, may partially explain the link between inflammation and tumorigenesis. Furthermore, this process may elucidate the therapeutic benefit of anti-inflammatory drugs in cancer treatment. The complexity of the tumor microenvironment has been revealed in the past decade. Currently, intense investigation is aimed at various aspects of the tumor microenvironment in addition to the tumor cells themselves. Here, we review the role of CAMs in extravasation of circulating cancer cells, a key step in metastasis.


Evaluation of the Efficacy of Different Cleaning Methods for Orthodontic Thermoplastic Retainers in terms of Bacterial Colonization.
This study investigated the antimicrobial efficacy of three different cleaning methods on Streptococcus mutans (SM) and Lactobacillus (LB) bacteria colonization in vivo. The three different cleaning methods were applied by volunteers on clear vacuum formed retainers (VFRs). In this prospective, cross-over study, a total of 21 volunteers were included. All VFRs used by the volunteers were cleaned using three different cleaning methods in a sequence. These methods were peroxide-based cleanser tablets (PBCTs) plus brushing, control (only brushing), and vinegar plus brushing, respectively. The obtained salivary, VFR material, and periodontal data were statistically compared by factorial design repeated measures analysis of variance. The SM and LB bacteria counts on VFRs after using both PBCTs and vinegar were statistically similar (p>0.05), but bacteria counts were statistically lower than the control method (p<0.01). There were no statistically significant differences between the SM and LB bacteria counts in saliva samples taken before and after the application of the cleaning methods (p>0.05). Similarly, there were no significant differences between periodontal data obtained from plaque and bleeding indices at all study times. The periodontal pocket depth gradually decreased in the successively performed cleaning applications (p<0.05). The application of PBCTs and vinegar to VFRs at sequential time intervals resulted in similar bacteria counts. The higher LB counts and similarly higher SM counts on the VFR samples indicate that mechanical cleaning only (control method) is not adequate to obtain hygiene. Salivary flora was not correlated with bacteria counts of VFRs.


Acute phencyclidine neurotoxicity in rat forebrain: induction of haem oxygenase-1 and attenuation by the antioxidant dimethylthiourea.
Phencyclidine and other N-methyl-D-aspartate receptor antagonists are toxic to pyramidal neurons in the posterior cingulate/retrosplenial cortex of rat brain. Previous studies have shown induction of heat shock protein 70 in affected neurons. In this study, expression of haem oxygenase-1, a heat shock protein induced by oxidative stress, was examined in rat forebrain after administration of a single intraperitoneal dose of phencyclidine (50 mg/kg). Northern and Western blot analyses of brain tissue extracts from phencyclidine-treated rats revealed a marked induction of haem oxygenase-1 mRNA and protein, respectively. Immunohistochemistry studies revealed that phencyclidine increased haem oxygenase-1 immunoreactivity primarily in posterior cingulate/retrosplenial, piriform and entorhinal cortices, striatum and hippocampus. Haem oxygenase-1 protein was induced in non-neuronal cells, mainly astrocytes. Some microglia expressing haem oxygenase-1 protein were also found in the posterior cingulate/retrosplenial cortex. Haem oxygenase-1 immunoreactive astrocytes and microglia were present in close proximity to the heat shock protein 70-positive neurons in the posterior cingulate/retrosplenial cortex following phencyclidine. Pretreatment of rats with 1,3-dimethylthiourea, an antioxidant, significantly reduced haem oxygenase-1 protein induction by phencyclidine. Thus, induction of haem oxygenase-1 in glia by phencyclidine appears to be mediated mostly by oxidative stress. Experiments with the amino cupric silver stain for neuronal degeneration revealed phencyclidine-induced neurotoxicity in the posterior cingulate/retrosplenial cortex. The number of affected neurons was significantly reduced after 1,3-dimethylthiourea pretreatment. This suggests that the neurotoxicity of N-methyl-D-aspartate antagonists is due in part to the oxidative stress and may be amenable to therapeutic interventions.


Instabilities and solitons in systems with spatiotemporal dispersion.
A simple model based on the 1D nonlinear Schrödinger equation is studied, which contains both spatially and temporally dispersive terms. Parametric instabilities for plane waves are analyzed in detail, and solitary waves (both bright and dark) are found. The model presented here is able to describe the non-trivial unstable dynamics of intense, nonlinear light propagation near a material resonance in presence of negative spatial dispersion. We provide as a practical example the light propagation near the tail of an exciton-polariton resonance in a specially designed semiconductor superlattice.


Compartmentalisation of enzymes for cascade reactions through biomimetic layer-by-layer mineralization.
Cellular metabolic pathways are paradigms for the rapid and waste-free conversion of molecules into useful products through multiple enzyme-catalyzed steps (cascade reactions). Attempts to establish efficient cascade reactions for technological applications have focused on mimicking nature's high degree of organization by controlling the positioning of enzymes through immobilization in tailor-made compartments. The present work utilized peptide-mediated layer-by-layer mineralization as a facile and generic method for the compartmentalisation of multi-enzyme systems in nanoscale silica layers. It is demonstrated that, in a multilayer system, the overall rate of the reaction cascade was primarily affected by the placement of the enzyme catalyzing the first step, with the placement of the enzyme possessing the lowest catalytic efficiency also being an important factor. As the rate-limiting enzymes were positioned closer to the external silica surface, the overall rate of cascade reactions increased. Furthermore, distributing the enzymes into different adjacent silica compartments yielded higher overall cascade reaction rates compared to placement of the enzymes into the same silica layer. The synthetic methods and kinetic analyses presented here provide guidance for improving the performance of immobilized multi-enzyme systems for a wide range of technological applications.


Effect of protein kinase C modulation on outcome of experimental CNS ischemia.
Protein kinase C (PKC) is an important intracellular regulator, and its activity may play a central role in the modulation of neuronal ischemic damage. Staurosporine and the compound H-7 are potent in vitro inhibitors of PKC, and 1,2-oleoylacetylglycerol (OAG) is an effective activator. We administered these compounds through a spinal subarachnoid catheter and demonstrated in vivo alteration of spinal cord PKC activity. We then tested the effects of altering PKC activity in a well-established rabbit model of reversible spinal cord ischemia. Animals within each experimental group were subjected to a range of spinal cord ischemic durations by temporary occlusion of the infrarenal abdominal aorta. Compared to control, both staurosporine and H-7 significantly shortened the duration of ischemia that the animals could tolerate, without developing permanent paraplegia. OAG resulted in an insignificant lengthening of the ischemic duration that the animals could withstand. The worsening of ischemic outcome by PKC inhibitors suggests that the enzyme is important for maintaining neurologic function under ischemic conditions, possibly secondary to modulation of intracellular calcium levels.


[Complex treatment of elderly patients with pain reflectory manifestations of lumbosacral osteochondrosis and peripheral circulation disorder].
Involutional age makes difficult the treatment of pain syndromes of vertebra origin. Concomitant diseases, age stress syndrome, hormone disbalance constrain the management of the disease. Combination of laser stimulation, reflexotherapy, psychodiagnostics and psychocorrection enhances the efficiency of the treatment of pain syndromes in patients with lumbo-sacral radiculitis and peripheral circulation disorder.


Monte Carlo study of the impact of a magnetic field on the dose distribution in MRI-guided HDR brachytherapy using Ir-192.
In the process of developing a robotic MRI-guided high-dose-rate (HDR) prostate brachytherapy treatment, the influence of the MRI scanner's magnetic field on the dose distribution needs to be investigated. A magnetic field causes a deflection of electrons in the plane perpendicular to the magnetic field, and it leads to less lateral scattering along the direction parallel with the magnetic field. Monte Carlo simulations were carried out to determine the influence of the magnetic field on the electron behavior and on the total dose distribution around an Ir-192 source. Furthermore, the influence of air pockets being present near the source was studied. The Monte Carlo package Geant4 was utilized for the simulations. The simulated geometries consisted of a simplified point source inside a water phantom. Magnetic field strengths of 0 T, 1.5 T, 3 T, and 7 T were considered. The simulation results demonstrated that the dose distribution was nearly unaffected by the magnetic field for all investigated magnetic field strengths. Evidence was found that, from a dose perspective, the HDR prostate brachytherapy treatment using Ir-192 can be performed safely inside the MRI scanner. No need was found to account for the magnetic field during treatment planning. Nevertheless, the presence of air pockets in close vicinity to the source, particularly along the direction parallel with the magnetic field, appeared to be an important point for consideration.


[Functional evaluation in patients with kidney calculi].
Nephrolithiasis is a common disorder and a significant problem because of incidence, recurrence and severe consequences. Stone disease is a surgical as well as a medical problem. Major progress has been made recently in understanding the pathophysiological disturbances responsible for stone formation as well as in the techniques of stone removal. The introduction of extracorporeal shock wave lithotripsy has considerably reduced the need for surgery. Improvements in methods of kidney stone removal have not diminished the need for the application of an effective prophylactic program. The internist should take a complete history of stone events (number, composition, location and outcome of stone event), family history of stones, dietary habits (focusing on the consumption of animal protein, salt and dairy products), medications and physical examination. Radiopaque stones should be documented by plane X-ray films. Ultrasonography should be used to image calculi that are nonopaque, and to easily distinguish them from masses such as tumour or blood clot. Computed tomography is also an excellent method for imaging nonopaque renal calculi but higher cost and radiation exposure are disadvantages [2]. Crystallographic analysis is the essential diagnostic procedure. If available, previous stones should also be examined. "In stone disease, everything is measurement. What the laboratory cannot tell you, you will not know; what it tells you in error, you will not correct by using your instincts, your medical experience, or your art [3]". Reliable diagnostic protocols are available for the identification of different causes of stones. The complexity of protocols depend on the severity of nephrolithiasis. Patients with a single stone episode undergo simple protocol, and extensive detailed protocol is used for patients with recurrent stone disease, or patients at increased risk. Simple protocol, besides the already mentioned history of stone events, radiographic investigation and crystallographic analysis, includes serum urea, creatinine, uric acid, sodium, calcium, phosphorus and protein levels, urinary pH and volume, urine samples for culture and urinary calcium, uric acid, oxalate and citrate. Extensive metabolic evaluation includes simple protocol, determination of serum levels of alkaline phosphatase, parathyroid hormone, thyroxin, magnesium. A 24-h collection of urine specimen is analysed for urea, creatinine, uric acid, calcium, phosphate, sodium, magnesium, oxalate and citrate. Extensive protocol includes specialized evaluation tests [5]. Urinary acidification test is important for detecting distal renal tubular acidosis. Two 24-h urine specimens are collected while the patient is on the regular diet. The patient is then placed on a restricted diet (400 mg of calcium and 100 mEq of sodium) for a week, and another 24-h urine sample is collected. After that fasting and calcium load tests are performed (Sheme 1). Fasting urinary calcium is used to detect renal calcium leak, and calciuric response to oral calcium load provides an indirect measure of intestinal calcium absorption. Diagnostic criteria for major forms of stone disease [8] are presented in Table 1. There are some still unsolved questions: does time after passage of stones or urological intervention influence the frequency of urine abnormalities that can be detected; are there differences in 24-h urine composition between weekdays and weekends: what is the prevalence of the most important urinary risk factors of recurrent idiopathic calcium nephrolithiasis: do male patients differ from females with respect to urinary risk factors or recurrent idiopathic calcium nephrolithiasis? [7].


Antimicrobial prospect of newly synthesized 1,3-thiazole derivatives.
A new series of 1,3-thiazole and benzo[d]thiazole derivatives 10-15 has been developed, characterized, and evaluated for in vitro antimicrobial activity at concentrations of 25-200 μg/mL against Gram+ve organisms such as methicillin-resistant Staphylococcus aureus (MRSA), Gram-ve organisms such as Escherichia coli (E. coli), and the fungal strain Aspergillus niger (A. niger) by the cup plate method. Ofloxacin and ketoconazole (10 μg/mL) were used as reference standards for antibacterial and antifungal activity, respectively. Compounds 11 and 12 showed notable antibacterial and antifungal activities at higher concentrations (125-200 μg/mL), whereas benzo[d]thiazole derivatives 13 and 14 were found to display significant antibacterial or antifungal activity (50-75 μg/mL) against the Gram+ve, Gram-ve bacteria, or fungal cells used in the present study. In addition, a correlation between calculated and determined partition coefficient (log P) was established which allows future development of compounds within this series to be carried out based on calculated log P values. Moreover, compounds 13 and 14 show that the optimum logarithm of partition coefficient (log P) should be around 4.


Child and Adult Factors Related to Quality of Life in Adults with Autism.
The WHO Quality of Life-Brief questionnaire was used to assess quality of life (QoL) among 52 adults with autism (mean age 49 years) followed-up since childhood. Overall, assessments of QOL were more positive than measures of objective social outcome (jobs, independence, relationships etc.) but correlations between caregiver and self-reports were low. Informant ratings indicated few correlations between current QoL and any child or adult factors. On self-report ratings, QoL was significantly negatively correlated with severity of repetitive behaviours in childhood; higher QoL was positively associated with better adult social outcomes. However, only a minority of adults (n = 22) could provide self-report data and findings highlight the need to develop valid measures for assessing the well-being of adults with autism.


Solid state mechanochemical processes for better electroceramics.
The present short overview focuses on the renovation of solid state processes toward phase pure and well-crystallized complex oxides centered on the electroceramic materials. Elevation of the reactivity and preservation of stoichiometry of the starting mixture or precursor are of universal importance. Mechanical activation, being considered as versatile, may also need reconsideration in view of contamination and process rationalization. After briefly reviewing mechanochemical processes for direct synthesis of complex oxides, solid state processes toward well crystallized fine particles of complex oxides are discussed by starting from mechanochemically derived precursors with subsequent optimized calcination. Case studies were cited from literatures for complex oxides, including author's own experimental studies mainly with BaBi(2)Ta(2)O(9) (BBT), Ba(Mg(1/3)Ta(2/3))O(3) (BMT) and KNbO(3) (KN). The substances discussed are mostly associated with ferroelectric materials, with a few exceptions of iron-containing magnetic materials.


Formation of nearly monodisperse In2O3 nanodots and oriented-attached nanoflowers: hydrolysis and alcoholysis vs pyrolysis.
Single crystalline and nearly monodisperse In2O3 nanocrystals with both dot and flower shapes were synthesized in a simple reaction system. This system used indium carboxylates as the precursors with or without alcohol as the activating reagents in a hydrocarbon solvent under elevated temperatures. Limited ligand protection (LLP) led to three-dimensional (3D) oriented attachment of nanodots, resulting in 3D nanoflowers. When the system had sufficient ligand protection for the nanocrystals, nanodots were found to be the stable products. The diameters of nearly monodisperse nanodots and nanoflowers were varied in a range from approximately 5 to approximately 15 nm and approximately 15 to approximately 60 nm, respectively. The simple reaction system made it possible to have a systematic study of the reaction mechanisms along with the growth kinetics of nanocrystals. Hydrolysis and alcoholysis were identified as the major paths for this system, as opposed to pyrolysis. Both nearly monodispersed nanodots and nanoflowers can be made through either of the reaction pathways. Hydrolysis was found as a reversible pathway, and alcoholysis was confirmed to be irreversible. Consequently, a sufficient amount of alcohol was able to force the yield of nanocrystals, both dots and flowers, to unity.


The nature of the relationship between neurocognition and theory of mind impairments in stroke patients.
Theory of mind (ToM) is a complex, high-level cognitive function that allows people to infer the cognitive and affective mental states of others. Previous studies have produced limited and frequently contradictory findings on the neuropsychological underpinnings of ToM performance in patients with stroke. The aim of the present study is to investigate neuropsychological mechanisms of cognitive and affective theory of mind dysfunctions in patients with stroke. Fifty-eight patients with stroke and 22 healthy controls matched in age, gender, and education level underwent robust neuropsychological examination of their pragmatic abilities, executive functions, attention, memory, psychomotor speed, and visuospatial abilities as well as a cognitive and affective ToM assessment. Patients with stroke demonstrated impaired performance in all ToM tasks. While pragmatic competence and, to a lesser degree, executive functions had the strongest contribution to ToM impairments, attention and general cognitive functioning did not directly affect mentalizing abilities, as demonstrated by a path analysis. Our study reveals the different roles of cognitive functions in cognitive and affective components of ToM. Executive functions contributed only to the cognitive components of ToM. Deficits in cognitive aspects of ToM are best explained by impairment of pragmatic competence and executive functions. In contrast, executive dysfunction does not affect the ability to understand the affective mental states of others. (PsycINFO Database Record


BRAF and KRAS Mutations in hyperplastic polyps and serrated adenomas of the colorectum: relationship to histology and CpG island methylation status.
The aim of this study was to test the hypothesis that mutations of the oncogenes BRAF or KRAS are early events in the putative serrated polyp neoplasia pathway and more advanced pathology is associated with acquired mutator and suppressor gene inactivation by CpG island methylation of promoter regions. We assayed 79 sporadic hyperplastic polyps (HPs) classified according to the schema of Torlakovic et al and 25 serrated adenomas (SAs) for BRAF and KRAS mutations and related the findings to histologic characteristics and CpG island methylation phenotype (CIMP). Mutations at exon 15, codon 599, of BRAF were assayed using an allele-specific PCR (AS-PCR) technique and confirmed in a sample of AS-PCR- positive cases by direct sequencing of exon 15. AS-PCR-negative HPs and SAs were also sequenced on exon 15 and exon 11 to detect additional mutations. PCR-RFLP was used to assay KRAS codon 12 and 13 mutations, and these mutations were further validated by direct sequencing of the KRAS gene. BRAF599 mutations were identified in a total of 55 HPs (69.6%) and KRAS mutations in a total of 13 (16.5%). BRAF599 mutations occurred with similar frequencies among microvesicular serrated polyp (76.3%) and serrated polyp with abnormal proliferation (82.1%) subtypes but less frequently in goblet cell serrated polyps (23.1%). Conversely, KRAS mutations were most frequent in goblet cell serrated polyp (46.2%) and less frequent in microvesicular serrated polyp (13.2%) and serrated polyp with abnormal proliferation (7.1%). BRAF599 and KRAS mutations were present in 15 (60.0%) and 7 (28.0%) of SAs, respectively. BRAF 599 mutation and KRAS were mutually exclusive findings in the polyps studied and one or the other occurred in 68 of 79 (86.1%) HPs and 22 of 25 (88.0%) SAs. CpG island methylation involving 2 or more genes (CIMP-H) was present in 80.0% of SAs, 75% serrated polyp with abnormal proliferations, 47.4% of microvesicular serrated polyps, and 15.4% of goblet cell serrated polyps. SAs were significantly more likely to be CIMP-H than HPs (odds ratio 3.7; 95% confidence interval, 1.27-10.86; P = 0.017). A similar high frequency of KRAS or BRAF mutations across the histologic spectrum of the serrated polyps assayed suggests that these are early events in the serrated polyp neoplasia pathway. In contrast, the association of higher levels of CpG island methylation with more advanced histologic changes suggests that CpG island methylation plays a role in serrated polyp progression toward colorectal carcinoma.


Nucleotide sequence of cloned unintegrated avian sarcoma virus DNA: viral DNA contains direct and inverted repeats similar to those in transposable elements.
We have determined the nucleotide sequence of portions of two circular avian sarcoma virus (ASV) DNA molecules cloned in a prokaryotic host--vector system. The region whose sequence was determined represents the circle junction site--i.e., the site at which the ends of the unintegrated linear DNA are fused to form circular DNA. The sequence from one cloned molecule, SRA-2, shows that the circle junction site is the center of a 330-base-pair (bp) tandem direct repeat, presumably representing the fusion of the long terminal repeat (LTR) units known to be present at the ends of the linear DNA. The circle junction site is also the center of a 15-bp imperfect inverted repeat, which thus appears at the boundaries of the LTR. The structure of ASV DNA--unique coding region flanked by a direct repeat that is, in turn, terminated with a short inverted repeat--is very similar to the structure of certain transposable elements. Several features of the sequence imply that circularization to form the SRA-2 molecule occurred without loss of information from the linear DNA precursor. Circularization of another cloned viral DNA molecule, SRA-1, probably occurred by a different mechanism. The circle junction site of the SRA-1 molecule has a 63-bp deletion, which may have arisen by a mechanism that is analogous to the integration of viral DNA into the host genome. Flanking one side of the tandem direct repeat is the binding site for tRNATrp, the previously described primer for synthesis of the first strand of viral DNA. The other side of the direct repeat is flanked by a polypurine tract, A-G-G-G-A-G-G-G-G-G-A, which may represent the position of the primer for synthesis of the second strand of viral DNA. An A+T-rich region, upstream from the RNA capping site, and the sequence A-A-T-A-A-A are present within the direct repeat sequence. These sequences may serve as a promoter site and poly(A) addition signal, respectively, as proposed for other eukaryotic transcription units.


Neolignan Constituents with Potential Beneficial Effects in Prevention of Type 2 Diabetes from Viburnum fordiae Hance Fruits.
Nine new neolignan glycosides (1-9), viburfordosides A-I, two new neolignans, fordianes A and B (10, 11), and seven known analogues (12-18) have been isolated and identified from the fruits of Viburnum fordiae Hance. The structures and absolute configurations of undescribed neolignan constituents were identified by chemical methods and spectroscopic analyses. The α-glucosidase inhibitory, ABTS•+ and DPPH• scavenging, and anti-inflammatory activities of these secondary metabolites were evaluated. Some of them exhibited significant potency in inhibiting α-glucosidase and scavenging free radicals. Among the 14 metabolites that were found to have the capacity to inhibit NO production in LPS-stimulated RAW264.7 macrophage cells, compounds 2, 4, 6, 10, 11, 14, 17, and 18 were potent with IC50 values of 10.88-41.10 μM. These results support that V. fordiae fruits possessing the neolignan compounds may serve as both a functional food and a medicinal resource to prevent and treat type 2 diabetes (T2D).


Dysfunctions in dopamine systems and ADHD: evidence from animals and modeling.
Animal models are useful for characterizing neural substrates of neuropsychiatric disorders. Several models have been proposed for the study of Attention Deficit Hyperactivity Disorder (ADHD). The models can be divided into various groups: (i) genetically derived hyperactivity/ inattention, (ii) animal models showing symptoms after pharmacological intervention, and (iii) those based on spontaneous variations in a random population. Spontaneously hypertensive (SHR) and Naples High Excitability (NHE) rats show behavioral traits featuring the main aspects of ADHD in humans but show different changes in dopamine (DA) systems. In fact, the enzyme tyrosine hydroxylase is hyperexpressed in NHE rats and hypoexpressed in SHR. The DA transporter is hyperexpressed in both lines, although in the SHR, DAT activity is low (reduced DA uptake). The DA levels in the striatum and prefrontal cortex are increased in the juvenile SHR, but are decreased in handled young and non-handled older animals. The mRNA of the D1 DA receptor is upregulated in the prefrontal cortex of SHR and down-regulated in NHE. The D2 DA receptors are likely to be hypofunctioning in SHR, although the experimental evidence is not univocal, whereas their mRNA is hyperexpressed in NHE. Thus, in SHR both the mesocortical and mesolimbic DA pathways appear to be involved, whereas in NHE only the mesocortical system. To understand the effects of methylphenidate, the elective ADHD drug treatment in humans, in a dysfunctioning DA system, we realized a simple mathematical model of DA regulation based on experimental data from electrophysiological, cyclic voltammetry, and microdialysis studies. This model allows the estimation of a higher firing frequency of DA neurons in SHR rats and suggests that methylphenidate increases attentive processes by regulating the firing rate of DA neurons.


Irritability in child and adolescent anxiety disorders.
Our objective was to compare self- and parent-reported irritability in youths with anxiety disorders, healthy youths, and those with mood disorders characterized by irritability. Irritability is a common but relatively understudied psychiatric symptom in child and adolescent anxiety disorders. In anxious youths, little is known about the severity of irritability, its impact on functioning, or the effect of informant source on reports of irritability. We compared parent- and self-report forms of the Affective Reactivity Index (ARI), a validated measure of irritability, in youths ages 8-17 years with no psychopathology (healthy comparison, HC; n = 38), anxiety disorders (ANX; n = 42), bipolar disorder (BD; n = 35), or severe mood dysregulation (SMD; n = 61; a phenotype characterized by chronic, severely impairing irritability). Irritability was significantly higher in ANX than HC youths by both parent and self-report (partial η(2) = 0.24 and 0.22, respectively, P's < 0.001). Informant effects differed among ANX, BD, and SMD. Overall, parent-reported irritability was higher in BD with comorbid anxiety disorders and SMD with or without comorbid anxiety disorders than ANX (P's < 0.007), but self-reported irritability was not significantly different among the three patient groups. By both parent and self-report, youths with anxiety disorders exhibit significantly more irritability and associated impairment than healthy subjects. Self-reported irritability in youths with anxiety disorders is comparable to that observed in youths with severe mood disorders, although parental reports of irritability differ among the disorders. Future research should examine the pathophysiology of anxiety-associated irritability, as well as its prognostic and treatment implications.


Rapidly Developed Multiple Face and Neck Skin Cancers in a Patient with Sjögren's Syndrome: A Case Report.
BACKGROUND Sjögren's syndrome is a chronic, systemic disorder of an autoimmune nature, and its primary etiopathogenetic events are not known. Previous studies have found elevated incidence of malignancies in patients with primary Sjögren's syndrome. However, there are few reports regarding the association of Sjögren's syndrome with skin cancers, especially with multiple skin cancers developed within a short time. CASE REPORT We reported an unusual case of a patient with primary Sjögren's syndrome who suffered from rapidly developed facial and neck skin cancers within two years. CONCLUSIONS Sjögren's syndrome associated with skin cancer is rare. Our case report suggests that Sjögren's syndrome patients require continuous follow-up with conventional cancer examination, including skin biopsy for suspected skin lesions.


Effects of balance training in elderly evaluated by clinical tests and dynamic posturography.
All persons aged 70 through 75 years (N = 457) in a Swedish community were invited to participate in a 9 week balance training study. Out of 55 interested subjects, 15 were chosen at random for a study group; 15 matched controls were also selected. Before and after the investigation period the balance function was assessed by clinical balance tests and dynamic posturography. In the clinical balance tests, the training group significantly improved their balance standing on one leg with eyes closed as well as standing on one leg while shaking the head; they also walked 15 m back and forth faster. In the dynamic posturography the training group significantly improved their equilibrium scores in the 3 most difficult test conditions. The results of the control group were unchanged except for one test condition in the dynamic posturography. The differences in one-leg standing with head shaking, walking 2 x 15 m, and the equilibrium score using sway-referenced platform in dynamic posturography were proved to be attributable to the training. The first investigation in all 29 subjects formed normative dynamic posturography data for the age group 70 through 75 years. The normative posturographic data of this age group differed from previously obtained data in the age groups 20 through 59 and 60 through 69 years. It is concluded that elderly may improve their balance by regular balance training exercises for as short a period as 9 weeks. This might prove to be of great value in improving balance and thereby preventing accidental falls and subsequent fractures in elderly.


[Neurophysiology of cannabis].
Various neurophysiological properties of cannabis are described in a review of the literature. The properties of the principal active cannabinoid, delta-9-tetrahydrocannabinol (delta-9-THC) are described, followed by a review of psychological and neuropsychiatric effects. Cannabis disturbs EEG, decreasing in particular REM sleep time. Cerebral atrophy is not evident, assuming no concurrent drug abuse, but cerebral perfusion is noted to decrease after THC absorption. Endocrine effects have been described in animal studies: inhibiting effects on sex hormone production and thyroid function, and a stimulating action on corticosteroid activity have been observed. These actions are thought to exert themselves at the hypothalamic level. There is also evidence of an effect on prostaglandin metabolism. At the cellular and subcellular level THC acts on membranes of neurones and synaptic vesicles, possibly affecting membrane transport systems. The morphology of the synapse is also altered. THC acts on neurone conduction, stimulating polysynaptic transmission. Various mechanisms of THC activity are considered: via specific lipoprotein receptors, via central benzodiazepine receptors and via opioid receptors. THC also exerts an effect on neurotransmitters: an increase in central cholinergic activity, a mode-rate increase in catecholaminergic activity and effects on the GABA and serotonin systems are postulated.


A Metabolomic Approach to Target Compounds from the Asteraceae Family for Dual COX and LOX Inhibition.
The application of metabolomics in phytochemical analysis is an innovative strategy for targeting active compounds from a complex plant extract. Species of the Asteraceae family are well-known to exhibit potent anti-inflammatory (AI) activity. Dual inhibition of the enzymes COX-1 and 5-LOX is essential for the treatment of several inflammatory diseases, but there is not much investigation reported in the literature for natural products. In this study, 57 leaf extracts (EtOH-H2O 7:3, v/v) from different genera and species of the Asteraceae family were tested against COX-1 and 5-LOX while HPLC-ESI-HRMS analysis of the extracts indicated high diversity in their chemical compositions. Using O2PLS-DA (R2 > 0.92; VIP > 1 and positive Y-correlation values), dual inhibition potential of low-abundance metabolites was determined. The O2PLS-DA results exhibited good validation values (cross-validation = Q2 > 0.7 and external validation = P2 > 0.6) with 0% of false positive predictions. The metabolomic approach determined biomarkers for the required biological activity and detected active compounds in the extracts displaying unique mechanisms of action. In addition, the PCA data also gave insights on the chemotaxonomy of the family Asteraceae across its diverse range of genera and tribes.


Cerebral vasculopathy is associated with severe vascular manifestations in systemic sclerosis.
To investigate brain involvement in patients with systemic sclerosis (SSc). Sixty-three patients with SSc fulfilling the American College of Rheumatology and/or Leroy and Medsger criteria were retrospectively studied, including 30 (47.6%) with limited cutaneous and 27 (42.9%) with diffuse cutaneous SSc. Forty-one patients underwent computed tomography (CT) scan and magnetic resonance imaging (MRI) of the brain, 11 patients only CT scan, and the remaining 11 patients only MRI. Cerebral vasculopathy on MRI and CT scan was defined as absent or mild (score < 1), moderate (1 <or= score < 2), or severe (score >or= 2) on a 4-point scale (0 to 3). Cerebral vasculopathy was identified on CT scan in 22 patients (moderate in 12 and severe in 10) and on MRI in 38 patients (moderate in 28 and severe in 10). Patients with severe cerebral vasculopathy seen on MRI were more likely to have pulmonary arterial hypertension (PAH; p = 0.003) and showed a tendency to have scleroderma renal crisis (SRC; p = 0.25, test for trend p = 0.097). A similar association was found between severe cerebral vasculopathy seen on CT scan and PAH (p = 0.026) or SRC (p = 0.04). After adjusting for age and hypertension, severe cerebral vasculopathy was still associated with increased risk of severe vascular manifestations [odds ratio (OR) 32, 95% confidence interval (CI) 3.45-297, p = 0.002 for CT scan; OR 26, 95% CI 1.71-394, p = 0.019 for MRI]. Severe cerebral vasculopathy is associated with severe vascular manifestations in SSc patients. SSc patients with severe vascular complications should undergo neuroradiological imaging assessment of brain involvement.


The MRX complex regulates Exo1 resection activity by altering DNA end structure.
Homologous recombination is triggered by nucleolytic degradation (resection) of DNA double-strand breaks (DSBs). DSB resection requires the Mre11-Rad50-Xrs2 (MRX) complex, which promotes the activity of Exo1 nuclease through a poorly understood mechanism. Here, we describe the Mre11-R10T mutant variant that accelerates DSB resection compared to wild-type Mre11 by potentiating Exo1-mediated processing. This increased Exo1 resection activity leads to a decreased association of the Ku complex to DSBs and an enhanced DSB resection in G1, indicating that Exo1 has a direct function in preventing Ku association with DSBs. Molecular dynamics simulations show that rotation of the Mre11 capping domains is able to induce unwinding of double-strand DNA (dsDNA). The R10T substitution causes altered orientation of the Mre11 capping domain that leads to persistent melting of the dsDNA end. We propose that MRX creates a specific DNA end structure that promotes Exo1 resection activity by facilitating the persistence of this nuclease on the DSB ends, uncovering a novel MRX function in DSB resection.


Generalization lags behind learning on an auditory perceptual task.
The generalization of learning from trained to untrained conditions is of great potential value because it markedly increases the efficacy of practice. In principle, generalization and the learning itself could arise from either the same or distinct neural changes. Here, we assessed these two possibilities in the realm of human perceptual learning by comparing the time course of improvement on a trained condition (learning) to that on an untrained condition (generalization) for an auditory temporal-interval discrimination task. While significant improvement on the trained condition occurred within 2 d, generalization to the untrained condition lagged behind, only emerging after 4 d. The different time courses for learning and generalization suggest that these two types of perceptual improvement can arise from at least partially distinct neural changes. The notably longer time course for generalization than learning demonstrates that increasing the duration of training can be an effective means to increase the number of conditions to which learning generalizes on perceptual tasks.


Melatonin-mediated cytoprotection against hyperglycemic injury in Müller cells.
Oxidative stress is a contributing factor to the development and progression of diabetic retinopathy, a leading cause of blindness in people at working age worldwide. Recent studies showed that Müller cells play key roles in diabetic retinopathy and produce vascular endothelial growth factor (VEGF) that regulates retinal vascular leakage and proliferation. Melatonin is a potent antioxidant capable of protecting variety of retinal cells from oxidative damage. In addition to the pineal gland, the retina produces melatonin. In the current study, we investigated whether melatonin protects against hyperglycemia-induced oxidative injury to Müller cells and explored the potential underlying mechanisms. Our results show that both melatonin membrane receptors, MT1 and MT2, are expressed in cultured primary Müller cells and are upregulated by elevated glucose levels. Both basal and high glucose-induced VEGF production was attenuated by melatonin treatment in a dose-dependent manner. Furthermore, we found that melatonin is a potent activator of Akt in Müller cells. Our findings suggest that in addition to functioning as a direct free radical scavenger, melatonin can elicit cellular signaling pathways that are protective against retinal injury during diabetic retinopathy.


AAV-expressed eCD4-Ig provides durable protection from multiple SHIV challenges.
Long-term in vivo expression of a broad and potent entry inhibitor could circumvent the need for a conventional vaccine for HIV-1. Adeno-associated virus (AAV) vectors can stably express HIV-1 broadly neutralizing antibodies (bNAbs). However, even the best bNAbs neutralize 10-50% of HIV-1 isolates inefficiently (80% inhibitory concentration (IC80) > 5 μg ml(-1)), suggesting that high concentrations of these antibodies would be necessary to achieve general protection. Here we show that eCD4-Ig, a fusion of CD4-Ig with a small CCR5-mimetic sulfopeptide, binds avidly and cooperatively to the HIV-1 envelope glycoprotein (Env) and is more potent than the best bNAbs (geometric mean half-maximum inhibitory concentration (IC50) < 0.05 μg ml(-1)). Because eCD4-Ig binds only conserved regions of Env, it is also much broader than any bNAb. For example, eCD4-Ig efficiently neutralized 100% of a diverse panel of neutralization-resistant HIV-1, HIV-2 and simian immunodeficiency virus isolates, including a comprehensive set of isolates resistant to the CD4-binding site bNAbs VRC01, NIH45-46 and 3BNC117. Rhesus macaques inoculated with an AAV vector stably expressed 17-77 μg ml(-1) of fully functional rhesus eCD4-Ig for more than 40 weeks, and these macaques were protected from several infectious challenges with SHIV-AD8. Rhesus eCD4-Ig was also markedly less immunogenic than rhesus forms of four well-characterized bNAbs. Our data suggest that AAV-delivered eCD4-Ig can function like an effective HIV-1 vaccine.


Transmission Infrared Microscopy and Machine Learning Applied to the Forensic Examination of Original Automotive Paint.
Alternate least squares (ALS) reconstructions of the infrared (IR) spectra of the individual layers from original automotive paint were analyzed using machine learning methods to improve both the accuracy and speed of a forensic automotive paint examination. Twenty-six original equipment manufacturer (OEM) paints from vehicles sold in North America between 2000 and 2006 served as a test bed to validate the ALS procedure developed in a previous study for the spectral reconstruction of each layer from IR line maps of cross-sectioned OEM paint samples. An examination of the IR spectra from an in-house library (collected with a high-pressure transmission diamond cell) and the ALS reconstructed IR spectra of the same paint samples (obtained at ambient pressure using an IR transmission microscope equipped with a BaF2 cell) showed large peak shifts (approximately 10 cm-1) with some vibrational modes in many samples comprising the cohort. These peak shifts are attributed to differences in the residual polarization of the IR beam of the transmission IR microscope and the IR spectrometer used to collect the in-house IR spectral library. To solve the problem of frequency shifts encountered with some vibrational modes, IR spectra from the in-house spectral library and the IR microscope were transformed using a correction algorithm previously developed by our laboratory to simulate ATR spectra collected on an iS-50 FT-IR spectrometer. Applying this correction algorithm to both the ALS reconstructed spectra and in-house IR library spectra, the large peak shifts previously encountered with some vibrational modes were successfully mitigated. Using machine learning methods to identify the manufacturer and the assembly plant of the vehicle from which the OEM paint sample originated, each of the twenty-six cross-sectioned automotive paint samples was correctly classified as to the "make" and model of the vehicle and was also matched to the correct paint sample in the in-house IR spectral library.


Common variable immunodeficiency presenting in a girl as lung infiltrates and mediastinal adenopathies leading to severe "superior vena caval" syndrome.
Common variable immunodeficiency (CVID) is a primary immunodeficiency disorder characterized by decreased levels of circulating immunoglobulins (Ig) and increased susceptibility to infections. We describe the case of a girl, progressively developing CVID, whose first clinical manifestations were noninfectious diffuse pulmonary infiltrates and rapidly developing hilar and mediastinal lymphadenopathies causing a severe "superior vena caval syndrome". Histological evaluation of surgical samples showed follicular and paracortical hyperplasia of the lymph nodes and poorly organized, non-necrotizing, noninfectious, "reactive" granulomata in lymph nodes and pulmonary tissue. Daily treatment with azathioprine and prednisone induced resolution of the intrathoracic abnormalities but was associated with a progressive decrease of circulating IgG and IgA levels and natural killer (NK) lymphocytes that was not related to treatment. This case demonstrates that granulomatous inflammatory changes may be the first manifestations of common variable immunodeficiency, and that this disorder must be included in the differential diagnosis of lymphoid interstitial pneumonitis and of bilateral mediastinal lymph node enlargement leading to superior vena caval syndrome.


How to punish cyber criminals: A study to investigate the target and consequence based punishments for malware attacks in UK, USA, China, Ethiopia & Pakistan.
Numerous research studies have highlighted the exponential growth of malware attacks worldwide, posing a significant threat to society. Cybercriminals are becoming increasingly merciless and show no signs of pity towards individuals or organizations. It is evident that cyber criminals will stop at nothing to gain unauthorized access to confidential information. To effectively combat malware attacks, strict cyber laws are necessary, and the use of malware is punishable in many countries. However, the literature has not addressed whether these penalties create deterrence or not. This research article has addressed this gap. In this study, the effectiveness of criminal laws related to malware-related crimes in various jurisdictions was analyzed using the doctrinal research methodology. The cyber laws of the USA, UK, Ethiopia, Pakistan, and China were examined to determine whether the penalties imposed for these crimes are appropriate given the severity of the harm caused. The study concludes that malware penalties should take into account the creation or use of malicious code, targeting individuals or organizations, and the magnitude of consequences, regardless of whether mens rea is present or not.


Postural control after unexpected external perturbation: Effects of Parkinson's disease subtype.
Different clinical subtypes of Parkinson's disease (PD) have long been recognized. Recent studies have focused on two PD subtypes: Postural Instability and Gait Difficulty (PIGD) and Tremor Dominant (TD). PIGD patients have greater difficulties in postural control in relation to TD. However, knowledge about the differences in reactive adjustment mechanisms following a perturbation in TD and PIGD is limited. This study aimed to compare reactive postural adjustments under unexpected external perturbation in TD, PIGD, and control group (CG) subjects. Forty-five individuals (15 TD, 15 PIGD, and 15 CG) participated in this study. Postural perturbation was applied by the posterior displacement of the support surface in an unexpected condition. The velocity (15 cm/s) and displacement (5 cm/s) of perturbation were the same for all participants. Center of pressure (CoP) and center of mass (CoM) were analyzed for two reactive windows after the perturbation (0-200 ms and 200-700 ms). The Bonferroni post hoc test indicated a higher range of CoP in the PIGD when compared to the CG (p = 0.021). The PIGD demonstrated greater time to recover the stable posture compared to the TD (p = 0.017) and CG (p = 0.003). Furthermore, the TD showed higher AP-acceleration peak of CoM when compared to the PIGD (p = 0.048) and CG (p = 0.013), and greater AP-acceleration range of CoM in relation to the CG (p = 0.022). These findings suggest that PD patients present worse reactive postural control after perturbation compared to healthy older individuals. CoP and CoM parameters are sensitive to understand and detect the differences in reactive postural mechanisms in PD subtypes.


Concurrent cervical and craniofacial pain. A review of empiric and basic science evidence.
Because many patients present themselves for treatment with both craniofacial and craniocervical pain, 2 questions arise: (1) What are the sensory and motor consequences of dysfunction in either of these areas on the other? (2) Do craniofacial and craniocervical pain have a similar cause? These questions formed the impetus for this review article. The phenomenon of concurrent pain in craniofacial and cervical structures is considered, and clinical reports and opinions are presented regarding theories of cervical-to-craniofacial and craniofacial-to-cervical pain referral. Because pain referral between these 2 areas requires anatomic and functional connectivity between trigeminally and cervically innervated structures, basic neurophysiologic and neuroanatomic literature is reviewed. The published data clearly demonstrate neurophysiologic and structural convergence of cervical sensory and muscle afferent inputs onto trigeminal subnucleus caudalis nociceptive and non-nociceptive neurons. Moreover, changes in metabolic activity and blood flow in the brainstem and cervical dorsal horn of the spinal cord in both monkeys and cats have been demonstrated after electric stimulation of the V1-innervated superior sagittal sinus. In conclusion, the animal experimental data support the findings of human empiric and experimental studies, which suggest that strong connectivity exists between trigeminal and cervical motor and sensory responses.


Human research subjects as human research workers.
Biomedical research involving human subjects has traditionally been treated as a unique endeavor, presenting special risks and demanding special protections. But in several ways, the regulatory scheme governing human subjects research is counter-intuitively less protective than the labor and employment laws applicable to many workers. This Article relies on analogical and legal reasoning to demonstrate that this should not be the case; in a number of ways, human research subjects ought to be fundamentally recast as human research workers. Like other workers protected under worklaw, biomedical research subjects often have interests that diverge from those in positions of control but little bargaining power for change. Bearing these important similarities in mind, the question becomes whether there is any good reason to treat subjects and protected workers differently as a matter of law. With regard to unrestricted payment, eligibility for a minimum wage, compensation for injury, and rights to engage in concerted activity, the answer is no and human subjects regulations ought to be revised accordingly.


The circular stapling device as a risk factor for anastomotic leakage.
To investigate the relation between the type of circular stapler and anastomotic leak in rectal cancer surgery. During the past decades results from rectal cancer surgery have improved considerably regarding risk of local recurrence and survival. Two main paradigm changes are considered to be the cause for this: the introduction of total mesorectal excision (TME) and the increasing use of radiotherapy. However, rectal cancer surgery is associated with an unacceptably high frequency of complications of which anastomotic leak is one of the most severe ones. The hypothesis was raised that the choice of stapler influenced the leakage rates. A questionnaire was sent to all departments of surgery (n = 66) performing rectal cancer surgery in Sweden to determine the choice of circular stapler when performing anterior resection for rectal cancer. These data were linked to the Swedish Rectal Cancer Registry for the period 1995-99. A total of 3316 patients had an anterior resection. The choice of circular stapling device was determined in 70% of the cases. When stapler A was used, the leakage rate was 11% whereas it was 7% when stapler B was used (P = 0.0039). In the cases where it was impossible to determine which stapler had been used the leakage rate was 8%. Quality control is an important part of medicine and the present study suggests that it also must include surgical instruments. A prospective randomised study is needed to confirm the results.


High levels of hyaluronan in idiopathic pulmonary arterial hypertension.
Hyaluronan (HA), a large glycosaminoglycan found in the ECM, has major roles in lung and vascular biology and disease. However, its role in idiopathic pulmonary arterial hypertension (IPAH) is unknown. We hypothesized that HA metabolism is abnormal in IPAH. We measured the plasma levels of HA in IPAH and healthy individuals. We also evaluated HA synthesis and the expression of HA synthases and hyaluronidases in pulmonary artery smooth muscle cells (PASMCs) from explanted lungs. Plasma HA levels were markedly elevated in IPAH compared with controls [HA (ng/ml, mean +/- SD): IPAH 325 +/- 80, control 28 +/- 9; P = 0.02]. In vitro, unstimulated IPAH PASMCs produced high levels of HA compared with control cells [HA in supernatant (microg/ml, mean +/- SD): IPAH 12 +/- 2, controls 6 +/- 0.9; P = 0.04]. HA levels were also higher in IPAH PASMC lysates. The increased HA was biologically relevant as shown by tissue staining and increased HA-specific binding of mononuclear cells to IPAH compared with control PASMCs [number of bound cells x 10(4) (mean +/- SD): IPAH 9.5 +/- 3, control 3.0 +/- 1; P = 0.01]. This binding was abrogated by the addition of hyaluronidase. HA synthase-2 and hyaluronidase-2 were predominant in control and IPAH PASMCs. Interestingly, the expressions of HA synthase-2 and hyaluronidase-2 were approximately 2-fold lower in IPAH compared with controls [HA synthase-2 (relative expression mean +/- SE): IPAH 4.3 +/- 0.02, control 7.8 +/- 0.1; P = 0.0004; hyaluronidase-2 (relative expression mean +/- SE): IPAH 4.2 +/- 0.06, control 7.6 +/- 0.07; P = 0.008]. Thus patients with IPAH have higher circulating levels of HA, and PASMCs derived from IPAH lungs produce more HA compared with controls. This is associated with increased tissue levels and increased binding of inflammatory cells suggesting a role for HA in remodeling and inflammation in IPAH.


Expression of nitric oxide, peroxynitrite, and apoptosis in loose total hip replacements.
Nitric oxide (NO) is an effector molecule associated with inflammation, immune function, bone metabolism, and the induction of apoptosis. This study examined the role of NO, peroxynitrite (ONOO(-)), and apoptosis in cases of revision total hip replacements (THRs). We hypothesized that apoptosis and excess production of NO contribute to the inflammatory reaction to orthopedic biomaterial wear debris that is associated with loosening and osteolysis. Periprosthetic membranous specimens were collected from revised cemented acetabular components with simple loosening and ballooning osteolysis. Synovial samples from patients undergoing primary THR were used as controls. The presence of macrophages (CD68(+)) and levels of inducible nitric oxide synthase (INOS), endothelial nitric oxide synthase (EcNOS), ONOO(-) (Nitro, assayed by the amount of nitrated tyrosine residues), and apoptosis (TUNEL staining) were examined using immunohistochemistry. Increased expression for INOS, EcNOS, and ONOO(-) in both the loose/osteolytic and the loose/non-osteolytic groups was observed when compared to the synovium group. There were no significant differences between the loose/osteolytic group and loose/non-osteolytic group for these biologic markers. TUNEL staining showed a significant increase in apoptosis in the loose/osteolytic group compared to the loose/non-osteolytic group and synovial tissues. These findings suggest that NO and NO-derived molecules, such as ONOO(-), may be involved in sustaining the foreign-body reaction to wear debris. NO and ONOO(-) may prove to be useful markers of prosthetic loosening whereas apoptosis may be a marker distinguishing ballooning from simple osteolysis.


Review of the toxicology of chlorpyrifos with an emphasis on human exposure and neurodevelopment.
This review examines the large body of toxicological and epidemiological information on human exposures to chlorpyrifos, with an emphasis on the controversial potential for chlorpyrifos to induce neurodevelopmental effects at low doses. The results of this review demonstrate that the use of urinary 3,5,6-trichlorpyridinol (TCPy), a metabolite of chlorpyrifos as a biomarker of nonoccupational exposure is problematic and may overestimate nonoccupational exposures to chlorpyrifos by 10-to 20-fold because of the widespread presence of both TCPy and chlorpyrifos-methyl in the food supply. Current "background" (nonoccupational) levels of exposure to chlorpyrifos are several orders of magnitude lower than those required to inhibit plasma cholinesterase activity, which is a more sensitive target than nervous system cholinesterase. However, several in vitro studies have identified putative neurodevelopmental mechanisms that are altered at concentrations of chlorpyrifos below those that inhibit cholinesterases. Although one human cohort study reported an association between maternal and cord blood chlorpyrifos levels and several measures of neurodevelopment, two other cohort studies that utilized urinary TCPy as a surrogate for chlorpyrifos exposure did not demonstrate an association. Although the weight of the scientific evidence demonstrates that current levels of chlorpyrifos exposure will not have any adverse effects on neurodevelopment that might result from inhibition of nervous system cholinesterases, several recent studies propose alternative mechanisms. Thus, further in vivo investigation on neurodevelopment in an appropriate animal model is needed; additional epidemiological studies may be warranted if a suitable, chlorpyrifos-exposed cohort can be identified and more rigorous measures of exposure are utilized.


The stress response protein REDD1 promotes diabetes-induced oxidative stress in the retina by Keap1-independent Nrf2 degradation.
The transcription factor nuclear factor erythroid-2-related factor 2 (Nrf2) plays a critical role in reducing oxidative stress by promoting the expression of antioxidant genes. Both individuals with diabetes and preclinical diabetes models exhibit evidence of a defect in retinal Nrf2 activation. We recently demonstrated that increased expression of the stress response protein regulated in development and DNA damage 1 (REDD1) is necessary for the development of oxidative stress in the retina of streptozotocin-induced diabetic mice. In the present study, we tested the hypothesis that REDD1 suppresses the retinal antioxidant response to diabetes by repressing Nrf2 function. We found that REDD1 ablation enhances Nrf2 DNA-binding activity in the retina and that the suppressive effect of diabetes on Nrf2 activity is absent in the retina of REDD1-deficient mice compared with WT. In human MIO-M1 Müller cell cultures, REDD1 deletion prevented oxidative stress in response to hyperglycemic conditions, and this protective effect required Nrf2. REDD1 suppressed Nrf2 stability by promoting its proteasomal degradation independently of Nrf2's interaction with Kelch-like ECH-associated protein 1 (Keap1), but REDD1-mediated Nrf2 degradation required glycogen synthase kinase 3 (GSK3) activity and Ser-351/Ser-356 of Nrf2. Diabetes diminished inhibitory phosphorylation of glycogen synthase kinase 3β (GSK3β) at Ser-9 in the retina of WT mice but not in REDD1-deficient mice. Pharmacological inhibition of GSK3 enhanced Nrf2 activity and prevented oxidative stress in the retina of diabetic mice. The findings support a model wherein hyperglycemia-induced REDD1 blunts the Nrf2 antioxidant response to diabetes by activating GSK3, which, in turn, phosphorylates Nrf2 to promote its degradation.


Anti-Abortion Exceptionalism after Dobbs.
The end of the constitutional right to abortion with Dobbs v. Jackson Women's Health stands to generate massive conflict between abortion regulation and the First Amendment. Abortion exceptionalism within constitutional doctrine -- which both treats abortion differently than other areas and favors anti-abortion over pro-choice viewpoints -- will not retreat but advance, unless confronted by the courts.


Corticospinal and spinal adaptations to motor skill and resistance training: Potential mechanisms and implications for motor rehabilitation and athletic development.
Optimal strategies for enhancing strength and improving motor skills are vital in athletic performance and clinical rehabilitation. Initial increases in strength and the acquisition of new motor skills have long been attributed to neurological adaptations. However, early increases in strength may be predominantly due to improvements in inter-muscular coordination rather than the force-generating capacity of the muscle. Despite the plethora of research investigating neurological adaptations from motor skill or resistance training in isolation, little effort has been made in consolidating this research to compare motor skill and resistance training adaptations. The findings of this review demonstrated that motor skill and resistance training adaptations show similar short-term mechanisms of adaptations, particularly at a cortical level. Increases in corticospinal excitability and a release in short-interval cortical inhibition occur as a result of the commencement of both resistance and motor skill training. Spinal changes show evidence of task-specific adaptations from the acquired motor skill, with an increase or decrease in spinal reflex excitability, dependant on the motor task. An increase in synaptic efficacy of the reticulospinal projections is likely to be a prominent mechanism for driving strength adaptations at the subcortical level, though more research is needed. Transcranial electric stimulation has been shown to increase corticospinal excitability and augment motor skill adaptations, but limited evidence exists for further enhancing strength adaptations from resistance training. Despite the logistical challenges, future work should compare the longitudinal adaptations between motor skill and resistance training to further optimise exercise programming.


Injection of Adipose-Derived Stromal Cells in the Knee of Patients with Severe Osteoarthritis has a Systemic Effect and Promotes an Anti-Inflammatory Phenotype of Circulating Immune Cells.
Rationale: Recent studies confirmed that osteoarthritis (OA) is associated with systemic inflammation. Adipose-derived stromal cells (ASCs) could become the most promising cell-based therapy in OA, based not only on their differentiation capacities and trophic and paracrine effects on the existing cartilage, but also on their immunomodulatory properties. Here, we wanted to determine the biological effect of autologous ASC intra-articular (IA) injection. Method: To this aim, we monitored the profile of immune cells in fresh peripheral blood after IA injection of autologous ASCs in the knee of 18 patients with severe OA (ADIPOA phase I study). Specifically, we used 8-color flow cytometry antibody panels to characterize the frequencies of innate and adaptive immune cell subsets (monocytes, dendritic cells, regulatory T cells and B cells) in blood samples at baseline (before injection) and one week, one month and three months after ASC injection. Results: We found that the percentage of CD4+CD25highCD127lowFOXP3+ regulatory T cells was significantly increased at 1 month after ASC injection, and this effect persisted for at least 3 months. Moreover, CD24highCD38high transitional B cells also were increased, whereas the percentage of classical CD14+ monocytes was decreased, at 3 months after ASC injection. These results suggest a global switch toward regulatory immune cells following IA injection of ASCs, underscoring the safety of ASC-based therapy. We did not find any correlation between the scores for the Visual Analogic Scale for pain, the Western Ontario and McMaster Universities Osteoarthritis Index (pain subscale and total score) at baseline and the immune cell profile changes, but this could be due to the small number of analyzed patients. Conclusion: ASCs may drive an immediate local response by releasing paracrine factors and cytokines, and our results suggest that ASCs could also initiate a cascade resulting in a long-lasting systemic immune modulation.


Parent and family perspectives on home-based newborn care practices in lower-income countries: a systematic review of qualitative studies.
To understand family and parent perspectives on newborn care provided at home to infants in the first 28 days of life, in order to inform behavioural interventions for improving care in low-income countries, where the majority of newborn deaths occur. A comprehensive, qualitative systematic review was conducted. MEDLINE/PubMed, Embase and Cumulative Index of Nursing and Allied Health databases were systematically searched for studies examining the views of parents and family members on newborn care at home. The search period included all studies published from 2006 to 2017. Studies using qualitative approaches or mixed-methods studies with substantial use of qualitative techniques in both the methods and analysis sections were included. Studies meeting the inclusion criteria were extracted and evaluated using Critical Appraisal Skills Programme guidelines. Following the initial selection and appraisal, barriers and facilitators to recommended care practices across several domains were synthesised. Of 411 results retrieved, 37 met both inclusion and quality appraisal criteria for methodology and reporting. Geographical representation largely reflected that of newborn health outcomes globally, with the majority of studies conducted in the region of Sub-Saharan Africa and South Asia. Specific barriers and facilitators were identified among a range of domains including: cord care, drying and wrapping, thermal control, skin to skin contact, hygiene, breast feeding, care-seeking for illness, and low birthweight recognition. Cross cutting facilitators, common to all domains were evident and includeddelivery at a health facility, inclusion of female relatives in care counselling, lower healthcare costs, and exposure to newborn care behaviour change messaging in the community. When designing behavioural interventions to address newborn mortality at scale, policy-makers and practitioners must include barriers and facilitators important to families in low-income settings. CRD42016035674.


Endocrine Resistance in Hormone Receptor Positive Breast Cancer-From Mechanism to Therapy.
The importance and role of the estrogen receptor (ER) pathway has been well-documented in both breast cancer (BC) development and progression. The treatment of choice in women with metastatic breast cancer (MBC) is classically divided into a variety of endocrine therapies, 3 of the most common being: selective estrogen receptor modulators (SERM), aromatase inhibitors (AI) and selective estrogen receptor down-regulators (SERD). In a proportion of patients, resistance develops to endocrine therapy due to a sophisticated and at times redundant interference, at the molecular level between the ER and growth factor. The progression to endocrine resistance is considered to be a gradual, step-wise process. Several mechanisms have been proposed but thus far none of them can be defined as the complete explanation behind the phenomenon of endocrine resistance. Although multiple cellular, molecular and immune mechanisms have been and are being extensively studied, their individual roles are often poorly understood. In this review, we summarize current progress in our understanding of ER biology and the molecular mechanisms that predispose and determine endocrine resistance in breast cancer patients.


Identification of residues crucially involved in soluble guanylate cyclase activation.
The ubiquitous heterodimeric nitric oxide (NO) receptor soluble guanylate cyclase (sGC) plays a key role in various signal transduction pathways. Binding of NO takes place at the prosthetic heme moiety at the N-terminus of the beta(1)-subunit of sGC. The induced structural changes lead to an activation of the catalytic C-terminal domain of the enzyme and to an increased conversion of GTP into the second messenger cyclic GMP (cGMP). In the present work we selected and substituted different residues of the sGC heme-binding pocket based on a sGC homology model. The generated sGC variants were tested in a cGMP reporter cell for their effect on the enzyme activation by heme-dependent (NO, BAY 41-2272) stimulators and heme-independent (BAY 58-2667) activators. The use of these experimental tools allows the enzyme's heme content to be explored in a non-invasive manner. Asp(44), Asp(45) and Phe(74) of the beta(1)-subunit were identified as being crucially important for functional enzyme activation. beta(1)Asp(45) may serve as a switch between different conformational states of sGC and point to a possible mechanism of action of the heme dependent sGC stimulator BAY 41-2272. Furthermore, our data shows that the activation profile of beta(1)IIe(145) Tyr is unchanged compared to the native enzyme, suggesting that Tyr(145) does not confer the ability to distinguish between NO and O(2). In summary, the present work further elucidated intramolecular mechanisms underlying the NO- and BAY 41-2272-mediated sGC activation and raises questions regarding the postulated role of Tyr(145) for ligand discrimination.


Study of coinfection with local strains of infectious bursal disease virus and infectious bronchitis virus in specific pathogen-free chickens.
Immunosuppressive diseases cause great losses in the poultry industry, increasing the susceptibility to infections by other pathogens and promoting a suboptimal response to vaccination. Among them, infectious bursal disease virus (IBDV) arises as one of the most important around the world. IBDV infects immature B lymphocytes, affecting the immune status of birds and facilitating infections by other pathogens such as avian infectious bronchitis virus (IBV). Although it has been reported that the interaction between these viruses increases IBV clinical signs, there are no actual studies about the interaction between regional circulating isolates that validate this statement. In this context, the objective of our work was to evaluate the effect of the interaction between local isolates of IBDV (belonging to genogroup 4) and IBV (lineage GI-16) in chickens. Thus, specific pathogen-free chickens were orally inoculated with IBDV genogroup (G) 4 or with PBS at 5 d of age. At 14-days postinoculation (dpi) the animals were intratracheally inoculated with a GI-16 IBV or with PBS. At multiple time points, groups of birds were euthanized and different parameters such as histological damage, viral load, lymphocyte populations and specific antibodies were evaluated. The success of IBDV infection was confirmed by the severity of bursal atrophy, viral detection, and presence of anti-IBDV antibodies. In IBV-infected animals, the presence of viral genome was detected in both kidney and bursa. The coinfected animals showed higher degree of lymphocyte infiltration in kidney, higher rate of animals with IBV viral genome in bursa at 28 dpi, and a clear decrease in antibody response against IBV at 28, 35, and 40 dpi. The results indicate that the infection with the local isolate of IBDV affects the immune status of the chickens, causing major severe damage, in response to IBV infection, which could consequently severely affect the local poultry industry.


Adoptive transfer of EBV specific CD8+ T cell clones can transiently control EBV infection in humanized mice.
Epstein Barr virus (EBV) infection expands CD8+ T cells specific for lytic antigens to high frequencies during symptomatic primary infection, and maintains these at significant numbers during persistence. Despite this, the protective function of these lytic EBV antigen-specific cytotoxic CD8+ T cells remains unclear. Here we demonstrate that lytic EBV replication does not significantly contribute to virus-induced B cell proliferation in vitro and in vivo in a mouse model with reconstituted human immune system components (huNSG mice). However, we report a trend to reduction of EBV-induced lymphoproliferation outside of lymphoid organs upon diminished lytic replication. Moreover, we could demonstrate that CD8+ T cells against the lytic EBV antigen BMLF1 can eliminate lytically replicating EBV-transformed B cells from lymphoblastoid cell lines (LCLs) and in vivo, thereby transiently controlling high viremia after adoptive transfer into EBV infected huNSG mice. These findings suggest a protective function for lytic EBV antigen-specific CD8+ T cells against EBV infection and against virus-associated tumors in extra-lymphoid organs. These specificities should be explored for EBV-specific vaccine development.


Varenicline and cytisine: two nicotinic acetylcholine receptor ligands reduce ethanol intake in University of Chile bibulous rats.
Neuronal nicotinic acetylcholine receptors (nAChRs) are pharmacological targets that have recently been implicated in the reinforcing effects of many drugs of abuse, including ethanol. Varenicline and cytisine are nAChR partial agonists in clinical use as smoking cessation aids. However, their efficacies to reduce alcohol consumption have not been fully studied. This study aims to compare the effects of varenicline and cytisine on ethanol consumption by rats bred for many generations as high ethanol drinkers (UChB). Repeated dosing (0.5 or 1.0 mg/kg/day i.p.) of varenicline or cytisine, for three consecutive days, to male UChB rats pre-exposed to 10 % (v/v) ethanol and water 24 h/day for 4 weeks, significantly reduced alcohol intake and preference of ethanol over water during 1- and 24-h ethanol access periods. This effect was specific for ethanol intake and was not observed for 0.2 % saccharin or water consumption. Varenicline appears to be more effective than cytisine, probably due to its more favorable pharmacokinetic and pharmacodynamic properties. Long-term use of both nAChRs ligands for more than 8-10 days induced tolerance to their effects on ethanol consumption. This preclinical study in UChB rats demonstrated that both varenicline and cytisine reduce alcohol intake, with varenicline producing a greater and longer-lasting reduction than cytisine. However, dose adjustment will have to be considered as a possible way to counter tolerance arising after continued use.


Regulation of the adhE gene, which encodes ethanol dehydrogenase in Escherichia coli.
The respiratory control of adhE, which encodes ethanol (alcohol) dehydrogenase in Escherichia coli, was examined at the transcriptional level by using various phi (adhE'-lacZ) adhE+ merodiploid strains. Expression of the adhE'-lacZ operon fusion was increased about eightfold by anaerobic growth. Under anaerobic growth conditions, provision of nitrate lowered the level of expression. Nitrate repression was more severe than aerobic repression. From analyses of various mutants with mutations related to nitrate reduction, nitrate repression appears to result from two effects. (i) When nitrate was present, NarL, the positive regulator of the nar operon, exerted a direct repression on adhE expression, which was demonstrable even aerobically. (ii) The chemical reduction of nitrate exerted an indirect effect by altering the cellular redox potential.


Stress echocardiography expert consensus statement: European Association of Echocardiography (EAE) (a registered branch of the ESC).
Stress echocardiography is the combination of 2D echocardiography with a physical, pharmacological or electrical stress. The diagnostic end point for the detection of myocardial ischemia is the induction of a transient worsening in regional function during stress. Stress echocardiography provides similar diagnostic and prognostic accuracy as radionuclide stress perfusion imaging, but at a substantially lower cost, without environmental impact, and with no biohazards for the patient and the physician. Among different stresses of comparable diagnostic and prognostic accuracy, semisupine exercise is the most used, dobutamine the best test for viability, and dipyridamole the safest and simplest pharmacological stress and the most suitable for combined wall motion coronary flow reserve assessment. The additional clinical benefit of myocardial perfusion contrast echocardiography and myocardial velocity imaging has been inconsistent to date, whereas the potential of adding - coronary flow reserve evaluation of left anterior descending coronary artery by transthoracic Doppler echocardiography adds another potentially important dimension to stress echocardiography. New emerging fields of application taking advantage from the versatility of the technique are Doppler stress echo in valvular heart disease and in dilated cardiomyopathy. In spite of its dependence upon operator's training, stress echocardiography is today the best (most cost-effective and risk-effective) possible imaging choice to achieve the still elusive target of sustainable cardiac imaging in the field of noninvasive diagnosis of coronary artery disease.


Feedback interventions for improving self-awareness after brain injury: a protocol for a pragmatic randomised controlled trial.
Occupational therapists working in brain injury rehabilitation use functional tasks as a means of providing feedback to improve self-awareness of people who have a brain injury and ultimately improve their occupational performance. To compare the effectiveness of video, verbal and experiential feedback for improving self-awareness in people with traumatic brain injury. A randomised controlled trial will be conducted to compare the efficacy of video and verbal feedback during occupational therapy. Fifty-four participants with traumatic brain injury will be randomly allocated into three intervention groups: (i) video plus verbal feedback, (ii) verbal feedback and (iii) experiential feedback (control condition). Participants will receive the allocated intervention based on performance of a meal preparation task. The intervention sessions will occur four times during a two-week period. Blinded assessment will occur at baseline, post-intervention, and two months follow up. The primary outcome will be a measure of on-line self-awareness (number of self-corrected and therapist corrected errors). Secondary outcomes to be assessed include levels of intellectual self-awareness, emotional distress, and acceptance of disability. Data will be analysed using an intention to treat approach. Linear mixed effects models will be used to investigate the intervention effects. Results will contribute to evidence-based guidelines to support therapists to choose the most effective form of feedback for people with decreased self-awareness after traumatic brain injury.


Paraspinal muscle imaging measurements for common spinal disorders: review and consensus-based recommendations from the ISSLS degenerative spinal phenotypes group.
Paraspinal muscle imaging is of growing interest related to improved phenotyping, prognosis, and treatment of common spinal disorders. We reviewed issues related to paraspinal muscle imaging measurement that contribute to inconsistent findings between studies and impede understanding. Three key contributors to inconsistencies among studies of paraspinal muscle imaging measurements were reviewed: failure to consider possible mechanisms underlying changes in paraspinal muscles, lack of control of confounding factors, and variations in spinal muscle imaging modalities and measurement protocols. Recommendations are provided to address these issues to improve the quality and coherence of future research. Possible pathophysiological responses of paraspinal muscle to various common spinal disorders in acute or chronic phases are often overlooked, yet have important implications for the timing, distribution, and nature of changes in paraspinal muscle. These considerations, as well as adjustment for possible confounding factors, such as sex, age, and physical activity must be considered when planning and interpreting paraspinal muscle measurements in studies of spinal conditions. Adoption of standardised imaging measurement protocols for paraspinal muscle morphology and composition, considering the strengths and limitations of various imaging modalities, is critically important to interpretation and synthesis of research. Study designs that consider physiological and pathophysiological responses of muscle, adjust for possible confounding factors, and use common, standardised measures are needed to advance knowledge of the determinants of variations or changes in paraspinal muscle and their influence on spinal health.


Left A2 Fusiform Aneurysm: 2-Dimensional Operative Video.
Anterior cerebral artery aneurysms within the A2 segment that are fusiform or giant A2 aneurysms with a wide neck often are not candidates for endovascular treatment and require surgical intervention. These lesions necessitate a bypass procedure to preserve distal flow along the anterior cerebral artery. This patient demonstrated a left-sided unruptured A2 fusiform aneurysm that necessitated a bypass procedure for management. The patient was positioned with the head rotated laterally to permit orientation of the interhemispheric fissure within the horizontal plane and achieve gravity retraction of the dependent hemisphere. The bilateral pericallosal branches were exposed and liberated from arachnoid adhesions to permit mobilization necessary for the side-to-side anastomosis. While the anastomosis was performed, the continuous suture loops were left loose to permit complete visualization of the inner and outer walls prior to the final tightening and tying of the anastomotic suture. Following the completion of the anastomosis, the temporary clips were removed. A permanent clip was placed on the distal A2 to prevent outflow from the parent artery and thereby allow for aneurysm thrombosis. The patient gave informed consent for surgery and video recording. Institutional review board approval was deemed unnecessary. Used with permission from Barrow Neurological Institute, Phoenix, Arizona.


[Reconstructive-plastic operations as one of the measures for the medical rehabilitation of lung cancer patients].
The follow-up result of reconstructive-plastic operations in 68 patients with pulmonary cancer are studied. Preoperative irradiation therapy (20 Gr the summary dose), using middle fractions, was conducted in 21 patients. While doing the operation, pleura was worked by polymethylsiloxane-400 in 32 patients for the postoperative adhesive pleuritis prophylaxis. The patients survival index after reconstructive-plastic operations did not differ from such in a control group.


Cell pH regulation in rabbit outer medullary collecting duct cells: mechanisms of HCO3(-)-independent processes.
To clarify mechanisms of intracellular pH (pHi) regulation in outer stripe of outer medullary collecting duct (OMCDOS), isolated perfused OMCDOS of the rabbit were loaded with 2',7'-bis(carboxyethyl)-5(6)-carboxyfluorescein (BCECF), and single cell pHi was monitored by an image processing system. Initial pHi recovery rates (dpHi/dt, pH unit/s x 10(3)) after intracellular acid load made by NH4Cl prepulse were determined. In the absence of exogenous CO2-HCO3-, dpHi/dt was 12.3 +/- 0.9 (means +/- SE) in principal cells (PC), and 11.5 +/- 1.0 in intercalated cells (IC). In PC, total ambient Na+ removal halted pHi recovery (dpHi/dt = 0.6 +/- 0.5), and pHi recovered when Na+ was added to the basolateral (dpHi/dt = 14.7 +/- 0.8) but not to the luminal (dpHi/dt = 0.9 +/- 0.5) solutions. This bath Na+ effect was amiloride inhibitable. In IC, pHi recovered (dpHi/dt = 6.4 +/- 0.3) in the absence of ambient Na+. This pHi recovery was significantly reduced by luminal 0.5 mM N-ethylmaleimide (NEM) or 0.5 mM N,N'-dicyclohexylcarbodiimide (DCCD). Basolateral NEM or DCCD had no significant effect. Basolateral addition of Na+ significantly accelerated the pHi recovery. These data suggest the presence of basolateral Na(+)-H+ exchange in both PC and IC, and luminal NEM- and DCCD-sensitive H+ pump in IC of rabbit OMCDOS.


Evaluation of the effect of a supratherapeutic dose of intravenous ceftaroline fosamil on the corrected QT interval.
A randomized, double-blind, placebo-controlled, 3-period crossover study was conducted in 54 healthy adults to assess the effect of ceftaroline fosamil on the corrected QT (QTc) interval. The QT interval, corrected for heart rate using an individual correction formula (QTcIb), was determined predose and at 1, 1.25, 1.5, 2, 4, 8, 12, and 24.5 h after intravenous dosing with a supratherapeutic dose (1,500 mg) of ceftaroline fosamil, 400 mg moxifloxacin (positive control), and placebo. The pharmacokinetic profile of ceftaroline was also evaluated. At each time point following ceftaroline fosamil administration, the upper limit of the 90% confidence interval (CI) for the placebo-corrected change from predose baseline in QTcIb (ΔΔQTcIb) was below 10 ms (maximum, 3.4 ms at 1.5 h after dosing), indicating an absence of clinically meaningful QTc increase. The lower limit of the 90% CI of ΔΔQTcIb for moxifloxacin versus placebo was greater than 5 ms at 5 time points (maximum, 12.8 ms at 1 h after dosing), demonstrating assay sensitivity. There was no apparent correlation between ceftaroline plasma concentrations and ΔΔQTcIb. The supratherapeutic dose of ceftaroline fosamil (1,500 mg) resulted in substantially greater systemic exposure to ceftaroline than previously observed with standard therapeutic doses. Ceftaroline fosamil was well tolerated after a single 1,500-mg intravenous dose, and no clinically meaningful abnormalities in laboratory values or vital signs were observed.


Cystatins.
Chicken egg white cystatin was first described in the late 1960s. Since then, our knowledge about a superfamily of similar proteins present in mammals, birds, fish, insects, plants and some protozoa has expanded, and their properties as potent peptidase inhibitors have been firmly established. Today, 12 functional chicken cystatin relatives are known in humans, but a few evolutionarily related gene products still remain to be characterized. The type 1 cystatins (A and B) are mainly intracellular, the type 2 cystatins (C, D, E/M, F, G, S, SN and SA) are extracellular, and the type 3 cystatins (L- and H-kininogens) are intravascular proteins. All true cystatins inhibit cysteine peptidases of the papain (C1) family, and some also inhibit legumain (C13) family enzymes. These peptidases play key roles in physiological processes, such as intracellular protein degradation (cathepsins B, H and L), are pivotal in the remodelling of bone (cathepsin K), and may be important in the control of antigen presentation (cathepsin S, mammalian legumain). Moreover, the activities of such peptidases are increased in pathophysiological conditions, such as cancer metastasis and inflammation. Additionally, such peptidases are essential for several pathogenic parasites and bacteria. Thus cystatins not only have capacity to regulate normal body processes and perhaps cause disease when down-regulated, but may also participate in the defence against microbial infections. In this chapter, we have aimed to summarize our present knowledge about the human cystatins.


Bidirectional Expression of Metabolic, Structural, and Immune Pathways in Early Myopia and Hyperopia.
Myopia (short-sightedness) affects 1.45 billion people worldwide, many of whom will develop sight-threatening secondary disorders. Myopic eyes are characterized by excessive size while hyperopic (long-sighted) eyes are typically small. The biological and genetic mechanisms underpinning the retina's local control of these growth patterns remain unclear. In the present study, we used RNA sequencing to examine gene expression in the retina/RPE/choroid across 3 days of optically-induced myopia and hyperopia induction in chick. Data were analyzed for differential expression of single genes, and Gene Set Enrichment Analysis (GSEA) was used to identify gene sets correlated with ocular axial length and refraction across lens groups. Like previous studies, we found few single genes that were differentially-expressed in a sign-of-defocus dependent manner (only BMP2 at 1 day). Using GSEA, however, we are the first to show that more subtle shifts in structural, metabolic, and immune pathway expression are correlated with the eye size and refractive changes induced by lens defocus. Our findings link gene expression with the morphological characteristics of refractive error, and suggest that physiological stress arising from metabolic and inflammatory pathway activation could increase the vulnerability of myopic eyes to secondary pathologies.


CoReumaPt protocol: the Portuguese cohort of rheumatic diseases.
Rheumatic diseases (RD) are conditions with a variety of clinical manifestations and prognosis influenced by several factors. Cohorts and registries have been already established in some countries and have contributed to important knowledge about the disease course and the long-term outcomes of RD. This paper introduces the CoReumaPt project and sets the first step towards the creation of a prospective cohort study including the main RD occurring in the Portuguese population. CoReumaPt will allow outcomes research of chronic RD and the assessment of factors influencing the development and progression of RD. It will also allow to further evaluate the economic impact and the burden of RD in Portugal. CoReumaPt will be linked to Reuma.pt, the National Register of Rheumatic Diseases from the Portuguese Society of Rheumatology. An open cohort will be created, initially composed by the randomly selected population of the crosssectional National Epidemiological Rheumatic Diseases study (EpiReumaPt) and afterwards by other sources, namely through self- and physician's referral. Follow-up with annual self-administered questionnaires will be performed, in order to systematically collect and analyze outcomes of interest, mainly patient-reported outcomes. Data concerning less frequent assessments, such as radiographs and biomarkers, will also be assembled. CoReumaPt will be a valuable resource for scientific research and will deliver pivotal information to improve public health policies concerning the prevention and the management of RD in Portugal.


Using Grain-Size Distribution Methods for Estimation of Air Permeability.
Knowledge of air permeability (ka ) at dry conditions is critical for the use of air flow models in porous media; however, it is usually difficult and time consuming to measure ka at dry conditions. It is thus desirable to estimate ka at dry conditions from other readily obtainable properties. In this study, the feasibility of using information derived from grain-size distributions (GSDs) for estimating ka at dry conditions was examined. Fourteen GSD-based equations originally developed for estimating saturated hydraulic conductivity were tested using ka measured at dry conditions in both undisturbed and disturbed river sediment samples. On average, the estimated ka from all the equations, except for the method of Slichter, differed by less than ± 4 times from the measured ka for both undisturbed and disturbed groups. In particular, for the two sediment groups, the results given by the methods of Terzaghi and Hazen-modified were comparable to the measured ka . In addition, two methods (e.g., Barr and Beyer) for the undisturbed samples and one method (e.g., Hazen-original) for the undisturbed samples were also able to produce comparable ka estimates. Moreover, after adjusting the values of the coefficient C in the GSD-based equations, the estimation of ka was significantly improved with the differences between the measured and estimated ka less than ±4% on average (except for the method of Barr). As demonstrated by this study, GSD-based equations may provide a promising and efficient way to estimate ka at dry conditions.


DNA-programmable nanoparticle crystallization.
It was first shown more than ten years ago that DNA oligonucleotides can be attached to gold nanoparticles rationally to direct the formation of larger assemblies. Since then, oligonucleotide-functionalized nanoparticles have been developed into powerful diagnostic tools for nucleic acids and proteins, and into intracellular probes and gene regulators. In contrast, the conceptually simple yet powerful idea that functionalized nanoparticles might serve as basic building blocks that can be rationally assembled through programmable base-pairing interactions into highly ordered macroscopic materials remains poorly developed. So far, the approach has mainly resulted in polymerization, with modest control over the placement of, the periodicity in, and the distance between particles within the assembled material. That is, most of the materials obtained thus far are best classified as amorphous polymers, although a few examples of colloidal crystal formation exist. Here, we demonstrate that DNA can be used to control the crystallization of nanoparticle-oligonucleotide conjugates to the extent that different DNA sequences guide the assembly of the same type of inorganic nanoparticle into different crystalline states. We show that the choice of DNA sequences attached to the nanoparticle building blocks, the DNA linking molecules and the absence or presence of a non-bonding single-base flexor can be adjusted so that gold nanoparticles assemble into micrometre-sized face-centred-cubic or body-centred-cubic crystal structures. Our findings thus clearly demonstrate that synthetically programmable colloidal crystallization is possible, and that a single-component system can be directed to form different structures.