Incidence of eating disorders in Danish psychiatric secondary healthcare 1970-2008.
Is an increased focus on eating disorders during the past few decades reflected by increasing occurrence in the psychiatric health service system. All first-time diagnoses of eating disorders identified in the Danish Central Psychiatric Research Register 1970-2008 constitute the present research database. Age-standardized rates per 100,000 inhabitants were calculated and autoregressive models were fitted for males and females separately as well as for in- and outpatients. The incidence of eating disorders diagnosed in Danish psychiatric secondary healthcare has increased considerably during a nearly 40-year period of observation both within the general category of eating disorders and also specifically for anorexia nervosa. The steepest increase is seen within females aged 15-19 years, where the highest incidences are also found. Anorexia nervosa constitutes the vast majority of all eating disorders. Throughout the time interval investigated, the number of males, however, is negligible compared to females. Most patients are seen in outpatient services, increasing towards recent years. However, the number of patients being treated as inpatients has increased linearly through the entire four decenniums investigated. Changes in registration principles, public awareness and higher acceptance of mental disorders in the public is the most obvious explanation for the incidence increase of anorexia nervosa and other eating disorders. This study does not answer whether there has been an increase in true incidence in the population.


Safety assessment of ocimum basilicum hydroalcoholic extract in wistar rats: acute and subchronic toxicity studies.
Ocimum basilicum L. is widely used in folk medicine of many countries including . Both O. basilicum and its oil extract have received considerable attention for their potential medicinal properties, but there are a few reports about possible toxicity of this plant. Therefore, in the present study, acute and subchronic toxicity of O. basilicum hydroalcohlic extract have been evaluated in Wistar rats. For the acute toxicity assessment, five groups of 10 animals (5 male, 5 female) received four different single dose of extract orally, the animals were, then, kept under observation for 14 days. For subchronic toxicity, the animals were divided into four groups (5 male, 5 female) and were gavaged daily by 50, 200 and 500 mg/kg of extract. Mortality, clinical signs, body weight changes, food and water consumption, and hematological and biochemical parameters were monitored during the study period. On the 45th day, animals were sacrificed and gross findings, weight of liver and left kidney and liver histological markers were assessed. The results of acute study indicated that LD50 of O. basilicum is higher than 5 mg/kg. In subchronic study, no adverse effects were observed on serum parameters in male and female rats. The hematological results showed a reduction in the hematocrit, platelets and RBC in both sexes. No abnormalities were observed in other parameters. Based on the results of this study, present data suggest that hematologic system could serve as a target organ in oral toxicity of this plant.


Bisphosphonate-polyaspartamide conjugates as bone targeted drug delivery systems.
Poly-hydroxy-aspartamide was used as a backbone to synthesize bisphosphonate derivatives thus achieving macromolecular carriers to be potentially used as targeting agents for bone drug delivery. Molecules bearing bisphosphonate groups, such as aminobisphosphonate (ABP) and neridronate (NRD), have been conjugated to polyaspartamide (α,β-poly(N-2-hydroxyethyl)-dl-aspartamide, PHEA), with or without a spacer (succinic acid or 6-aminocaproic acid) thus obtaining PHEA-succinate-ABP and PHEA-caproylcarbamate-ABP and PHEA-ABP and PHEA-NRD, respectively. Bisphosphonate-polymer conjugates were physico-chemically characterized using size exclusion chromatography and 1H-NMR; and their in vitro and ex vivo affinity for bone tissue has been further tested using the hydroxylapatite and rabbit bone binding assays, respectively. In vivo studies were carried out using rats to evaluate the biodistribution features of bisphosphonate-polymer conjugates in comparison with the starting PHEA. In vivo findings evidenced a suitable selectivity of bisphosphonate-polymer conjugates toward the bone tissues also as a function of time.


Induced Pluripotent Stem Cell-Derived Endothelial Cells in Insulin Resistance and Metabolic Syndrome.
Insulin resistance leads to a number of metabolic and cellular abnormalities including endothelial dysfunction that increase the risk of vascular disease. Although it has been particularly challenging to study the genetic determinants that predispose to abnormal function of the endothelium in insulin-resistant states, the possibility of deriving endothelial cells from induced pluripotent stem cells generated from individuals with detailed clinical phenotyping, including accurate measurements of insulin resistance accompanied by multilevel omic data (eg, genetic and genomic characterization), has opened new avenues to study this relationship. Unfortunately, several technical barriers have hampered these efforts. In the present review, we summarize the current status of induced pluripotent stem cell-derived endothelial cells for modeling endothelial dysfunction associated with insulin resistance and discuss the challenges to overcoming these limitations.


Smoking cessation in severe mental illness: what works?
The physical health of people with severe mental illness (SMI) is poor. Smoking-related illnesses are a major contributor to excess mortality and morbidity. An up-to-date review of the evidence for smoking cessation interventions in SMI is needed to inform clinical guidelines. We searched bibliographic databases for relevant studies and independently extracted data. Included studies were randomized controlled trials (RCTs) of smoking cessation or reduction conducted in adult smokers with SMI. Interventions were compared to usual care or placebo. The primary outcome was smoking cessation and secondary outcomes were smoking reduction, change in weight, change in psychiatric symptoms and adverse events. We included eight RCTs of pharmacological and/or psychological interventions. Most cessation interventions showed moderate positive results, some reaching statistical significance. One study compared behavioural support and nicotine replacement therapy (NRT) to usual care and showed a risk ratio (RR) of 2.74 (95% CI 1.10-6.81) for short-term smoking cessation, which was not significant at longer follow-up. We pooled five trials that effectively compared bupropion to placebo giving an RR of 2.77 (95% CI 1.48-5.16), which was comparable to Hughes et al.'s 2009 figures for general population data; RR = 1.69 (95% CI 1.53-1.85). Smoking reduction data were too heterogeneous for meta-analysis, but results were generally positive. Trials suggest few adverse events. All trials recorded psychiatric symptoms and the most significant changes favoured the intervention groups over the control groups. Treating tobacco dependence is effective in patients with SMI. Treatments that work in the general population work for those with severe mental illness and appear approximately equally effective. Treating tobacco dependence in patients with stable psychiatric conditions does not worsen mental state.


Acute bacterial meningitis caused by acute otitis media in adults: A series of 12 patients.
We conducted a retrospective chart review to characterize the outcomes of 12 patients-9 men and 3 women, aged 21 to 79 years (mean: 49)-who had been treated at our tertiary care center for acute bacterial meningitis caused by acute otitis media (ABMAO). Fever was the most common presenting sign/symptom, observed in 8 patients, followed by otalgia, neck stiffness, headache, and confusion. An opaque and bulging tympanic membrane was observed in 8 patients. Cultures were positive for Streptococcus pneumoniae in the cerebrospinal fluid, ear, and blood in 7, 5, and 3 patients, respectively. Immediate treatment included tympanocentesis, with aspirates sent for bacteriologic cultures. Seven patients (58.3%) underwent surgery; 5 were operated on early, and 2 underwent surgery at a later stage because of a suspected defect in the mastoid bone. A cortical mastoidectomy was performed in 6 of the 7 surgical patients; the remaining patient underwent a canal-wall-down procedure. Ten patients experienced a full recovery, 1 died, and 1 had a poor neurologic outcome (vegetative state); both of the latter 2 patients were older than 60 years. We conclude that early diagnosis, administration of antibiotics, and myringotomy are crucial for control of ABMAO. A cortical mastoidectomy with ventilation tube insertion can be reserved for patients who do not respond, which is common.


Phytochemical Profile and Biological Properties of Colchicum triphyllum (Meadow Saffron).
In this work, the phytochemical profile and the biological properties of Colchicum triphyllum (an unexplored Turkish cultivar belonging to Colchicaceae) have been comprehensively investigated for the first time. Herein, we focused on the evaluation of the in vitro antioxidant and enzyme inhibitory effects of flower, tuber, and leaf extracts, obtained using different extraction methods, namely maceration (both aqueous and methanolic), infusion, and Soxhlet. Besides, the complete phenolic and alkaloid untargeted metabolomic profiling of the different extracts was investigated. In this regard, ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UHPLC-QTOF-MS) allowed us to putatively annotate 285 compounds when considering the different matrix extracts, including mainly alkaloids, flavonoids, lignans, phenolic acids, and tyrosol equivalents. The most abundant polyphenols were flavonoids (119 compounds), while colchicine, demecolcine, and lumicolchicine isomers were some of the most widespread alkaloids in each extract analyzed. In addition, our findings showed that C. triphyllum tuber extracts were a superior source of both total alkaloids and total polyphenols, being on average 2.89 and 10.41 mg/g, respectively. Multivariate statistics following metabolomics allowed for the detection of those compounds most affected by the different extraction methods. Overall, C. triphyllum leaf extracts showed a strong in vitro antioxidant capacity, in terms of cupric reducing antioxidant power (CUPRAC; on average 96.45 mg Trolox Equivalents (TE)/g) and ferric reducing antioxidant power (FRAP) reducing power (on average 66.86 mg TE/g). Interestingly, each C. triphyllum methanolic extract analyzed (i.e., from tuber, leaf, and flower) was active against the tyrosinase in terms of inhibition, recording the higher values for methanolic macerated leaves (i.e., 125.78 mg kojic acid equivalent (KAE)/g). On the other hand, moderate inhibitory activities were observed against AChE and α-amylase. Strong correlations (p < 0.01) were also observed between the phytochemical profiles and the biological activities determined. Therefore, our findings highlighted, for the first time, the potential of C. triphhyllum extracts in food and pharmaceutical applications.


Pregabalin and pain after total knee arthroplasty: a double-blind, randomized, placebo-controlled, multidose trial.
Pregabalin may reduce postoperative pain and opioid use. Higher doses may be more effective, but may cause sedation and confusion. This prospective, randomized, blinded, placebo-controlled study tested the hypothesis that pregabalin reduces pain at 2 weeks after total knee arthroplasty, but increases drowsiness and confusion. Patients (30 per group) received capsules containing pregabalin (0, 50, 100, or 150 mg); two capsules before surgery, one capsule twice a day until postoperative day (POD) 14, one on POD15, and one on POD16. Multimodal analgesia included femoral nerve block, epidural analgesia, oxycodone-paracetamol, and meloxicam. The primary outcome was pain with flexion (POD14). Pregabalin did not reduce pain at rest, with ambulation, or with flexion at 2 weeks (P=0.69, 0.23, and 0.90, respectively). Pregabalin increased POD1 drowsiness (34.5, 37.9, 55.2, and 58.6% in the 0, 50, 100, and 150 mg arms, respectively; P=0.030), but did not increase confusion (0, 3.5, 0, and 3.5%, respectively; P=0.75). Pregabalin had no effect on acute or chronic pain, opioid consumption, or analgesic side-effects. Pregabalin reduced POD14 patient satisfaction [1-10 scale, median (first quartile, third quartile): 9 (8, 10), 8 (7, 10), 8 (5, 9), and 8 (6, 9.3), respectively; P=0.023). Protocol compliance was 63% by POD14 (50.0, 70.0, 76.7, and 56.7% compliance, respectively), with no effect of dose on compliance. Per-protocol analysis of compliant patients showed no effect of pregabalin on pain scores. Pregabalin had no beneficial effects, but increased sedation and decreased patient satisfaction. This study does not support routine perioperative pregabalin for total knee arthroplasty patients. ClinicalTrials.gov: http://www.clinicaltrials.gov/ct2/show/study/NCT01333956.


Pharmaceutical co-crystals: A green way to enhance drug stability and solubility for improved therapeutic efficacy.
Pharmaceutical co-crystals have gained significant attention in recent years as a promising green and sustainable method for poorly soluble drugs to improve their solubility, stability, and bioavailability. In the drug development research field, it is an extremely useful technique as it does not require a large number of synthetic steps as well a minimum amount of solvent is utilized or sometimes without solvent. This review presents a comprehensive investigation into the design, synthesis, characterization, and evaluation of pharmaceutical co-crystals. The study focuses on exploring different strategies for co-crystal formation, including co-grinding, solvent evaporation, and liquid-assisted grinding. Various characterization techniques such as SCXRD, PXRD, FTIR, and DSC were employed to confirm the formation and structural features of the co-crystals. The article also highlights the significance of understanding the intermolecular interactions within co-crystals and their influence on physicochemical properties. Furthermore, the article discusses the potential applications of pharmaceutical co-crystals in enhancing drug solubility, dissolution rate, and oral bioavailability, leading to improved therapeutic efficacy. Overall, this review provides valuable insights into the design and development of pharmaceutical co-crystals, offering a promising avenue for overcoming the difficulties brought on by poorly soluble drugs.


Effects of Digested Onion Extracts on Intestinal Gene Expression: An Interspecies Comparison Using Different Intestine Models.
Human intestinal tissue samples are barely accessible to study potential health benefits of nutritional compounds. Numbers of animals used in animal trials, however, need to be minimalized. Therefore, we explored the applicability of in vitro (human Caco-2 cells) and ex vivo intestine models (rat precision cut intestine slices and the pig in-situ small intestinal segment perfusion (SISP) technique) to study the effect of food compounds. In vitro digested yellow (YOd) and white onion extracts (WOd) were used as model food compounds and transcriptomics was applied to obtain more insight into which extent mode of actions depend on the model. The three intestine models shared 9,140 genes which were used to compare the responses to digested onions between the models. Unsupervised clustering analysis showed that genes up- or down-regulated by WOd in human Caco-2 cells and rat intestine slices were similarly regulated by YOd, indicating comparable modes of action for the two onion species. Highly variable responses to onion were found in the pig SISP model. By focussing only on genes with significant differential expression, in combination with a fold change > 1.5, 15 genes showed similar onion-induced expression in human Caco-2 cells and rat intestine slices and 2 overlapping genes were found between the human Caco-2 and pig SISP model. Pathway analyses revealed that mainly processes related to oxidative stress, and especially the Keap1-Nrf2 pathway, were affected by onions in all three models. Our data fit with previous in vivo studies showing that the beneficial effects of onions are mostly linked to their antioxidant properties. Taken together, our data indicate that each of the in vitro and ex vivo intestine models used in this study, taking into account their limitations, can be used to determine modes of action of nutritional compounds and can thereby reduce the number of animals used in conventional nutritional intervention studies.


PCSK9 activation promotes early atherosclerosis in a vascular microphysiological system.
Atherosclerosis is a primary precursor of cardiovascular disease (CVD), the leading cause of death worldwide. While proprotein convertase subtilisin/kexin 9 (PCSK9) contributes to CVD by degrading low-density lipoprotein receptors (LDLR) and altering lipid metabolism, PCSK9 also influences vascular inflammation, further promoting atherosclerosis. Here, we utilized a vascular microphysiological system to test the effect of PCSK9 activation or repression on the initiation of atherosclerosis and to screen the efficacy of a small molecule PCSK9 inhibitor. We have generated PCSK9 over-expressed (P+) or repressed (P-) human induced pluripotent stem cells (iPSCs) and further differentiated them to smooth muscle cells (viSMCs) or endothelial cells (viECs). Tissue-engineered blood vessels (TEBVs) made from P+ viSMCs and viECs resulted in increased monocyte adhesion compared to the wild type (WT) or P- equivalents when treated with enzyme-modified LDL (eLDL) and TNF-α. We also found significant viEC dysfunction, such as increased secretion of VCAM-1, TNF-α, and IL-6, in P+ viECs treated with eLDL and TNF-α. A small molecule compound, NYX-1492, that was originally designed to block PCSK9 binding with the LDLR was tested in TEBVs to determine its effect on lowering PCSK9-induced inflammation. The compound reduced monocyte adhesion in P+ TEBVs with evidence of lowering secretion of VCAM-1 and TNF-α. These results suggest that PCSK9 inhibition may decrease vascular inflammation in addition to lowering plasma LDL levels, enhancing its anti-atherosclerotic effects, particularly in patients with elevated chronic inflammation.


Posterior cortical atrophy: visuomotor deficits in reaching and grasping.
Posterior Cortical Atrophy (PCA) is a rare clinical syndrome characterized by the predominance of higher-order visual disturbances such as optic ataxia, a characteristic of Balint's syndrome. Deficits result from progressive neurodegeneration of occipito-temporal and occipito-parietal cortices. The current study sought to explore the visuomotor functioning of four individuals with PCA by testing their ability to reach out and grasp real objects under various viewing conditions. Experiment 1 had participants reach out and grasp simple, rectangular blocks under visually- and memory-guided conditions. Experiment 2 explored participants' abilities to accurately reach for objects located in their visual periphery. This investigation revealed that PCA patients demonstrate many of the same deficits that have been previously reported in other individuals with optic ataxia, such as "magnetic misreaching"-a pathological reaching bias toward the point of visual fixation when grasping peripheral targets. Unlike many other individuals with optic ataxia, however, the patients in the current study also show symptoms indicative of damage to the more perceptual stream of visual processing, including abolished grip scaling during memory-guided grasping and deficits in face and object identification. These investigations are the first to perform a quantitative analysis of the visuomotor deficits exhibited by patients with PCA. Critically, this study helps characterize common symptoms of PCA, a vital first step for generating effective diagnostic criteria and therapeutic strategies for this understudied neurodegenerative disorder.


Measuring and understanding treatment effectiveness in hand surgery.
Incorporating evidence-based medicine into practice is now an expectation for hand surgeons. Hand surgeons need to be able to assess associated benefits, risks, cost, and applicability of a treatment option when providing care to their patients. Using a clinical example, this article takes the reader through the three-step approach when using a publication from the medical literature on therapy. The focus of this article is primarily the second and third steps, which involve measuring and understanding treatment effectiveness.


New species in the genus Monoecocestus (Cestoda: Anoplocephalidae) from neotropical rodents (Caviidae and Sigmodontinae).
Anoplocephalid cestodes have a worldwide distribution, but relatively few species are known from South American rodents. By examining the collections of the Harold W. Manter Laboratory of Parasitology and the United States National Parasite Collection, 6 new species of Monoecocestus Beddard, 1914, are described, along with a redescription of Monoecocestus mackiewiczi Schmidt and Martin, 1978, based on the type specimens. The discussion includes commentary about uterine development, an important taxonomic character of the family, the vaginal dilation in immature segments (a character of potential taxonomic importance), and the implication of host usage to the evolutionary history and biogeography of species in this genus.


Contamination and sensitivity issues with a real-time universal 16S rRNA PCR.
A set of universal oligonucleotide primers specific for the conserved regions of the eubacterial 16S rRNA gene was designed for use with the real-time PCR Applied Biosystems 7700 (TaqMan) system. During the development of this PCR, problems were noted with the use of this gene as an amplification target. Contamination of reagents with bacterial DNA was a major problem exacerbated by the highly sensitive nature of the real-time PCR chemistry. This was compounded by the use of a small amplicon of approximately 100 bases, as is necessary with TaqMan chemistry. In an attempt to overcome this problem, several methodologies were applied. Certain treatments were more effective than others in eliminating the contaminating DNA; however, to achieve this there was a decrease in sensitivity. With UV irradiation there was a 4-log reduction in PCR sensitivity, with 8-methoxypsoralen activity facilitated by UV there was between a 5- and a 7-log reduction, and with DNase alone and in combination with restriction digestion there was a 1.66-log reduction. Restriction endonuclease treatment singly and together did not reduce the level of contaminating DNA. Without the development of ultrapure Taq DNA polymerase, ultrapure reagents, and plasticware guaranteed to be free of DNA, the implementation of a PCR for detection of eubacterial 16S rRNA with the TaqMan system will continue to be problematical.


Pedicular stress fracture in lumbar spine.
The purpose of this article is to report two cases of pedicular stress fracture of the lumbar spine, which is an uncommon cause of low back and leg pain. The relevant literature is reviewed and features of the cases that differ from those already reported are highlighted. One of our cases of pediculolysis is the first in the literature for that is caused by rotational instability induced by laminectomy. The remodeling of the fractured pedicle was striking when compared with its normal counterpart. To the authors' knowledge, this is the first report of a pediculolysis with T2-weighted imaging findings. In addition, we report the computed tomographic (CT) and magnetic resonance (MR) imaging findings of a healed pediculolysis that has not been reported previously.


Probabilistic category learning in developmental dyslexia: Evidence from feedback and paired-associate weather prediction tasks.
Developmental dyslexia is presumed to arise from specific phonological impairments. However, an emerging theoretical framework suggests that phonological impairments may be symptoms stemming from an underlying dysfunction of procedural learning. We tested procedural learning in adults with dyslexia (n = 15) and matched-controls (n = 15) using 2 versions of the weather prediction task: feedback (FB) and paired-associate (PA). In the FB-based task, participants learned associations between cues and outcomes initially by guessing and subsequently through feedback indicating the correctness of response. In the PA-based learning task, participants viewed the cue and its associated outcome simultaneously without overt response or feedback. In both versions, participants trained across 150 trials. Learning was assessed in a subsequent test without presentation of the outcome, or corrective feedback. The dyslexia group exhibited impaired learning compared with the control group on both the FB and PA versions of the weather prediction task. The results indicate that the ability to learn by feedback is not selectively impaired in dyslexia. Rather it seems that the probabilistic nature of the task, shared by the FB and PA versions of the weather prediction task, hampers learning in those with dyslexia. Results are discussed in light of procedural learning impairments among participants with dyslexia.


The transition from mental hospital based to community based psychiatry.
The reformation of psychiatry has been a central topic in health planning and care in most industrialized countries during the last decades. A short review of the political, professional and other forces behind this development is done. A short history of the development in Sweden is given as an example of the general tendencies in this change and the actors involved. There is now a rapidly growing scientific basis for the psychiatric service with a definite shift into a community based orientation. It is concluded that early intervention and prevention will be in focus for the development during the next decades.


Chromosomal instability and bladder cancer: the UroVysion(TM) test in the UroScreen study.
What's known on the subject? and what does the study add?: UroVysion™ is a multicolour fluorescence in situ hybridisation assay that detects DNA gain at chromosomes 3, 7 and 17 and loss at the 9p21 locus in exfoliated urothelial cells. This cell-based test is time-consuming and costly compared with voided urine cytology or other molecular markers for the early detection of bladder cancer. We determined copy number changes at chromosomes 3, 7 and 17 and at the 9p21 locus with UroVysion in a prospective screening study among chemical workers. Strong correlations between DNA gains yield a similar performance in detecting bladder cancer with just one of the probes for chromosomes 3, 7 or 17 instead of all, supporting the development of a simpler and cheaper assay. To explore changes at chromosomes 3, 7, 17 and 9p21 in order to assess associations with bladder cancer for possible improvements of the UroVysion™ assay regarding screening. In all, 1609 men took part in the prospective study UroScreen. Annual screening for bladder cancer was offered to male chemical workers with former exposure to aromatic amines as a voluntary surveillance programme between 2003 and 2010. In all, 191 434 cells in 6517 UroVysion tests were analysed for copy number variations (CNV) at chromosome 3, 7, 17 (gains) and 9p21 (deletions) in 1595 men. We assessed CNVs at single or multiple loci using polysomy indices (PIs, called multiple PI and PI 3, PI 7 and PI 17). We calculated Spearman's rank correlation coefficients (rs ) between these PIs and receiver operating characteristic (ROC) curves with areas under the curves (AUCs). We applied Cox regression to estimate hazard ratios (HRs) to assess the risk of developing bladder cancer. Nine out of 21 bladder tumours detected in 20 participants ('cases') had a positive UroVysion test, including seven high-grade carcinomas and seven overlapping results with a positive cytology. Four cases with negative test results did not attend screening annually. No case was found because of a complete loss of 9p21 in at least 12 cells. There were strong correlations between pairwise combinations of gains at chromosome 3, 7 or 17, ranging between rs = 0.98 and rs = 0.99 in cases and between rs = 0.84 and rs = 0.88 in non-cases (P < 0.001). Associations were less pronounced with CNVs at 9p21 among cases and were lacking in non-cases. Estimates of the relative risk of DNA gain for developing a bladder tumour assessed with PIs (threshold 10% of cells) were 47.7 (95% confidence interval [CI] 18.3-124.1) for the multiple PI, 44.5 (95%CI 16.5-119.9) for PI 3, 34.7 (95%CI 13.1-92.1) for PI 7 and 52.4 (95%CI 20.7-132.6) for PI 17, as well as 7.9 (95%CI 3.0-20.6) for a complete loss of 9p21 (threshold 2.5% of cells), respectively. ROC analyses showed similar AUCs for multiple PI compared with PIs of single chromosomes 3, 7 and 17 (all AUCs between 0.79 and 0.80) and a lower AUC for a homozygous loss of 9p21 (AUC 0.72). The UroVysion assay showed a reasonable performance in detecting bladder cancer in the present study population and shared positive test results with cytology, which is much cheaper. A simpler, faster and cheaper version of the UroVysion assay might rely on the very strong correlations between gains at chromosomes 3, 7 and 17, resulting in a similar performance in detecting bladder cancer with single-probe PIs compared with the full set of these probes. Loss of 9p21 was less predictive for developing bladder cancer in UroScreen.


Analysis of broadly tunable coupled-cavity semiconductor lasers.
We report on the spectral properties for above-threshold operation of broadly tunable, asymmetric multiple quantum well (AMQW), coupled-cavity InGaAsP/InP semiconductor diode lasers. We developed a traveling wave model to understand the mode selection that the lasers exhibit. We find that a weak, short external cavity (SXC) can be used to obtain single frequency operation on each longitudinal mode over the ∼100 nm tuning range of the uncoated AMQW coupled-cavity lasers. We measured the spectral properties of AMQW coupled-cavity lasers with and without an SXC. In a synthesized optical coherent optical tomography application, the use of an SXC with an AMQW coupled-cavity laser reduces the coherence length and hence enhances the performance of the AMQW coupled-cavity laser for optical coherence tomography applications.


Rituximab induces clonal expansion of IgG memory B-cells in patients with inflammatory central nervous system demyelination.
Rituximab, a monoclonal B-cell cytolytic antibody, has beneficial effects in patients with inflammatory demyelinating diseases. So far, little data exists on B-cell subset recovery after rituximab treatment in inflammatory demyelinating diseases of the central nervous system (CNS). To elucidate whether rituximab promotes qualitative changes in the IgG memory B-cell repertoire we performed a single cell analysis in three patients with CNS demyelination. We did not observe any qualitative changes but detected an increased clonal expansion in the IgG memory B-cell compartment after treatment, indicating that a single course of rituximab does not eliminate specific IgG memory B-cells.


3D Neuronal Cell Culture Modeling Based on Highly Porous Ultra-High Molecular Weight Polyethylene.
Cell culturing methods in its classical 2D approach have limitations associated with altered cell morphology, gene expression patterns, migration, cell cycle and proliferation. Moreover, high throughput drug screening is mainly performed on 2D cell cultures which are physiologically far from proper cell functions resulting in inadequate hit-compounds which subsequently fail. A shift to 3D culturing protocols could solve issues with altered cell biochemistry and signaling which would lead to a proper recapitulation of physiological conditions in test systems. Here, we examined porous ultra-high molecular weight polyethylene (UHMWPE) as an inexpensive and robust material with varying pore sizes for cell culturing. We tested and developed culturing protocols for immortalized human neuroblastoma and primary mice hippocampal cells which resulted in high rate of cell penetration within one week of cultivation. UHMWPE was additionally functionalized with gelatin, poly-L-lysine, BSA and chitosan, resulting in increased cell penetrations of the material. We have also successfully traced GFP-tagged cells which were grown on a UHMWPE sample after one week from implantation into mice brain. Our findings highlight the importance of UHMWPE use as a 3D matrix and show new possibilities arising from the use of cheap and chemically homogeneous material for studying various types of cell-surface interactions further improving cell adhesion, viability and biocompatibility.


[Endotheliopathy in the genesis of fetus loss syndrome as observed in pregnant women with chronic viral infection].
The main reasons of fetus loss as observed in pregnant women with chronic viral infection are discussed in the paper. The role of the immune system and different mechanisms of endothelial lesions inherent in the genesis of the discussed phenomenon are analyzed. An approach to management of pregnancy in patients with viral infections targeted at the prevention of pregnancy complications is suggested on the basis of the analysis and independent authors' observations.


Deceptive meaning of oxygen uptake measured at the anaerobic threshold in patients with systolic heart failure and atrial fibrillation.
Oxygen uptake at the anaerobic threshold (VO2AT), a submaximal exercise-derived variable, independent of patients' motivation, is a marker of outcome in heart failure (HF). However, previous evidence of VO2AT values paradoxically higher in HF patients with permanent atrial fibrillation (AF) than in those with sinus rhythm (SR) raised uncertainties. We tested the prognostic role of VO2AT in a large cohort of systolic HF patients, focusing on possible differences between SR and AF. Altogether 2976 HF patients (2578 with SR and 398 with AF) were prospectively followed. Besides a clinical examination, each patient underwent a maximal cardiopulmonary exercise test (CPET). The follow-up was analysed for up to 1500 days. Cardiovascular death or urgent cardiac transplantation occurred in 303 patients (250 (9.6%) patients with SR and 53 (13.3%) patients with AF, p = 0.023). In the entire population, multivariate analysis including peak oxygen uptake (VO2) showed a prognostic capacity (C-index) similar to that obtained including VO2AT (0.76 vs 0.72). Also, left ventricular ejection fraction, ventilation vs carbon dioxide production slope, β-blocker and digoxin therapy proved to be significant prognostic indexes. The receiver-operating characteristic (ROC) curves analysis showed that the best predictive VO2AT cut-off for the SR group was 11.7 ml/kg/min, while it was 12.8 ml/kg/min for the AF group. VO2AT, a submaximal CPET-derived parameter, is reliable for long-term cardiovascular mortality prognostication in stable systolic HF. However, different VO2AT cut-off values between SR and AF HF patients should be adopted.


A Modified Triangular Osteosynthesis Protocol for the Rod and Pedicle Screw Fixation of Vertical Unstable Sacral Fractures.
The stabilization of vertical unstable sacral fractures has been a complex problem that is a challenge to current standard approaches. Here, we attempted to evaluate a modified technique for sacral fractures. In the modified triangular osteosynthesis technique, we adopted a vertical and transverse fixation with a rod and pedicle screw system to reduce and fix sacral fractures in 28 subjects. The postsurgery effect of this technique was evaluated by physical examination and radiography. In the postoperative day 1, the patients were able to move body position passively from the lateral side to supine and exercise their legs by themselves. After a follow up of 20 months, radiological evaluation showed that fracture fragment reduction was excellent in 18 (64%), good (displacement 5-10 mm) in 8 (29%), and fair (displacement 10-15 mm) in 2 (7%) patients. Three patients with a preoperative perineal neurological impairment had a complete recovery after surgical decompression. All patients had achieved bone union of fractures, and no loss of fracture reduction was detected. The modified procedures offered an easier approach to fix vertical unstable sacral fractures, thereby achieving quicker and stable functionality. This suggests an alternative approach to manage unstable sacral fractures. 4. The stabilization of vertical unstable sacral fractures has been a complex problem that is a challenge to current standard approaches. We attempted to introduce a modified technique for sacral fractures.


IL-27 mediates HLA class I up-regulation, which can be inhibited by the IL-6 pathway, in HLA-deficient Small Cell Lung Cancer cells.
Recently, immunotherapy with anti-PD-1 antibodies has shown clinical benefit in recurrent Small Cell Lung Cancer (SCLC). Since anti-PD-1 re-activates anti-tumor Cytotoxic T Lymphocyte (CTL) responses, it is crucial to understand the mechanisms regulating HLA class I, and PD-L1 expression in HLA-negative SCLC. Here we addressed the role of IL-27, a cytokine related to both IL-6 and IL-12 families. The human SCLC cell lines NCI-N592, -H69, -H146, -H446 and -H82 were treated in vitro with different cytokines (IL-27, IFN-γ, IL-6 or a soluble IL-6R/IL-6 chimera [sIL-6R/IL-6]) at different time points and analyzed for tyrosine-phosphorylated STAT proteins by Western blot, for surface molecule expression by immunofluorescence and FACS analyses or for specific mRNA expression by QRT-PCR. Relative quantification of mRNAs was calculated by the ΔΔCT method. The Student's T test was used for the statistical analysis of experimental replicates. IL-27 triggered STAT1/3 phosphorylation and up-regulated the expression of surface HLA class I antigen and of TAP1 and TAP2 mRNA in four out of five SCLC cell lines tested. The IL-27-resistant NCI-H146 cells showed up-regulation of HLA class I by IFN-γ. IFN-γ also induced expression of PD-L1 in SCLC cells, while IL-27 was less potent in this respect. IL-27 failed to activate STAT1/3 phosphorylation in NCI-H146 cells, which display a low expression of the IL-27RA and GP130 receptor chains. As GP130 is shared in IL-27R and IL-6R complexes, we assessed its functionality in response to sIL-6R/IL-6. sIL-6R/IL-6 failed to trigger STAT1/3 signaling in NCI-H146 cells, suggesting low GP130 expression or uncoupling from signal transduction. Although both sIL-6R/IL-6 and IL-27 triggered STAT1/3 phosphorylation, sIL-6R/IL-6 failed to up-regulate HLA class I expression, in relationship to the weak activation of STAT1. Finally sIL-6R/IL-6 limited IL-27-effects, particularly in NCI-H69 cells, in a SOCS3-independent manner, but did not modify IFN-γ induced HLA class I up-regulation. In conclusion, IL-27 is a potentially interesting cytokine for restoring HLA class I expression for SCLC combined immunotherapy purposes. However, the concomitant activation of the IL-6 pathway may limit the IL-27 effect on HLA class I induction but did not significantly alter the responsiveness to IFN-γ.


Engaging children and parents in service design and delivery.
The involvement of all user groups, including children, young people (CYP) and their parents, encourages people to take responsibility for healthier lifestyle behaviours, improves treatment compliance and leads to more appropriate use of healthcare resources. Initiatives to engage CYP in the UK are gathering momentum, but significant improvements are still needed. There is a national drive from the department of health (DH) and NHS England, strategic clinical networks, operational delivery networks (including newborn networks), charities, parent groups and a number of other bodies to embed CYP involvement in service design and delivery. User engagement and patient choice, is underpinned by the NHS outcomes framework, and a myriad of other DH and NHS England policies and practice frameworks. It is now everybody's business.


Stem Cell Paracrine Signaling for Treatment of Premature Ovarian Insufficiency.
Premature ovarian insufficiency is a common disorder affecting young women and represents the worst-case ovarian scenario due to the substantial impact on the reproductive lifespan of these patients. Due to the complexity of this condition, which is not fully understood, non-effective treatments have yet been established for these patients. Different experimental approaches are being explored and strategies based on stem cells deserve special attention. The regenerative and immunomodulatory properties of stem cells have been successfully tested in different tissues, including ovary. Numerous works point out to the efficacy of stem cells in POI treatment, and a wide range of clinical trials have been developed in order to prove safety and effectiveness of stem cells therapy-in diminished ovarian reserve and POI women. The main purpose of this review is to describe the state of the art of the treatment of POI involving stem cells, especially those that use mobilization of stem cells or paracrine signaling.


Detection of Alcohol Intoxication Using Voice Features: A Controlled Laboratory Study.
Devices such as mobile phones and smart speakers could be useful to remotely identify voice alterations associated with alcohol intoxication that could be used to deliver just-in-time interventions, but data to support such approaches for the English language are lacking. In this controlled laboratory study, we compare how well English spectrographic voice features identify alcohol intoxication. A total of 18 participants (72% male, ages 21-62 years) read a randomly assigned tongue twister before drinking and each hour for up to 7 hours after drinking a weight-based dose of alcohol. Vocal segments were cleaned and split into 1-second windows. We built support vector machine models for detecting alcohol intoxication, defined as breath alcohol concentration > .08%, comparing the baseline voice spectrographic signature to each subsequent timepoint and examined accuracy with 95% confidence intervals (CIs). Alcohol intoxication was predicted with an accuracy of 98% (95% CI [97.1, 98.6]); mean sensitivity = .98; specificity = .97; positive predictive value = .97; and negative predictive value = .98. In this small, controlled laboratory study, voice spectrographic signatures collected from brief recorded English segments were useful in identifying alcohol intoxication. Larger studies using varied voice samples are needed to validate and expand models.


Leadership, job satisfaction and organizational commitment in healthcare sector: proposing and testing a model.
none declared. Employees' job satisfaction and commitment depends upon the leadership style of managers. This study clarifies further the relationships between leadership behaviors of managers and two employees' work-related attitudes-job satisfaction and organizational at public hospitals in Iran. A better understanding of these issues and their relationships can pinpoint better strategies for recruiting, promotion, and training of future hospital managers and employees, particularly in Iran but perhaps in other societies as well. This cross-sectional study was conducted using self-administered questionnaires distributed among 814 hospital employees and managers through a stratified random sampling. The dominant leadership style of hospital managers was participative style. Hospital employees were moderately satisfied with their jobs and committed to their organization. Salaries, benefits, promotion, contingent rewards, interpersonal relationships and working conditions were the best predictors of job satisfaction among hospitals employees. Leadership, job satisfaction and commitment were closely interrelated. The leadership behavior of managers explained 28% and 20% of the variations in job satisfaction and organizational commitment respectively. This study clarifies the causal relations of job satisfaction and commitment, and highlights the crucial role of leadership in employees' job satisfaction and commitment. Nevertheless, participative management is not always a good leadership style. Managers should select the best leadership style according to the organizational culture and employees' organizational maturity.


Neighborhood- and individual-level socioeconomic variation in perceptions of racial discrimination.
In approaching the study of racial discrimination and health, the neighborhood- and individual-level antecedents of perceived discrimination need further exploration. We investigated the relationship between neighborhood- and individual-level socioeconomic position (SEP), neighborhood racial composition, and perceived racial discrimination in a cohort of African-American and White women age 40-79 from Connecticut, USA. The logistic regression analysis included 1249 women (39% African-American and 61% White). Neighborhood-level SEP and racial composition were determined using 1990 census tract information. Individual-level SEP indicators included income, education, and occupation. Perceived racial discrimination was measured as lifetime experience in seven situations. For African-American women, living in the most disadvantaged neighborhoods was associated with fewer reports of racial discrimination (odds ratio (OR) 0.44; 95% confidence interval (CI) 0.26, 0.75), with results attenuated after adjustment for individual-level SEP (OR 0.54, CI: 0.29, 1.03), and additional adjustment for neighborhood racial composition (OR 0.70, CI: 0.30, 1.63). African-American women with 12 years of education or less were less likely to report racial discrimination, compared with women with more than 12 years of education (OR 0.57, CI: 0.33, 0.98 (12 years); OR 0.51, CI: 0.26, 0.99 (less than 12 years)) in the fully adjusted model. For White women, neither neighborhood-level SEP nor individual-level SEP was associated with perceived racial discrimination. Individual- and neighborhood-level SEP may be important in understanding how racial discrimination is perceived, reported, processed, and how it may influence health. In order to fully assess the role of racism in future studies, inclusion of additional dimensions of discrimination may be warranted.


Aldose reductase activity in retinal and cerebral microvessels and cultured vascular cells.
Isolated microvessels (primarily capillaries) from bovine retina and cerebral cortex, as well as cultured bovine retinal capillary pericytes and porcine and canine retinal capillary endothelial cells contain apparent aldose reductase activity. This conclusion is based on the ability of these cultured cells and vessel fragments to reduce DL-glyceraldehyde in preference to D-glucuronate at low (0.1 mM) substrate concentrations, in the presence of NADPH, and in the accumulation of high levels of sorbitol or galactitol when retinal pericytes and endothelial cells are cultured in media enriched in glucose or galactose. The quantitative similarities of these activities in bovine retinal and cerebral microvessels, as well as the quantitatively similar ability of these two sets of microvessels to oxidize 14C-labeled glucose with the label either in the C-1 or the C-6 position, suggests that aldose reductase may not be a major causal factor in diabetic retinopathy. This conclusion is suggested because, while these metabolic activities are similar in bovine retinal and cerebral microvessels, only the retinal microvasculature suffers major anatomic and functional damage in diabetes. This conclusion must be viewed with caution, however, because other metabolic pathways that we have not investigated may be altered by an excess of sugar alcohols, and be present in differing activities in retinal and cerebral microvessels; species differences may exist; and similar experiments have not been conducted using human microvessels.


Comparison of spectral-domain and time-domain optical coherence tomography in the detection of neovascular age-related macular degeneration activity.
To compare the sensitivity of commonly used time-domain (TD-OCT) and spectral-domain optical coherence tomography platforms and scanning modalities in the management of neovascular age-related macular degeneration in a population with a high prevalence of exudative disease activity. Fifty consecutive patients within the prospective SAVE (Super-dose Anti-Vascular Endothelial growth factor) trial, which analyzed the utility of 2.0 mg intravitreal ranibizumab for the treatment of recalcitrant neovascular age-related macular degeneration, were enrolled in a comparison trial of 3 different optical coherence tomography (OCT) platforms. Stratus TD-OCT radial scan (Carl Zeiss Meditec, Inc) was compared with 3 Heidelberg Spectralis Heidelberg Retinal Angiograph+OCT (Heidelberg Engineering) acquisition settings (radial, 7-line raster, volumetric) and 2 Cirrus high definition (HD)-OCT (Carl Zeiss Meditec, Inc) acquisition settings (5-line raster, volumetric). Using every imaging platform and acquisition setting, evidence of exudative disease activity was positively identified in 163 of 191 patient visits (85.3%). Intraretinal cysts were identified in 83 of 191 visits (43.5%), and subretinal fluid was identified in 116 of 191 visits (60.7%). Of these positive visits, the Stratus TD-OCT radial scanning technology demonstrated a significantly lower rate of detection (71.8%) when compared with the Spectralis HRA+OCT spectral domain scanning modalities (radial 87.1%, P < 0.001; 7-line raster 92.0%, P < 0.001; volumetric 94.5%, P < 0.001) or the Cirrus HD-OCT spectral domain scanning modalities (5-line raster 81.6%, P = 0.001; volumetric 92.0%, P < 0.001). Intraretinal cysts and subretinal fluid were identified in 83 visits (43.5%) and 116 visits (60.7%), respectively, with 36 eyes (18.8%) having fluid in both locations. No individual imaging modality demonstrated a diagnostic advantage for detecting subretinal fluid versus intraretinal cysts (e.g., Cirrus volume detected 86.7% of intraretinal cysts and 88.8% of subretinal fluid, P = 0.33). In this neovascular age-related macular degeneration patient population, spectral-domain ocular coherence tomography was a superior diagnostic tool when compared with TD-OCT, with each spectral domain platform and acquisition setting identifying significantly more exudative disease activity. The two spectral domain platforms (Cirrus and Spectralis) were not directly compared because identical image acquisition parameters were not used. No individual imaging modality demonstrated a diagnostic advantage for detecting subretinal fluid versus intraretinal cysts.


Mechanism of hypersensitive transport in tilted sharp ratchets.
The noise-flatness-induced hypersensitive transport of overdamped Brownian particles in a tilted ratchet system driven by multiplicative nonequilibrium three-level Markovian noise and additive white noise is considered. At low temperatures, the enhancement of current is very sensitive to the applied small static tilting force. It is established that the enhancement of mobility depends nonmonotonically on the parameters (flatness, correlation time) of multiplicative noise. The optimal values of noise parameters maximizing the mobility are found.


Prevalence of diabetes estimated by plasma glucose criteria combined with standardized measurement of HbA1c among health checkup participants on Miyako Island, Japan.
To estimate the prevalence of diabetes in participants of an annual health checkup in the district of the Miyako Public Health Center (Okinawa, Japan) by using the revised criteria of the Japan Diabetes Society (JDS). The subjects studied here were all Japanese and 45-75 years of age at the time of the health examination in 1998. Diagnosis of diabetes was based on the following: 1) fasting plasma glucose > or =7.0 mmol/l, 2) casual plasma glucose > or =11.1 mmol/l, 3) HbA(1c) > or =6.1%, and 4) self-report on a special questionnaire given at the examination. The HbA(1c) value was standardized by the measurement of 2 standard samples provided by the JDS. Among the 2,621 subjects, 59.7% had their fasting blood glucose levels measured. Of the subjects diagnosed as having diabetes, 154 (12.6%) were men and 115 (8.6%) women. Among the subjects newly diagnosed with diabetes from their fasting blood glucose levels. 27.5% of the men and 21.9% of the women had diagnoses based on HbA(1c) alone. Overall, 34.9% of the subjects with newly diagnosed diabetes were identified by plasma glucose (PG) alone and 33.0% were diagnosed by HbA(1c) alone. The combination of PG and HbA(1c) resulted in a considerable increase in newly diagnosed diabetes as compared with the use of only one of these parameters. Considering the convenience and correlation with vascular complications, use of the 2 tests may be beneficial in epidemiological studies of the Japanese population to identify high-risk groups for micro- and macrovascular diseases.


Extrinsic and intrinsic factors control the genesis of amacrine and cone cells in the rat retina.
The seven major classes of cells of the vertebrate neural retina are generated from a pool of multipotent progenitor cells. Recent studies suggest a model of retinal development in which both the progenitor cells and the environment change over time (Cepko, C. L., Austin, C. P., Yang, X., Alexiades, M. and Ezzeddine, D. (1996). Proc. Natl. Acad. Sci. USA 93, 589-595). We have utilized a reaggregate culture system to test this model. A labeled population of progenitors from the embryonic rat retina were cultured with an excess of postnatal retinal cells and then assayed for their cell fate choices. We found that the postnatal environment had at least two signals that affected the embryonic cells' choice of fate; one signal inhibited the production of amacrine cells and a second affected the production of cone cells. No increase in cell types generated postnatally was observed. The source of the inhibitor of the amacrine cell fate appeared to be previously generated amacrine cells, suggesting that amacrine cell number is controlled by feedback inhibition. The progenitor cell lost its ability to be inhibited for production of an amacrine cell as it entered M phase of the cell cycle. We suggest that postmitotic cells influence progenitor cell fate decisions, but that they do so in a manner restricted by the intrinsic biases of progenitor cells.


Successful Rescue Intervention of Internal Mammary Artery Anastomotic Site Acute Graft Failure With Direct New Generation Covered Stenting.
Acute, perioperative myocardial infarction (MI) from acute left internal mammary artery (LIMA) to left anterior descending (LAD) graft failure immediately following coronary artery bypass grafting (CABG) surgery is associated with significantly increased in-hospital mortality. The leading etiology of such acute graft failure is acute thrombosis, dissection, spasm, anastomosis failure or no-reflow phenomenon. Repeat bypass surgery carries incremental risk and may not be feasible in hemodynamically unstable patients. Traditional percutaneous coronary intervention (PCI), with or without stent placement is sometimes used in such cases; however, graft anatomy and lesion location increase procedural complexity and challenge technical feasibility. This is particularly true of the LIMA to LAD graft anastomosis, where PCI carries the risk of anastomotic site perforation or avulsion. Therefore, the best revascularization strategy for such a lesion involving the LIMA to LAD graft anastomosis in the immediate perioperative period remains unknown. We present a case of 75-year-old male who suffered an acute MI complicated by cardiogenic shock less than 24 h after two-vessel CABG. Selective angiography revealed acute LIMA to LAD anastomotic site closure, posing a risk for perforation if treated with traditional angioplasty or stenting. We successfully performed rescue PCI, by directly deploying a PK Papyrus covered stent (Biotronik, Berlin, Germany) across the anastomosis. Our case report describes the upfront (rather than a bail out) use of the new covered stent as a novel revascularization strategy to treat "perforation prone" LIMA to LAD anastomotic site acute graft failure.


Experiences of U.S. military nurses in the Iraq and Afghanistan wars, 2003-2009.
The purpose of this study is to describe the lived experience of U.S. military nurses who served in Iraq or Afghanistan during the war years 2003 to 2009, and life after returning from war. Colaizzi's phenomenological method guided discovery. This method includes elements of both descriptive and interpretive phenomenology. The sample consisted of 37 military nurses who served in the Army, Navy, or Air Force in the Iraq or Afghanistan wars. Four data-generating questions guided the interview process. Most interviews were face-to-face and conducted in naturalistic settings chosen by the participants. Several interviews were conducted telephonically due to geographic constraints. Data analysis followed Colaizzi's method of analysis. Seven themes emerged from the data, including "deploying to war;""remembrance of war: most chaotic scene;""nurses in harm's way: more than I bargained for;""kinship and bonding: my military family;""my war stress: I'm a different person now;""professional growth: expanding my skills;" and "listen to me: advice to deploying nurses." Analysis continued until data saturation was achieved. Results indicated that wartime deployment was a difficult challenge, lessons learned should be shared with nurses deploying in future years, homecoming was more difficult than most nurses anticipated, and reintegration after coming home takes time and effort. Nursing in war is a unique experience regardless of education, preparation and training. There are a myriad of variables that enter into the experience and effect outcomes, both personal and professional. Wartime nursing is a reality in the current clinical practice arena. War takes its toll on everyone involved, including the caregivers. Nurses returning from war can provide valuable insights to those that follow.


Targeting Cancer Stem Cells to Overcome Chemoresistance.
Cancers are heterogeneous at the cell level, and the mechanisms leading to cancer heterogeneity could be clonal evolution or cancer stem cells. Cancer stem cells are resistant to most anti-cancer treatments and could be preferential targets to reverse this resistance, either targeting stemness pathways or cancer stem cell surface markers. Gold nanoparticles have emerged as innovative tools, particularly for photo-thermal therapy since they can be excited by laser to induce hyperthermia. Gold nanoparticles can be functionalized with antibodies to specifically target cancer stem cells. Preclinical studies using photo-thermal therapy have demonstrated the feasibility of targeting chemo-resistant cancer cells to reverse clinical chemoresistance. Here, we review the data linking cancer stem cells and chemoresistance and discuss the way to target them to reverse resistance. We particularly focus on the use of functionalized gold nanoparticles in the treatment of chemo-resistant metastatic cancers.


Effects of ligand density on hydrophobic charge induction chromatography: molecular dynamics simulation.
High ligand density is usually required in hydrophobic charge induction chromatography (HCIC) for high adsorption capacity. However, it is not clear to what extent the ligand density alters the adsorption and desorption behaviors, or if this leads to the protein conformational transition within adsorbent pores. In the present study, molecular dynamics simulation is performed to examine the effects of ligand density in HCIC using a 46-bead beta-barrel coarse-grained model protein and a coarse-grained adsorbent pore model established in our earlier work. Four ligand densities (1.474, 1.769, 2.212, and 2.949 micromol/m(2)) are simulated at 298.15 K. The simulations indicate that both the capacity and irreversibility of adsorption increase with ligand density. However, it is found that the fastest adsorption occurs at a ligand density of 2.212 micromol/m(2) rather than at the highest density studied. Analyses of adsorption trajectories, protein-ligand interaction energy, and the free energy map indicate that there is repulsion of protein when unfavorable contacts of the protein and ligands occur. There is an enhanced repulsion at 2.949 micromol/m(2), which increases the energy barrier to the transition region and reduces the opportunities to get stable adsorption, thus leading to the decreased adsorption rate. At 2.212 micromol/m(2), however, the repulsion is mild and the high ligand coverage provides abundant opportunities for the protein to get the fastest adsorption and thus causes the maximum unfolding. In the following simulations, complete and irreversible desorption is observed at all ligand densities, in agreement with the easy pH-induced elution behavior of HCIC observed experimentally. It is found that there is a suitable balance between hydrophobic attraction and electrostatic repulsion at 2.212 micromol/m(2), which leads to the slowest desorption kinetics and causes the maximum unfolding. Moreover, analysis of unfolded protein distribution indicates that unfolding occurs mainly on the ligand surface in both adsorption and desorption. The behaviors have been comprehensively elucidated by molecular and thermodynamic analyses.


Increase in hepatic blood flow during early sepsis is due to increased portal blood flow.
Although hepatic blood flow increases significantly during early sepsis [as produced by cecal ligation and puncture (CLP)], it is not known whether this is due to the increase in portal or hepatic arterial blood flows. To study this, rats were subjected to CLP, after which they and sham-operated rats received either 3 or 6 ml normal saline/100 g body wt subcutaneously (i.e., all rats received crystalloid therapy). Blood flow in various organs was determined by using a radioactive microsphere technique at 5 and 20 h after CLP or sham operation. Portal blood flow was calculated as the sum of blood flows to the spleen, pancreas, gastrointestinal tract, and mesentery. Total hepatic blood flow was the sum of portal blood flow and hepatic arterial blood flow. A significant increase in portal blood flow and in total hepatic blood flow was observed at 5 h after CLP (i.e., early sepsis), and this was not altered by doubling the volume of crystalloid resuscitation after the induction of sepsis. In contrast, hepatic arterial blood flow during early sepsis was found to be similar to control; however, it was significantly reduced in late sepsis (i.e., 20 h after CLP). Cardiac output was significantly higher than the control in early sepsis. However, even in late sepsis, cardiac output and total hepatic blood flow were not significantly different from controls. These results indicate that the increased total hepatic blood flow during early hyperdynamic sepsis is solely due to the increased portal blood flow.


Prediction of the structure of a silk-like protein in oligomeric states using explicit and implicit solvent models.
We perform Replica Exchange Molecular Dynamics (REMD) simulations on a silk-like protein design with amino-acid sequence [(Gly-Ala)3-Gly-Glu]5 to investigate the stability of a single protein, a dimer, a trimer and a tetramer made up of these proteins starting from β-roll and β-sheet structures in both explicit (TIP3P) and implicit (GBSA) solvent models. Our simulation results for the implicit solvent model agree with those for the explicit solvent model for simulation times up to the longest tested, being 30 ns per replica. From this we infer that the implicit solvent model that we use is reliable, allowing us to reach much longer time scales (up to 200 ns per replica). We find that the self-assembly of fibers of these proteins in solution must be a nucleated process, involving nuclei made up of at least three monomers. We also find that the conformation of the protein changes upon assembly, i.e., there is a transition from a disordered globular state to an ordered β-sheet structure in the self-assembled state of aggregates containing more than two monomers. This indicates that autosteric effects must be important in the polymerization of this protein, reminiscent of what is observed for β-amyloids. Our findings are consistent with recent experimental results on a protein with an amino acid sequence similar to that of the protein we study.


Sedative-hypnotics are widely abused by drivers apprehended for driving under the influence of drugs.
Sedative-hypnotics are commonly encountered in drivers apprehended for driving under the influence of drugs (DUID). Previous research has mainly concentrated on the residual effects of the drugs. In this study, the extent of sleep medication use and abuse among drivers apprehended on suspicion of DUID was assessed. Additionally, the prevalence and concentrations of the drugs, concomitant use of other drugs of abuse, and the age and sex of the drivers positive for the most commonly prescribed sedative-hypnotics (temazepam, midazolam, nitrazepam, zopiclone, and zolpidem) in DUID cases in Finland in 2009 to 2011 were examined. Sedative-hypnotics were found in 3155 samples of the 13,248 that were analyzed. Temazepam was present in over half of the cases (57.9%), along with other benzodiazepines such as midazolam (13.1%) and nitrazepam (7.0%) and the non-benzodiazepine hypnotics zopiclone (12.2%) and zolpidem (9.8%). The mean age of the drivers using the studied sedative-hypnotics was 33.5 years. Many of the drivers were polydrug users; concomitant stimulant use was found in nearly half of the cases. Cannabis and alcohol were also very common co-findings. In nearly 20% of the cases, the driver had taken more than 1 of the studied sedative-hypnotics; only 2.5% had no findings other than a single sedative-hypnotic in their blood. The drug use pattern of those positive for zopiclone and zolpidem was somewhat different from that of users of benzodiazepine sedative-hypnotics; their age was higher and the concomitant use of illegal stimulants was markedly less prevalent than among the users of temazepam, midazolam, and nitrazepam. There were very few cases in our study population where the positive sedative-hypnotic finding could have been due to appropriate medical use. The extremely prevalent concomitant use of other psychoactive drugs and the high median serum concentrations of the studied sedative-hypnotics suggest their widespread abuse among apprehended drivers.


Expression profile of proteins involved in scar formation in the healing process of full-thickness excisional wounds in the porcine model.
Scar formation in deep dermal wounds is associated with excessive collagen deposition and contraction. Increased collagen synthesis and decreased collagen degradation are the mechanisms through which this form of fibrosis can occur. Another factor might be a different kind of collagen cross-linking seen in fibrotic skin diseases. This type of cross-linking is dependent on the enzyme lysyl hydroxylase-2b. In this study, we examined the expression profile of the potential key players in scar formation in time in healing of acute wounds. Collagen types I and III, lysyl hydroxylase-2b, alpha-smooth muscle actin, transforming growth factor betas, and the matrix metalloproteinases and their inhibitor mRNA levels were determined. All genes examined show distinct expression patterns over time. The expression of lysyl hydroxylase-2b peaks at day 7, and precedes collagen types I and III expression. Eight weeks after wounding, the scars showed an increased level of lysyl hydroxylase-2b-mediated collagen cross-linking. This study shows that the fibrosis-specific type of cross-linking of collagen seen in human hypertrophic scarring also plays a role in this animal model of wound healing. Moreover, the expression of the putative gene responsible for this type of cross-linking, the lysyl hydroxylase-2b, is elevated during wound healing.


Applying the Theoretical Domains Framework to identify police, fire, and paramedic preferences for accessing mental health care in a First Responder Operational Stress Injury Clinic: a qualitative study.
First responders and other public safety personnel (PSP; e.g. correctional workers, firefighters, paramedics, police, public safety communicators) are often exposed to events that have the potential to be psychologically traumatizing. Such exposures may contribute to poor mental health outcomes and a greater need to seek mental health care. However, a theoretically driven, structured qualitative study of barriers and facilitators of help-seeking behaviours has not yet been undertaken in this population. This study used the Theoretical Domains Framework (TDF) to identify and better understand critical barriers and facilitators of help-seeking and accessing mental health care for a planned First Responder Operational Stress Injury (OSI) clinic. We conducted face-to-face, one-on-one semistructured interviews with 24 first responders (11 firefighters, five paramedics, and eight police officers), recruited using purposive and snowball sampling. Interviews were analyzed using deductive content analysis. The TDF guided study design, interview content, data collection, and analysis. The most reported barriers included concerns regarding confidentiality, lack of trust, cultural competency of clinicians, lack of clarity about the availability and accessibility of services, and stigma within first responder organizations. Key themes influencing help-seeking were classified into six of the TDF's 14 theoretical domains: environmental context and resources; knowledge; social influences; social/professional role and identity; emotion; and beliefs about consequences. The results identified key actions that can be utilized to tailor interventions to encourage attendance at a First Responder OSI Clinic. Such approaches include providing transparency around confidentiality, policies to ensure greater cultural competency in all clinic staff, and clear descriptions of how to access care; routinely involving families; and addressing stigma.


From stereotype threat to stereotype threats: implications of a multi-threat framework for causes, moderators, mediators, consequences, and interventions.
More than 100 articles have examined the construct of stereotype threat and its implications. However, stereotype threat seems to mean different things to different researchers and has been employed to describe and explain processes and phenomena that appear to be fundamentally distinct. Complementing existing models, the authors posit a Multi-Threat Framework in which six qualitatively distinct stereotype threats arise from the intersection of two dimensions--the target of the threat (the self/one's group) and the source of the threat (the self/outgroup others/ingroup others). The authors propose that these threats constitute the core of the broader stereotype threat construct and provide the foundation for understanding additional, as of yet uncharacterized, stereotype threats. The proposed threats likely differentially peril those with different stigmatizable characteristics, have different eliciting conditions and moderators, are mediated by somewhat different processes, are coped with and compensated for in different ways, and require different interventions to overcome.


Polyphenols in human nutrition: from the in vitro antioxidant capacity to the beneficial effects on cardiometabolic health and related inter-individual variability - an overview and perspective.
Oxidative damage of cells and tissues is broadly implicated in human pathophysiology, including cardiometabolic diseases. Polyphenols, as important constituents of the human diet and potent in vitro free radical scavengers, have been extensively studied for their beneficial effects on cardiometabolic health. However, it has been demonstrated that the in vivo antioxidant activity of polyphenols is distinct from their in vitro free radical-scavenging capacity. Indeed, bioavailability of nutritional polyphenols is low and conditioned by complex mechanisms of absorption, distribution, metabolism and excretion. Nowadays, it is commonly accepted that the cellular antioxidant activity of polyphenols is mainly carried out via modification of transcription of genes involved in antioxidant defence. Importantly, polyphenols also contribute to cardiometabolic health by modulation of a plethora of cellular processes that are not directly associated with antioxidant enzymes, through nutri(epi)genomic mechanisms. Numerous human intervention studies have demonstrated beneficial effects of polyphenols on the key cardiometabolic risk factors. However, inconsistency of the results of some studies led to identification of the inter-individual variability in response to consumption of polyphenols. In perspective, a detailed investigation of the determinants of this inter-individual variability will potentially lead us towards personalised dietary recommendations. The phenomenon of inter-individual variability is also of relevance for supplementation with antioxidant (pro)vitamins.


Pharmacokinetic evaluation of twice-a-week micafungin for prophylaxis of invasive fungal disease in children with acute lymphoblastic leukaemia: a prospective observational cohort study.
To determine the pharmacokinetics of twice-a-week micafungin prophylaxis in paediatric leukaemic patients to provide the rationale for this approach. Twice-a-week micafungin at a dose of 9 mg/kg (maximum 300 mg) was given during the leukaemic induction treatment with at least one pharmacokinetic assessment. Non-linear mixed-effects modelling was used for analysis. For model building, our paediatric data were strengthened with existing adult data. Monte Carlo simulations were performed with twice-a-week dosing regimens of 5, 7 and 9 mg/kg and flat dosing per weight band. Simulated paediatric exposures were compared with the exposure in adults after a once-daily 100 mg regimen. Sixty-one paediatric patients were included with a median age and weight of 4.0 years (range 1.0-17) and 19.5 kg (range 8.60-182), respectively. A two-compartment model best fitted the data. CL and central Vd were lower (P < 0.01) in paediatric patients compared with adults. Predicted exposures (AUC0-168 h) for the 5, 7 and 9 mg/kg and flat dosing per weight band regimens exceeded the adult reference exposure. All twice-a-week regimens appeared to result in adequate exposure for Candida therapy, with simulated exposures well above the adult reference exposure. These findings provide the rationale for the pharmacokinetic equivalence of twice-a-week and once-daily micafungin regimens. The greater micafungin exposures seem to be caused by a slower-than-anticipated CL in our paediatric leukaemic patients. The generalizability of our results for Aspergillus prophylaxis cannot be provided without assumptions on target concentrations and within-class identical efficacy.


[Oxidative damage to E. coli membranes caused by superoxide radicals].
The effects of bacterial membranes of active oxygen species photochemically generated by riboflavin-histidine systems were studied. According to SDS-PAAG data, the formation of high molecular weight protein aggregates and the appearance of fluorochromes whose fluorescence is seen in the longwave length region of the spectrum (lambda excit = 350 nm, lambda emis = 400-500 nm) and which are bound to the proteins, are suggestive of membrane oxidation consisting in the chemical modification of protein components. The presence in E. coli membranes of endogenous photosensitizers which upon illumination with visible light induce the oxidation of membrane proteins, was established.


[Factors influencing visual acuity following vitrectomy for exudative age-related macular degeneration].
We investigated the influence of various factors on visual results in patients undergoing surgical removal of choroidal neovascular membranes (CNVM) caused by age-related macular degeneration (AMD). This study was performed in 55 eyes of 55 patients who underwent surgical removal of CNVM for AMD and followed them for 6 months or more. The criterion for surgical eligibility was active subfoveal choroidal neovascular membrane of 0.5 or more disc diameter above the retinal pigment epithelium with visual acuity of 0.3 or worse. We investigated the influence of various factors on the logarithm of the minimum angle of resolution (log MAR) final visual acuity. The factors were age, symptom duration, preoperative log MAR visual acuity, CNVM diameter, mean deviation with visual field analyzer, previous laser treatment, posterior vitreous detachment, findings of indocyanine green angiography, operative complications, and recurrence of CNVM. Better preoperative visual acuity, shorter symptom duration, and smaller CNVM diameter were correlated with better postoperative final visual acuity. Surgical excision of subfoveal CNVM may be the better therapeutic choice in selected cases with AMD.


Performance and adsorption mechanism of a magnetic calcium silicate hydrate composite for phosphate removal and recovery.
A novel magnetic calcium silicate hydrate composite (Fe3O4@CSH) was proposed for phosphorus (P) removal and recovery from a synthetic phosphate solution, facilitated by a magnetic separation technique. The Fe3O4@CSH material was characterized by transmission electron microscopy (TEM), scanning electron microscopy (SEM), powder Fourier transform infrared (FTIR) spectroscopy, X-ray diffraction (XRD), X-ray photoelectron spectroscopy (XPS), zeta-potential and magnetic curves. The chemical composition and structure of Fe3O4@CSH and the successful surface loading of hydroxyl functional groups were confirmed. Phosphate adsorption kinetics, isotherm, and thermodynamic experiments showed that adsorption reaches equilibrium at 24 h, with a maximum adsorption capacity of 55.84 mg P/g under optimized experimental conditions. Adsorption kinetics fitted well to the pseudo second-order model, and equilibrium data fit the Freundlich isotherm model. Thermodynamic analysis provided a positive value for ΔH° (129.84 KJ/mol) and confirmed that phosphate adsorption on these materials is endothermic. The P-laden Fe3O4@CSH materials could be rapidly separated from aqueous solution by a magnetic separation technique within 1 min. A removal rate of more than 60% was still obtained after eight adsorption/desorption cycles, demonstrating the excellent reusability of the particles. The results demonstrated that the Fe3O4@CSH materials had high P-adsorption efficiency and were reusable.


Prevalence of type 2 diabetes among acute inpatients and its impact on length of hospital stay in Japan.
The goal of this study was to determine the prevalence of type 2 diabetes among acute inpatients and evaluate its impact on the length of hospital stay in Japan. The discharge records of 2,120,170 acute inpatients who were 30 years old or older and discharged between July and December of 2008 were obtained from the Japanese administrative case-mix system, Diagnosis Procedure Combination (DPC), and allocated for analysis. Type 2 diabetes was defined by E11 of the ICD-10 coding system on patient records. Other types of diabetes, including type 1 diabetes and diabetes in pregnancy (defined by ICD-10 codes E10 E12-14 and O24, respectively) were excluded from the analyses. Type 2 diabetes was observed among 11.4% of the records, 9.9% of which were cases of diabetes as a comorbidity. Total length of hospital stay was 33,468,152 days, with diabetes patients occupying 13.9% of the total bed days. Patients with type 2 diabetes as a comorbidity had prolonged lengths of hospital stay compared to patients free from diabetes, and stratification by sex, age, surgical treatment, and disease category did not alter these results. The median length of hospital stay was 9 days among patients without diabetes and 13 days among patients with diabetes. Japanese acute health care is experiencing an epidemic of type 2 diabetes, with 1 out of 10 acute inpatients suffering from the disease. Complication of type 2 diabetes is associated with prolonged length of hospital stay.


Phylogenetic analysis of avian influenza viruses of H11 subtype isolated in Kazakhstan.
Avian influenza viruses A/turkey/Almaty/535/04 (H11N9) and A/herring gull/Atyrau/2186/07 (H11N2) isolated in Kazakhstan were characterized as low pathogenic in biological and genetic studies. Putative glycosylation sites were identical to the putative sites in published H11, N2, and N9 isolates sequences. Compared with published data no additional basic amino acid residues were found in the hemagglutinin (HA) cleavage site of these Kazakhstan strains. Phylogenetic analysis revealed a rare case of Eurasian-American reassortment in the HA gene of A/herring gull/Atyrau/2186/07 (H11N2) virus and significant sequence difference of the HA and the neuraminidase genes of the virus A/turkey/Almaty/535/04 (H11N9) from the previously published GenBank viruses.


Psychological considerations in submarine escape training: brief overview and future directions.
The inability of a submarine to surface must rate as one of greatest risks to sailors in peacetime. To prepare for such emergencies, many navies provide training to master the procedures required to escape successfully from disabled submarines. This paper provides a brief overview of some of the psychological principles in simulated submarine escape training. It further discusses applicable psychological constructs such as positive outcome expectancies, the role of anxiety, and other personal factors mediating outcomes of such training. It concludes with recommendations for future research aimed at enhancing the safety and impact of submarine escape training. These include enhanced detection of psychological risk factors such as anxiety, as well as investigating the relative contribution of personality variables to in-training safety and positive outcome expectancies. These recommendations do not only apply to submarine escape training, but may also be applicable to high fidelity safety training in other off-shore survival contexts, such as helicopter underwater escape training, freefall lifeboat training and smoke diving.


Effects of traditional vs. alternating whole-body strength training on squat performance.
Traditional strength training with 80% of 1 repetition maximum (1RM) uses 2- to 5-minute rest periods between sets. These long rest periods minimize decreases in volume and intensity but result in long workouts. Performing upper-body exercises during lower-body rest intervals may decrease workout duration but may affect workout performance. Therefore, the purpose of this study was to compare the effects of traditional vs. alternating whole-body strength training on squat performance. Twenty male (24 ± 2 years) volunteers performed 2 workouts. The traditional set (TS) workout consisted of 4 sets of squats (SQ) at 80% of 1RM on a force plate with 3-minute rest between sets. The alternating set (AS) workout also consisted of 4 sets of SQ at 80% of 1RM but with bench press, and bench pull exercises performed between squat sets 1, 2 and 3 with between-exercise rest of 50 seconds, resulting in approximately 3-minute rest between squat sets. Sets 1-3 were performed for 4 repetitions, whereas set 4 was performed to concentric failure. Total number of completed repetitions of the fourth squat set to failure was recorded. Peak ground reaction force (GRF), peak power (PP), and average power (AP) of every squat repetition were recorded and averaged for each set. There was no significant interaction for GRF, PP, or AP. However, volume-equated AP was greater during the TS condition (989 ± 183) than the AS condition (937 ± 176). During the fourth squat set, the TS condition resulted in more repetitions to failure (7.5 ± 2.2) than the AS condition (6.5 ± 2.2). Therefore, individuals who aim to optimize squat AP should refrain from performing more than 3 ASs per exercise. Likewise, those who aim to maximize squat repetitions to failure should refrain from performing upper-body multijoint exercises during squat rest intervals.


[Open aortic valvuloplasty in surgical treatment of associated heart diseases].
The article presents indications to surgical correction of aortal stenosis and its variants in patients with rheumatic diseases combined with a lesion of other valves of the heart. The possibility to fulfill valve-saving plastic operations on the stenosed aortal valve was shown in 110 operations. The results obtained proved advantages of plastic operations over prostheses of the stenosed aortas without calcinosis universalis. Such operations allowed the intrahospital lethality to be considerably decreased. In later terms only two patients needed prostheses of the valve. The functional class was improved in the overwhelming majority of the patients.


Interleukin-10 and the interleukin-10 receptor.
Interleukin-10 (IL-10), first recognized for its ability to inhibit activation and effector function of T cells, monocytes, and macrophages, is a multifunctional cytokine with diverse effects on most hemopoietic cell types. The principal routine function of IL-10 appears to be to limit and ultimately terminate inflammatory responses. In addition to these activities, IL-10 regulates growth and/or differentiation of B cells, NK cells, cytotoxic and helper T cells, mast cells, granulocytes, dendritic cells, keratinocytes, and endothelial cells. IL-10 plays a key role in differentiation and function of a newly appreciated type of T cell, the T regulatory cell, which may figure prominently in control of immune responses and tolerance in vivo. Uniquely among hemopoietic cytokines, IL-10 has closely related homologs in several virus genomes, which testify to its crucial role in regulating immune and inflammatory responses. This review highlights findings that have advanced our understanding of IL-10 and its receptor, as well as its in vivo function in health and disease.


lncRNA DLG1-AS1 Promotes Cell Proliferation by Competitively Binding with miR-107 and Up-Regulating ZHX1 Expression in Cervical Cancer.
Recent studies have revealed that long non-coding RNAs (lncRNAs) are involved in the occurrence and development of various tumors, thereby attracting increasing attention from researchers. The important biological functions of lncRNAs have been recognized gradually, but their mechanism in cervical cancer remains unclear. Differentially expressed lncRNAs in cervical cancer and para-carcinoma tissues were identified by screening using an lncRNA array, and candidate lncRNAs were verified by quantitative real-time PCR. A series of bioinformatics and molecular biological methods were adopted to investigate the interactions among lncRNAs, microRNAs (miRNAs), and miRNA target genes in cervical cancer. Cell viability was measured using a Cell Counting Kit-8 assay. DLG1-AS1 was the most significantly up-regulated lncRNA in cervical cancer tissues, and it was confirmed that cervical cancer patients with high DLG1-AS1 expression had a poor prognosis. Down-regulation of DLG1-AS1 expression suppressed the proliferation of cervical cancer cells. Further investigation revealed that DLG1-AS1 eliminated the inhibition of miR-107 on the expression of its target gene ZHX1 by competitively binding to miR-107. Moreover, rescue assays proved that the effect of DLG1-AS1 on the proliferation of cervical cancer cells was dependent on miR-107. DLG1-AS1/miR-107/ZHX1 can form a competitive endogenous RNA network that regulates the proliferation of cervical cancer cells, resulting in tumor progression.


A brain-specific cytochrome P450 responsible for the majority of deltamethrin resistance in the QTC279 strain of Tribolium castaneum.
Cytochrome P450-mediated detoxification is one of the most important mechanisms involved in insecticide resistance. However, the molecular basis of this mechanism and the physiological functions of P450s associated with insecticide resistance remain largely unknown. Here, we exploited the functional genomics and reverse genetic approaches to identify and characterize a P450 gene responsible for the majority of deltamethrin resistance observed in the QTC279 strain of Tribolium castaneum. We used recently completed whole-genome sequence of T. castaneum to prepare custom microarrays and identified a P450 gene, CYP6BQ9, which showed more than a 200-fold higher expression in the deltamethrin-resistant QTC279 strain when compared with its expression in the deltamethrin-susceptible Lab-S strain. Functional studies using both double-strand RNA (dsRNA)-mediated knockdown in the expression of CYP6BQ9 and transgenic expression of CYP6BQ9 in Drosophila melanogaster showed that CYP6BQ9 confers deltamethrin resistance. Furthermore, CYP6BQ9 enzyme expressed in baculovirus metabolizes deltamethrin to 4-hydroxy deltamethrin. Strikingly, we also found that unlike many P450 genes involved in insecticide resistance that were reported previously, CYP6BQ9 is predominantly expressed in the brain, a part of the central nervous system (CNS) containing voltage-gated sodium channels targeted by deltamethrin. Taken together, the current studies on the brain-specific insect P450 involved in deltamethrin resistance shed new light on the understanding of the molecular basis and evolution of insecticide resistance.


Quantitative determination of fluorescence labeling implemented in cell cultures.
Labeling efficiency is a crucial parameter in fluorescence applications, especially when studying biomolecular interactions. Current approaches for estimating the yield of fluorescent labeling have critical drawbacks that usually lead them to be inaccurate or not quantitative. We present a method to quantify fluorescent-labeling efficiency that addresses the critical issues marring existing approaches. The method operates in the same conditions of the target experiments by exploiting a ratiometric evaluation with two fluorophores used in sequential reactions. We show the ability of the protocol to extract reliable quantification for different fluorescent probes, reagents concentrations, and reaction timing and to optimize labeling performance. As paradigm, we consider the labeling of the membrane-receptor TrkA through 4'-phosphopantetheinyl transferase Sfp in living cells, visualizing the results by TIRF microscopy. This investigation allows us to find conditions for demanding single and multi-color single-molecule studies requiring high degrees of labeling. The developed method allows the quantitative determination and the optimization of staining efficiency in any labeling strategy based on stable reactions.


A visual mechanism tuned to black.
Chubb et al. [Journal of the Optical Society of America A 11 (1994) 2350] investigated preattentive discrimination of achromatic textures comprising random mixtures of 17 Weber contrasts ranging linearly from -1 to 1. They showed that only a single mechanism B is used to discriminate between textures whose histograms are equated in mean and in variance. Although they provided a partial characterization of B, their methods did not allow them to measure the sensitivity of B to texture mean and variance. Here, additional measurements are performed to complete the functional characterization of B. The results reveal that B (i) is strongly activated by texture elements of the lowest contrast (near -1), (ii) is slightly activated by texture elements of contrast -0.875, and (iii) barely distinguishes the 15 contrasts ranging from -0.75 all the way up to 1. To reflect the sharpness of its tuning to very dark, sparse elements in a predominantly bright scene, we call B the blackshot mechanism.


Amplitude encoded phase-only filters.
The phase-only filter (POF) and binary phase-only filter (BPOF) have been shown to have high optical efficiency, good target discrimination, and virtually no sidelobes. We present a simple method of amplitude encoding the signal phase information. These new filters have most of the same advantages of a POF or BPOF so that amplitude modulating spatial light modulators or photographic film can be used.


Dynamic Structural Effects on the Second-Harmonic Generation of Tryptophane-Rich Peptides and Gramicidin A.
Peptide chains can model endogenous biotags for applications in second-harmonic imaging microscopy. Such structures are flexible which may strongly affect their structure-property relationship. Here, we explore quantum-mechanically the conformational space of a set of tryptophan-rich model peptides. This has become feasible because of the recently proposed meta-dynamics method based on efficient tight-binding (TB) calculations. The TB version of the simplified time-dependent density functional theory (sTD-DFT-xTB) method is used to evaluate the first hyperpolarizability (β). These new tools enable us to calculate nonlinear optical properties for systems with several thousand atoms and/or to screen large structure ensembles. First, we show that the indole chromophore in tryptophan residues dominates the β response of these systems. Their relative orientation mostly determines the global β tensor and affects the static β response. The results underline the importance of finding low-energy conformers for modeling β of flexible molecules. Additionally, we compare calculated and extrapolated experimental static β. The sTD-DFT-xTB method is capable of providing reliable second-harmonic generation values for tryptophan-rich systems at a fraction of the computational cost of the commonly used TD-DFT/TD-HF levels of theory.


Metabolic imaging of patients with cardiomyopathy.
The cardiomyopathies comprise a diverse group of illnesses that can be characterized functionally by several techniques. However, the delineation of derangements of regional perfusion and metabolism have been accomplished only relatively recently with positron emission tomography (PET). Regional myocardial accumulation and clearance of 11C-palmitate, the primary myocardial substrate under most conditions, demonstrate marked spatial heterogeneity when studied under fasting conditions or with glucose loading. PET with 11C-palmitate permits the noninvasive differentiation of patients with nonischemic from ischemic dilated cardiomyopathy, since patients with ischemic cardiomyopathy demonstrate large zones of intensely depressed accumulation of 11C-palmitate, probably reflecting prior infarction. Patients with hypertrophic cardiomyopathy and Duchenne's muscular dystrophy demonstrate relatively unique patterns of myocardial abnormalities of perfusion and metabolism. The availability of new tracers and techniques for the evaluation of myocardial metabolism (11C-acetate), perfusion (H2(15)O), and autonomic tone (11-C-hydroxyephedrine) should facilitate further understanding of the pathogenesis of the cardiomyopathies.


Comparative sequence analysis of the P-, M- and L-coding region of the measles virus CAM-70 live attenuated vaccine strain.
Measles virus is a highly contagious agent which causes a major health problem in developing countries. The viral genomic RNA is single-stranded, nonsegmented and of negative polarity. Many live attenuated vaccines for measles virus have been developed using either the prototype Edmonston strain or other locally isolated measles strains. Despite the diverse geographic origins of the vaccine viruses and the different attenuation methods used, there was remarkable sequence similarity of H, F and N genes among all vaccine strains. CAM-70 is a Japanese measles attenuated vaccine strain widely used in Brazilian children and produced by Bio-Manguinhos since 1982. Previous studies have characterized this vaccine biologically and genomically. Nevertheless, only the F, H and N genes have been sequenced. In the present study we have sequenced the remaining P, M and L genes (approximately 1.6, 1.4 and 6.5 kb, respectively) to complete the genomic characterization of CAM-70 and to assess the extent of genetic relationship between CAM-70 and other current vaccines. These genes were amplified using long-range or standard RT-PCR techniques, and the cDNA was cloned and automatically sequenced using the dideoxy chain-termination method. The sequence analysis comparing previously sequenced genotype A strains with the CAM-70 Bio-Manguinhos strain showed a low divergence among them. However, the CAM-70 strains (CAM-70 Bio-Manguinhos and a recently sequenced CAM-70 submaster seed strain) were assigned to a specific group by phylogenetic analysis using the neighbor-joining method. Information about our product at the genomic level is important for monitoring vaccination campaigns and for future studies of measles virus attenuation.


Fenofibrate attenuates tubulointerstitial fibrosis and inflammation through suppression of nuclear factor-κB and transforming growth factor-β1/Smad3 in diabetic nephropathy.
Fibrates, the ligands of peroxisome proliferator-activated receptor-alpha, have been shown to have a renal protective action in diabetic models of renal disease, but the mechanisms underlying this effect are unknown. In the present study, we sought to investigate in greater detail the effect of fenofibrate and its mechanism of action on renal inflammation and tubulointerstitial fibrosis in an animal model of type 2 diabetes mellitus. Twelve-week-old non-diabetic Zucker lean (ZL) and Zucker diabetic fatty (ZD) rats were treated with vehicle or fenofibrate for 10 weeks. mRNA and protein analyses were performed by real-time polymerase chain reaction, Western blot and immunostaining. The diabetic condition of ZD rats was associated with an increase in collagen and alpha-smooth muscle actin accumulation in the kidney, which was significantly reduced by fenofibrate. Chronic treatment of ZD rats with fenofibrate attenuated renal inflammation and tubular injury as evidenced by a decrease in mRNA and protein expression of secreted phosphoprotein-1, monocyte chemotactic protein-1 and kidney injury molecule-1 in the kidneys. Renal interstitial macrophage infiltration was also significantly reduced in the kidneys of fenofibrate-treated diabetic animals. Moreover, renal nuclear factor (NF)-kappaB DNA-binding activity, transforming growth factor (TGF)-beta1 and phospho-Smad3 proteins were significantly higher in ZD animals compared with ZL ones. This increase in NF-kappaB activity, TGF-beta1 expression and Smad3 phosphorylation was greatly attenuated by fenofibrate in the diabetic kidneys. Taken together, fenofibrate suppressed NF-kappaB and TGF-beta1/Smad3 signaling pathways and reduced renal inflammation and tubulointerstitial fibrosis in diabetic ZD animals.