Dynamic manipulation by light and electric fields: micrometer particles to microliter droplets.
We demonstrate a new hybrid optoelectric technique that can manipulate objects across several length scales. The technique leverages a variety of different physical mechanisms to achieve the dynamic manipulation of droplets and also the in situ concentration of colloidal particles suspended in the droplets. Various physical mechanisms such as optoelectrowetting, electrothermal flows, and ac electroosmosis are leveraged through different modes of operation of the device. Each operational mode, which is activated through the proper combination of an applied ac bias and the illumination used, is characterized by the ability to manipulate objects on a certain length scale. We also demonstrate that the device lends itself to the active control of microstructure patterns that emerge from a droplet evaporation process.


Postnatal growth of the human pons: a morphometric and immunohistochemical analysis.
Despite its critical importance to global brain function, the postnatal development of the human pons remains poorly understood. In the present study, we first performed magnetic resonance imaging (MRI)-based morphometric analyses of the postnatal human pons (0-18 years; n = 6-14/timepoint). Pons volume increased 6-fold from birth to 5 years, followed by continued slower growth throughout childhood. The observed growth was primarily due to expansion of the basis pontis. T2-based MRI analysis suggests that this growth is linked to increased myelination, and histological analysis of myelin basic protein in human postmortem specimens confirmed a dramatic increase in myelination during infancy. Analysis of cellular proliferation revealed many Ki67(+) cells during the first 7 months of life, particularly during the first month, where proliferation was increased in the basis relative to tegmentum. The majority of proliferative cells in the postnatal pons expressed the transcription factor Olig2, suggesting an oligodendrocyte lineage. The proportion of proliferating cells that were Olig2(+) was similar through the first 7 months of life and between basis and tegmentum. The number of Ki67(+) cells declined dramatically from birth to 7 months and further decreased by 3 years, with a small number of Ki67(+) cells observed throughout childhood. In addition, two populations of vimentin/nestin-expressing cells were identified: a dorsal group near the ventricular surface, which persists throughout childhood, and a parenchymal population that diminishes by 7 months and was not evident later in childhood. Together, our data reveal remarkable postnatal growth in the ventral pons, particularly during infancy when cells are most proliferative and myelination increases.


Inflammatory microcrystals induce murine macrophage survival and DNA synthesis.
The interaction of particulates with resident macrophages is a consistent feature in certain forms of crystal-induced inflammation, for example, in synovial tissues, lung, and the peritoneum. The mitogenic activity of basic calcium phosphate (BCP) crystals and calcium pyrophosphate dihydrate (CPPD) crystals on synovial fibroblasts has been considered relevant to the synovial hyperplasia observed in crystal-induced arthritis. The aim of the study was to determine whether microcrystals such as these could enhance macrophage survival and induce DNA synthesis, thus indicating that they may contribute to the tissue hyperplasia. Murine bone-marrow-derived macrophages were treated in vitro with microcrystals, the cell numbers were monitored over time, and DNA synthesis was measured as the incorporation of [methyl-(3)H]thymidine (TdR). We report here that BCP, monosodium urate, talc, and, to a lesser extent, CPPD crystals promote macrophage survival and DNA synthesis; the latter response is particularly striking in the presence of low concentrations of macrophage-colony stimulating factor (M-CSF, CSF-1). Enhanced macrophage survival or proliferation may contribute to the synovial hyperplasia noted in crystal-associated arthropathies, as well as to talc-induced inflammation and granuloma formation. The crystals studied join the list of particulates having these effects on macrophages, indicating the generality of this type of response.


Elevated monoclonal and polyclonal serum immunoglobulin free light chain as prognostic factors in B- and T-cell non-Hodgkin lymphoma.
The serum immunoglobulin free light chain (FLC) assay quantitates free kappa (κ) and lambda (λ) light chains. FLC elevations in patients with diffuse large B-cell lymphoma (DLBCL), Hodgkin lymphoma (HL), and chronic lymphocytic leukemia (CLL) are associated with an inferior survival. These increases in FLC can be monoclonal (as in myeloma) or polyclonal. The goal was to estimate the frequency of these elevations within distinct types of B-cell and T-cell non-Hodgkin lymphoma (NHL) and whether the FLC measurements are associated with event-free survival (EFS). We studied serum for FLC abnormalities using normal laboratory reference ranges to define an elevated κ or λ FLC. Elevations were further classified as polyclonal or monoclonal. Four hundred ninety-two patients were studied: 453 B-cell and 34 T-cell NHL patients. Twenty-nine % (142/453) of patients had an elevated FLC of which 10% were monoclonal elevations. Within B-cell NHL, FLC abnormalities were most common in lymphoplasmacytic (79%), mantle cell (68%), and lymphomas of mucosa associated lymphoid tissue (31%); they were least common in follicular (15%). The hazard ratio (HR) for EFS in all patients was 1.41 (95% CI; 1.11-1.81); in all B-cell NHL the HR was 1.44 (95% CI 1.11-1.96); in all T-cell NHL the HR was 1.17 (95% CI 0.55-2.49). FLC abnormalities predicted an inferior OS (HR = 2.75, 95% CI: 1.93-3.90, P < 0.0001). The serum FLC assay is useful for prognosis in both B-cell and T-cell types of NHL. In B-cell NHL further discrimination between a monoclonal and polyclonal elevation may be helpful and should be analyzed in prospective clinical trials.


Copy number variations are progressively associated with the pathogenesis of colorectal cancer in ulcerative colitis.
To evaluate the association of known copy number variations (CNVs) in ulcerative colitis (UC) progressing to colorectal cancer. Microsatellite instability analysis using the National Cancer Institute's panel of markers, and CNV association studies using Agilent 2 × 105 k arrays were done in tissue samples from four patient groups with UC: those at low risk (LR) or high risk of developing colorectal cancer, those with premalignant dysplastic lesions, and those with colitis-associated colorectal cancer (CAC). DNA from tissue samples of these groups were independently hybridized on arrays and analyzed. The data obtained were further subjected to downstream bioinformatics enrichment analysis to examine the correlation with CAC progression. Microarray analysis highlighted a progressive increase in the total number of CNVs [LR (n = 178) vs CAC (n = 958), 5.3-fold], gains and losses [LR (n = 37 and 141) vs CAC (n = 495 and 463), 13.4- and 3.3-fold, respectively], size [LR (964.2 kb) vs CAC (10540 kb), 10.9-fold] and the number of genes in such regions [LR (n = 119) vs CAC (n = 455), 3.8-fold]. Chromosome-wise analysis of CNVs also showed an increase in the number of CNVs across each chromosome. There were 38 genes common to all four groups in the study; 13 of these were common to cancer genes from the Genetic Disease Association dataset. The gene set enrichment analysis and ontology analysis highlighted many cancer-associated genes. All the samples in the different groups were microsatellite stable. Increasing numbers of CNVs are associated with the progression of UC to CAC, and warrant further detailed exploration.


Adaptations and mechanisms of human heat acclimation: Applications for competitive athletes and sports.
Exercise heat acclimation induces physiological adaptations that improve thermoregulation, attenuate physiological strain, reduce the risk of serious heat illness, and improve aerobic performance in warm-hot environments and potentially in temperate environments. The adaptations include improved sweating, improved skin blood flow, lowered body temperatures, reduced cardiovascular strain, improved fluid balance, altered metabolism, and enhanced cellular protection. The magnitudes of adaptations are determined by the intensity, duration, frequency, and number of heat exposures, as well as the environmental conditions (i.e., dry or humid heat). Evidence is emerging that controlled hyperthermia regimens where a target core temperature is maintained, enable more rapid and complete adaptations relative to the traditional constant work rate exercise heat acclimation regimens. Furthermore, inducing heat acclimation outdoors in a natural field setting may provide more specific adaptations based on direct exposure to the exact environmental and exercise conditions to be encountered during competition. This review initially examines the physiological adaptations associated with heat acclimation induction regimens, and subsequently emphasizes their application to competitive athletes and sports.


Markers of bone resorption predict hip fracture in elderly women: the EPIDOS Prospective Study.
Increased bone turnover has been suggested as a potential risk factor for osteoporotic fractures. We investigated this hypothesis in a prospective cohort study performed on 7598 healthy women more than 75 years of age. One hundred and twenty-six women (mean years 82.5) who sustained a hip fracture during a mean 22-month follow-up were age-matched with three controls who did not fracture. Baseline samples were collected prior to fracture for the measurement of two markers of bone formation and three urinary markers of bone resorption: type I collagen cross-linked N- (NTX) or C-telopeptide (CTX) and free deoxypyridinoline (free D-Pyr). Elderly women had increased bone formation and resorption compared with healthy premenopausal women. Urinary excretion of CTX and free D-Pyr, but not other markers, was higher in patients with hip fracture than in age-matched controls (p = 0.02 and 0.005, respectively). CTX and free D-Pyr excretion above the upper limit of the premenopausal range was associated with an increased hip fracture risk with an odds ratio (95% confidence interval) of 2.2 (1.3-3.6) and 1.9 (1.1-3.2), respectively, while markers of formation were not. Increased bone resorption predicted hip fracture independently of bone mass, i.e., after adjustment for femoral neck bone mineral density (BMD) and independently of mobility status assessed by the gait speed. Women with both a femoral BMD value of 2.5 SD or more below the mean of young adults and either high CTX or high free D-Pyr levels were at greater risk of hip fracture, with an odds ratio of 4.8 and 4.1, respectively, than those with only low BMD or high bone resorption. Elderly women are characterized by increased bone turnover, and some markers of bone resorption predict the subsequent risk of hip fracture independently of hip BMD. Combining the measurement of BMD and bone resorption may be useful to improve the assessment of the risk of hip fracture in elderly women.


Simultaneous interpreters as a model for neuronal adaptation in the domain of language processing.
In the context of language processing, proficiency and age of acquisition have reliably been shown to have a strong influence on the functional and structural architecture of the human brain. The aim of the present EEG study was to examine the impact of language training as experienced by simultaneous interpreters (SI) on auditory word processing and to disentangle its influence from that of proficiency and age of acquisition. Eleven native German SI and controls matched in L2 proficiency and age of acquisition were asked to judge whether auditory presented disyllabic noun pairs both within and across the German (L1) and English (L2) languages were either semantically congruent or incongruent. We revealed enlarged N400 responses in SI while they detected incongruent trials both within the native (L1) and non-native (L2) language and also while they performed the task in the opposite direction as specifically trained (L1 to L2). These enlarged N400 responses in SI suggest a training-induced altered sensitivity to semantic processing within and across L1 and L2. The enlarged N400 responses we revealed in SI to congruent noun pairs during the German-English condition (L1 to L2) may indicate that SI could not benefit from an L1 prime when the target was a L2 word, suggesting additional processing resulting from long-term backwards (L2 to L1) training.


Viltolarsen: First Approval.
Viltolarsen (Viltepso® in Japan) is a phosphorodiamidate morpholino antisense oligonucleotide being developed by Nippon Shinyaku, in collaboration with the National Center of Neurology and Psychiatry (NCNP), for the treatment of Duchenne muscular dystrophy (DMD). Viltolarsen binds to exon 53 of the dystrophin mRNA precursor and restores the amino acid open-reading frame by skipping exon 53, resulting in the production of a shortened dystrophin protein that contains essential functional portions. In March 2020, intravenous viltolarsen received its first global approval in Japan for the treatment of DMD in patients with confirmed deletion of the dystrophin gene that is amenable to exon 53 skipping. Viltolarsen is under regulatory review in the USA and clinical trials continue in the USA, Canada and globally. This article summarizes the milestones in the development of viltolarsen leading to the first approval for DMD.


Polyspecies biofilm formation on implant surfaces with different surface characteristics.
To investigate the microbial adherence and colonization of a polyspecies biofilm on 7 differently processed titanium surfaces. Six-species biofilms were formed anaerobically on 5-mm-diameter sterilized, saliva-preconditioned titanium discs. Material surfaces used were either machined, stained, acid-etched or sandblasted/acid-etched (SLA). Samples of the latter two materials were also provided in a chemically modified form, with increased wettability characteristics. Surface roughness and contact angles of all materials were determined. The discs were then incubated anaerobically for up to 16.5 h. Initial microbial adherence was evaluated after 20 min incubation and further colonization after 2, 4, 8, and 16.5 h using non-selective and selective culture techniques. Results at different time points were compared using ANOVA and Scheffé post hoc analysis. The mean differences in microorganisms colonizing after the first 20 min were in a very narrow range (4.5 to 4.8 log CFU). At up to 16.5 h, the modified SLA surface exhibited the highest values for colonization (6.9±0.2 log CFU, p<0.05) but increasing growth was observed on all test surfaces over time. Discrepancies among bacterial strains on the differently crafted titanium surfaces were very similar to those described for total log CFU. F. nucleatum was below the detection limit on all surfaces after 4 h. Within the limitations of this in vitro study, surface roughness had a moderate influence on biofilm formation, while wettability did not seem to influence biofilm formation under the experimental conditions described. The modified SLA surface showed the highest trend for bacterial colonization.


The impact of school nonresponse on substance use prevalence estimates - Germany as a case study.
The European School Survey Project on Alcohol and Other Drugs (ESPAD) is a survey study that collects comparable data on substance use of students aged 15-16 years old in European countries. The present study aims at investigating the impact of school refusal to participate in ESPAD on substance use prevalence estimates. Data came from the 2007 German ESPAD study; the sample consisted of 12,246 students in 552 schools within seven German federal states. A simulation approach was used in order to study the effects of systematic exclusion of participating schools on prevalence estimates of key ESPAD outcomes including the use of tobacco, alcohol, cannabis, and other illegal drugs. The systematic exclusion of schools based on city-, school-, and class size, school environment, and schools' substance use policies resulted in significant changes in prevalence estimates in 23 of 25 examined combinations of selection criterion and outcome. Yet, these effects were small, with differences remaining below three percentage points around the original estimates. This simulation approach suggests that nonparticipation of schools in surveys on students' substance use in Germany does not largely affect the validity of resulting prevalence estimates. Even a reduced number of schools may be sufficient to gain valid prevalence figures.


Antibiotics May Trigger Mitochondrial Dysfunction Inducing Psychiatric Disorders.
Clinical usage of several classes of antibiotics is associated with moderate to severe side effects due to the promotion of mitochondrial dysfunction. We contend that this may be due to perturbation of unique evolutionary relationships that link selective biochemical and molecular aspects of mitochondrial biology to conserved enzymatic processes derived from bacterial progenitors. Operationally, stereo-selective conformational matching between mitochondrial respiratory complexes, cytosolic and nuclear signaling complexes appears to support the conservation of a critically important set of chemical messengers required for existential regulation of homeostatic cellular processes. Accordingly, perturbation of normative mitochondrial function by select classes of antibiotics is certainly reflective of the high degree of evolutionary pressure designed to maintain ongoing bidirectional signaling processes between cellular compartments. These issues are of critical importance in evaluating potentially severe side effects of antibiotics on complex behavioral functions mediated by CNS neuronal groups. The CNS is extremely dependent on delivery of molecular oxygen for maintaining a required level of metabolic activity, as reflected by the high concentration of neuronal mitochondria. Thus, it is not surprising to find several distinct behavioral abnormalities conforming to established psychiatric criteria that are associated with antibiotic usage in humans. The manifestation of acute and/or chronic psychiatric conditions following antibiotic usage may provide unique insights into key etiological factors of major psychiatric syndromes that involve rundown of cellular bioenergetics via mitochondrial dysfunction. Thus, a potential window of opportunity exists for development of novel therapeutic agents targeting diminished mitochondrial function as a factor in severe behavioral disorders.


Pharmacological treatment strategies in obsessive compulsive disorder: A cross-sectional view in nine international OCD centers.
It is unknown what next-step strategies are being used in clinical practice for patients with obsessive-compulsive disorder (OCD) who do not respond to first-line treatment. As part of a cross-sectional study of OCD, treatment and symptom information was collected. Consecutive OCD out-patients in nine international centers were evaluated by self-report measures and clinical/structured interviews. OCD symptom severity was evaluated by the Yale Brown Obsessive Compulsive Scale (YBOCS) and Clinical Global Impression-Severity Scale (CGI-S). Clinical response to current treatment was evaluated by the CGI-Improvement Scale (CGI-I ≤ 2). In total, 361 participants reported taking medication; 77.6% were taking a selective serotonin reuptake inhibitor; 50% reported use of at least one augmentation strategy. Antipsychotics were most often prescribed as augmenters (30.3%), followed by benzodiazepines (24.9%) and antidepressants (21.9%). No differences in OCD symptom severity were found between patients taking different classes of augmentation agents. Results from this international cross-sectional study indicate that current OCD treatment is in line with evidence-based treatment guidelines. Although augmentation strategies are widely used, no significant differences in OCD symptom severity were found between monotherapy and augmentation or between different therapeutic agents.


Analysis of positional candidate genes in the AAA1 susceptibility locus for abdominal aortic aneurysms on chromosome 19.
Abdominal aortic aneurysm (AAA) is a complex disorder with multiple genetic risk factors. Using affected relative pair linkage analysis, we previously identified an AAA susceptibility locus on chromosome 19q13. This locus has been designated as the AAA1 susceptibility locus in the Online Mendelian Inheritance in Man (OMIM) database. Nine candidate genes were selected from the AAA1 locus based on their function, as well as mRNA expression levels in the aorta. A sample of 394 cases and 419 controls was genotyped for 41 SNPs located in or around the selected nine candidate genes using the Illumina GoldenGate platform. Single marker and haplotype analyses were performed. Three genes (CEBPG, PEPD and CD22) were selected for DNA sequencing based on the association study results, and exonic regions were analyzed. Immunohistochemical staining of aortic tissue sections from AAA and control individuals was carried out for the CD22 and PEPD proteins with specific antibodies. Several SNPs were nominally associated with AAA (p < 0.05). The SNPs with most significant p-values were located near the CCAAT enhancer binding protein (CEBPG), peptidase D (PEPD), and CD22. Haplotype analysis found a nominally associated 5-SNP haplotype in the CEBPG/PEPD locus, as well as a nominally associated 2-SNP haplotype in the CD22 locus. DNA sequencing of the coding regions revealed no variation in CEBPG. Seven sequence variants were identified in PEPD, including three not present in the NCBI SNP (dbSNP) database. Sequencing of all 14 exons of CD22 identified 20 sequence variants, five of which were in the coding region and six were in the 3'-untranslated region. Five variants were not present in dbSNP. Immunohistochemical staining for CD22 revealed protein expression in lymphocytes present in the aneurysmal aortic wall only and no detectable expression in control aorta. PEPD protein was expressed in fibroblasts and myofibroblasts in the media-adventitia border in both aneurysmal and non-aneurysmal tissue samples. Association testing of the functional positional candidate genes on the AAA1 locus on chromosome 19q13 demonstrated nominal association in three genes. PEPD and CD22 were considered the most promising candidate genes for altering AAA risk, based on gene function, association evidence, gene expression, and protein expression.


Genome-wide profiling of H3K56 acetylation and transcription factor binding sites in human adipocytes.
The growing epidemic of obesity and metabolic diseases calls for a better understanding of adipocyte biology. The regulation of transcription in adipocytes is particularly important, as it is a target for several therapeutic approaches. Transcriptional outcomes are influenced by both histone modifications and transcription factor binding. Although the epigenetic states and binding sites of several important transcription factors have been profiled in the mouse 3T3-L1 cell line, such data are lacking in human adipocytes. In this study, we identified H3K56 acetylation sites in human adipocytes derived from mesenchymal stem cells. H3K56 is acetylated by CBP and p300, and deacetylated by SIRT1, all are proteins with important roles in diabetes and insulin signaling. We found that while almost half of the genome shows signs of H3K56 acetylation, the highest level of H3K56 acetylation is associated with transcription factors and proteins in the adipokine signaling and Type II Diabetes pathways. In order to discover the transcription factors that recruit acetyltransferases and deacetylases to sites of H3K56 acetylation, we analyzed DNA sequences near H3K56 acetylated regions and found that the E2F recognition sequence was enriched. Using chromatin immunoprecipitation followed by high-throughput sequencing, we confirmed that genes bound by E2F4, as well as those by HSF-1 and C/EBPα, have higher than expected levels of H3K56 acetylation, and that the transcription factor binding sites and acetylation sites are often adjacent but rarely overlap. We also discovered a significant difference between bound targets of C/EBPα in 3T3-L1 and human adipocytes, highlighting the need to construct species-specific epigenetic and transcription factor binding site maps. This is the first genome-wide profile of H3K56 acetylation, E2F4, C/EBPα and HSF-1 binding in human adipocytes, and will serve as an important resource for better understanding adipocyte transcriptional regulation.


Nutrient and sediment removal by a restored wetland receiving agricultural runoff.
Few studies have measured removal of pollutants by restored wetlands that receive highly variable inflows. We used automated flow-proportional sampling to monitor the removal of nutrients and suspended solids by a 1.3-ha restored wetland receiving unregulated inflows from a 14-ha agricultural watershed in Maryland, USA. Water entered the wetland mainly in brief pulses of runoff, which sometimes exceeded the 2500-m3 water holding capacity of the wetland. Half of the total water inflow occurred in only 24 days scattered throughout the two-year study. Measured annual water gains were within 5% of balancing water losses. Annual removal of nutrients differed greatly between the two years of the study. The most removal occurred in the first year, which included a three-month period of decreasing water level in the wetland. In that year, the wetland removed 59% of the total P, 38% of the total N, and 41% of the total organic C it received. However, in the second year, which lacked a drying period, there was no significant (p > 0.05) net removal of total N or P, although 30% of the total organic C input was removed. For the entire two-year period, the wetland removed 25% of the ammonium, 52% of the nitrate, and 34% of the organic C it received, but there was no significant net removal of total suspended solids (TSS) or other forms of N and P. Although the variability of inflow may have decreased the capacity of the wetland to remove materials, the wetland still reduced nonpoint-source pollution.


Plant biotechnology for lignocellulosic biofuel production.
Lignocelluloses from plant cell walls are attractive resources for sustainable biofuel production. However, conversion of lignocellulose to biofuel is more expensive than other current technologies, due to the costs of chemical pretreatment and enzyme hydrolysis for cell wall deconstruction. Recalcitrance of cell walls to deconstruction has been reduced in many plant species by modifying plant cell walls through biotechnology. These results have been achieved by reducing lignin content and altering its composition and structure. Reduction of recalcitrance has also been achieved by manipulating hemicellulose biosynthesis and by overexpression of bacterial enzymes in plants to disrupt linkages in the lignin-carbohydrate complexes. These modified plants often have improved saccharification yield and higher ethanol production. Cell wall-degrading (CWD) enzymes from bacteria and fungi have been expressed at high levels in plants to increase the efficiency of saccharification compared with exogenous addition of cellulolytic enzymes. In planta expression of heat-stable CWD enzymes from bacterial thermophiles has made autohydrolysis possible. Transgenic plants can be engineered to reduce recalcitrance without any yield penalty, indicating that successful cell wall modification can be achieved without impacting cell wall integrity or plant development. A more complete understanding of cell wall formation and structure should greatly improve lignocellulosic feedstocks and reduce the cost of biofuel production.


Extruded blend of soybean meal and sunflower seeds for dairy cattle in early lactation.
An extruded blend of 44% crude protein soybean meal (50%), whole sunflower seeds (45%), and premix (5%) was evaluated as a protein and energy source for dairy cows in early lactation. Thirty Holstein cows (24 multiparous and 6 primiparous) were assigned to either a corn-oats-soybean meal concentrate or a concentrate where soy-sunflower blend replaced all soybean meal and portions of corn and oats. Dry matter of total mixed diets was 36% corn silage, 21% alfalfa haylage, and 43% concentrate. Yields of milk (33.6 and 33.8 kg/d) and 4% fat-corrected milk (30.9 and 30.5 kg/d) were similar, and percentages of total solids (11.92 and 11.38), fat (3.55 and 3.30), and protein (2.91 and 2.74) were lower in milk from cows fed soy-sunflower blend. Milk from cows fed soy-sunflower blend contained fewer short- and medium-chain fatty acids, more 18-carbon fatty acids, and was more unsaturated than from cows fed soybean meal. Intakes of dry matter and changes in body weight were not different among diets. Ruminal fluid pH and molar ratio of acetate to propionate were higher, and concentrations of total volatile fatty acids and ammonia were lower in cows fed soy-sunflower blend. Concentrations of essential amino acids in arterial serum, calculated mammary uptakes, and transfer efficiencies indicated more desirable amino acid balance in cows fed soy-sunflower blend. Methionine appeared to be most limiting with both diets.


Effectiveness of a multifactorial cardiovascular risk reduction clinic for diabetes patients with depression.
Depression may attenuate the effects of diabetes interventions. Our ongoing Cardiovascular Risk Reduction Clinic simultaneously addresses hyperglycemia, hypertension, smoking, and hyperlipidemia. We examined the relationship between depression diagnosis and responsiveness to the Cardiovascular Risk Reduction Clinic. We studied Cardiovascular Risk Reduction Clinic participants with diabetes who had a depression diagnosis and those with no mental health diagnosis. Our outcome measure was change in 20-year cardiovascular mortality risk according to the United Kingdom Prospective Diabetes Study (UKPDS) score. Of 231 participants, 36 (15.6%) had a depression diagnosis. Participants with a depression diagnosis had a higher baseline UKPDS score (56.8 [SD 21.3]) than participants with no mental health diagnosis (49.5 [SD 18.7], P = .04). After Cardiovascular Risk Reduction Clinic participation, mean UKPDS scores did not differ significantly (37.8 [SD 15.9] for no mental health diagnosis and 39.4 [SD 18.6] for depression diagnosis). Mean UKPDS score reduction was 11.6 [SD 15.6] for no mental health diagnosis compared with 18.4 [SD 15.9] for depression diagnosis (P = .03). Multivariable linear regression that controlled for baseline creatinine, number of Cardiovascular Risk Reduction Clinic visits, sex, and history of congestive heart failure showed significantly greater improvement in UKPDS score among participants with a depression diagnosis (beta = 6.0, P = .04) and those with more Cardiovascular Risk Reduction Clinic visits (beta = 2.1, P < .001). The Cardiovascular Risk Reduction Clinic program reduced cardiovascular disease risk among patients with diabetes and a diagnosis of depression. Further work should examine how depressive symptom burden and treatment modify the effect of this collaborative multifactorial program and should attempt to determine the durability of the effect.


Interleaved and partial transmission interleaved optical coherent orthogonal frequency division multiplexing.
A novel group of interleaved orthogonal frequency division multiplexing (IL-OFDM) and partial transmission IL-OFDM (PT-IL-OFDM) schemes are proposed for optical coherent detection. In the interleaved operation, the odd (or even) subcarriers of an OFDM symbol are reserved (closed). Such an interleaved arrangement can gain the tolerance toward phase noise induced interchannel interference (PN-ICI). Meanwhile, the peak to average power ratio (PAPR) of an OFDM signal can be significantly reduced. However, half of the capacity is sacrificed for the interleaved operation. By exploring the time domain symmetry property of IL-OFDM, the concept of PT-IL-OFDM is proposed to double the capacity of IL-OFDM. Consequently, PT-IL-OFDM can gain the merits of low PAPR and a high tolerance toward PN-ICI without capacity sacrifice. Numerical simulation verified the advanced properties of IL-OFDM and PT-IL-OFDM.


Rapid learning of associations between sound and action through observed movement. A TMS study.
Research has established that there is a cognitive link between perception and production of the same movement. However, there has been relatively little research into the relevance of this for non-expert perceivers, such as music listeners who do not play instruments themselves. In two experiments we tested whether participants can quickly learn new associations between sounds and observed movement without performing those movements themselves. We measured motor evoked potentials (MEPs) in the first dorsal interosseous muscle of participants' right hands while test tones were heard and single transcranial magnetic stimulation (TMS) pulses were used to trigger motor activity. In Experiment 1 participants in a 'human' condition (n=4) learnt to associate the test tone with finger movement of the experimenter, while participants in a 'computer' condition (n=4) learnt that the test tone was triggered by a computer. Participants in the human condition showed a larger increase in MEPs compared with those in the computer condition. In a second experiment pairing between sounds and movement occurred without participants repeatedly observing the movement and we found no such difference between the human (n=4) and computer (n=4) conditions. These results suggest that observers can quickly learn to associate sound with movement, so it should not be necessary to have played an instrument to experience some motor resonance when hearing that instrument.


Endothelial damage with phacoemulsification and intraocular lens implantation.
Specular microscopy was used to study the corneal endothelium in a series of patients undergoing phacoemulsification with and without intraocular lens implantation by one experienced surgeon. Cell loss after phacoemulsification alone was variable, but the mean was 18%, with a median of 11%. The patients undergoing phacoemulsification plus lens implantation showed a mean cell loss of 26%, with a median of 17%. This is considerably lower than has been reported with standard intracapsular extraction plus lens implantation. The decreased endothelial damage is attributed to the technique of inserting the lens through a small incision and maintaining an air bubble between the lens and the cornea throughout lens insertion and wound closure.


Nanoscale voxel spectroscopy by simultaneous EELS and EDS tomography.
Extending the capabilities of electron tomography with advanced imaging techniques and novel data processing methods, can augment the information content in three-dimensional (3D) reconstructions from projections taken in the transmission electron microscope (TEM). In this work we present the application of simultaneous electron energy-loss spectroscopy (EELS) and energy-dispersive X-ray spectroscopy (EDS) to scanning TEM tomography. Various tools, including refined tilt alignment procedures, multivariate statistical analysis and total-variation minimization enable the 3D reconstruction of analytical tomograms, providing 3D analytical metrics of materials science samples at the nanometer scale. This includes volumetric elemental maps, and reconstructions of EDS, low-loss and core-loss EELS spectra as four-dimensional spectrum volumes containing 3D local voxel spectra. From these spectra, compositional, 3D localized elemental analysis becomes possible opening the pathway to 3D nanoscale elemental quantification.


Postoperative venous thromboembolism predicts survival in cancer patients.
To determine whether a postoperative venous thromboembolism (VTE) is associated with a worse prognosis and/or a more advanced cancer stage and to evaluate the association between a postoperative VTE and cancer-specific survival when known prognostic factors, such as age, stage, cancer type, and type of surgery, are controlled. It is unknown whether oncology patients who develop a venous thromboembolism after a complete curative resection are at the same survival disadvantage as oncology patients with a spontaneous VTE. A retrospective case control study was conducted at Memorial Sloan-Kettering Cancer Center. Years of study: January 1, 2000, to December 31, 2005. Median follow-up: 24.9 months (Interquartile range 13.0, 43.0). All cancer patients who underwent abdominal, pelvic, thoracic, or soft tissue procedures and those who developed a VTE within 30 days of the procedure were identified from a prospective morbidity and mortality database. Overall survival (OS) was calculated for the entire cohort. In the matched cohort, OS and disease-specific survival (DSS) were calculated for stages 0 to 3 and stages 0 to 2. A total of 23,541 cancer patients underwent an invasive procedure and 474 (2%) had a postoperative VTE. VTE patients had a significantly worse 5-year OS compared to no-VTE patients (43.8% vs 61.2%; P < 0.0001); 205 VTE patients (stages 0-3) were matched to 2050 controls by age, sex, cancer type, stage, and surgical procedure. In this matched analysis, VTE patients continued to demonstrate a significantly worse prognosis with an inferior 5-year OS (54.7% vs 66.3%; P < 0.0001) and DSS (67.8% vs 79.5%; P = 0.0007) as compared to controls. The survival difference persisted in early stage disease (stage 0-2), with 5-year DSS of 82.9% versus 87.3% (P = 0.01). Postoperative VTE in oncology patients with limited disease and a complete surgical resection is associated with an inferior cancer survival. A postoperative VTE remains a poor prognostic factor, even when controlling for age, stage, cancer type, and surgical procedure further supporting an independent link between hypercoagulability and cancer survival.


Video assisted right mini-thoracotomy for aortic valve replacement.
Aortic valve replacement through minimally invasive access is increasing. These procedures have several advantages over conventional sternotomy including decreased intensive care unit and hospital length of stay and decreased ventilation time. The right anterior mini-thoracotomy (RAM) approach is potentially attractive in that it completely spares the sternum leading to improved cosmesis, reduced blood loss, and improved patient satisfaction. However, this approach is underutilized due to anticipated technical challenges including difficulty with visualization and annular suture placement. We employ a camera and automated suturing technology for supra-annular valve implant. A RAM is performed via a 5 cm incision in the right second intercostal space with a camera port placed lateral to the incision. Peripheral venous cannulation is performed along with central arterial cannulation. Aortic cross clamp is placed through a 5 mm incision in the third interspace anterior to mid axillary line. Histidine tryptophan ketoglutarate (HTK) cardioplegia is administered. After aortic leaflet removal, annular and prosthetic sutures are placed with shafted instruments or with automated suturing technology. Aortic valve replacement can be performed safely through a RAM. The use of a camera and automated suturing technology facilitates this procedure, potentially enabling more surgeons to offer this less invasive approach to patients.


Evaluation of microelectrode materials for direct-current electrocorticography.
Direct-current electrocorticography (DC-ECoG) allows a more complete characterization of brain states and pathologies than traditional alternating-current recordings (AC-ECoG). However, reliable recording of DC signals is challenging because of electrode polarization-induced potential drift, particularly at low frequencies and for more conducting materials. Further challenges arise as electrode size decreases, since impedance is increased and the potential drift is augmented. While microelectrodes have been investigated for AC-ECoG recordings, little work has addressed microelectrode properties for DC-signal recording. In this paper, we investigated several common microelectrode materials used in biomedical application for DC-ECoG. Five of the most common materials including gold (Au), silver/silver chloride (Ag/AgCl), platinum (Pt), Iridium oxide (IrOx), and platinum-iridium oxide (Pt/IrOx) were investigated for electrode diameters of 300 μm. The critical characteristics such as polarization impedance, AC current-induced polarization, long-term stability and low-frequency noise were studied in vitro (0.9% saline). The two most promising materials, Pt and Pt/lrOx were further investigated in vivo by recording waves of spreading depolarization, one of the most important applications for DC-ECoG in clinical and basic science research. Our experimental results indicate that IrOx-based microelectrodes, particularly with composite layers of nanostructures, are excellent in all of the common evaluation characteristics both in vitro and in vivo and are most suitable for multimodal monitoring applications. Pt electrodes suffer high current-induced polarization, but have acceptable long-term stability suitable for DC-ECoG. Major significance. The results of this study provide quantitative data on the electrical properties of microelectrodes with commonly-used materials and will be valuable for development of neural recordings inclusive of low frequencies.


Prevention of osteocyte and osteoblast apoptosis by bisphosphonates and calcitonin.
Glucocorticoid-induced osteoporosis may be due, in part, to increased apoptosis of osteocytes and osteoblasts, and bisphosphonates (BPs) are effective in the management of this condition. We have tested the hypothesis that BPs suppress apoptosis in these cell types. Etidronate, alendronate, pamidronate, olpadronate, or amino-olpadronate (IG9402, a bisphosphonate that lacks antiresorptive activity) at 10(-9) to 10(-6) M prevented apoptosis of murine osteocytic MLO-Y4 cells, whether it was induced by etoposide, TNF-alpha, or the synthetic glucocorticoid dexamethasone. BPs also inhibited apoptosis of primary murine osteoblastic cells isolated from calvaria. Similar antiapoptotic effects on MLO-Y4 and osteoblastic cells were seen with nanomolar concentrations of the peptide hormone calcitonin. The antiapoptotic effect of BPs and calcitonin was associated with a rapid increase in the phosphorylated fraction of extracellular signal regulated kinases (ERKs) and was blocked by specific inhibitors of ERK activation. Consistent with these in vitro results, alendronate abolished the increased prevalence of apoptosis in vertebral cancellous bone osteocytes and osteoblasts that follows prednisolone administration to mice. These results suggest that the therapeutic efficacy of BPs or calcitonin in diseases such as glucocorticoid-induced osteoporosis may be due, in part, to their ability to prevent osteocyte and osteoblast apoptosis.


MID Max: LC-MS/MS Method for Measuring the Precursor and Product Mass Isotopomer Distributions of Metabolic Intermediates and Cofactors for Metabolic Flux Analysis Applications.
The analytical challenges to acquire accurate isotopic data of intracellular metabolic intermediates for stationary, nonstationary, and dynamic metabolic flux analysis (MFA) are numerous. This work presents MID Max, a novel LC-MS/MS workflow, acquisition, and isotopomer deconvolution method for MFA that takes advantage of additional scan types that maximizes the number of mass isotopomer distributions (MIDs) that can be acquired in a given experiment. The analytical method was found to measure the MIDs of 97 metabolites, corresponding to 74 unique metabolite-fragment pairs (32 precursor spectra and 42 product spectra) with accuracy and precision. The compounds measured included metabolic intermediates in central carbohydrate metabolism and cofactors of peripheral metabolism (e.g., ATP). Using only a subset of the acquired MIDs, the method was found to improve the precision of flux estimations and number of resolved exchange fluxes for wild-type E. coli compared to traditional methods and previously published data sets.


Social support and health-related quality of life in women with breast cancer: a longitudinal study.
A breast cancer diagnosis is a distressing event that impacts on physical and psychological functioning. This study examined the longitudinal relationships among a diagnosis of breast cancer, social support, and health-related quality of life (HRQOL). Participants were 412 women from the 1946-1951 birth cohort of the Australian Longitudinal Study on Women's Health who self-reported a new diagnosis of breast cancer between 1998 and 2007. The three surveys of longitudinal data analyzed included data 3 years before diagnosis, at diagnosis (baseline), and 3 years after diagnosis (follow-up). Social support was measured using the 19-item Medical Outcomes Study Social Support Survey; HRQOL was measured using the Medical Outcomes Study 36-item Short-Form Health Survey. Compared with pre-diagnosis HRQOL, women newly diagnosed with breast cancer reported significantly poorer HRQOL in subscales related to pain, physical functioning, and health and vitality. At 3-year follow-up, HRQOL had improved in most domains to levels consistent with pre-diagnosis. Levels of social support remained stable across time. The structural equation model showed that social support was positively predictive of better physical and mental HRQOL at 3-year follow-up. Longitudinal analyses indicate that social support appears to be an important predictor of HRQOL in women diagnosed with breast cancer. In particular, positive emotional and informational support that may normally be provided by a partner is important in maintaining HRQOL. Identification of those lacking social support, especially patients without partners, will enable them to be guided to appropriate support networks and programs.


Allostasis and allostatic load: implications for neuropsychopharmacology.
The primary hormonal mediators of the stress response, glucocorticoids and catecholamines, have both protective and damaging effects on the body. In the short run, they are essential for adaptation, maintenance of homeostasis, and survival (allostasis). Yet, over longer time intervals, they exact a cost (allostatic load) that can accelerate disease processes. The concepts of allostasis and allostatic load center around the brain as interpreter and responder to environmental challenges and as a target of those challenges. In anxiety disorders, depressive illness, hostile and aggressive states, substance abuse, and post-traumatic stress disorder (PTSD), allostatic load takes the form of chemical imbalances as well as perturbations in the diurnal rhythm, and, in some cases, atrophy of brain structures. In addition, growing evidence indicates that depressive illness and hostility are both associated with cardiovascular disease (CVD) and other systemic disorders. A major risk factor for these conditions is early childhood experiences of abuse and neglect that increase allostatic load later in life and lead individuals into social isolation, hostility, depression, and conditions like extreme obesity and CVD. Animal models support the notion of lifelong influences of early experience on stress hormone reactivity. Whereas, depression and childhood abuse and neglect tend to be more prevalent in individuals at the lower end of the socioeconomic ladder, cardiovascular and other diseases follow a gradient across the full range of socioeconomic status (SES). An SES gradient is also evident for measures of allostatic load. Wide-ranging SES gradients have also been described for substance abuse and affective and anxiety disorders as a function of education. These aspects are discussed as important, emerging public health issues where the brain plays a key role.


Intraocular Pressure Changes and Relationship With Corneal Biomechanics After SMILE and FS-LASIK.
The purpose of this article was to evaluate intraocular pressure (IOP) changes and investigate the relationship with corneal biomechanics after small-incision lenticule extraction (SMILE) and femtosecond laser-assisted laser in situ keratomileusis (FS-LASIK). A total of 193 eyes of 193 patients who underwent SMILE and FS-LASIK procedures were included in this retrospective study. Data were collected preoperatively and postoperatively, including Goldmann-correlated IOP (IOPg), corneal-compensated IOP (IOPcc), corneal hysteresis (CH), and corneal resistance factor (CRF) by ocular response analyzer, noncontact intraocular pressure (IOPNCT) by noncontact tonometer, and Ehlers, Shah, Dresden, Kohlhaas, Orssengo/Pye by the Pentacam corrected system. Changes in both groups and differences between groups were evaluated. Multiple linear regression models were constructed to explore factors influencing IOP changes. In SMILE, the IOPg, IOPcc, IOPNCT, and Kohlhaas decreased significantly at 1 month postoperatively (P < 0.01), whereas with the Ehlers formula they significantly increased (P < 0.01). IOPs decreased at 3 and 6 months compared with all preoperative values except Ehlers values (P < 0.01), but there was no significant difference between 3 and 6 months (P > 0.05). In FS-LASIK, the IOPg, IOPcc, and IOPNCT decreased significantly at 1 month (P < 0.01), whereas in the Ehlers and Shah formulas they significantly increased (P < 0.01). Compared with preoperative values, the IOPs decreased at 3 and 6 months except in the Ehlers and Shah formulas (P < 0.01). Only IOPg and IOPcc differed between 3 and 6 months (P < 0.05). The Ehlers and Shah formulas were closer to the preoperative IOP for both groups, with variation approximately 1 mm Hg at 6 months postoperatively. Preoperative IOP, postoperative corneal resistance factor, corneal hysteresis, and flat keratometry were enrolled into the regression equations. IOP underestimation after SMILE or FS-LASIK was related to corneal biomechanics as well as preoperative IOP and flat keratometry. IOP after SMILE seem to remain more stable. Accordingly, the Ehlers and Shah formulas were closer to the preoperative IOP. It may be useful to estimate future IOP with the best-fit models after surgery.


Efficient nematode swimming in a shear thinning colloidal suspension.
The swimming behavior of a nematode Caenorhabditis elegans (C. elegans) is investigated in a non-Newtonian shear thinning colloidal suspension. At the onset value (ϕ∼ 8%), the suspension begins to exhibit shear thinning behavior, and the average swimming speed of worms jumps by approximately 12% more than that measured in a Newtonian solution exhibiting no shear dependent viscosity. In the shear thinning regime, we observe a gradual yet significant improvement in swimming efficiency with an increase in ϕ while the swimming speed remains nearly constant. We postulate that this enhanced swimming can be explained by the temporal change in the stroke form of the nematode that is uniquely observed in a shear thinning colloidal suspension: the nematode features a fast and large stroke in its head to overcome the temporally high drag imposed by the viscous medium, whose effective viscosity (ηs) is shown to drop drastically, inversely proportional to the strength of its stroke. Our results suggest new insights into how nematodes efficiently maneuver through the complex fluid environment in their natural habitat.


Imaging agents for in vivo molecular profiling of disseminated prostate cancer--targeting EGFR receptors in prostate cancer: comparison of cellular processing of [111In]-labeled affibody molecule Z(EGFR:2377) and cetuximab.
Expression of receptor tyrosine-kinase (RTK) EGFR is low in normal prostate, but increases in prostate cancer. This receptor is significantly up-regulated as tumors progress into higher grade, androgen-insensitive and metastatic lesions. The up-regulated receptors could serve as targets for novel selective anti-cancer drugs, e.g. antibodies and tyrosine kinase inhibitors. Radionuclide imaging of RTK can facilitate patient stratification and monitoring of anti-RTK therapy of prostate cancer. The goal of the study was to evaluate binding and cellar processing of radiolabeled EGFR-targeting conjugates by prostate cancer cell lines. Receptor expression of EGFR was studied in three prostate cancer cell lines: DU145 (brain metastasis of PC, hormone insensitive), PC3 (bone metastasis of PC) and LNCaP (lymph node metastasis of PC, androgen and estrogen receptor positive). Uptake and internalization of anti-EGFR mAbs (cetuximab) and affibody molecule (Z2377) labeled with indium-111 was investigated. EGFR expression on prostate cancer cell lines was clearly demonstrated. Both labelled conjugates 111In-Z2377 and 111In-cetuximab bound to prostate cancer cells in the receptor mediated model. Expression levels were modest but correlate with degree of hormone independence. Internalization of Affibody molecules was relatively slow in all cell lines. Internalization of mAbs was more rapid. The level of EGFR expression in these cell lines is sufficient for in vivo molecular imaging. Slow internalization indicates possibility of the use of non-residualizing labels for affibody molecules.


delta-Opioid receptor-stimulated Akt signaling in neuroblastoma x glioma (NG108-15) hybrid cells involves receptor tyrosine kinase-mediated PI3K activation.
delta-Opioid receptor (DOR) agonists possess cytoprotective properties, an effect associated with activation of the "pro-survival" kinase Akt. Here we delineate the signal transduction pathway by which opioids induce Akt activation in neuroblastomaxglioma (NG108-15) hybrid cells. Exposure of the cells to both [D-Pen(2,5)]enkephalin and etorphine resulted in a time- and dose-dependent increase in Akt activity, as measured by means of an activation-specific antibody recognizing phosphoserine-473. DOR-mediated Akt signaling is blocked by the opioid antagonist naloxone and involves inhibitory G(i/o) proteins, because pre-treatment with pertussis toxin, but not over-expression of the G(q/11) scavengers EBP50 and GRK2-K220R, prevented this effect. Further studies with Wortmannin and LY294002 revealed that phophoinositol-3-kinase (PI3K) plays a central role in opioid-induced Akt activation. Opioids stimulate Akt activity through transactivation of receptor tyrosine kinases (RTK), because pre-treatment of the cells with inhibitors for neurotrophin receptor tyrosine kinases (AG879) and the insulin-like growth factor receptor IGF-1 (AG1024), but not over-expression of the Gbetagamma scavenger phosducin, abolished this effect. Activated Akt translocates to the nuclear membrane, where it promotes GSK3 phosphorylation and prevents caspase-3 cleavage, two key events mediating inhibition of cell apoptosis and enhancement of cell survival. Taken together, these results demonstrate that in NG108-15 hybrid cells DOR agonists possess cytoprotective properties mediated by activation of the RTK/PI3K/Akt signaling pathway.


'Clean athlete status' cannot be certified: Calling for caution, evidence and transparency in 'alternative' anti-doping systems.
Athletes, sponsors and sport organisations all have a vested interest in upholding the values of clean sport. Despite the considerable and concerted efforts of the global anti-doping system over two decades, the present system is imperfect. Capitalising upon consequent frustrations of athletes, event organisers and sponsors, alternative anti-doping systems have emerged outside the global regulatory framework. The operating principles of these systems raise several concerns, notably including accountability, legitimacy and fairness to athletes. In this paper, we scrutinise the Clean Protocol™, which is the most comprehensive alternative system, for its shortcomings through detailed analysis of its alleged logical and scientific merits. Specifically, we draw the attention of the anti-doping community - including researchers and practitioners - to the potential pitfalls of using assessment tools beyond the scope for which they have been validated, and implementing new approaches without validation. Further, we argue that whilst protecting clean sport is critically important to all stakeholders, protocols that put athletes in disadvantageous positions and/or pose risks to their professional and personal lives lack legitimacy. We criticise the use of anti-doping data and scientific research out of context, and highlight unintended harms that are likely to arise from the widespread implementation of such protocols in parallel with - or in place of - the existing global anti-doping framework.


Adoptive transfer of insulitis and diabetes in neonates of diabetes-prone and -resistant rats. Tissue localization of injected blasts.
Intravenous transfusion of concanavalin A-activated splenic cells from acutely diabetic BB or diabetic BB/hooded hybrid donor rats into 6- to 36-h-old neonate recipients of diabetes-prone and -resistant rat lines induced insulitis and in some severe diabetes. These effects were observed 10-20 days after the injection of the blasts. Focal lesions of insulitis were absent in neonates killed 1 and 3 days after the blast injection but were observed in neonates killed on the 5th and 8th day. As determined by autoradiography after the injection of [3H]thymidine-labeled blasts, numerous blast cells migrated and settled in various immature lymph nodes and in the spleen within 24 h after injection. Focal mononuclear infiltrations in the islets containing labeled and unlabeled cells were again observed on the 5th and 8th day but not on the 1st and 3rd day after injection. These experiments indicate that target-specific blasts undergo a short phase of proliferation and maturation in lymphoid organs of the recipients, before initiating the autoimmune process in the pancreatic islets. They further suggest that specific immune cells rather than humoral anti-islet antibodies are more likely to play the major role in this autoimmune animal model of diabetes.


Functional relaxation as complementary therapy in irritable bowel syndrome: a randomized, controlled clinical trial.
Irritable bowel syndrome (IBS) is a frequently disabling and almost invariably distressing disease with a high overall prevalence. Numerous trials identified the importance of psychogenic and emotional etiological factors, and this is obvious in clinical practice. Although relaxation techniques are frequently recommended, there is still a lack of evidence for their efficacy in the management of IBS. This study therefore aims to determine the efficacy of functional relaxation (FR) in IBS. The subjects were 80 patients with IBS. Participants were randomly allocated either to FR or to enhanced medical care (EMC: treatment as usual plus two counseling interviews) as control intervention with 2 weekly sessions over the 5-week trial each. Thirty-nine (39) patients completed FR and 39 received EMC. An impairment-severity score (IS) was employed as the primary outcome parameter with assessment at baseline, after treatment, and again after 3-month follow-up. FR was significantly superior to EMC with a standardized effect size of 0.85. The achieved effects through FR remained stable in terms of psychic and bodily impairment after 3-month follow-up. The results of our trial suggest a positive effect of FR training on subjective functional impairment in the IS, if provided in addition to treatment as usual (TAU). There appears to be a clinically relevant long-term benefit of FR as a nonpharmacological and complementary therapy approach in IBS.


Neuroprotective effect of Celastrus paniculatus on chronic stress-induced cognitive impairment.
Several studies report that chronic stress results in impaired spatial learning and working memory and enhanced anxiety-like behavior. However, not many studies have looked into the possible ways of reversing stress-induced deficits. Celastrus paniculatus (CP), a traditional ayurvedic herbal medicine, was used to treat cognitive deficits in mentally retarded children. CP oil has been reported to have neuroprotective and antioxidant activities. However, the effects of CP oil on chronic stress-induced cognitive deficits are unclear. In the present study, we intended to analyze the neuroprotective effects of CP oil on stress-associated cognitive dysfunctions. Chronic stress was induced by subjecting rats to restrainers for 6 h a day for 21 days. CP oil (400, 600 mg/kg) or vehicle was administered intraperitoneally (i.p.) after stress protocol once a day over the next 14 days. Groups used in the present study: normal control, stress, stress + vehicle, stress + CP oil at 2 different doses (400 and 600 mg/kg, i.p.). After the drug treatment, open field and elevated plus maze (EPM) were used to analyze anxiety-like behavior, and partially baited radial arm maze (RAM) and T-maze were used to evaluate spatial learning and memory capabilities. Analysis has been done using two-way ANOVA followed by Bonferroni's post hoc test and one-way ANOVA followed by Tukey's post hoc test. Stressed rats showed enhanced anxiety-like behavior in EPM (P < 0.001) and impaired performance in RAM (P < 0.001) and T-maze tasks (P < 0.001) compared to normal animals. In contrast, CP oil treatment to these rats improved their performance in both RAM (P < 0.001) and T-maze (P < 0.001). In addition, CP oil significantly reduced stress-induced anxiety behavior (P < 0.001). Chronic treatment with CP oil is to improve cognitive abilities in chronically stressed rats. The current study provides a novel perspective on beneficial effect of herbal therapy on stress-induced cognitive dysfunctions.


Decoupling the Variances of Heterosis and Inbreeding Effects Is Evidenced in Yeast's Life-History and Proteomic Traits.
Heterosis (hybrid vigor) and inbreeding depression, commonly considered as corollary phenomena, could nevertheless be decoupled under certain assumptions according to theoretical population genetics works. To explore this issue on real data, we analyzed the components of genetic variation in a population derived from a half-diallel cross between strains from Saccharomyces cerevisiae and S. uvarum, two related yeast species involved in alcoholic fermentation. A large number of phenotypic traits, either molecular (coming from quantitative proteomics) or related to fermentation and life history, were measured during alcoholic fermentation. Because the parental strains were included in the design, we were able to distinguish between inbreeding effects, which measure phenotypic differences between inbred and hybrids, and heterosis, which measures phenotypic differences between a specific hybrid and the other hybrids sharing a common parent. The sources of phenotypic variation differed depending on the temperature, indicating the predominance of genotype-by-environment interactions. Decomposing the total genetic variance into variances of additive (intra- and interspecific) effects, of inbreeding effects, and of heterosis (intra- and interspecific) effects, we showed that the distribution of variance components defined clear-cut groups of proteins and traits. Moreover, it was possible to cluster fermentation and life-history traits into most proteomic groups. Within groups, we observed positive, negative, or null correlations between the variances of heterosis and inbreeding effects. To our knowledge, such a decoupling had never been experimentally demonstrated. This result suggests that, despite a common evolutionary history of individuals within a species, the different types of traits have been subject to different selective pressures.


Anti-oxidized low-density lipoprotein antibodies in chronic heart failure.
Oxidative stress may play a significant role in the pathogenesis of heart failure (HF). Antibodies to oxidized low-density lipoprotein (oxLDL Abs) reflect an immune response to LDL over a prolonged period and may represent long-term oxidative stress in HF. The oxLDL plasma level is a useful predictor of mortality in HF patients, and measurement of the oxLDL Abs level may allow better management of those patients. Antibodies to oxLDL also significantly correlate with the New York Heart Association score. Hypercholesterolemia, smoking, hypertension, and obesity are risk factors for atherosclerotic coronary heart disease (CHD) leading to HF, but these factors account for only one-half of all cases, and understanding of the pathologic process underlying HF remains incomplete. Nutrients with antioxidant properties can reduce the susceptibility of LDL to oxidation. Antioxidant therapy may be an adjunct to lipid-lowering, angiotensin converting enzyme inhibition and metformin (in diabetes) therapy for the greatest impact on CHD and HF. Observational data suggest a protective effect of antioxidant supplementation on the incidence of HD. This review summarizes the data on oxLDL Abs as a predictor of morbidity and mortality in HF patients.


The Effects of Tree Nut and Peanut Consumption on Energy Compensation and Energy Expenditure: A Systematic Review and Meta-Analysis.
Nut consumption is not associated with a higher body weight, and potential energy-regulating mechanisms may include a reduced subsequent energy intake and increased EE. The aim of this study was to examine the effect of tree nut and peanut consumption on energy intake, compensation, and expenditure. PubMed, MEDLINE, CINAHL, Cochrane, and Embase databases were searched from inception to June 2, 2021. Human studies with adults aged ≥18 y older were included. Energy intake and compensation studies were restricted to acute effects (intervention duration of ≤24 h), whereas intervention duration was not limited for EE studies. Random effects meta-analyses were conducted to explore weighted mean differences in REE. Twenty-eight articles from 27 studies (16 energy intake studies, 10 EE studies, and 1 study investigating both) with 1121 participants were included in this review, with a variety of nut types addressed (almonds, Brazil nuts, cashews, chestnuts, hazelnuts, peanuts, pistachios, walnuts, and mixed nuts). Energy compensation occurred after nut-containing loads (range: -280.5% to +176.4%) and the degree of compensation varied depending on the form (whole and chopped) and how they were consumed (alone and within a meal). The meta-analyses identified a nonsignificant increase in REE associated with nut consumption (weighted mean difference: 28.6 kcal/d; 95% CI: -10.7, 67.8 kcal/d). This study provided support for energy compensation as a potential mechanism for a lack of association between nut consumption and body weight, whereas no evidence was found for EE as an energy-regulating mechanism of nuts. This review was registered at PROSPERO as CRD42021252292.


Exponential stability analysis for delayed neural networks with switching parameters: average dwell time approach.
This paper is concerned with the problem of exponential stability analysis of continuous-time switched delayed neural networks. By using the average dwell time approach together with the piecewise Lyapunov function technique and by combining a novel Lyapunov-Krasovskii functional, which benefits from the delay partitioning method, with the free-weighting matrix technique, sufficient conditions are proposed to guarantee the exponential stability for the switched neural networks with constant and time-varying delays, respectively. Moreover, the decay estimates are explicitly given. The results reported in this paper not only depend upon the delay but also depend upon the partitioning, which aims at reducing the conservatism. Numerical examples are presented to demonstrate the usefulness of the derived theoretical results.


Advantages of extended bottom-up proteomics using Sap9 for analysis of monoclonal antibodies.
Despite the recent advances in structural analysis of monoclonal antibodies with bottom-up, middle-down, and top-down mass spectrometry (MS), further improvements in analysis accuracy, depth, and speed are needed. The remaining challenges include quantitatively accurate assignment of post-translational modifications, reduction of artifacts introduced during sample preparation, increased sequence coverage per liquid chromatography (LC) MS experiment, and ability to extend the detailed characterization to simple antibody cocktails and more complex antibody mixtures. Here, we evaluate the recently introduced extended bottom-up proteomics (eBUP) approach based on proteolysis with secreted aspartic protease 9, Sap9, for analysis of monoclonal antibodies. Key findings of the Sap9-based proteomics analysis of a single antibody include: (i) extensive antibody sequence coverage with up to 100% for the light chain and up to 99-100% for the heavy chain in a single LC-MS run; (ii) connectivity of complementarity-determining regions (CDRs) via Sap9-produced large proteolytic peptides (3.4 kDa on average) containing up to two CDRs per peptide; (iii) reduced artifact introduction (e. g., deamidation) during proteolysis with Sap9 compared to conventional bottom-up proteomics workflows. The analysis of a mixture of six antibodies via Sap9-based eBUP produced comparable results. Due to the reasons specified above, Sap9-produced proteolytic peptides improve the identification confidence of antibodies from the mixtures compared to conventional bottom-up proteomics dealing with shorter proteolytic peptides.


Nickel ions selectively inhibit lipopolysaccharide-induced interleukin-6 production by decreasing its mRNA stability.
Nickel (Ni) ions easily elute from many alloys and elicit inflammation and allergies. Previous studies have shown that infections due to the implantation of medical devices cause inflammation and enhance the elution of Ni ions (Ni²⁺). However, cross-talk between infection- and Ni²⁺-induced signaling pathways has not yet been elucidated in detail. In the present study, we investigated the effects of Ni2+ on the lipopolysaccharide (LPS)-induced production of cytokines in a LPS-induced air pouch-type inflammation model in BALB/c mice and the murine macrophage cell line RAW264. We demonstrated that Ni²⁺ inhibited the LPS-induced production of interleukin (IL)-6, but not that of tumor necrosis factor (TNF)-α both in vivo and in vitro. This inhibitory effect was also observed with cobalt ion (Co²⁺), but not with chloride ion (Cl⁻), zinc ion (Zn²⁺), or palladium ion (Pd²⁺), and was highly selective to the production of IL-6. Ni²⁺ did not inhibit the activation of ERK1/2, p38 MAPK, or JNK. Although Ni²⁺ decreased IL-6 mRNA levels, it failed to inhibit the LPS-induced activation of the IL-6 promoter. An experiment using actinomycin D, a transcription inhibitor, revealed that Ni²⁺ decreased the stability of IL-6 mRNA. Moreover, Ni²⁺ inhibited the LPS-induced expression of Arid5a, but not regnase-1. These results demonstrated that Ni²⁺ may have selectively inhibited the LPS-induced production of IL-6 by decreasing the Arid5a-dependent stabilization of IL-6 mRNA.


Control of human trophoblast function.
The trophoblast, i.e. the peripheral part of the human conceptus, exerts a crucial role in implantation and placentation. Both processes properly occur as a consequence of an intimate dialogue between fetal and maternal tissues, fulfilled by membrane ligands and receptors, as well as by hormone and local factor release. During blastocyst implantation, generation of distinct trophoblast cell types begins, namely the villous and the extravillous trophoblast, the former of which is devoted to fetal-maternal exchanges and the latter binds the placental body to the uterine wall. Physiological placentation is characterized by the invasion of the uterine spiral arteries by extravillous trophoblast cells arising from anchoring villi. Due to this invasion, the arterial structure is replaced by amorphous fibrinoid material and endovascular trophoblastic cells. This transformation establishes a low-resistance, high-capacity perfusion system from the radial arteries to the intervillous space, in which the villous tree is embedded. The physiology of pregnancy depends upon the orderly progress of structural and functional changes of villous and extravillous trophoblast, whereas a derangement of such processes can lead to different types of complications of varying degrees of gravity, including possible pregnancy loss and maternal life-threatening diseases. In this review we describe the mechanisms which regulate trophoblast differentiation, proliferation, migration and invasiveness, and the alterations in these mechanisms which lead to pathological conditions. Furthermore, based on the growing evidence that proper inflammatory changes and oxidative balance are needed for successful gestation, we explain the mechanisms by which agents able to influence such processes may be useful in the prevention and treatment of pregnancy disorders.


Chondroitin sulfate-E is a negative regulator of a pro-tumorigenic Wnt/beta-catenin-Collagen 1 axis in breast cancer cells.
Expression of the glycosaminoglycan chondroitin sulfate-E (CS-E) is misregulated in many human cancers, including breast cancer. Cell-surface associated CS-E has been shown to have pro-tumorigenic functions, and pharmacological treatment with exogenous CS-E has been proposed to interfere with tumor progression mediated by endogenous CS-E. However, the effects of exogenous CS-E on breast cancer cell behavior, and the molecular mechanisms deployed by CS-E are not well understood. We show here that treatment with CS-E, but not other chondroitin forms, could interfere with the invasive protrusion formation and migration of breast cancer cells in three-dimensional organotypic cultures. Microarray analysis identified transcriptional programs controlled by CS-E in these cells. Importantly, negative regulation of the pro-metastatic extracellular matrix gene Col1a1 was required for the anti-migratory effects of exogenous CS-E. Knock-down of Col1a1 gene expression mimics the effects of CS-E treatment, while exposing cells to a preformed collagen I matrix interfered with the anti-migratory effects of CS-E. In addition, CS-E specifically interfered with Wnt/beta-catenin signaling, a known pro-tumorigenic pathway. Lastly, we demonstrate that Col1a1 is a positively regulated target gene of the Wnt/beta-catenin pathway in breast cancer cells. Together, our data identify treatment with exogenous CS-E as negative regulatory mechanism of breast cancer cell motility through interference with a pro-tumorigenic Wnt/beta-catenin-Collagen I axis.


FK506 (tacrolimus) compared with cyclosporine for primary immunosuppression after pediatric liver transplantation. Results from the U.S. Multicenter Trial.
We report on the efficacy and safety of FK506 (tacrolimus) compared with a cyclosporine (CsA)-based immunosuppressive regimen after 1 year of treatment in pediatric liver allograft recipients (< 12 years) participating in a multicenter U.S. randomized trial. Patients received either FK506 or CsA as primary immunosuppression following a first ABO-compatible liver transplant. Intravenous FK506 was initiated at 0.1 mg/kg per day, followed by oral FK506 beginning at 0.3 mg/kg per day. The dose was adjusted to maintain plasma trough levels of 0.5-2.0 ng/ml. The CsA group was treated according to each center's usual protocol. Both groups received the same initial doses of corticosteroids. All rejection episodes were biopsy-proven and a standardized algorithm was adopted for the treatment of rejection. Thirty patients were randomized to the FK506 group and 20 to the CsA group. After twelve months of follow-up 20 patients remained in the FK506 group and 13 in the CsA group. Patient survivals were 80% and graft survival 70% in the FK506 group compared with 81% and 71% respectively, in the CsA group. 48% of the FK506 group remained rejection-free compared with 21% of the CsA group, and 79% of FK506-treated patients did not require OKT3 compared with 68% of CsA treated patients. The cumulative corticosteroid dose was less at each time point throughout the first year in the FK506 group. The incidence of serious and minor infections was similar in both groups. Nephrotoxicity, neurotoxicity, and gastrointestinal disturbances were the major toxicities reported. Differences did not reach statistical significance between the two groups although major neurologic events, diarrhea and dyspepsia were more often reported in the FK506 group. There was no difference in mean serum creatinine at 12 months between the two groups. There was a tendency toward lower mean serum cholesterol in the FK506 group. There was no hirsuitism in the FK506 group compared with a 30% incidence in the CsA group. In conclusion, compared with CsA, there is a trend toward less rejection in FK506-treated pediatric allograft recipients, while both drugs have a similar spectrum of side effects.


Two-component signal transduction pathways regulating growth and cell cycle progression in a bacterium: a system-level analysis.
Two-component signal transduction systems, comprised of histidine kinases and their response regulator substrates, are the predominant means by which bacteria sense and respond to extracellular signals. These systems allow cells to adapt to prevailing conditions by modifying cellular physiology, including initiating programs of gene expression, catalyzing reactions, or modifying protein-protein interactions. These signaling pathways have also been demonstrated to play a role in coordinating bacterial cell cycle progression and development. Here we report a system-level investigation of two-component pathways in the model organism Caulobacter crescentus. First, by a comprehensive deletion analysis we show that at least 39 of the 106 two-component genes are required for cell cycle progression, growth, or morphogenesis. These include nine genes essential for growth or viability of the organism. We then use a systematic biochemical approach, called phosphotransfer profiling, to map the connectivity of histidine kinases and response regulators. Combining these genetic and biochemical approaches, we identify a new, highly conserved essential signaling pathway from the histidine kinase CenK to the response regulator CenR, which plays a critical role in controlling cell envelope biogenesis and structure. Depletion of either cenK or cenR leads to an unusual, severe blebbing of cell envelope material, whereas constitutive activation of the pathway compromises cell envelope integrity, resulting in cell lysis and death. We propose that the CenK-CenR pathway may be a suitable target for new antibiotic development, given previous successes in targeting the bacterial cell wall. Finally, the ability of our in vitro phosphotransfer profiling method to identify signaling pathways that operate in vivo takes advantage of an observation that histidine kinases are endowed with a global kinetic preference for their cognate response regulators. We propose that this system-wide selectivity insulates two-component pathways from one another, preventing unwanted cross-talk.


Photocarcinogenesis study of glycolic acid and salicylic acid (CAS Nos. 79-14-1 and 69-72-7) in SKH-1 mice (simulated solar light and topical application study).
Acidic solutions have been used for decades to treat a variety of skin conditions. Many of these solutions consist of organic acids with a hydroxy group on a carbon adjacent to the carbonyl carbon and are referred to as alpha-hydroxy acids (AHA). Organic acids with hydroxy groups on the second carbon from the carbonyl carbon are referred to as beta-hydroxy acids (BHA). Both AHA and BHA are used to treat various skin conditions. One of the most widely used AHA is glycolic acid, while salicylic acid is a commonly used BHA. Chemical peels containing 20% to 70% glycolic acid have been used by dermatologists to treat ichthyosis, acne, xerosis, actinic keratosis, seborrheic keratoses, warts, and psoriasis. AHA have recently been used to treat photoaged skin and are now included in many commercially available cosmetic skin treatments. When used in a formulation for a chemical peel, topical treatment of skin with AHA and BHA can result in removal of the stratum corneum, alteration of the skin's histology, and increased cell proliferation in the basal layer of the epidermis. Since AHA and BHA are used to correct photoaged skin, and since exposure to sunlight of skin treated with AHA or BHA is likely, studies were designed to determine the effects of topical application of creams containing AHA (0%, 4%, or 10% glycolic acid, pH 3.5) or BHA (0%, 2%, or 4% salicylic acid, pH 4.0) on the photocarcinogenesis of simulated solar radiation using a filtered 6.5 kW xenon arc light source [simulated solar light (SSL)]. Male and female Crl:SKH-1 (hr-/hr-) hairless mice were exposed to glycolic acid or salicylic acid alone or in combination with SSL for 40 weeks, and the mice were followed for an additional 12 weeks. 1-YEAR STUDY IN MICE: Groups of 36 male and 36 female mice were exposed to 0.0, 0.3, 0.6, or 0.9 minimal erythema dose (MED) of SSL during the afternoon (1200 to 1600 hours) 5 days per week for 40 weeks. Groups of 18 male and 18 female mice were treated in the morning (0800 to 1100 hours) with 2 mg/cm2 control cream, 4% glycolic acid cream, 10% glycolic acid cream, 2% salicylic acid cream, or 4% salicylic acid cream on the dorsal skin, and in the afternoon (1200 to 1600 hours) with 0.3 MED of SSL 5 days per week for 40 weeks. Additional groups of 18 male and 18 female mice were treated in the morning (0800 to 1100 hours) with 2 mg/cm2 control cream, 4% glycolic acid cream, 10% glycolic acid cream, 2% salicylic acid cream, or 4% salicylic acid cream on the dorsal skin, and in the afternoon (1200 to 1600 hours) with 0.6 MED of SSL 5 days per week for 40 weeks. All mice were held an additional 12 weeks following the end of treatment. There were no effects of SSL exposure or topical treatment on the body weights of the mice. Increasing doses of SSL resulted in an SSL-dose trend in survival, with the greatest dose of SSL causing the earliest removal. This effect was present in both the untreated and control cream treated mice. The only consistent effect of glycolic acid on survival was a dose-dependent increase in survival of females at 0.3 MED SSL. Survival was increased in mice exposed to 0.6 MED of SSL and treated with 2% and 4% salicylic acid compared to mice treated with 0.6 MED and treated only with the vehicle. This effect was not observed in the mice treated with 0.0 and 0.3 MED of SSL and salicylic acid compared to the control groups. The mean or median time to first skin tumor of at least 1 mm decreased with increasing SSL exposure concentration in mice that were not treated with cream. Addition of the control cream resulted in a decrease in the time to tumor at 0.3 and 0.6 MED of SSL in male and female mice. The addition of glycolic acid (4% or 10%) did not affect the time to tumor in male or female mice at either SSL dose when compared to mice receiving the control cream. When compared to mice receiving control cream, the inclusion of 4% salicylic acid in the cream increased the time to tumor for male mice receiving 0.3 or 0.6 MED of SSL and female mice receiving 0.3 MED of SSL. The results indicate that inclusion of glycolic acid in the topical cream had no effect on the time required to induce tumors by SSL; however, inclusion of salicylic acid at 4% in the cream was photoprotective, increasing the time required to achieve median tumor incidence at a corresponding dose of SSL and control cream. The skin tumors induced by SSL in mice were squamous cell papilloma, carcinoma in situ, and squamous cell carcinoma. Except for papilloma in male mice, the tumors were induced in a dose-dependent manner by SSL in male and female mice. In male and female mice treated with control cream, the exposure to SSL caused significant increases in the incidences of carcinoma in situ, squamous cell carcinoma, and the combined incidence of carcinoma in situ and squamous cell carcinoma. When male or female mice were exposed to 0.3 or 0.6 MED SSL, the inclusion of 4% or 10% glycolic acid did not affect the induction of skin neoplasms over the incidence detected when the control cream was used, with the single exception of a glycolic acid dose-trend in squamous cell carcinoma incidence in male mice receiving 0.3 MED SSL. The inclusion of salicylic acid in the cream that was topically applied to female mice did not affect squamous cell papilloma formation at either SSL dose. The incidence of carcinoma in situ was decreased in male and female mice at 0.3 MED SSL when treated with 4% salicylic acid. A salicylic acid dose-trend was also observed in both sexes at 0.3 MED SSL. These experiments investigated the impact of topical application of a cosmetic formulation containing 4% or 10% glycolic acid (pH 3.5) or 2% or 4% salicylic acid (pH 4) on the photocarcinogenesis of filtered 6.5 kW xenon arc simulated solar light (SSL) in SKH-1 hairless mice. Taking into consideration the survival data, time to tumor data, and the pathology results, glycolic acid did not alter the photocarcinogenesis of SSL, and salicylic acid was photoprotective, reducing the carcinogenicity of 0.3 MED SSL.


The maintenance of traditions in marmosets: individual habit, not social conformity? A field experiment.
Social conformity is a cornerstone of human culture because it accelerates and maintains the spread of behaviour within a group. Few empirical studies have investigated the role of social conformity in the maintenance of traditions despite an increasing body of literature on the formation of behavioural patterns in non-human animals. The current report presents a field experiment with free-ranging marmosets (Callithrix jacchus) which investigated whether social conformity is necessary for the maintenance of behavioural patterns within groups or whether individual effects such as habit formation would suffice. Using a two-action apparatus, we established alternative behavioural patterns in six family groups composed of 36 individuals. These groups experienced only one technique during a training phase and were thereafter tested with two techniques available. The monkeys reliably maintained the trained method over a period of three weeks, despite discovering the alternative technique. Three additional groups were given the same number of sessions, but those 21 individuals could freely choose the method to obtain a reward. In these control groups, an overall bias towards one of the two methods was observed, but animals with a different preference did not adjust towards the group norm. Thirteen of the fifteen animals that discovered both techniques remained with the action with which they were initially successful, independent of the group preference and the type of action (Binomial test: exp. proportion: 0.5, p<0.01). The results indicate that the maintenance of behavioural patterns within groups 1) could be explained by the first rewarded manipulation and subsequent habit formation and 2) do not require social conformity as a mechanism. After an initial spread of a behaviour throughout a group, this mechanism may lead to a superficial appearance of conformity without the involvement of such a socially and cognitively complex mechanism. This is the first time that such an experiment has been conducted with free-ranging primates.


Chromatin methylation activity of Dnmt3a and Dnmt3a/3L is guided by interaction of the ADD domain with the histone H3 tail.
Using peptide arrays and binding to native histone proteins, we show that the ADD domain of Dnmt3a specifically interacts with the H3 histone 1-19 tail. Binding is disrupted by di- and trimethylation of K4, phosphorylation of T3, S10 or T11 and acetylation of K4. We did not observe binding to the H4 1-19 tail. The ADD domain of Dnmt3b shows the same binding specificity, suggesting that the distinct biological functions of both enzymes are not related to their ADD domains. To establish a functional role of the ADD domain binding to unmodified H3 tails, we analyzed the DNA methylation of in vitro reconstituted chromatin with Dnmt3a2, the Dnmt3a2/Dnmt3L complex, and the catalytic domain of Dnmt3a. All Dnmt3a complexes preferentially methylated linker DNA regions. Chromatin substrates with unmodified H3 tail or with H3K9me3 modification were methylated more efficiently by full-length Dnmt3a and full-length Dnmt3a/3L complexes than chromatin trimethylated at H3K4. In contrast, the catalytic domain of Dnmt3a was not affected by the H3K4me3 modification. These results demonstrate that the binding of the ADD domain to H3 tails unmethylated at K4 leads to the preferential methylation of DNA bound to chromatin with this modification state. Our in vitro results recapitulate DNA methylation patterns observed in genome-wide DNA methylation studies.


Comparison of molecular strategies for breast cancer virotherapy using oncolytic adenovirus.
Oncolytic viruses are regulated by the tumor phenotype to replicate and lyse cancer cells selectively. To identify optimal strategies for breast cancer we compared five adenoviruses with distinct regulatory mechanisms: Ad-dl922-947 (targets G1-S checkpoint); Ad-Onyx-015 and Ad-Onyx-017 (target p53/mRNA export); Ad-vKH1 (targets Wnt pathway), and AdEHE2F (targets estrogen receptor/G1-S checkpoint/hypoxic signaling). The quantity of virus required to kill 50% of breast cancer cells after 6 days (EC(50), plaque-forming units per cell) was measured. The most potent virus was Ad-dl922-947 (EC(50), 0.01-5.4 in SkBr3, MDA-231, MDA-468, MCF7, and ZR75.1 cells), followed by wild-type (Ad-WT; EC(50), 0.3-5.5) and AdEHE2F (EC(50), 1.4-3.9). Ad-vKH1 (EC(50), 7.2-72.1), Ad-Onyx-017 (EC(50), 8.4-167), and Ad-Onyx-015 (EC(50), 17.7-377) showed less activity. Most viruses showed limited cytotoxicity in normal human cells, including breast epithelium MCF10A (EC(50), >722) and fibroblasts (EC(50), >192) and only moderate cytotoxicity in normal microvascular endothelial cells (HMVECs; EC(50), 42.8-149), except Ad-dl922-947, which was active in HMVECs (EC(50), 1.6). After injection into MDA-231 xenografts, Ad-WT, AdEHE2F, and Ad-dl922-947 showed replication, assessed by hexon staining and quantitative polymerase chain reaction measurement of viral DNA, and significantly inhibited tumor growth, leading to extended survival. After intravenous injection Ad-dl922-947 showed DNA replication (233% of the injected dose was measured in liver after 3 days) whereas AdEHE2F did not. Overall, AdEHE2F showed the best combination of low toxicity in normal cells and high activity in breast cancer in vitro and in vivo, suggesting that molecular targeting using estrogen response elements, hypoxia response elements, and a dysregulated G1-S checkpoint is a promising strategy for virotherapy of breast cancer.


[NUTRIENTS AND RADIOTHERAPY; REVIEW OF THE LITERATURE].
nutrition is an important influence on treatments and quality of life of cancer patients. The relationship between different nutritional components and radiotherapy is today a topic of growing interest. to evaluate the influence of macro and micronutrients on tolerance and effectiveness of radiotherapy and their role in modulating chronic toxicity. we performed a research of the published literature by consulting the MEDLINE database and Cochrane Library online between 1995 and 2015, relevant publications based on impact factor were selected. Data from the analyzed studies were exposed in sections by type of nutrient. most of the studies showed common features: small sample sizes, high heterogeneity, underpowered results and few prospective randomized studies. In the section of fiber, its use in prophylaxis and treatment of radiation enteritis has been successfully evaluated in some studies, although evidence of its recommendation is still weak. Omega 3 and omega 6 fatty acids have a high metabolic potential, however the evidence regarding this benefit is limited to observational studies in certain tumors. Among the amino acids, glutamine is the most studied, and controversial results of its effect on mucositis, esophagitis and radiation enteritis were found. Vitamins and minerals are a heterogeneous group of substances that showed potential benefit due to their antioxidant activity and their supposed protector effect against toxicity secondary to radiotherapy. Ketogenic diets are beginning to be clinically studied after promising preclinical results. the analyzed studies show controversial or inconclusive results regarding the influence of nutrients in the radiotherapy. It has been not found Sorong evidence about their role in patients receiving ionizing radiation. Well-designed, prospective and randomized studies are needed to establish recommendations.


3D microlasers from self-assembled cholesteric liquid-crystal microdroplets.
We demonstrate a tunable and omnidirectional microlaser in the form of a microdroplet of a dye-doped, cholesteric liquid crystal in a carrier fluid. The cholesteric forms a Bragg-onion optical microcavity and the omnidirectional 3D lasing is due to the stimulated emission of light from the dye molecules in the liquid crystal. The lasing wavelength depends solely on the natural helical period of the cholesteric and can be tuned by varying the temperature. Millions of microlasers can be formed simply by mixing a liquid crystal, a laser dye and a carrier fluid, thus providing microlasers for soft-matter photonic devices.


Differential prevalence and diversity of haemosporidian parasites in two sympatric closely related non-migratory passerines.
Haemosporidian parasites infecting birds show distinct heterogeneity in their distribution among host species. However, despite numerous studies on the prevalence and diversity of parasite communities across species, very little is known on patterns of differences between them. Such data is lacking because up to date the majority of studies explored the patterns of variation in infections in different years, different time of sampling within a year or a breeding cycle, different study sites or was based on a small sample size, all of which may affect the estimates of prevalence and parasite diversity. Here, the prevalence, richness and diversity of haemosporidian parasites from the genera Plasmodium and Haemoproteus were studied in two closely related non-migratory hole-nesting passerines: Great Tits and Blue Tits. Birds were sampled in sympatrically breeding populations during two seasons at the same stage of their breeding cycle - late nestling care. Great Tits were more prevalently infected with Plasmodium and Haemoproteus parasites (97·1 vs 71·2%), harboured a higher proportion of multiple infections (26·2 vs 3·2%) and had a more diverse parasite community (11 vs 5 parasite lineages) than Blue Tits. Observed differences between two host species are discussed with reference to their breeding densities and immunological and behavioural characteristics.


17 Beta-estradiol increases VEGF receptor-2 and promotes DNA synthesis in retinal microvascular endothelial cells.
Estrogen is known to promote angiogenesis in gonads. The presence of estrogen receptors in the vascular endothelium of organs other than gonads has been reported. The goal of this study was to determine whether estrogen promotes the proliferation of retinal microvascular endothelial cells and to explore the mechanism of it. DNA was quantitated using primary cultures of bovine retinal endothelial cells that were incubated with different doses of 17 beta-estradiol (E2), VEGF, or both. The changes in expression level of VEGF and VEGF receptor-2 (VEGFR2) were measured using northern blot analysis after treatment with E2. The presence of estrogen receptors in the endothelial cells was studied by immunohistochemistry and western blot analysis. 17 Beta-estradiol (E2) increased the DNA level in bovine retinal capillary endothelial cells (BRECs) by 177% at 1 nM (P < 0.05) and 150% at 10 nM (P < 0.05) by comparison with unstimulated BREC. One hundred nanomole tamoxifen completely blocked the E2-induced DNA synthesis in BRECs. Ten nanomole E2 augmented vascular endothelial growth factor (VEGF)-induced DNA synthesis in BRECs significantly (160%, P < 0.01). Ten nanomole E2 also increased VEGF mRNA expression, which peaked after 24 hours (6.7 times, P < 0.05), and VEGF receptor-2 (VEGFR2) mRNA expression, which peaked after 9 hours (2.4 times, P < 0.05). The mRNA expression level of VEGFR2 peaked with 10 nM E2 (P < 0.05) and that of VEGF reached maximum with 1 nM E2 (15 times, P < 0.001). VEGFR2 and VEGF proteins increased in parallel with their mRNA levels. Immunocytochemistry showed estrogen receptor expression in BRECs, and western blot analysis indicated the presence of a 67-kDa protein that was compatible with the estrogen receptor. These findings suggest that E2 may stimulate BREC growth by the receptor-mediated pathway and that E2 may augment the VEGF-dependent angiogenesis partly through the upregulation of VEGFR2.


On Design Mining: Coevolution and Surrogate Models.
Design mining is the use of computational intelligence techniques to iteratively search and model the attribute space of physical objects evaluated directly through rapid prototyping to meet given objectives. It enables the exploitation of novel materials and processes without formal models or complex simulation. In this article, we focus upon the coevolutionary nature of the design process when it is decomposed into concurrent sub-design-threads due to the overall complexity of the task. Using an abstract, tunable model of coevolution, we consider strategies to sample subthread designs for whole-system testing and how best to construct and use surrogate models within the coevolutionary scenario. Drawing on our findings, we then describe the effective design of an array of six heterogeneous vertical-axis wind turbines.