Activin regulates betaA-subunit and activin receptor messenger ribonucleic acid and cellular proliferation in activin-responsive testicular tumor cells.
Activin, a member of the transforming growth factor-beta superfamily of growth and differentiation factors, has a number of actions in embryonic as well as adult tissues. These actions are mediated via a family of receptors containing two subtypes and at least two members of each subtype. Recent evidence demonstrates that activin-responsive cell lines containing different subsets of these receptors are valuable models for dissecting functional relationships among receptor subtype, signal transduced, and response obtained. TT cells, derived from a p53(-/-)/alpha-inhibin(-/-) mouse testicular tumor, respond to activin by proliferating, a response that can be inhibited by follistatin (FS) treatment. Using semiquantitative RT-PCR methods, we characterized steady state messenger RNA (mRNA) levels for the inhibin/activin subunits, FS, and activin receptor subtypes under basal conditions and in the presence of activin or FS. These cells produced ample immunoreactive activin A and FS, necessitating higher treatment doses to observe any modulation of cellular proliferation. Furthermore, in the presence of exogenous activin, mRNA levels for activin receptor type IIA (ACTRIIA) and betaA were significantly and profoundly suppressed. In addition, both ACTR1B and ACTRIIB were detectable and down-regulated by exogenous activin, although not to the degree observed for ACTRIIA and betaA. Finally, activin treatment at the higher doses, which decreased activin receptor mRNA levels, resulted in inhibition of cellular proliferation. Taken together with previous observations, our results support the model that these tumor cells respond to an autocrine activin signal by proliferating, whereas exogenous or excess activin results in down-regulation of activin receptor and activin biosynthesis, suggesting a potential autocrine/paracrine mechanism by which activin can modulate its own signal.


Incidence of drug-induced liver injury in medical inpatients.
Drug-induced liver injury (DILI) is a common concern. However, data on DILI epidemiology in inpatients are sparse. To investigate the incidence of DILI, we screened all patients in the pharmacoepidemiological inpatient database according to the CIOMS (Council for International Organisation of Medical Science) criteria, which consist of the evaluation of some clinical chemistry laboratory liver parameters (CIOMS laboratory criteria) and the exclusion of any disease-related causes for the liver injury. Thus, only cases with probable or certain causality according to the World Health Organization criteria were included. Among a total of 6383 patients, liver parameters were determined in 4610, and 489 among them fulfilled the CIOMS laboratory criteria. However, 401 patients had to be excluded because of disease-related liver injury and, thus, the study cohort consisted of 4209 patients at risk for DILI. Among a total of 88 DILI cases, 31 had no documented normal baseline liver parameters and, thus, represented prevalent cases. The remaining 57 represented incident DILI cases. Thus, the incidence of DILI was 1.4% (95% CI 1.0, 1.7). The drug classes most frequently causing DILI were heparins, antibacterials, tuberculostatics and antineoplastic agents. Among those, antineoplastic agents and tuberculostatics showed the highest incidence. Liver injury was not mentioned among the diagnoses or in the physician's discharge letter in about 52-68% of all cases. Approximately 1 in 100 patients develops DILI during hospitalisation in a department of medicine. Incidences of DILI were highest for antineoplastic agents and tuberculostatics. DILI is frequently missed and, therefore, DILI detection by diagnoses will result in misleadingly low incidence rates.


Influence of Prior and Intercurrent Brain Injury on 5-Year Outcome Trajectories After Moderate to Severe Traumatic Brain Injury.
To characterize the influence of additional (both prior and subsequent) traumatic brain injuries (TBIs) on recovery after a moderate to severe index TBI. Traumatic Brain Injury Model Systems centers. Persons with moderate to severe TBI (N = 5054) enrolled in the TBI Model Systems National Database with complete outcome data for the outcomes of interest at 1-, 2-, and 5-year follow-up. Secondary analysis of a prospective longitudinal data set. Prior and intercurrent TBI from the Ohio State University TBI Identification Method (OSU TBI-ID), Disability Rating Scale (DRS), and Functional Independence Measure (FIM). Prior moderate-severe TBIs significantly predicted overall level of functioning on the DRS, FIM Cognitive, and FIM Motor for participants with less severe index injuries. Moderate-severe intercurrent TBIs (TBIs subsequent to the index injury) were predictive of poorer functioning for both Index Severity groups, reflected in higher mean scores on the DRS in participants with less severe index injuries and lower mean Cognitive FIM in participants with more severe index injuries. Multiple brain injuries, particularly those of moderate or greater severity, have a significantly greater impact on patients' level of functioning compared with a single injury, but not the rate or shape of recovery.


Absence of microsomal triglyceride transfer protein in individuals with abetalipoproteinemia.
Abetalipoproteinemia is a human genetic disease that is characterized by a defect in the assembly or secretion of plasma very low density lipoproteins and chylomicrons. The microsomal triglyceride transfer protein (MTP), which is located in the lumen of microsomes isolated from the liver and intestine, has been proposed to function in lipoprotein assembly. MTP activity and the 88-kilodalton component of MTP were present in intestinal biopsy samples from eight control individuals but were absent in four abetalipoproteinemic subjects. This finding suggests that a defect in MTP is the basis for abetalipoproteinemia and that MTP is indeed required for lipoprotein assembly.


Non-cell autonomous impairment of oligodendrocyte differentiation precedes CNS degeneration in the Zitter rat: implications of macrophage/microglial activation in the pathogenesis.
The zitter (zi/zi) rat, a loss-of-function mutant of the glycosylated transmembrane protein attractin (atrn), exhibits widespread age-dependent spongiform degeneration, hypomyelination, and abnormal metabolism of reactive oxygen species (ROS) in the brain. To date, the mechanisms underlying these phenotypes have remained unclear. Here, we show differentiation defects in zi/zi oligodendrocytes, accompanied by aberrant extension of cell-processes and hypomyelination. Axonal bundles were relatively preserved during postnatal development. With increasing in age, the injured oligodendrocytes in zi/zi rats become pathological, as evidenced by the accumulation of iron in their cell bodies. Immunohistochemical analysis revealed that atrn expression was absent from an oligodendrocyte lineage, including A2B5-positive progenitors and CNPase-positive differentiated cells. The number and distribution of Olig2-positive oligodendrocyte progenitors was unchanged in the zi/zi brain. Furthermore, an in vitro differentiation assay of cultured oligodendrocyte progenitors prepared from zi/zi brains revealed their normal competence for proliferation and differentiation into mature oligodendrocytes. Interestingly, we demonstrated the accelerated recruitment of ED1-positive macrophages/microglia to the developing zi/zi brain parenchyma prior to the onset of hypomyelination. Semiquantitative RT-PCR analysis revealed a significant up-regulation of CD26 and IL1-beta in the zi/zi brain during this early postnatal stage. We demonstrated that the onset of the impairment of oligodendrocyte differentiation occurs in a non-cell autonomous manner in zi/zi rats. Hypomyelination of oligodendrocytes was not due to a failure of the intrinsic program of oligodendrocytes, but rather, was caused by extrinsic factors that interrupt oligodendrocyte development. It is likely that macrophage/microglial activation in the zi/zi CNS leads to disturbances in oligodendrocyte differentiation via deleterious extrinsic factors, such as the cytokine IL1-beta or ROS. Atrn might be involved in the activation of brain macrophages/microglia by suppressing excessive migration of monocytes into the CNS, or by accelerating the transformation of brain monocytes into resting microglia. Understanding the pathogenesis of the zi/zi rat may provide novel insights into the developmental interaction betweens macrophages/microglia and cells of an oligodendrocyte lineage.


Constitutive genetic deletion of the growth regulator Nogo-A induces schizophrenia-related endophenotypes.
The membrane protein Nogo-A, which is predominantly expressed by oligodendrocytes in the adult CNS and by neurons mainly during development, is well known for limiting neurite outgrowth and regeneration in the injured mammalian CNS. In addition, it has recently been proposed that abnormal Nogo-A expression or Nogo receptor (NgR) mutations may confer genetic risks for neuropsychiatric disorders of presumed neurodevelopmental origin, such as schizophrenia. We therefore evaluated whether Nogo-A deletion may lead to schizophrenia-like abnormalities in a mouse model of genetic Nogo-A deficiency. Here, we show that systemic, lifelong knock-out of the Nogo-A gene can lead to specific behavioral abnormalities resembling schizophrenia-related endophenotypes: deficient sensorimotor gating, disrupted latent inhibition, perseverative behavior, and increased sensitivity to the locomotor stimulating effects of amphetamine. These behavioral phenotypes were accompanied by altered monoaminergic transmitter levels in specific striatal and limbic structures, as well as changes in dopamine D2 receptor expression in the same brain regions. Nogo-A deletion was further associated with elevated expression of growth-related markers. In contrast, acute antibody-mediated Nogo-A neutralization in adult wild-type mice failed to produce such phenotypes, suggesting that the phenotypes observed in the knock-out mice might be of developmental origin, and that Nogo-A normally subserves critical functions in neurodevelopment. This study provides the first experimental demonstration that Nogo-A bears neuropsychiatric relevance, and alterations in its expression may be one etiological factor in schizophrenia and related disorders.


Expression Patterns and Levels of All Tubulin Isotypes Analyzed in GFP Knock-In C. elegans Strains.
Most organisms have multiple α- and β-tubulin isotypes that likely contribute to the diversity of microtubule (MT) functions. To understand the functional differences of tubulin isotypes in Caenorhabditis elegans, which has nine α-tubulin isotypes and six β-tubulin isotypes, we systematically constructed null mutants and GFP-fusion strains for all tubulin isotypes with the CRISPR/Cas9 system and analyzed their expression patterns and levels in adult hermaphrodites. Four isotypes-α-tubulins TBA-1 and TBA-2 and β-tubulins TBB-1 and TBB-2-were expressed in virtually all tissues, with a distinct tissue-specific spectrum. Other isotypes were expressed in specific tissues or cell types at significantly lower levels than the broadly expressed isotypes. Four isotypes (TBA-5, TBA-6, TBA-9, and TBB-4) were expressed in different subsets of ciliated sensory neurons, and TBB-4 was inefficiently incorporated into mitotic spindle MTs. Taken together, we propose that MTs in C. elegans are mainly composed of four broadly expressed tubulin isotypes and that incorporation of a small amount of tissue-specific isotypes may contribute to tissue-specific MT properties. These newly constructed strains will be useful for further elucidating the distinct roles of tubulin isotypes.Key words: tubulin isotypes, microtubules, C. elegans.


Localization of tenascin in uterine sarcomas and partially transformed endometrial stromal cells.
Normal mesenchymal cells within developing embryonic organs and transformed stromal cells in organs undergoing spontaneous carcinogenesis have the capacity for normal or altered expression of the extracellular matrix glycoprotein tenascin (Tn). Mesenchymal cell constituents of normal adult organs show only a very limited tendency to deposit Tn in their extracellular matrix. In the present study, we investigated whether malignant human mesenchymal cells derived from uterine sarcomas or normal human endometrial stromal cells partially transformed via transfection with selected oncogenes have the capacity to produce and deposit Tn. We reached the following conclusions: (1) compared with normal endometrial tissues, uterine sarcomas show heterogeneous, but increased, immunoreactive staining patterns exclusively within the extracellular compartment, regardless of the histologic subtype of the tumor; (2) in vitro, all normal and transfected stromal cells and cell lines examined secreted Tn into the tissue culture medium; (3) this secretory ability was not translated into morphologic uniformity, since immunoreactivity detected by confocal laser scanning microscopy was observed in only selected cell populations; (4) also, the deposition and the incorporation of Tn depended upon the density of transfected cells, and (5) double-staining experiments revealed that Tn could always be localized in close proximity to fibronectin. In summary, the production of Tn is increased in most cases of human uterine sarcoma. The capacity of stromal cells to deposit Tn in a matrix-like structure in vitro, rather than increase production of Tn, is correlated with the degree of neoplastic progression.


High biological species diversity in the arctic flora.
The arctic flora is considered to be impoverished, but estimates of species diversity are based on morphological assessments, which may not provide accurate counts of biological species. Here we report on crossing relationships within three diploid circumpolar plant species in the genus Draba (Brassicaceae). Although 99% of parental individuals were fully fertile, the fertility of intraspecific crosses was surprisingly low. Hybrids from crosses within populations were mostly fertile (63%), but only 8% of the hybrids from crosses within and among geographic regions (Alaska, Greenland, Svalbard, and Norway) were fertile. The frequent occurrence of intraspecific crossing barriers is not accompanied by significant morphological or ecological differentiation, indicating that numerous cryptic biological species have arisen within each taxonomic species despite their recent (Pleistocene) origin.


Identification of a conserved RNA replication element (cre) within the 3Dpol-coding sequence of hepatoviruses.
Internally located, cis-acting RNA replication elements (cre) have been identified within the genomes of viruses representing each of the major picornavirus genera (Enterovirus, Rhinovirus, Aphthovirus, and Cardiovirus) except Hepatovirus. Previous efforts to identify a stem-loop structure with cre function in hepatitis A virus (HAV), the type species of this genus, by phylogenetic analyses or thermodynamic predictions have not succeeded. However, a region of markedly suppressed synonymous codon variability was identified in alignments of HAV sequences near the 5' end of the 3D(pol)-coding sequence of HAV, consistent with noncoding constraints imposed by an underlying RNA secondary structure. Subsequent MFOLD predictions identified a 110-nucleotide (nt) complex stem-loop in this region with a typical AAACA/G cre motif in its top loop. A potentially homologous RNA structure was identified in this region of the avian encephalitis virus genome, despite little nucleotide sequence relatedness between it and HAV. Mutations that disrupted secondary RNA structure or the AAACA/G motif, without altering the amino acid sequence of 3D(pol), ablated replication of a subgenomic HAV replicon in transfected human hepatoma cells. Replication competence could be rescued by reinsertion of the native 110-nt stem-loop structure (but not an abbreviated 45-nt stem-loop) upstream of the HAV coding sequence in the replicon. These results suggest that this stem-loop is functionally similar to cre elements of other picornaviruses and likely involved in templating VPg uridylylation as in other picornaviruses, despite its significantly larger size and lower free folding energy.


Beta-cyclodextrin modified mesoporous bioactive glass nanoparticles/silk fibroin hybrid nanofibers as an implantable estradiol delivery system for the potential treatment of osteoporosis.
Osteoporosis, a systemic skeletal disease prevalent in elderly women, is associated with post-menopausal estrogen deficiency. Although systemic administration of exogenous estradiol (E2) reduced fragility fractures, the treatment has adverse effects. Localized delivery technologies of E2 could be utilized to circumvent the systemic adverse effects of systemic administration. In this study, a localized E2 delivery system is developed. Mesoporous bioactive glass nanoparticles (MBGNPs) with inherent osteogenic properties are modified with β-cyclodextrin (CD-MBGNPs) to enhance their affinity for E2. To ensure mechanical stability and integrity, E2 loaded CD-MBGNPs are further electrospun with silk fibroin (SF) to produce a nanofibrous mesh (E2@CD-MBGNPs/SF). The incorporation of MBGNPs in SF enhances in vitro apatite formation and sustains the constant release of E2. Moreover, osteoblast proliferation and differentiation markers such as alkaline phosphatase activity, collagen 1 and osteocalcin expression of MC3T3-E1 are augmented in CD-MBGNPs/SF and E2@CD-MBGNPs/SF as compared to SF nanofibers. On the other hand, osteoclast DNA, tartrate resistant acid phosphatase activity and multinucleated cell formation are reduced in E2@CD-MBGNPs/SF as compared to CD-MBGNPs/SF and SF. Hence the presence of CD-MBGNPs in SF stimulates osteoblast function whereas E2 incorporation in CD-MBGNPs/SF reduces osteoclast activity. This is the first report to develop CD-MBGNPs/SF as a localized delivery system for hydrophobic molecules such as estradiol to treat osteoporosis.


Reassessing the role of protein-carbohydrate complementarity during sperm-egg interactions in the mouse.
Despite years of intense study by many investigators, it may appear that we have made little progress towards a molecular understanding of mammalian sperm binding to the egg zona pellucida. An abundance of evidence derived from in vitro assays suggests that sperm-zona pellucida binding is dependent upon sperm recognition of specific glycan moieties on the zona pellucida glycoproteins. However, there is considerable disagreement regarding the identity of the zona pellucida sugars thought to mediate sperm binding, as well as disagreement over the identity of the sperm receptors themselves. Moreover, results from in vivo gene-targeting strategies fail to support a role for many, if not all, of the sperm receptors and their zona pellucida ligands implicated from in vitro assays. Nevertheless, a retrospective view of the literature suggests that some common principles are emerging regarding the molecular basis of mammalian sperm-zona binding, both with respect to the nature of the components that mediate binding, as well as the involvement of distinct receptor-ligand interactions, that involve both protein- and carbohydrate-dependent mechanisms of binding.


DNA effects of low level occupational exposure to extremely low frequency electromagnetic fields (50/60 Hz).
Exposure to extremely low frequency magnetic fields (ELF-MF) occurs from natural and artificial sources. Although ELF-MF has been classified as a suspected humans carcinogen agent by the International Agency for Research on Cancer, little is known of the effects of ELF-MF at lower exposure levels of the recommended range. In the present study, DNA damage in the peripheral blood cells of power line workers was investigated. Occupational exposure to ELF-MF in a power plant was measured using the National Institute for Occupational Safety and Health (NIOSH) manual. Single-strand breaks (SSBs) in DNA were evaluated in 29 male utility workers as the exposed population and 28 male support personnel as the control subjects using the comet assay. Effects of ELF-MF on subjects were evaluated using DNA percent in tails, tail length, olive length, and tail moment. Occupational exposure levels to ELF-MF in the utility workers were less than the threshold limit values (TLV) recommended by the American Conference of Government Industrial Hygienist (ACGIH). The median value of the magnetic field at the working sites was 0.85 µT. Induction of DNA damage was observed for the exposed workers compared with the controls. Olive length, tail moment, and tail DNA percent increased significantly (p < 0.05) in the utility workers. Exposure to ELF-MF at levels less than the ACGIH exposure limit can produce DNA strand breaks.


Bidirectional chaos communication between two outer semiconductor lasers coupled mutually with a central semiconductor laser.
Based on a linear chain composed of a central semiconductor laser and two outer semiconductor lasers, chaos synchronization and bidirectional communication between two outer lasers have been investigated under the case that the central laser and the two outer lasers are coupled mutually, whereas there exists no coupling between the two outer lasers. The simulation results show that high-quality and stable isochronal synchronization between the two outer lasers can be achieved, while the cross-correlation coefficients between the two outer lasers and the central laser are very low under proper operation condition. Based on the high performance chaos synchronization between the two outer lasers, message bidirectional transmissions of bit rates up to 20 Gbit/s can be realized through adopting a novel decoding scheme which is different from that based on chaos pass filtering effect. Furthermore, the security of bidirectional communication is also analyzed.


Characterization of sequence-specific errors in various next-generation sequencing systems.
Next-generation sequencing (NGS) is a popular method for assessing the molecular diversity of microbial communities without cultivation, for identifying polymorphisms in populations, and for comparing genomes and transcriptomes. However, sequence-specific errors (SSEs) by NGS systems can result in genome mis-assembly, overestimation of diversity in microbial community analyses, and false polymorphism discovery. SSEs can be particularly problematic due to rich microbial biodiversity and genomes containing frequent repeats. In this study, SSEs in public data from all popular NGS systems were discovered using a Markov chain model and hotspots for sequence errors were identified. Deletion errors were frequently preceded by homopolymers in non-Illumina NGS systems, such as GS FLX+. Substitution errors were often related to high GC contents and long G/C homopolymers in Illumina sequencing systems such as HiSeq. After removal of long G/C homopolymers in HiSeq, the average lengths of contigs and average SNP quality increased. SSEs were selectively removed from our mock community data by quality filtering, and a bias against specific microbes was identified. Our findings provide a scientific basis for filtering poor-quality reads, correcting deletion errors, preventing genome mis-assembly, and accurately assessing microbial community compositions and polymorphisms.


Violent and nonviolent suicide in veterans with substance-use disorders.
Substance-use disorders (SUDs) increase the risk of suicide. Examining risk factors for violent or nonviolent suicide in those with SUDs could improve suicide-prevention efforts in these individuals. We examined the demographic and clinical predictors of violent and nonviolent suicide among patients with SUDs. This study examined Veterans Affairs (VA) patients with SUDs who were alive at the beginning of Fiscal Year (FY) 2002 and died by suicide during FYs 2002 through 2006 (n = 854) and a random sample of 4,228 SUD patients who did not die by suicide during this period. Data were obtained from VA medical records and the National Death Index. Patients were considered to have an SUD if they received two or more diagnoses of the same SUD from FY 1997 to FY 2001. Of those who died by suicide, 70% (n = 600) used violent means. Diagnoses of depressive, other anxiety, bipolar and posttraumatic stress disorders, schizophrenia, and personality disorders were associated with increased risk of both violent and nonviolent suicide; however, these associations were stronger for nonviolent than violent suicide deaths. Risk of nonviolent suicide was greater among patients with opioid-use disorders or a clinical diagnosis of abuse or dependence (according to the International Classification of Diseases, Tenth Revision) on multiple substances. Most VA patients with SUDs who died from suicide used violent means. No specific SUD was associated with increased risk of violent suicide, but several SUD diagnoses were associated with increased risk of nonviolent suicide. Concurrent psychopathology increased risk for both suicide types; however, the relationship was strongest for nonviolent suicide.


Ketotifen Fumarate and Salbutamol Sulphate Combined Transdermal Patch Formulations: In vitro release and Ex vivo Permeation Studies.
The present work was performed to develop and evaluate transdermal patches of combined antiasthmatic drugs (salbutamol sulphate and ketotifen fumarate). Polyvinyl alcohol membrane was used as backing membrane and eudragit RL-100 was used as matrix material to suspend the drugs in the continuous thickness of the patch. Methanol was solvent and propylene glycol was used as plasticizer. Tween 20, isopropyl myristate, eucalyptus oil, castor oil and span-20 were used as permeability enhancers. Thickness, weight variation and drug uniformity were investigated. The patch formulations were also subjected to drug release in dissolution media and permeation through rabbit skin. Effects of different enhancers were evaluated on release and permeation of drugs. F3 formulations having isopropyl myristate as permeation enhancer, showed maximum amounts of drugs release (88.11% of salbutamol sulphate and 88.33% of ketotifen fumarate) at the end of 24 h dissolution study. F3 also showed maximum permeation of both drugs (4.235 mg salbutamol sulphate and 1.057 mg ketotifen fumarate) after 24 h permeation study through rabbit skin mounted in Franz cell. The patches having no enhancer in the formulation also showed some drug release and permeation due to the presence of plasticizer. The results of the study suggested that new controlled release transdermal formulations of combined antiasthmatic drugs can be suitably developed as an alternate to conventional dosage forms.


Evaluation of a low-cost 3D sound system for immersive virtual reality training systems.
Since Head Mounted Displays (HMD), datagloves, tracking systems, and powerful computer graphics resources are nowadays in an affordable price range, the usage of PC-based "Virtual Training Systems" becomes very attractive. However, due to the limited field of view of HMD devices, additional modalities have to be provided to benefit from 3D environments. A 3D sound simulation can improve the capabilities of VR systems dramatically. Unfortunately, realistic 3D sound simulations are expensive and demand a tremendous amount of computational power to calculate reverberation, occlusion, and obstruction effects. To use 3D sound in a PC-based training system as a way to direct and guide trainees to observe specific events in 3D space, a cheaper alternative has to be provided, so that a broader range of applications can take advantage of this modality. To address this issue, we focus in this paper on the evaluation of a low-cost 3D sound simulation that is capable of providing traceable 3D sound events. We describe our experimental system setup using conventional stereo headsets in combination with a tracked HMD device and present our results with regard to precision, speed, and used signal types for localizing simulated sound events in a virtual training environment.


Constitutive soxR mutations contribute to multiple-antibiotic resistance in clinical Escherichia coli isolates.
The soxRS regulon of Escherichia coli and Salmonella enterica is induced by redox-cycling compounds or nitric oxide and provides resistance to superoxide-generating agents, macrophage-generated nitric oxide, antibiotics, and organic solvents. We have previously shown that constitutive expression of soxRS can contribute to quinolone resistance in clinically relevant S. enterica. In this work, we have carried out an analysis of the mechanism of constitutive soxS expression and its role in antibiotic resistance in E. coli clinical isolates. We show that constitutive soxS expression in three out of six strains was caused by single point mutations in the soxR gene. The mutant SoxR proteins contributed to the multiple-antibiotic resistance phenotypes of the clinical strains and were sufficient to confer multiple-antibiotic resistance in a fresh genetic background. In the other three clinical isolates, we observed, for the first time, that elevated soxS expression was not due to mutations in soxR. The mechanism of such increased soxS expression remains unclear. The same E. coli clinical isolates harbored polymorphic soxR and soxS DNA sequences, also seen for the first time.


Small-scale spatial variability of atrazine and dinoseb adsorption parameters in an alluvial soil.
Soil sorption processes largely control the environmental fate of herbicides. Therefore, accuracy of sorption parameters is crucial for accurate prediction of herbicide mobility in agricultural soils. A combined experimental and statistical study was performed to investigate the small-scale spatial variability of sorption parameters for atrazine and dinoseb in soils and to establish the number of samples needed to provide a value of the distribution coefficient (K(d)) next to the mean, with a given precision. The study explored sorption properties of the two herbicides in subsurface samples collected from four pits distributed along a transect of an alluvial soil; two to four samples were taken at about 30 cm apart at each sampling location. When considering all the data, the distribution coefficients were found to be normally and log-normally distributed for atrazine and dinoseb, respectively; the CVs were relatively high (close to 50% for dinoseb and 40% for atrazine). When analyzed horizon by horizon, the data revealed distribution coefficients normally distributed for both herbicides, whatever the soil layer, with lower CVs. The K(d) values were shown to vary considerably between samples collected at very short distance (a few centimeters), suggesting that taking a single soil sample to determine sorption properties through batch experiments can lead to highly unrepresentative results and to poor sorption/mobility predictions.


Management of early-stage lung cancer: past, present, and future adjuvant trials.
The standard of care with regard to adjuvant chemotherapy of lung cancer has changed remarkably over the past 3 years. Until the initial report of the International Adjuvant Lung Trial in 2003, there was no real evidence from any individual randomized clinical trial (RCT) that adjuvant chemotherapy improves survival in resectable non-small-cell lung cancer. However, five RCTs that have now been reported indicate that adjuvant chemotherapy is effective, at least in certain subgroups of resectable patients. Moreover, numerous meta-analyses have also reported a positive effect from adjuvant treatment. Nonetheless, because of methodologic. issues and conflicting results, the question of who should be treated and what constitutes optimal adjuvant therapy remains controversial. This article reviews the recent randomized trials that have contributed to a change in the state of the art, as well as some of the methodologic problems that may have confounded their proper interpretation. It also considers newer approaches to adjuvant therapy, with a particular focus on strategies that incorporate our growing knowledge of molecular medicine and predictive factors to the field of adjuvant chemotherapy of lung cancer.


Coffee Consumption and Pancreatic Cancer Risk: An Update Meta-analysis of Cohort Studies.
The results of epidemiologic studies on the relationship between the coffee consumption and pancreatic cancer risk were inconsistent. Thus, we performed an update meta-analysis of cohort studies to quantitatively summarize the association between coffee consumption and pancreatic cancer risk. We searched CBM (China Biology Medicine disc) and MEDLINE for studies of coffee consumption and pancreatic cancer risk up to June 2015. A total of 20 cohort studies were identified in this meta-analysis, and we analyzed these studies using random effects model. The dose-response analysis was conducted too. The overall relative risk (RR) for highest coffee consumption versus lowest coffee consumption was 0.75 (95% Confidence Interval (CI), 0.63-0.86). Statistic significant heterogeneity was found among these studies (I (2) =37.8%, P for heterogeneity =0.045). The pooled RR for increment of 1 cup/day of coffee consumption was 0.99 (95%CI, 0.96-1.03) for the nine studies, without statistically significant. High coffee consumption is associated with a reduced pancreatic cancer risk. However, the result should be accepted with caution, due to the potential confounder and bias could not be excluded. Further well designed studies are needed to confirm the finding.


Transcriptional responses to glucose in Saccharomyces cerevisiae strains lacking a functional protein kinase A.
The pattern of gene transcripts in the yeast Saccharomyces cerevisiae is strongly affected by the presence of glucose. An increased activity of protein kinase A (PKA), triggered by a rise in the intracellular concentration of cAMP, can account for many of the effects of glucose on transcription. In S. cerevisiae three genes, TPK1, TPK2, and TPK3, encode catalytic subunits of PKA. The lack of viability of tpk1 tpk2 tpk3 triple mutants may be suppressed by mutations such as yak1 or msn2/msn4. To investigate the requirement for PKA in glucose control of gene expression, we have compared the effects of glucose on global transcription in a wild-type strain and in two strains devoid of PKA activity, tpk1 tpk2 tpk3 yak1 and tpk1 tpk2 tpk3 msn2 msn4. We have identified different classes of genes that can be induced -or repressed- by glucose in the absence of PKA. Representative examples are genes required for glucose utilization and genes involved in the metabolism of other carbon sources, respectively. Among the genes responding to glucose in strains devoid of PKA some are also controlled by a redundant signalling pathway involving PKA activation, while others are not affected when PKA is activated through an increase in cAMP concentration. On the other hand, among genes that do not respond to glucose in the absence of PKA, some give a full response to increased cAMP levels, even in the absence of glucose, while others appear to require the cooperation of different signalling pathways. We show also that, for a number of genes controlled by glucose through a PKA-dependent pathway, the changes in mRNA levels are transient. We found that, in cells grown in gluconeogenic conditions, expression of a small number of genes, mainly connected with the response to stress, is reduced in the strains lacking PKA. In S. cerevisiae, the transcriptional responses to glucose are triggered by a variety of pathways, alone or in combination, in which PKA is often involved. Redundant signalling pathways confer a greater robustness to the response to glucose, while cooperative pathways provide a greater flexibility.


Random amplification of polymorphic DNA reveals serotype-specific clonal clusters among enterotoxigenic Escherichia coli strains isolated from humans.
The genetic diversity of 47 enterotoxigenic Escherichia coli (ETEC) strains of serotypes O6:H16, O27:H7, O29:H21, O128ac:H12, and O153:H45, previously isolated from diarrheic patients in Brazil over a period of 15 years, was investigated by random amplification of polymorphic DNA (RAPD). Informative band arrays were obtained with three 10-mer primers with G+C contents of 50, 60, and 70%. Based on the combination of the band profiles generated by the three primers 22 RAPD types were detected, and 5 major clonal clusters, each one with at least 80% identical bands, were established. The clonal clusters corresponded to strains having the same serotype which, in most cases, also had the same virulence factors (colonization factors and toxin types) and outer membrane protein and lipopolysaccharide sodium dodecyl sulfate-polyacrylamide gel electrophoresis profiles. The results suggested a correlation between phenotypic properties and genetic relatedness of ETEC isolates of human origin and indicated that a reduced number of clonally related strains are found in areas of ETEC endemicity in Brazil. Moreover, the RAPD technique revealed intraserotype-specific variations, undetectable by the combination of several phenotypic typing methods, among the ETEC strains analyzed. These results show that RAPD typing represents a useful tool for population genetics as well as for epidemiological studies of ETEC.


Gender-specific estimates of sleep problems during the COVID-19 pandemic: Systematic review and meta-analysis.
The outbreak of the novel coronavirus disease 2019 (COVID-19) changed lifestyles worldwide and subsequently induced individuals' sleep problems. Sleep problems have been demonstrated by scattered evidence among the current literature on COVID-19; however, little is known regarding the synthesised prevalence of sleep problems (i.e. insomnia symptoms and poor sleep quality) for males and females separately. The present systematic review and meta-analysis aimed to answer the important question regarding prevalence of sleep problems during the COVID-19 outbreak period between genders. Using the Preferred Reporting Items for Systematic Reviews and Meta-Analyses guideline and Newcastle-Ottawa Scale checklist, relevant studies with satisfactory methodological quality searched for in five academic databases (Scopus, PubMed Central, ProQuest, Web of Science , and EMBASE) were included and analysed. The protocol of the project was registered in the International Prospective Register of Systematic Reviews (PROSPERO; identification code CRD42020181644). A total of 54 papers (N = 67,722) in the female subgroup and 45 papers (N = 45,718) in the male subgroup were pooled in the meta-analysis. The corrected pooled estimated prevalence of sleep problems was 24% (95% confidence interval [CI] 19%-29%) for female participants and 27% (95% CI 24%-30%) for male participants. Although in both gender subgroups, patients with COVID-19, health professionals and general population showed the highest prevalence of sleep problems, it did not reach statistical significance. Based on multivariable meta-regression, both gender groups had higher prevalence of sleep problems during the lockdown period. Therefore, healthcare providers should pay attention to the sleep problems and take appropriate preventive action.


Testing an APA practice guideline: symptom-targeted medication utilization for patients with borderline personality disorder.
The goal of this study was to test whether the use of psychotropic medication treatment for borderline personality disorder (BPD) was consistent with proposals in the recently published American Psychiatric Association's Practice Guideline for the Treatment of Patients with Borderline Personality Disorder. Medication utilization by patients with BPD was assessed prospectively over a 2-year period prior to the publication of the Guideline. Three BPD symptom clusters--cognitive-perceptual, affective dysregulation, and impulsive-behavioral dyscontrol--along with demographic and functioning variables were used to predict the use of five classes of medication. Symptoms of impulsive-behavioral dyscontrol significantly predicted use of both neuroleptics and anticonvulsants. Cognitive-perceptual symptoms were inversely related to anticonvulsant use. While some pre-Guideline medication practices with BPD patients were consistent with the recommended algorithms, physicians may also have been influenced by a variety of considerations, including level of functional impairment and the presence of comorbid Axis I conditions.


SU-E-J-150: To Design a Methodology Based on Numerical Phantom for Reconstruction of Dose Delivered to Moving Lung Tumors.
To design a methodology based on numerical phantom for reconstruction of dose delivered to moving lung tumors. MatlabTM 7.6 was used to generate a 4D numerical lung phantom (NLP). Customer parameter files were used as input to this NLP, which consists of multiple ellipsoids representing body, lung, cord and tumor. In this study, we studied the impact of varying breathing pattern on a left lower lobe tumor, where the tumor motion was simulated on the daily breathing pattern of the patient acquired using real time positioning management (RPMTM) system from Varian Medical Systems. Based on the daily breathing pattern, the original RPM signal and the original tumor trajectory, 5 sets of motion trajectories were simulated. This was then used to build 10 different phases of the numerical phantom. Average Intensity Projection (AIP) was then generated from the different phases. The actual delivered dose on the 5 AIP sets were compared to the intended dose on the original planning AIP image set. The mean target coverage (TC) recomputed on the 5 AIP sets was approximately 18% lower than the TC for the planning AIP image set. The mean homogeneity index (HI) recomputed on the 5 sets, was approximately 5 times higher than HI for the planning AIP image set. The lung NTDmean dose was approximately 9.5 Gy3 and did not differ much. The presented numerical simulation framework may assist in monitoring the changes in dose accumulation due to changes in the patient's breathing on a daily basis. This can also be used for validation of new motion tracking algorithms and its impact of dose coverage.


Acute and 2 days delayed effects of exhaustive stretch-shortening cycle exercise on barefoot walking and running patterns.
This study investigated the acute and 2 days delayed influences of exhaustive stretch-shortening cycle exercise (SSC) on barefoot walking and running gait patterns. The SSC exercise was performed on a sledge apparatus, on which the subjects (N = 10) repeated until exhaustion intermittent series of 25 bilateral submaximal rebounds. Maximal drop-jumps and submaximal barefoot treadmill walking and running were performed before (PRE) and after (POST) the exhaustive exercise and repeated 48 h (D2) later. Electromyographic activity and 3D kinematics of the right lower limb and foot were recorded for 15 s at gait initiation (BEG) and at the end (END: at 3 min of walk and 5 min of run). The exhaustive SSC exercise resulted in 6% reductions in maximal drop jump performance at POST and D2, and affected mostly both gait patterns at D2. The walking pattern presented compensatory neural adjustments within the triceps surae muscle group. This expected pain-induced protective strategy of the soleus muscle was sufficient to preserve the kinematics pattern. The running condition revealed a major knee strategy, which might support the concept of pain protective strategy of knee extensor muscles at the expense of impact cushioning. Regardless the testing session, most parameters showed fatigue-induced changes at gait initiation (BEG), which were opposite to subsequent BEG to END adjustments. This is likely to support anticipatory strategies rather than progressive adjustments during the exercise.


Quality assessment of a sample of mobile app-based health behavior change interventions using a tool based on the National Institute of Health and Care Excellence behavior change guidance.
To quality assess a sample of health behavior change apps from the NHS Apps Library using a rating tool based on the 2014 National Institute for Health and Care Excellence behavior change guidance (NICE BCG). A qualitative analysis of the NICE BCG identified themes and questions for a quality assessment of health behavior change apps. These were refined by further discussion and piloting, and applied by two independent raters to a sample of NHS Library apps (N=49). Disagreements were resolved following discussions with a third rater. Themes identified were; purpose, planning, usability, tailoring, behavior change technique (BCT), maintenance, evaluation, data security and documentation. Overall, purpose of the apps was clear, but evidence for collaboration with users or professionals was lacking. Usability information was poor and tailoring disappointing. Most used recognized BCTs but paid less attention to behavior maintenance than initiation. Information on app evaluation and documentation was sparse. This study furthers the work of the NHS Apps Library, adapting the NICE (2014) behavior change guidance for quality assessment of behavior change apps. This study helps lay the foundations for development of a quality assurance tool for mobile health apps aimed at health behavior change.


A Net Mold-based Method of Scaffold-free Three-Dimensional Cardiac Tissue Creation.
This protocol describes a novel and easy net mold-based method to create three-dimensional (3-D) cardiac tissues without additional scaffold material. Human-induced pluripotent stem-cell-derived cardiomyocytes (iPSC-CMs), human cardiac fibroblasts (HCFs), and human umbilical vein endothelial cells (HUVECs) are isolated and used to generate a cell suspension with 70% iPSC-CMs, 15% HCFs, and 15% HUVECs. They are co-cultured in an ultra-low attachment "hanging drop" system, which contains micropores for condensing hundreds of spheroids at one time. The cells aggregate and spontaneously form beating spheroids after 3 days of co-culture. The spheroids are harvested, seeded into a novel mold cavity, and cultured on a shaker in the incubator. The spheroids become a mature functional tissue approximately 7 days after seeding. The resultant multilayered tissues consist of fused spheroids with satisfactory structural integrity and synchronous beating behavior. This new method has promising potential as a reproducible and cost-effective method to create engineered tissues for the treatment of heart failure in the future.


Genetic association between GHSR1a 5'UTR-microsatellite and nt-7(C>A) loci and growth and carcass traits in Japanese Black cattle.
We carried out a genetic association study between five nucleotide polymorphisms (5'UTR microsatellite ((TG)(n)), nt-7(C>A), L24V, DelR242 and Intron 1 microsatellite) of the GHSR1a gene and growth and carcass traits in 1285 steers sired by 117 Japanese Black bulls in a progeny testing program. We report herein, a significant association between the 5'UTR microsatellite and nt-7(C>A) loci and growth and carcass traits. We also propose a translational hypothesis that the association is due to differences in the secondary structure of GHSR1b mRNA (the non-spliced type with the 5'UTR microsatellite) among the GHSR1a gene haplotypes. Furthermore, we predicted the potential increase in profitability due to increased carcass weight in cow-calf fattening enterprises through planned matings based on DNA testing of the 5'UTR microsatellite. Statistical analysis revealed that the 5'UTR microsatellite locus had a significant additive effect on carcass weight (CW) and average daily gain (ADG), but not on beef marbling score (BMS). One of the four major microsatellite alleles (19-TG allele) with an allele frequency of 0.145, had a significantly (P < 0.0007) desirable effect on CW and ADG. We concluded that the 19-TG allele could potentially be economically useful nucleotide markers for growth and carcass traits in Japanese Black cattle.


Quantitative morphology of B, A, D, and PP cells in infants of diabetic mothers.
Pancreatic specimens from 34 infants of diabetic mothers (IDM) and 32 control infants of gestational ages 26-44 wk were examined histologically using immunocytochemical stains for insulin, glucagon, somatostatin, and pancreatic polypeptide (PP). Each section was divided into PP-rich and PP-poor regions that are thought to be derived from the ventral and dorsal lobes of the gland, respectively. In some of these, the fractional area (%) occupied by positively stained B, A, and PP cells was determined by automatic image analysis, and the area occupied by D cells was determined by conventional point counting. The B cell fractional area was significantly higher in the IDM in both PP-poor and PP-rich areas (P less than 0.02). the fractional area of A cells in PP-poor areas and of PP cells in PP-rich areas was also significantly greater in IDM (P less than 0.02). The total endocrine cell fractional area was significantly greater in IDM in PP-poor but not in PP-rich regions of the pancreas. These results are not compatible with the hypothesis that maternal hyperglycemia results in specific fetal B cell hyperplasia and raise the possibility that hyperplasia of B, A, and PP cells in IDM may result from a variety of stimuli or that one stimulus acts on a pluripotential stem cell.


Comparative analysis of autofocus functions in digital in-line phase-shifting holography.
Numerical reconstruction of digital holograms relies on a precise knowledge of the original object position. However, there are a number of relevant applications where this parameter is not known in advance and an efficient autofocusing method is required. This paper addresses the problem of finding optimal focusing methods for use in reconstruction of digital holograms of macroscopic amplitude and phase objects, using digital in-line phase-shifting holography in transmission mode. Fifteen autofocus measures, including spatial-, spectral-, and sparsity-based methods, were evaluated for both synthetic and experimental holograms. The Fresnel transform and the angular spectrum reconstruction methods were compared. Evaluation criteria included unimodality, accuracy, resolution, and computational cost. Autofocusing under angular spectrum propagation tends to perform better with respect to accuracy and unimodality criteria. Phase objects are, generally, more difficult to focus than amplitude objects. The normalized variance, the standard correlation, and the Tenenbaum gradient are the most reliable spatial-based metrics, combining computational efficiency with good accuracy and resolution. A good trade-off between focus performance and computational cost was found for the Fresnelet sparsity method.


Switchgrass (Panicum virgatum L.) cell suspension cultures: Establishment, characterization, and application.
Switchgrass (Panicum virgatum L.) is a warm-season perennial grass that has received considerable attention as a potential dedicated biofuel and bioproduct feedstock. Genetic improvement of switchgrass is needed for better cellulosic ethanol production, especially to improve cellulose-to-lignin ratios. Cell suspension cultures offer an in vitro system for mutant selection, mass propagation, gene transfer, and cell biology. Toward this end, switchgrass cell suspension cultures were initiated from embryogenic callus obtained from genotype Alamo 2. They have been established and characterized with different cell type morphologies: sandy, fine milky, and ultrafine cultures. Characterization includes histological analysis using scanning electron microscopy, and utility using protoplast isolation. A high protoplast isolation rate of up to 10(6) protoplasts/1.0g of cells was achieved for the fine milky culture, whereas only a few protoplasts were isolated for the sandy and ultrafine cultures. These results indicate that switchgrass cell suspension type sizably impacts the efficiency of protoplast isolation, suggesting its significance in other applications. The establishment of different switchgrass suspension culture cell types provides the opportunity to gain insights into the versatility of the system that would further augment switchgrass biology research.


Comprehensive approach to understand the association between diurnal temperature range and mortality in East Asia.
An adverse association between diurnal temperature range (DTR) and mortality has been suggested, but with variable relationships in different cities. Comprehensive approaches to understanding the health effects of DTR using multinational data are required. We investigated the association between DTR and cause-specific mortality in an age-specific population and assessed the dependency of the health effects of DTR on geographic and climatic factors. Poisson generalized linear regression analyses with allowances for over-dispersion were applied to daily DTR and cause-specific mortality data from 30 cities in China, Japan, Korea, and Taiwan between 1979 and 2010, adjusted for various climatic and environmental factors. City-specific effects of DTR were estimated and summarized for the overall effects using geographic and climatic determinants in a meta-analysis. For all-cause, circulatory, and respiratory mortality, the greatest city-specific effects per 1°C DTR were found in Tianjin, China (1.80%; 95% confidence interval [CI]: 0.48, 3.14); Tangshan, China (2.25%; 95% CI: 0.65, 3.87); and Incheon, Korea (2.84%; 95% CI: 0.04, 5.73), respectively, and overall effects across 30 cities were 0.58% (95% CI: 0.44, 0.72), 0.81% (95% CI: 0.60, 1.03), and 0.90% (95% CI: 0.63, 1.18), respectively. Using quartile cutoff values for climatic (DTR, and mean temperature) and geographic (latitude, and longitude) characteristics, we divided the 30 cities into 4 different groups and conducted a meta-analysis within the groups using either a random or fixed effects model. Adverse effects of DTR were more pronounced for those aged ≥65years and varied according to geographic, longitudinal (0.07%; 95% CI: 0.05, 0.10), and climatic characteristics and the scale of DTR (0.33%; 95% CI: 0.12, 0.55) for overall all-cause mortality. The DTR is a risk factor affecting human health, depending on geographic location and the temperature variation, with particular vulnerability in aged populations.


Genistein inhibits radiation-induced activation of NF-kappaB in prostate cancer cells promoting apoptosis and G2/M cell cycle arrest.
New cancer therapeutic strategies must be investigated that enhance prostate cancer treatment while minimizing associated toxicities. We have previously shown that genistein, the major isoflavone found in soy, enhanced prostate cancer radiotherapy in vitro and in vivo. In this study, we investigated the cellular and molecular interaction between genistein and radiation using PC-3 human prostate cancer cells. Tumor cell survival and progression was determined by clonogenic analysis, flow cytometry, EMSA analysis of NF-kappaB, and western blot analysis of cyclin B1, p21WAF1/Cip1, and cleaved PARP protein. Genistein combined with radiation caused greater inhibition in PC-3 colony formation compared to genistein or radiation alone. Treatment sequence of genistein followed by radiation and continuous exposure to genistein showed optimal effect. Cell cycle analysis demonstrated a significant dose- and time-dependent G2/M arrest induced by genistein and radiation that correlated with increased p21WAF1/Cip1 and decreased cyclin B1 expression. NF-kappaB activity was significantly decreased by genistein, yet increased by radiation. Radiation-induced activation of NF-kappaB activity was strongly inhibited by genistein pre-treatment. A significant and striking increase in cleaved PARP protein was measured following combined genistein and radiation treatment, indicating increased apoptosis. A mechanism of increased cell death by genistein and radiation is proposed to occur via inhibition of NF-kappaB, leading to altered expression of regulatory cell cycle proteins such as cyclin B and/or p21WAF1/Cip1, thus promoting G2/M arrest and increased radiosensitivity. These findings support the important and novel strategy of combining genistein with radiation for the treatment of prostate cancer.


Mouse liver thiol methyltransferase. Assay conditions, biochemical properties, and strain variation.
Thiol methyltransferase (TMT) catalyzes the S-methylation of aliphatic sulfhydryl drugs and xenobiotic compounds. It would be useful if there were an experimental animal model in which the regulation and function of TMT could be studied. Therefore, TMT activity was measured in hepatic microsomes from A/J mice. Substrate kinetics for mouse liver microsomal TMT, like those for the enzyme in human red blood cell membranes and human kidney microsomes, were biphasic, with apparent "high" and "low" affinity forms of TMT with 2-mercaptoethanol as the methyl acceptor substrate. Apparent Michaelis (Km) constants of the high and low affinity activities for 2-mercaptoethanol were 30 microM and 12 mM, respectively. Apparent Km values of the high and low affinity activities for S-adenosyl-L-methionine, the methyl donor for the reaction, were 47 microM and 61 microM, respectively. The optimal pH for the high affinity activity was between 7.2 and 8.1, whereas the optimal pH for the low affinity activity was approximately 9.2. Differential centrifugation showed that more than 85% of both activities was associated with membrane fractions. SKF 525A, a potent inhibitor of TMT in human tissues, inhibited mouse liver high and low affinity TMT activities by 88% and 46%, respectively, at a concentration of 0.5 mM. TMT activities were then measured in hepatic microsomes from nine additional inbred strains of mice. High affinity TMT activities varied 1.7-fold, whereas low affinity activities varied 2.1-fold among these strains. Since the properties of TMT in mouse liver are similar to those of the enzyme in human tissue, the inbred mouse will be a useful experimental animal model in which to study the regulation and function of TMT.


A genomic map of a 6-Mb region at 13q21-q22 implicated in cancer development: identification and characterization of candidate genes.
Chromosomal region 13q21-q22 harbors a putative breast cancer susceptibility gene and has been implicated as a common site for somatic deletions in a variety of malignant tumors. We have built a complete physical clone contig for a region between D13S1308 and AFM220YE9 based on 18 yeast artificial chromosome and 81 bacterial artificial chromosome (BAC) clones linked together by 22 genetic markers and 61 other sequence tagged sites. Combining data from 47 sequenced BACs (as of June 2001), we have assembled in silico an integrated 5.7-Mb genomic map with 90% sequence coverage. This area contains eight known genes, two hypothetical proteins, 24 additional Unigene clusters, and approximately 100 predicted genes and exons. We have determined the cDNA and genomic sequence, and tissue expression profiles for the KIAA1008 protein (homologous to the yeast mitotic control protein dis3+), KLF12 (AP-2 repressor), progesterone induced blocking factor 1, zinc finger transcription factor KLF5, and LIM domain only-7, and for the hypothetical proteins FLJ22624 and FLJ21869. Mutation screening of the five known genes in 19 breast cancer families has revealed numerous polymorphisms, but no deleterious mutations. These data provide a basis and resources for further analyses of this chromosomal region in the development of cancer.


Radiology department strategies to protect radiologic technologists against COVID19: Experience from Wuhan.
The outbreak of Coronavirus Disease 2019 (COVID-19) is a huge threat to global public health security. In the absence of specific antiviral medicines to prevent or treat COVID-19, it is essential to detect the infected patients at an early stage and immediately isolate them from the healthy population. In view of the advantages of sensitivity and high spatial resolution, CT imaging has played an important role in screening and diagnosing of COVID-19 in China. The radiologic technologists performing CT scans for the infected patients become high-risk medical care personnel. It is critical for the radiology department to ensure the personal safety of radiologic technologists and avoid cross-infection. In this review article, we describe the systematic strategies to combat COVID-19 from the radiology department in Tongji hospital in Wuhan, P.R. China, including personnel arrangements, environmental modification, protection levels and configurations, radiological imaging (CT and radiography), and disinfection methods. It can provide guidance to other radiology departments faced with COVID-19 to reduce infection risk for radiologic technologists.


[Mitral valve insufficiency and surgical repair].
Management of mitral regurgitation have benefited over the last 10 years from the better understanding of its natural history and from the advent of new echocardiographic quantitative methods. The dismal prognosis displayed by patients with flail leaflet and severe mitral regurgitation medically treated in one hand and the demonstration of the dramatic consequences of impaired pre-operative left ventricular function in the other, have been a strong incentive for early surgical correction of the disease. In the same time, mitral valve repair developed because of the improvement in the surgical techniques, of changes in aetiology and because of the widespread use of intra-operative transoesophageal echocardiography. Mitral repair has been shown to be an independent and beneficial predictor of overall survival, operative mortality and late survival and consequently became the support of early surgical strategies. But it is not the only factor to predict mortality and morbidity, and one must not forget the decisive and independent part played by age, preoperative symptoms and above all pre-operative left ventricular function. Therefore, mitral valve repair must not be considered as a pretext to postpone intervention but should be an other reason to intervene earlier.


Breast primary epithelial cells that escape p16-dependent stasis enter a telomere-driven crisis state.
Breast cancer is the most common malignant disease in women, but some basic questions remain in breast cancer biology. To answer these, several cell models were developed. Recently, the use of improved cell-culture conditions has enabled the development of a new primary cell model with certain luminal characteristics. This model is relevant because, after the introduction of a specific set of genetic elements, the transformed cells yielded tumors resembling human adenocarcinomas in mice. The use of improved cell-culture conditions supporting the growth of these breast primary epithelial cells was expected to delay or eliminate stress-induced senescence and lead to the propagation of normal cells. However, no studies have been carried out to investigate these points. Propagation of breast primary epithelial cells was performed in WIT medium on Primaria plates. Immunofluorescence, western blot and qRT-PCR were used to detect molecular markers, and to determine the integrity of DNA damage-response pathways. Promoter methylation of p16 (INK4a) was assessed by pyrosequencing. In order to obtain a dynamic picture of chromosome instability over time in culture, we applied FISH methodologies. To better link chromosome instability with excessive telomere attrition, we introduced the telomerase reverse transcriptase human gene using a lentiviral vector. We report here that breast primary epithelial cells propagated in vitro with WIT medium on Primaria plates express some luminal characteristics, but not a complete luminal lineage phenotype. They undergo a p16-dependent stress-induced senescence (stasis), and the cells that escape stasis finally enter a crisis state with rampant chromosome instability. Chromosome instability in these cells is driven by excessive telomere attrition, as distributions of chromosomes involved in aberrations correlate with the profiles of telomere signal-free ends. Importantly, ectopic expression of the human TERT gene rescued their chromosomal instability phenotype. Essentially, our data show that contrary to what was previously suggested, improved culture conditions to propagate in vitro mammary epithelial cells with some luminal characteristics do not prevent stress-induced senescence. This barrier is overcome by spontaneous methylation of the p16 (INK4a) promoter, allowing the proliferation of cells with telomere dysfunction and ensuing chromosome instability.


Mitochondrial targeting of vitamin E succinate enhances its pro-apoptotic and anti-cancer activity via mitochondrial complex II.
Mitochondrial complex II (CII) has been recently identified as a novel target for anti-cancer drugs. Mitochondrially targeted vitamin E succinate (MitoVES) is modified so that it is preferentially localized to mitochondria, greatly enhancing its pro-apoptotic and anti-cancer activity. Using genetically manipulated cells, MitoVES caused apoptosis and generation of reactive oxygen species (ROS) in CII-proficient malignant cells but not their CII-dysfunctional counterparts. MitoVES inhibited the succinate dehydrogenase (SDH) activity of CII with IC(50) of 80 μM, whereas the electron transfer from CII to CIII was inhibited with IC(50) of 1.5 μM. The agent had no effect either on the enzymatic activity of CI or on electron transfer from CI to CIII. Over 24 h, MitoVES caused stabilization of the oxygen-dependent destruction domain of HIF1α fused to GFP, indicating promotion of the state of pseudohypoxia. Molecular modeling predicted the succinyl group anchored into the proximal CII ubiquinone (UbQ)-binding site and successively reduced interaction energies for serially shorter phytyl chain homologs of MitoVES correlated with their lower effects on apoptosis induction, ROS generation, and SDH activity. Mutation of the UbQ-binding Ser(68) within the proximal site of the CII SDHC subunit (S68A or S68L) suppressed both ROS generation and apoptosis induction by MitoVES. In vivo studies indicated that MitoVES also acts by causing pseudohypoxia in the context of tumor suppression. We propose that mitochondrial targeting of VES with an 11-carbon chain localizes the agent into an ideal position across the interface of the mitochondrial inner membrane and matrix, optimizing its biological effects as an anti-cancer drug.


Detection and molecular characterization of porcine circovirus type 2 from piglets with porcine circovirus associated diseases in Colombia.
The porcine circovirus-associated disease (PCVAD) has been known since 1991 in Canada, but the first outbreak of PCVAD in Colombia was reported in 2007. In order to understand the molecular epidemiology of the disease and to establish the origin of the virus in the country, the study presented here intended to evaluate the presence of PCV2-associated systemic infection in piglets from different geographical regions over a period of 9-years (2002-2010). The analysis included samples collected before, during and after outbreaks of PCVAD in pigs from Colombia. The PCV2 ORF2 from the positive samples was sequenced and used to determine the genotypes of the strains and to study the dynamic of these genotypes throughout the time. PCV2 DNA was detected in cases related to PCV2-associated systemic infection as well as in healthy pigs with a presumable persistent infection. The analysis of the ORF2 nucleotide full length sequence of twenty-three strains allowed to divide them into two groups: PCV2a and PCV2b. At the amino acid level the main variations in the sequence of the capsid protein were found in regions located within the immunoreactive areas. The results of this study demonstrated for the first time, that the two subgroups: PCV2a and PCV2b have been circulating in swine from Colombia. In addition, the study showed that genotype PCV2b is present in Colombian pigs suffering from both clinical and presumable persistent infection and that the PCV2b genotype was present in the Colombian pig population even before recognition of the disease in the country and it became predominant through time.


Unaltered size of right ventricular infarct in dogs with right ventricular hypertrophy induced by pressure overload.
We investigated whether right ventricular infarct size is increased in the hypertrophied right ventricle. In dogs, we produced chronic pressure overload-induced right ventricular hypertrophy (RVH) and acute pulmonary stenosis (APS) with a similar right ventricular systolic pressure to RVH. Then, in an open-chest model, right coronary artery occlusion for 4 h followed by 1 h of reperfusion was performed in animals with RVH or APS or in controls. Regional myocardial blood flow was measured with nonradioactive colored microspheres. Compared with the control group, regional myocardial oxygen consumption in the right ventricle was increased in the APS but not in the RVH group. The risk area and collateral blood flow were comparable in the three groups. However, the infarct size relative to risk area was increased in the APS group (42.3 +/- 4.9%), but that in the RVH group (21.1 +/- 4.4%) was not different from controls (26.1 +/- 4.2%). Thus right ventricular infarction in the heart with RVH was not increased compared with the normal heart.


Two Bistable Switches Govern M Phase Entry.
The abrupt and irreversible transition from interphase to M phase is essential to separate DNA replication from chromosome segregation. This transition requires the switch-like phosphorylation of hundreds of proteins by the cyclin-dependent kinase 1 (Cdk1):cyclin B (CycB) complex. Previous studies have ascribed these switch-like phosphorylations to the auto-activation of Cdk1:CycB through the removal of inhibitory phosphorylations on Cdk1-Tyr15 [1, 2]. The positive feedback in Cdk1 activation creates a bistable switch that makes mitotic commitment irreversible [2-4]. Here, we surprisingly find that Cdk1 auto-activation is dispensable for irreversible, switch-like mitotic entry due to a second mechanism, whereby Cdk1:CycB inhibits its counteracting phosphatase (PP2A:B55). We show that the PP2A:B55-inhibiting Greatwall (Gwl)-endosulfine (ENSA) pathway is both necessary and sufficient for switch-like phosphorylations of mitotic substrates. Using purified components of the Gwl-ENSA pathway in a reconstituted system, we found a sharp Cdk1 threshold for phosphorylation of a luminescent mitotic substrate. The Cdk1 threshold to induce mitotic phosphorylation is distinctly higher than the Cdk1 threshold required to maintain these phosphorylations-evidence for bistability. A combination of mathematical modeling and biochemical reconstitution show that the bistable behavior of the Gwl-ENSA pathway emerges from its mutual antagonism with PP2A:B55. Our results demonstrate that two interlinked bistable mechanisms provide a robust solution for irreversible and switch-like mitotic entry.


The Dalton quantum chemistry program system.
Dalton is a powerful general-purpose program system for the study of molecular electronic structure at the Hartree-Fock, Kohn-Sham, multiconfigurational self-consistent-field, Møller-Plesset, configuration-interaction, and coupled-cluster levels of theory. Apart from the total energy, a wide variety of molecular properties may be calculated using these electronic-structure models. Molecular gradients and Hessians are available for geometry optimizations, molecular dynamics, and vibrational studies, whereas magnetic resonance and optical activity can be studied in a gauge-origin-invariant manner. Frequency-dependent molecular properties can be calculated using linear, quadratic, and cubic response theory. A large number of singlet and triplet perturbation operators are available for the study of one-, two-, and three-photon processes. Environmental effects may be included using various dielectric-medium and quantum-mechanics/molecular-mechanics models. Large molecules may be studied using linear-scaling and massively parallel algorithms. Dalton is distributed at no cost from http://www.daltonprogram.org for a number of UNIX platforms.


Pilot study of the differences among the Puerto Rican aged by gender in opinion, attitude and exposure to abuse, mistreatment and neglect.
Contribute to the knowledge base of the risk factor literature by gender for abuse, mistreatment and neglect of the Puerto Rican aged. A review of the literature in Puerto Rico and the United States related to gender impacting upon elder abuse, mistreatment and neglect report conflicting findings. A 107 item questionnaire was developed. The definitions of abuse, mistreatment and neglect were based on those set out by the National Center on Elder Abuse in the United States (1998). The questionnaire has been preliminarily validated by Cronbach's Alpha .879. The sample of respondents was based upon availability. This method was selected in the face of practical considerations in spite of criticism to the external validity that this type of sample represents. Structured face to face interviews were carried out from August to December 2004 with 430 aged residing in community. Data were entered into the SPSS program whereby cross tabulations were carried out for gender. Elderly Puerto Ricans regardless of gender are knowledgeable concerning what constitutes attributes of abuse, mistreatment and neglect. Females were found to report a high percentage of yes responses than males to opinion statements related to situations of abuse, mistreatment and neglect. Males reported a higher percentage of yes responses to situations of exposure to abuse and neglect than females. Findings here are similar to some reported in the United States but are dissimilar to those reported in Puerto Rico. Further research must be carried out to pinpoint the living arrangements situations placing the aged in greatest risk for abuse and neglect. Such findings have implications for social policy, prevention and intervention strategies.


The multiple effects of aspirin in prostate cancer patients.
Aspirin is a commonly used medication with anti-inflammatory and analgesic properties, and it is widely used to reduce the risk of ischaemic heart disease-related events and/or cerebrovascular accidents. However, there is also evidence from epidemiological and interventional studies to suggest that regular aspirin use can reduce the risk of prostate cancer development and progression, and can reduce the risk of disease recurrence following anti-prostate cancer therapy. Aspirin use in African-American men is associated with a reduced incidence of advanced PCa and reduced disease recurrence, and there is evidence from other studies of an association between regular aspirin use and decreased PCa-related mortality. The cyclooxygenase-2 enzyme inhibited by Aspirin and other NSAIDs, and which catalyses prostaglandin synthesis and mediates inflammation, is overexpressed in prostate cancer, therefore inhibition of cyclooxygenase-2 may have direct, and indirect, therapeutic effects. This review explores the evidence suggesting that aspirin use can modify prostate cancer biology and disease characteristics, and explores the potential mechanisms underpinning the observed associations between aspirin use and modification of prostate cancer risk. It also summarises the potential for adjuvant aspirin use to combine with other therapeutic approaches such as radical surgery and radiotherapy.


Earthquakes and the Earth's Wobble.
Observational evidence is presented in support of the hypothesis that large earthquakes excite the earth's natural wobble and produce the observed secular polar shift. Previous theoretical calculations based on elasticity theory and earthquake statistics had predicted a significant effect. There appear to be some premonitory signs of large earthquakes in the pole path.


Applying a global justice lens to health systems research ethics: an initial exploration.
Recent scholarship has considered what, if anything, rich people owe to poor people to achieve justice in global health and the implications of this for international research. Yet this work has primarily focused on international clinical research. Health systems research is increasingly being performed in low and middle income countries and is essential to reducing global health disparities. This paper provides an initial description of the ethical issues related to priority setting, capacity-building, and the provision of post-study benefits that arise during the conduct of such research. It presents a selection of issues discussed in the health systems research literature and argues that they constitute ethical concerns based on their being inconsistent with a particular theory of global justice (the health capability paradigm). Issues identified include the fact that priority setting for health systems research at the global level is often not driven by national priorities and that capacity-building efforts frequently utilize one-size-fits-all approaches.


Gait retraining versus foot orthoses for patellofemoral pain: a pilot randomised clinical trial.
To determine the feasibility of a clinical trial that compares a 6-week, physiotherapist-guided gait retraining program with a foot orthoses intervention in runners with patellofemoral pain. Pilot randomised controlled trial. Runners aged 18-40 years with clinically diagnosed patellofemoral pain were randomly allocated to either a 6-week gait retraining intervention of increasing cadence and use of a minimalist shoe or prefabricated foot orthoses. Outcomes at baseline and 12-weeks included recruitment, retention, adherence, adverse events, global improvement, anterior knee pain scale, worst and average pain on a 100mm visual analogue scale. Of the 16 randomised participants, two withdrew prior to commencing treatment due to non-trial related matters (n=1 from each group) and 14 completed the pilot trial. Minor calf muscle soreness was reported by 3 participants in the gait retraining group while no adverse events were reported in the foot orthoses group. There were no deviations from the treatment protocols. There was a large between-group difference favouring gait retraining at 12-weeks in the anterior knee pain scale and the worst pain in the past week, which was reflected in the number needed-to-treat of 2. This study supports the feasibility of a trial comparing gait retraining with foot orthoses and provides point estimates of effect that informs the design and planning of a larger clinical trial. It appears that a 6-week gait retraining program has a clinically meaningful effect on runners with patellofemoral pain when compared to an evidence-based treatment of foot orthoses.


Brain health: Key to health, productivity, and well-being.
Brain health is essential for physical and mental health, social well-being, productivity, and creativity. Current neurological research focuses mainly on treating a diseased brain and preventing further deterioration rather than on developing and maintaining brain health. The pandemic has forced a shift toward virtual working environments that accelerated opportunities for transdisciplinary collaboration for fostering brain health among neurologists, psychiatrists, psychologists, neuro and socio-behavioral scientists, scholars in arts and humanities, policymakers, and citizens. This could shed light on the interconnectedness of physical, mental, environmental, and socioeconomic determinants of brain disease and health. We advocate making brain health the top priority worldwide, developing common measures and definitions to enhance research and policy, and finding the cause of the decline of incidence of stroke and dementia in some countries and then applying comprehensive customized cost-effective prevention solutions in actionable implementation units. Life cycle brain health offers the best single individual, communal, and global investment.


Normalization and variance stabilization of single-cell RNA-seq data using regularized negative binomial regression.
Single-cell RNA-seq (scRNA-seq) data exhibits significant cell-to-cell variation due to technical factors, including the number of molecules detected in each cell, which can confound biological heterogeneity with technical effects. To address this, we present a modeling framework for the normalization and variance stabilization of molecular count data from scRNA-seq experiments. We propose that the Pearson residuals from "regularized negative binomial regression," where cellular sequencing depth is utilized as a covariate in a generalized linear model, successfully remove the influence of technical characteristics from downstream analyses while preserving biological heterogeneity. Importantly, we show that an unconstrained negative binomial model may overfit scRNA-seq data, and overcome this by pooling information across genes with similar abundances to obtain stable parameter estimates. Our procedure omits the need for heuristic steps including pseudocount addition or log-transformation and improves common downstream analytical tasks such as variable gene selection, dimensional reduction, and differential expression. Our approach can be applied to any UMI-based scRNA-seq dataset and is freely available as part of the R package sctransform, with a direct interface to our single-cell toolkit Seurat.


Cardiovascular manifestations of the emerging dengue pandemic.
Dengue is one of the most important emerging viral diseases globally. The majority of symptomatic infections result in a relatively benign disease course. However, a small proportion of patients develop severe clinical manifestations, including bleeding, organ impairment, and endothelial dysfunction with increased capillary permeability causing hypovolaemic shock that can lead to cardiovascular collapse. Evidence is increasing that dengue can also cause myocardial impairment, arrhythmias and, occasionally, fulminant myocarditis. No antiviral agents or vaccines are licensed for dengue, and treatment remains supportive with judicious fluid replacement for patients with severe disease. Defining the role of cardiac dysfunction in the haemodynamic compromise of severe dengue has potentially important management implications. In this Review, we will outline the current understanding of the cardiovascular manifestations of dengue, including myocardial and vascular involvement, and conclude with a discussion of the available therapeutic options and potential future research directions.


The effects of a loading dose followed by constant rate infusion of xylazine compared with romifidine on sedation, ataxia and response to stimuli in horses.
To compare xylazine and romifidine constant rate infusion (CRI) protocols regarding degree of sedation, and effects on postural instability (PI), ataxia during motion (A) and reaction to different stimuli. Blinded randomized experimental cross-over study. Ten adult horses. Degree of sedation was assessed by head height above ground (HHAG). Effects on PI, A and reaction to visual, tactile and acoustic stimulation were assessed by numerical rating scale (NRS) and by visual analogue scale (VAS). After baseline measurements, horses were sedated by intravenous loading doses of xylazine (1 mg kg(-1) ) or romifidine (80 μg kg(-1) ) administered over 3 minutes, immediately followed by a CRI of xylazine (0.69 mg kg(-1 ) hour(-1) ) or romifidine (30 μg kg(-1 ) hour(-1) ) which was administered for 120 minutes. Degree of sedation, PI, A and reaction to the different stimuli were measured at different time points before, during and for one hour after discontinuing drug administration. Data were analysed using two-way repeated measures anova, a Generalized Linear Model and a Wilcoxon Signed Rank Test (p < 0.05). Significant changes over time were seen for all variables. With xylazine HHAG was significantly lower 10 minutes after the loading dose, and higher at 150 and 180 minutes (i.e. after CRI cessation) compared to romifidine. Reaction to acoustic stimulation was significantly more pronounced with xylazine. Reaction to visual stimulation was greater with xylazine at 145 and 175 minutes. PI was consistently but not significantly greater with xylazine during the first 30 minutes. Reaction to touch and A did not differ between treatments. Compared to romifidine, horses were more responsive to metallic noise with xylazine. Time to maximal sedation and to recovery were longer with romifidine than with xylazine. With romifidine sufficient time should be allowed for complete sedation before manipulation.


Analysis of bcl-2 expression in normal, inflamed, dysplastic nasopharyngeal epithelia, and nasopharyngeal carcinoma: association with p53 expression.
To further characterize bcl-2 expression in nasopharyngeal carcinoma (NPC), the authors analyzed bcl-2 expression immunohistochemically in biopsy specimens from 101 cases of NPC, of which 65 had the component of normal nasopharyngeal epithelium (NPE), 24 with dysplastic lesions adjacent to carcinoma, and 14 with both primary and metastatic lesions. An additional 25 nasopharyngeal biopsies of NPE from patients with chronic inflammation of nasopharynx were also included. The percentage of detectable bcl-2 expression shown in NPC (80%) and adjacent dysplastic lesions (71%) was significantly higher than in adjacent NPE (37%) and NPE from patients with chronic inflammation of the nasopharynx (30%) (P < .05). In both normal and inflamed NPE, the detectable bcl-2 expression was restricted to the basal cells; however, in dysplastic lesions, the bcl-2 staining distribution was increased with the dysplastic cell layers, and in entire layers of epithelium in severe dysplasia or carcinoma in situ. In addition, the staining intensity of bcl-2 in carcinomas and adjacent dysplastic lesions was generally stronger than that of adjacent NPE. These observations suggest that the expression of bcl-2 in dysplasia and carcinoma is enhanced relative to that of adjacent NPE. Enhanced bcl-2 expression to prevent apoptosis seems to occur from the early stages and may play an important role in the carcinogenesis of NPC. Furthermore, up to 77% of NPC with the coexpression of bcl-2 and p53 was observed and suggested that the association of bcl-2 and p53 expression seems to occur from the early stages of the development of NPC. The overexpression of p53 protein in NPC suggests that the mutation of p53 gene or altered function of wild-type p53 protein may contribute to the pathogenesis. It is conceivable that the presence of both enhanced bcl-2 expression and altered p53 functions may play a crucial synergistic effect in the carcinogenesis of NPC.


Strategies to design antimicrobial contact lenses and contact lens cases.
Contact lens wear is a primary risk factor for developing ocular complications, such as contact lens acute red eye (CLARE), contact lens-induced peripheral ulcer (CLPU) and microbial keratitis (MK). Infections occur due to microbial contamination of contact lenses, lens cases and lens care solution, which are exacerbated by extended lens wear and unsanitary lens care practices. The development of microbial biofilms inside lens cases is an additional complication, as the developed biofilms are resistant to conventional lens cleaning solutions. Ocular infections, particularly in the case of MK, can lead to visual impairment or even blindness, so there is a pressing need for the development of antimicrobial contact lenses and cases. Additionally, with the increasing use of bandage contact lenses and contact lenses as drug depots and with the development of smart contact lenses, contact lens hygiene becomes a therapeutically important issue. In this review, we attempt to compile and summarize various chemical strategies for developing antimicrobial contact lenses and lens cases by using silver, free-radical producing agents, antimicrobial peptides or by employing passive surface modification approaches. We also evaluated the advantages and disadvantages of each system and tried to provide input to future directions. Finally, we summarize the developing technologies of therapeutic contact lenses to shed light on the future of contact lens applications.


Optimization of a synthetic receptor for dimethyllysine using a biphenyl-2,6-dicarboxylic acid scaffold: insights into selective recognition of hydrophilic guests in water.
In the design of small molecule receptors for polar guests, much inspiration has been taken from proteins that have adapted effective ways to selectively bind polar molecules in aqueous environments. Nonetheless, molecular recognition of hydrophilic guests in water by synthetic receptors remains a challenging task. Here we report a new synthetic receptor, A2I, with improved affinity and selectivity for a biologically important polar guest, dimethyllysine (Kme2). A2I was prepared via redesign of a small molecule receptor (A2B) that preferentially binds trimethyllysine (Kme3) using dynamic combinatorial chemistry (DCC). We designed a new biphenyl-2,6-dicarboxylate monomer, I, with the goal of creating a buried salt bridge with Kme2 inside a synthetic receptor. Indeed, incorporation of I into the receptor A2I resulted in a receptor with 32-fold enhancement in binding affinity, which represents the highest affinity receptor for Kme2 in the context of a peptide to date and is tighter than most Kme2 reader proteins. It also exhibits a ∼2.5-fold increase in preference for Kme2 vs. Kme3 relative to the parent receptor, A2B. This work provides insight into effective strategies for binding hydrophilic, cationic guests in water and is an encouraging result toward a synthetic receptor that selectively binds Kme2 over other methylation states of lysine.


250 nm period grating transferred by proximity i-line mask-aligner lithography.
This Letter, describes a fabrication method based on a high refractive index binary phase mask combined with a suitable illumination setup, which produces a close to normal incidence illumination, to fabricate sub-micrometer diffraction gratings. The method uses the i-line (365 nm) of a mercury lamp spectrum in a mask-aligner in proximity mode, to avoid any contact between the mask and the wafer, which is normally used to produce high resolution structures. The transfer of the structure in a fused silica wafer demonstrates that mask-aligner lithography can produce high aspect ratio sub-wavelength structures without resorting to any contact between mask and wafer.


Interaction of gold nanosurfaces/nanoparticles with collagen-like peptides.
Nanotechnology has quickly emerged as a promising research field with potential effects in disease treatments. For example, gold nanoparticles (AuNPs) have been extensively used in diagnostics and therapeutics. When administrated into human tissues, AuNPs first encounter extracellular matrix (ECM) molecules. Amongst all the ECM components, collagen is the main tension-resisting constituent, whose biofunctional and mechanical properties are strongly dependent on its hierarchical structure. Therefore, an in-depth understanding of the structural response of collagen to the presence of gold nanosurfaces (AuNS) and AuNPs is crucial in terms of clinical applications of AuNPs. However, detailed understanding of the molecular-level and atomic-level interaction between AuNS/AuNPs and collagen in the ECM is elusive. In this study, comprehensive molecular dynamics (MD) simulations have been performed to investigate the molecular behaviour of a collagen molecule segment (CMS) in the presence of AuNS/AuNPs in explicit water, aiming to explore the interaction of AuNS/AuNPs with collagen triple helices at the molecular and atomic levels. The results show that the CMS forms a rapid association with AuNS/AuNPs and undergoes a severe unfolding upon adsorption on AuNS/AuNPs, indicating an unfolding propensity of gold surfaces. We conclude that collagen triple helices unfold readily on AuNS and bare AuNPs, due to the interaction of gold surfaces with the protein backbone. The revealed clear unfolding nature and the unravelled atomic-level unfolding mechanism of collagen triple helices onto AuNPs contribute to the development of AuNPs for biomedical and therapeutic applications, and the design of gold-binding proteins.


Cdkn1a deletion improves stem cell function and lifespan of mice with dysfunctional telomeres without accelerating cancer formation.
Telomere shortening limits the proliferative lifespan of human cells by activation of DNA damage pathways, including upregulation of the cell cycle inhibitor p21 (encoded by Cdkn1a, also known as Cip1 and Waf1)) (refs. 1-5). Telomere shortening in response to mutation of the gene encoding telomerase is associated with impaired organ maintenance and shortened lifespan in humans and in mice. The in vivo function of p21 in the context of telomere dysfunction is unknown. Here we show that deletion of p21 prolongs the lifespan of telomerase-deficient mice with dysfunctional telomeres. p21 deletion improved hematolymphopoiesis and the maintenance of intestinal epithelia without rescuing telomere function. Moreover, deletion of p21 rescued proliferation of intestinal progenitor cells and improved the repopulation capacity and self-renewal of hematopoietic stem cells from mice with dysfunctional telomeres. In these mice, apoptotic responses remained intact, and p21 deletion did not accelerate chromosomal instability or cancer formation. This study provides experimental evidence that telomere dysfunction induces p21-dependent checkpoints in vivo that can limit longevity at the organismal level.


Staphylococcus aureus sepsis induces early renal mitochondrial DNA repair and mitochondrial biogenesis in mice.
Acute kidney injury (AKI) contributes to the high morbidity and mortality of multi-system organ failure in sepsis. However, recovery of renal function after sepsis-induced AKI suggests active repair of energy-producing pathways. Here, we tested the hypothesis in mice that Staphyloccocus aureus sepsis damages mitochondrial DNA (mtDNA) in the kidney and activates mtDNA repair and mitochondrial biogenesis. Sepsis was induced in wild-type C57Bl/6J and Cox-8 Gfp-tagged mitochondrial-reporter mice via intraperitoneal fibrin clots embedded with S. aureus. Kidneys from surviving mice were harvested at time zero (control), 24, or 48 hours after infection and evaluated for renal inflammation, oxidative stress markers, mtDNA content, and mitochondrial biogenesis markers, and OGG1 and UDG mitochondrial DNA repair enzymes. We examined the kidneys of the mitochondrial reporter mice for changes in staining density and distribution. S. aureus sepsis induced sharp amplification of renal Tnf, Il-10, and Ngal mRNAs with decreased renal mtDNA content and increased tubular and glomerular cell death and accumulation of protein carbonyls and 8-OHdG. Subsequently, mtDNA repair and mitochondrial biogenesis was evidenced by elevated OGG1 levels and significant increases in NRF-1, NRF-2, and mtTFA expression. Overall, renal mitochondrial mass, tracked by citrate synthase mRNA and protein, increased in parallel with changes in mitochondrial GFP-fluorescence especially in proximal tubules in the renal cortex and medulla. Sub-lethal S. aureus sepsis thus induces widespread renal mitochondrial damage that triggers the induction of the renal mtDNA repair protein, OGG1, and mitochondrial biogenesis as a conspicuous resolution mechanism after systemic bacterial infection.


UK National Registry Study of Kidney Donation After Circulatory Death for Pediatric Recipients.
Donation after circulatory death (DCD) kidney transplantation has acceptable renal allograft survival in adults but there are few data in pediatric recipients. The aim of this study was to determine renal allograft outcomes for pediatric recipients of a DCD kidney. Data were collected from the UK Transplant Registry held by National Health Service Blood and Transplant. Kidney transplants performed for pediatric recipients (age, <18 years) in the United Kingdom from 2000 to 2014 were separated into DCD, donation after brain death (DBD), and living donor (LD) transplants, analyzing 3-year patient and renal allograft survival. One thousand seven hundred seventy-two kidney only transplants were analyzed. Twenty-one (1.2%) of these were from DCD donors, 955 (53.9%) from DBD donors, and 796 (44.9%) from LDs. Patient survival is 100% in the DCD group, 98.7% in the DBD group, and 98.9% in the LD group. Three-year renal allograft survival was 95.2% in the DCD group, 87.1% in the DBD group, and 92.9% in the LD group. There was no significant difference in 3-year renal allograft survival between the DCD and DBD groups (P = 0.42) or DCD and LD groups (P = 0.84). For DCD, the primary nonfunction rate was 5% and delayed graft function was 25%. Children receiving a DCD kidney transplant have good renal allograft survival at 3-year follow-up, comparable to those receiving a kidney from a DBD donor or a LD. This limited evidence encourages the use of selected DCD kidneys in pediatric transplantation, and DCD allocation algorithms may need to be reviewed in view of this.