[Retroperitoneal lymph node metastasis in epithelial cancer of the ovary: a report of 58 cases].
To analyze the retroperitoneal lymph node metastasis in epithelial cancer of the ovary and offer scientific indications for lymph node radical dissection. Fifty-eight patients with ovarian cancer treated from January 1990 to December 2000 were retrospectively reviewed. Single-factor and multifactor analysis with Logistic regression model were performed by SPSS 10.0 statistic software. The metastasis rates of overall lymph nodes, pelvic nodes and para-aortic nodes were 48.3%, 37.9% and 25.9% respectively, among which no significant difference was noted (P > 0.05). Single-factor analysis showed that tumor location, ascitic condition, clinical stage and the size of residual tumor were associated with retroperitoneal lymph node metastasis. Multifactor analysis revealed that clinical stage and size of residual tumor were independent risk factors for metastasis of retroperitoneal lymph nodes. For early ovarian cancer patients, it is extremely important to perform radical dissection of the retroperitoneal lymph nodes. For advanced or residual lesions, radical dissection of pelvic nodes and para-aortic nodes could be considered in the second exploration.


Transfer of carbohydrate-active enzymes from marine bacteria to Japanese gut microbiota.
Gut microbes supply the human body with energy from dietary polysaccharides through carbohydrate active enzymes, or CAZymes, which are absent in the human genome. These enzymes target polysaccharides from terrestrial plants that dominated diet throughout human evolution. The array of CAZymes in gut microbes is highly diverse, exemplified by the human gut symbiont Bacteroides thetaiotaomicron, which contains 261 glycoside hydrolases and polysaccharide lyases, as well as 208 homologues of susC and susD-genes coding for two outer membrane proteins involved in starch utilization. A fundamental question that, to our knowledge, has yet to be addressed is how this diversity evolved by acquiring new genes from microbes living outside the gut. Here we characterize the first porphyranases from a member of the marine Bacteroidetes, Zobellia galactanivorans, active on the sulphated polysaccharide porphyran from marine red algae of the genus Porphyra. Furthermore, we show that genes coding for these porphyranases, agarases and associated proteins have been transferred to the gut bacterium Bacteroides plebeius isolated from Japanese individuals. Our comparative gut metagenome analyses show that porphyranases and agarases are frequent in the Japanese population and that they are absent in metagenome data from North American individuals. Seaweeds make an important contribution to the daily diet in Japan (14.2 g per person per day), and Porphyra spp. (nori) is the most important nutritional seaweed, traditionally used to prepare sushi. This indicates that seaweeds with associated marine bacteria may have been the route by which these novel CAZymes were acquired in human gut bacteria, and that contact with non-sterile food may be a general factor in CAZyme diversity in human gut microbes.


Evidence of Cross-Regulation in Two Closely Related Pyruvate-Sensing Systems in Uropathogenic Escherichia coli.
Two-component systems (TCSs) dictate many bacterial responses to environmental change via the activation of a membrane-embedded sensor kinase, which has molecular specificity for a cognate response regulator protein. However, although the majority of TCSs operate through seemingly strict cognate protein-protein interactions, there have been several reports of TCSs that violate this classical model of signal transduction. Our group has recently demonstrated that some of these cross-interacting TCSs function in a manner that imparts a fitness advantage to bacterial pathogens. In this study, we describe interconnectivity between the metabolite-sensing TCSs YpdA/YpdB and BtsS/BtsR in uropathogenic Escherichia coli (UPEC). The YpdA/YpdB and BtsS/BtsR TCSs have been previously reported to interact in K12 E. coli, where they alter the expression of putative transporter genes yhjX and yjiY, respectively. These target genes are both upregulated in UPEC during acute and chronic murine models of urinary tract infection, as well as in response to pyruvate and serine added to growth media in vitro. Here, we show that proper regulation of yhjX in UPEC requires the presence of all components from both of these TCSs. By utilizing plasmid-encoded luciferase reporters tracking the activity of the yhjX and yjiY promoters, we demonstrate that deletions in one TCS substantially alter transcriptional activity of the opposing system's target gene. However, unlike in K12 E. coli, single gene deletions in the YpdA/YpdB system do not alter yjiY gene expression in UPEC, suggesting that niche and lifestyle-specific pressures may be selecting for differential cross-regulation of TCSs in pathogenic and non-pathogenic E. coli.


Efimov physics: a review.
This article reviews theoretical and experimental advances in Efimov physics, an array of quantum few-body and many-body phenomena arising for particles interacting via short-range resonant interactions, that is based on the appearance of a scale-invariant three-body attraction theoretically discovered by Vitaly Efimov in 1970. This three-body effect was originally proposed to explain the binding of nuclei such as the triton and the Hoyle state of carbon-12, and later considered as a simple explanation for the existence of some halo nuclei. It was subsequently evidenced in trapped ultra-cold atomic clouds and in diffracted molecular beams of gaseous helium. These experiments revealed that the previously undetermined three-body parameter introduced in the Efimov theory to stabilise the three-body attraction typically scales with the range of atomic interactions. The few- and many-body consequences of the Efimov attraction have been since investigated theoretically, and are expected to be observed in a broader spectrum of physical systems.


Antitumorigenic Effects of ZAKβ, an Alternative Splicing Isoform of ZAK.
Sterile alpha motif (SAM)- and leucine-zipper-containing kinase (ZAK) plays a role in the regulation of cell cycle progression and oncogenic transformation. The ZAK gene generates two transcript variants, ZAKα and ZAKβ, through alternative splicing. In this study, we identified that ZAKα proteins were upregulated in tumor tissues, whereas ZAKβ proteins were mostly expressed in corresponding normal tissues. The ectopically expressed ZAKβ proteins in cancer cells inhibited cancer cell proliferation as well as anchorage-independent growth. The ZAKβ:ZAKα protein ratio played a role in the regulation of the cyclic adenosine monophosphate (cAMP) signaling pathway, whereas high ZAKβ protein levels led to the activation of cAMP response element binding protein 1 (CREB1) and exerted antitumor properties. Overexpression of ZAKβ or CREB1 cDNAs in cancer cells inhibited anchorage-independent growth and also reduced the levels of cyclooxygenase 2 (Cox2) and β-catenin proteins. Cancer cells treated with doxorubicin (Doxo) resulted in the switching from the expression of ZAKα to ZAKβ and also inhibited cancer cell growth in soft agar, demonstrating that pharmacological drugs could be used to manipulate endogenous reprogramming splicing events and resulting in the activation of endogenous antitumorigenic properties. We showed that the two ZAK transcript variants, ZAKα and ZAKβ, had opposite biological functions in the regulation of tumor cell proliferation in that ZAKβ had powerful antitumor properties and that ZAKα could promote tumor growth.


G-quartet self-assembly under osmotic pressure: remote control by vesicle shrinking rather than stress.
Does osmotic pressure stimulate assembly or disassembly of supramolecules in vesicles? Self-assembly was conceivable as intravesicular response to osmotic shrinking upon application of extravesicular overpressure, whereas disassembly was conceivable as a response to bilayer stress in hyperosmotic vesicles. Self-assembly of guanosine 5'-monophosphates (GMPs) into G-quartets was selected to investigate the nature of remote control of supramolecular chemistry within vesicles by osmotic pressure. Using circular dichroism spectroscopy to selectively detect G-quartets, we found that extravesicular overpressure stimulates intravesicular self-assembly, whereas underpressure stimulates disassembly. G-quartet self-assembly by osmotic pressure exhibited ion-selective metal-cation templation, as expected. The key conclusions are that supramolecular chemistry within vesicles is governed by vesicle shape rather than vesicle stress and that detection of osmotic pressure by CD spectroscopy is an interesting alternative to the commonly used methods based on fluorescence self-quenching.


Cl-channel blockers inhibit cell proliferation and arrest the cell cycle of human ovarian cancer cells.
To investigate the role of chloride channels in cell proliferation and cell cycles of human ovarian cancer cell line A2780. Chloride channel blockers were used to observe the effects of chloride channels on A2780 cells with MTT assay and flow cytometry. NPPB (100 microM) significantly inhibited the cell proliferation and affected the cell cycle, which increased the percentage of cells in the G1 phase, and reduced it in the S phase. NFA (100 microM) and TAM (30 microM) had similar inhibitory effects. Glibenclamide (100 microM), however, had no effect on cell proliferation or cycle. Moreover, chloride channel blockers could inhibit Ca2+ influx in these cells. Chloride channels, voltage-gated chloride channels, and volume-sensitive chloride channels especially, play an important role in the cell proliferation and cycle of A2780 cells. It is likely that the influence of chloride channels on cell proliferation and cell cycle is mediated by a Ca2+-dependent mechanism.


Public health messages on arboviruses transmitted by Aedes aegypti in Brazil.
The outbreak of Zika virus in Brazil in 2015 followed the arrival of chikungunya in 2014 and a long history of dengue circulation. Vital to the response to these outbreaks of mosquito-borne pathogens has been the dissemination of public health messages, including those promoted through risk communication posters. This study explores the content of a sample of posters circulated in Brazil towards the end of the Zika epidemic in 2017 and analyses their potential effectiveness in inducing behaviour change. A content analysis was performed on 37 posters produced in Brazil to address outbreaks of mosquito-borne pathogens. The six variables of the Health Belief Model were used to assess the potential effectiveness of the posters to induce behaviour change. Three overarching key messages emerged from the posters. These included (i) the arboviruses and their outcomes, (ii) a battle against the mosquito, and (iii) a responsibility to protect and prevent. Among the six variables utilised through the Health Belief Model, cues to action were most commonly featured, whilst the perceived benefits of engaging in behaviours to prevent arbovirus transmission were the least commonly featured. The posters largely focused on mosquito-borne transmission and the need to eliminate breeding sites, and neglected the risk of the sexual and congenital transmission of Zika and the importance of alternative preventive actions. This, we argue, may have limited the potential effectiveness of these posters to induce behaviour change.


[Atrioventricular septal defect. Anatomopathologic study and embryological correlation].
Atrioventricular septal defect is a complex congenital heart disease in which cardiac septation is deficient especially at atrioventricular and ventricular levels. The anatomopathologic experience at the National Institute of Cardiology "Ignacio Chávez" is described; an anatomo-embryological correlation was made to gain an insight of the pathogenesis. Seventy hearts were analyzed morphologically with the sequential segmentary system. Atrial situs, connections of cardiac segments, cardiac septa, septal defects, ventricular geometry, fibrous skeleton and associated anomalies were determined. Histologic sections of chick embryonic hearts were analyzed and compared with stages of human heart development, serving as basis to elaborate the anatomo-embryological correlation. Seventy hearts had a common atrioventricular valve and five had two separated atrioventricular valves. The ventricular septal defect was open in the first type and closed in the second; the other features were the same in both. This cardiopathy is shown as a broad spectrum of anatomical variations. The morphological knowledge of this cardiac malformation is useful to understand the physiopathology, to make a precise clinical diagnosis, and to give the best orientation to the surgeon. Delay in the development of the atrioventricular cushions is emphasized as the pathogenetic explanation for this type of congenital heart disease.


Improving Parenting Practices Among Fathers Who Misuse Opioids: Fathering Through Change Intervention.
Fathers have been largely neglected in the parenting literature though there is a critical need to improve parenting practices among fathers who misuse opioids in the midst of the opioid epidemic. Urgency is critical to rapidly intervene in the lives of fathers and children to reduce misuse and interrupt intergenerational cycles of substance misuse. Thus, we provide an overview of solutions to adapt existing parenting interventions for fathers who misuse opioids to accelerate the pace of science for this population.


Effects of motivationally significant stimuli on the regulation of dominant responses.
In 4 experiments, Ss responded as quickly as possible to indicate whether character strings consisted of letters or numbers. Following a warning stimulus that was or was not designed to trigger a motivational-emotional response in particular subgroups (viz., those with high anxiety, discrepant self-concepts, and eating disorders), strings were presented in a central (dominant) location on 75% of the trials or in 1 of 4 peripheral (nondominant) locations. Consistent with hypotheses, response times to peripherally located strings were significantly slower following motivationally significant than following neutral warning stimuli. Contrary to hypotheses, such stimuli did not facilitate responding to centrally located strings. It is proposed that motivationally significant stimuli engender a temporary reduction in control processing that hinders regulation of dominant responses.


Smoking exposure, loss of forced expiratory volume in one second and the risk of lung cancer among patients with malignant disease who present with cardiac or pulmonary symptoms: a cross-sectional study.
Smokers with airway obstruction are at a higher risk of lung cancer than smokers without airway obstruction. Inflammation plays a key role in lung carcinogenesis. This single-center study prospectively assessed (i) the relationship between smoking exposure and the loss of forced expiratory volume in 1 s (FEV1) in determining lung cancer risk and (ii) the effect of lung cancer on systemic inflammation. The study group comprised 475 consecutively enrolled patients with cancer who presented with pulmonary or cardiac symptoms. The effects of smoking exposure and FEV1 loss on the predicted lung cancer risk were assessed using multiple logistic regression analysis. C-reactive protein (CRP) was used as a marker of inflammation. The prevalence of lung cancer was 0.23. The lung cancer risk increased with the number of pack years and FEV1 loss (p < 0.01). Moving from the 5th (-22% of the predicted value) to the 95th percentile of FEV1 loss (56% of the predicted value) increased lung cancer risk from 0.07 to 0.23 (Δ = 0.16) at 0 pack years and from 0.39 to 0.73 (Δ = 0.34) at 70 pack years (95th percentile). The values for Δ peaked at 61 pack years (0.34) and then decreased with a further increase in smoking exposure, without reaching the zero mark. Patients with lung cancer were more likely to have a CRP level above the median (4.05 mg/L) than patients with other cancers (adjusted odds ratio = 2.67). Systemic inflammation is more pronounced in patients with lung cancer than in patients with other cancers. The effect of FEV1 loss on the patients' predicted risks of lung cancer increases with increasing smoking exposure. Measurements of FEV1 loss are useful to identify patients facing an increased risk of developing lung cancer.


Erythrocytes and platelet adhesion to endothelium are mediated by specialized molecules.
Erythrocytes in normal conditions have weak interactions with other blood cells and endothelial cells while in pathological circumstances they can adhere to endothelium and aggregate or agglutinate to blood cells. Erythrocyte adhesion was found to be abnormal in sickle cell anemia and diabetes mellitus and correlated to the vascular complications. Further studies demonstrated that VLA-4 adhesion molecule (alpha4beta1) present on erythrocytes bound to vascular cell adhesion molecule (VCAM-1) of the endothelium. In addition, the blood group Lutheran molecule (LU) overexpressed on sickle erythrocytes bind to laminin present on cells or in the intercellular space. In diabetes mellitus the formation of advanced glycation end products (AGE) by reaction between carbohydrates and free aminogroups of lysine is responsible for red blood cell membrane glycation. AGEs present on RBCs bind to the receptor for AGE (RAGE) on endothelium, activating endothelial cells. A molecule related to blood group Rhesus was demonstrated to belong to the intercellular adhesion molecule (ICAM) family. ICAM-4 binds to integrins present on leukocytes (CD11-CD18) and on platelets (alpha2beta4) offering a surface, which can be involved in thrombosis. The identification of erythrocytes adhesion molecules open a new way to understand thrombotic processes and vascular dysfunction.


Low-grade sebaceous carcinoma presenting on the leg.
We present herein the case of an 80-year-old woman with a keratotic plaque measuring 1.4x1.3 cm on the left popliteal fossa. The plaque was resected and diagnosed on microscopic examination as low-grade sebaceous carcinoma. Sebaceous carcinoma on the leg seems extremely rare.


Alzheimer's disease as proteolytic disorders: anabolism and catabolism of beta-amyloid.
Recent studies on the familial Alzheimer's disease (FAD)-linked mutations in three independent genes have established the pathogenic role of beta-amyloid (Abeta) deposition as a common pathway leading to neurodegeneration. Most of these mutations seem to contribute to Abeta deposition by directly causing the overproduction of Abeta1-42, a form of Abeta with high insolubility attributed to its carboxyl-terminal structure, through secretory proteolysis. In contrast, the mechanism of Abeta deposition in sporadic Alzheimer's disease (SAD), which accounts for more than 90% of disease cases, is unclear. Because Abeta overproduction is rarely observed in SAD, a possible candidate mechanism is a decreased degradation, or dyscatabolism, of Abeta. It is notable that a reduction in catabolism of only 30-50% is estimated to exert an equivalent effect on Abeta metabolism as the overproduction seen in FAD. Identification of the in vivo catabolic processes responsible for Abeta disposition would provide a new basis for the development of preventive and therapeutic measures against the disease. I hypothesized recently that aminopeptidase-catalyzed proteolysis of Abeta may limit the rate of Abeta catabolism and that the reduction of a certain aminopeptidase activity would lead to Abeta dyscatabolism and thus to deposition (Aminopeptidase Hypothesis), based on the structural properties of Abeta deposited in human brain. Experimental and clinical observations supporting this hypothesis are accumulating although further work is necessary to fully evaluate its relevance. If the assumption proves to be true, both the familial and sporadic forms of AD may be referred to as "proteolytic disorders" in anabolic and catabolic terms, respectively.


Peripheral neuropathy with gammopathy responding to plasmapheresis.
Two patients with subacute sensorimotor peripheral neuropathy were found to have benign gammopathies. They both responded to treatment by plasmapheresis. We suggest that a vigorous search for a paraprotein should be made in all undiagnosed patients presenting with peripheral neuropathy, including electrophoresis with isoelectric focusing. Plasmapheresis may be a treatment modality that should be explored in such cases.


Investigation of the chiral recognition role of cyclodextrin hydroxyl moieties via high performance liquid chromatography.
Cyclodextrin (CD) is known to afford excellent enantioselectivities due to its hydrophobic cavity and external H-bonding sites from hydroxyl moieties. However, there is still a lack of direct and comprehensive evidence clearly illustrating the origin of the important H-bonding effect. Regarding this issue, herein, four allylimidazole CD derivatives by selective substitution of the primary (6-position) and/or secondary (2,3-position) CD were synthesized and clicked onto silica surfaces to afford the corresponding chiral stationary phases (CSPs). The chiral chromatographic performances were systematically evaluated by separating 35 racemic analytes including isoxazolines, dansyl-amino acids, flavonoids and other racemates under reversed-phase HPLC. The chiral selection factors (α) and retention times (k) of the analytes on the as-prepared CSPs were comprehensively compared and it reveals that the enantioseparation ability was significantly altered due to the selective substituents of CD hydroxyl groups. The natural allylimidazole CD CSP (AICDCSP) was superior to the 6-O-tert-butyldimethylsilyl AICDCSP (6-TBDMAICDCSP) for most analytes. Dansyl amino acids and Ar-Pys were well separated on AICDCSP and 6-TBDMAICDCSP, where dansyl amino leucine gained the highest resolution up to 4.72 on AICDCSP, and flavonoids and Ar-Oprs were only separated on AICDCSP. These interesting separation results demonstrate that the secondary hydroxyl groups play a pivotal role in the separation of chiral compounds. In addition, the size of the CD cavity and the choice of solute also have an effect on the separation of substances. The mechanism involved in enantioselective discrimination of the selectively substituted CDs was further investigated by the molecular docking simulation.


Nanocrystal Engineering of Thin-Film Yttria-Stabilized Zirconia Electrolytes for Low-Temperature Solid-Oxide Fuel Cells.
To overcome significantly sluggish oxygen-ion conduction in the electrolytes of low-temperature solid-oxide fuel cells (SOFCs), numerous researchers have devoted considerable effort to fabricating the electrolytes as thin as possible. However, thickness is not the only factor that affects the electrolyte performance; roughness, grain size, and internal film stress also play a role. In this study, yttria-stabilized zirconia (YSZ) was deposited via a reactive sputtering process to fabricate high-performance thin-film electrolytes. Various sputtering chamber pressures (5, 10, and 15 mTorr) were investigated to improve the electrolytes. As a result, high surface area, large grain size, and residual tensile stress were successfully obtained by increasing the sputtering pressure. To clarify the correlation between the microstructure and electrolyte performance, a YSZ thin-film electrolyte was applied to anodized aluminum oxide-supported SOFCs composed of conventional electrode materials which are Ni and Pt as the anode and the cathode, respectively. The thin-film SOFC with YSZ deposited at 15 mTorr exhibited the lowest ohmic resistance and, consequently, the highest maximum power density (493 mW/cm2) at 500 °C whose performance is more than five times higher than that of the cell with YSZ deposited at 5 mTorr (94.1 mW/cm2). Despite the basic electrode materials, exceptionally high performance at low operating temperature was achieved via controlling the single fabrication condition for the electrolyte.


Blood pressure and heart rate variability in workers of 8-hour shifts.
This study examined the effects of shiftwork on the cardiovascular system. The blood pressure (BP) and heart rate variability (HRV) of 134 male workers, who worked 8-hour shifts with rapid rotation of shifts at 3-day intervals, were examined for all the three shifts. In addition, the job stress was measured by Karasek's JCQ 49-item questionnaire and the circadian type was assessed by the morningness-eveningness questionnaire. The smoking and alcohol drinking habits, marital status and past medical history were also obtained. The method of analyzing the measured data based on a mixed model was used to illustrate the association between the shiftwork duration and the BP or HRV. The average age of workers was 29 years (between 25-44). Among them, 77.9% were current smokers, 50% showed the passive type of job strain in Karasek's model. The mean shiftwork duration was 5.21 years (range 5.4 months--10 years). In the circadian type, none of them belonged to a definitely morning type or a definitely evening type. In the multivariate analysis adjusted by age, job strain, shift, circadian rhythm and smoking, the blood pressure showed significantly increasing trends according to shiftwork duration in both the systolic and diastolic BP. The heart rate variability also showed a significantly decreasing trend according to the shiftwork duration in both the parasympathetic and sympathetic functions (p < 0.05). These results suggests that there are negative health effects arising from shiftwork on the cardiovascular system.


suPAR remains uninfluenced by surgery in septic patients with bloodstream infection.
Surgical trauma induces activation of the immune system and may cause an increase of inflammatory biomarkers tested postoperatively in septic patients treated for bloodstream infection. The aim of this study was to determine the impact of surgical interventions on the novel sepsis biomarker soluble urokinase plasminogen activator receptor (suPAR) and to compare results with those of routine laboratory parameters CRP, PCT, and IL-6 in patients with culture-proven bloodstream infection. Forty-six adult patients with positive blood culture undergoing minor or major surgical intervention were investigated, 12 blood culture positive patients served as control group. Blood was collected 24 hours before and after surgical intervention for determination of the sepsis biomarkers suPAR, CRP, PCT, and IL-6. Within the surgical study cohort, a non-significant increase of suPAR, CRP, and PCT was observed postoperatively (p 0.642; p 0.773; p 0.087). In contrast, a slight decrease of IL-6 (p 0.599) was observed. A significant correlation was calculated for the pre- and postoperative difference of CRP (p 0.028) and PCT (p 0.008) and type of surgical intervention received: after minor surgical intervention only PCT decreased significantly (p<0.001), while after major surgical interventions no significant differences were observed for all biomarkers evaluated. In the control group, a significant decrease of CRP (p 0.005) and PCT (p 0.005) was observed. In patients treated adequately for bloodstream infections, postoperative suPAR levels remained uninfluenced of the surgical trauma and might therefore be a reliable parameter for postoperative infectious monitoring. After minor surgical intervention, PCT seems to be the most reliable parameter.


Herpes simplex virus keratitis: an update of the pathogenesis and current treatment with oral and topical antiviral agents.
Ophthalmic herpes simplex viral keratitis is responsible for a range of ocular manifestations from superficial epithelial disease to stromal keratitis and endotheliitis. The Herpetic Eye Disease Study has guided the management of herpetic eye disease for almost twenty years, but newer medications such as valacyclovir are now available and are considered to have better bioavailability than acyclovir. In this review, we examine the existing evidence on the pathogenesis of different ophthalmic herpes simplex viral keratitis disease modalities and the role of oral and topically administered antiviral drugs in the treatment of herpes simplex viral keratitis.


Protein inter-domain linker prediction using Random Forest and amino acid physiochemical properties.
Protein chains are generally long and consist of multiple domains. Domains are distinct structural units of a protein that can evolve and function independently. The accurate prediction of protein domain linkers and boundaries is often regarded as the initial step of protein tertiary structure and function predictions. Such information not only enhances protein-targeted drug development but also reduces the experimental cost of protein analysis by allowing researchers to work on a set of smaller and independent units. In this study, we propose a novel and accurate domain-linker prediction approach based on protein primary structure information only. We utilize a nature-inspired machine-learning model called Random Forest along with a novel domain-linker profile that contains physiochemical and domain-linker information of amino acid sequences. The proposed approach was tested on two well-known benchmark protein datasets and achieved 68% sensitivity and 99% precision, which is better than any existing protein domain-linker predictor. Without applying any data balancing technique such as class weighting and data re-sampling, the proposed approach is able to accurately classify inter-domain linkers from highly imbalanced datasets. Our experimental results prove that the proposed approach is useful for domain-linker identification in highly imbalanced single- and multi-domain proteins.


Inflammatory monocytes regulate Th1 oriented immunity to CpG adjuvanted protein vaccines through production of IL-12.
Due to their capacity to skew T cell responses towards Th1 oriented immunity, oligonucleotides containing unmethylated CpG motifs (CpG) have emerged as interesting adjuvants for vaccination. Whereas the signalling pathways in response to CpG mediated TLR9 activation have been extensively documented at the level of the individual cell, little is however known on the precise identity of the innate immune cells that govern T cell priming and polarisation to CpG adjuvanted protein antigens in vivo. In this study, we demonstrate that optimal induction of Th1 oriented immunity to CpG adjuvanted protein vaccines requires the coordinated actions of conventional DCs and of monocytes. Whilst conventional DCs were required for antigen presentation and initial T cell priming, monocytes constitute the main source of the Th1 polarising cytokine IL-12.


Quantitative assessment of the vascularity of the proximal part of the humerus.
The current consensus in the literature is that the anterolateral branch of the anterior humeral circumflex artery provides the main blood supply to the humeral head. While the artery is disrupted in association with 80% of proximal humeral fractures, resultant osteonecrosis is infrequent. This inconsistency suggests a greater role for the posterior humeral circumflex artery than has been previously described. We hypothesized that the posterior humeral circumflex artery provides a greater percentage of perfusion to the humeral head than the anterior humeral circumflex artery does. In twenty-four fresh-frozen cadaver shoulders (twelve matched pairs), we cannulated the axillary artery proximal to the thoracoacromial branch and ligated the brachial artery in the forearm. In each pair, one shoulder served as a control with intact vasculature and, in the contralateral shoulder, either the anterior humeral circumflex artery or the posterior humeral circumflex artery was ligated. Gadolinium was injected through the cannulated axillary arteries, and magnetic resonance imaging was performed. After imaging, a urethane polymer was injected, and specimens were dissected. For volumetric analysis, the gadolinium uptake on the magnetic resonance imaging was quantified in each quadrant of the humeral head with use of a custom automated program. The gadolinium uptake was compared between the control and ligated sides and between the ligated anterior humeral circumflex artery and ligated posterior humeral circumflex artery groups. The posterior humeral circumflex artery provided 64% of the blood supply to the humeral head overall, whereas the anterior humeral circumflex artery supplied 36%. The posterior humeral circumflex artery also provided significantly more of the blood supply in three of the four quadrants of the humeral head. The finding that the posterior humeral circumflex artery provides 64% of the blood supply to the humeral head provides a possible explanation for the relatively low rates of osteonecrosis seen in association with displaced fractures of the proximal part of the humerus. In addition, protecting the posterior humeral circumflex artery during the surgical approach and fracture fixation may minimize loss of the blood supply to the humeral head.


[Pulp diseases in the deciduous teeth. I. Histopathological changes (correlation between microscopic picture and clinical diagnosis)].
Clinical and histopathological examinations of 85 deciduous molars from children aged between 2.5 and 8 years show considerable disagreement between the microscopical picture and the clinical diagnosis according to the classification of WILGA. This calls in question the appropriateness of the histopathological classification for clinical diagnosis. For this reason, the present study was undertaken to elaborate a new classification of pulpal affections in the deciduous dentition with special regard to the various clinical pictures.


Diversity of the osteoblastic phenotype.
Studies of bone cells in culture have raised two salient questions: are the findings representative of the in vivo situation and can the conflicting data from different cell models be reconciled? Review of the literature indicates that all osteoblastic cells, defined by their origin or by their ability to produce mineralized matrix, have a few common properties: production of type I collagen; increased alkaline phosphatase activity; and parathyroid hormone-stimulated adenylate cyclase. Other features, such as osteocalcin and prostaglandin E production and the response to prostaglandin E, are selectively expressed by certain cell types. Pilot studies on mRNA levels of 'bone proteins' in developing calvaria suggest that such differences may reflect stages in osteoblastic differentiation. Immortalization of calvaria-derived cells using a SV40 large T antigen vector, which may freeze the cells in their particular state of differentiation (as proposed for leukaemia cells), yields phenotypes consistent with that hypothesis. Immortal cell lines may thus help to characterize osteoblastic differentiation. The diversity of osteoblast responses in culture to hormones and growth factors could be due to these phenotype differences but could also represent a subspecialization of differentiated cells. In addition, in the organism regulatory agents act in concert on a heterogeneous interactive cell population. Nonetheless cell cultures can be useful in screening for and predicting in vivo responses, as was shown by the 1,25-(OH)2D3 stimulation of osteocalcin, and for studying the molecular mechanisms of regulatory effects. Cell lines are also convenient for the production of specific proteins and cDNA libraries, and for the expression of specific genes.


Hypertriglyceridemia and Cardiovascular Outcomes.
Cardiovascular disease, particularly ischemic heart disease, is one of the most common causes of morbidity and mortality in the United States. Atherosclerosis, the root cause of ischemic heart disease, is promoted by risk factors like elevated plasma low-density lipoprotein, low plasma high-density lipoprotein, smoking, hypertension, and diabetes mellitus. Even 66 years after a relation between triglycerides (TG) and cardiovascular disease was first suspected, TGs still continue to be a controversial risk factor and target for therapy. Some previous studies did not show any significant positive relationship between TG and cardiovascular mortality; however, recent meta-analyses found otherwise. The role of elevated TG in patients with low low-density lipoprotein and interventions to lower TG to reduce cardiovascular mortality and morbidity is an area of active research.


Assessment of knowledge and perception about silver diamine fluoride (SDF) for treating older adults among graduating dental students.
Silver diamine fluoride (SDF) is a promising therapy for arresting and preventing caries in difficult to treat, high-risk populations, including institutionalized older adults. This study investigates the knowledge and perceptions about SDF of graduating dental students in multiple U.S. dental schools, as well as their willingness to use SDF in their practices. A survey was designed consisting of 21 total questions: 6 questions regarding students' demographic information and their SDF content exposure contextualization, 8 questions tailored to investigate 2 domains regarding students' knowledge about SDF (properties and indication), and 7 questions aiming to investigate 3 domains regarding students' perceptions about SDF (SDF usefulness, appropriateness of using SDF, and willingness to use SDF when in private practice). The survey was then distributed to graduating dental students at 7 U.S. dental schools. A total of 386 surveys (response rate of 55%) was collected from 7 schools in the Spring of 2019. The median score resulting from the SDF content exposure questions was 3 (SD = 1.43) from a range of 0-5. The median score from knowledge about SDF properties was 4 (SD = 1.18) from a range of 0 to 6. In the multivariate analysis, a linear model found that the covariates "SDF Usefulness", "SDF Appropriateness" and "SDF Patient Willingness to Use" were significantly associated with higher student willingness to use SDF (R2 = 0.395). The results indicated that the graduating students have a positive perception of SDF regarding its usefulness and appropriateness. Graduating students appear inclined to utilize SDF upon entering private practice.


A carcinogenicity study of diphenylarsinic acid in F344 rats in drinking water for 104 weeks.
Diphenylarsinic acid (DPAA), a neurotoxic organic arsenical used as a chemical warfare agent, is present in the groundwater and soil in some regions of Japan due to illegal dumping after World War II. We previously demonstrated that DPAA promotes diethylnitrosamine-induced liver carcinogenesis in a medium-term rat liver bioassay. The purpose of the present study was to evaluate the potential carcinogenicity of DPAA, including investigation of whether the bile duct hyperplasia in the liver that was observed in a previous 52 week rat chronic study develops into a tumor, when administered to rats in their drinking water for 104 weeks. DPAA was administered to groups 1-4 at concentrations of 0, 5, 10, and 20 ppm in their drinking water for 104 weeks. A significant decrease in survival rate was found for females in the 20 ppm DPAA group. Body weights of males in the 20 ppm and females in the 10 and 20 ppm DPAA groups were significantly decreased compared to the controls. Overall histopathological evaluation of neoplasms in all tissues showed no significant increase of tumor incidence in any organ or tissue of the 5, 10, or 20 ppm DPAA-treated male or female F344 rats. In conclusion, the present study demonstrated that DPAA is not a complete carcinogen in male or female F344 rats.


Patient, sufferer, victim, casualty or person with cervical myelopathy: let us decide our identifier.
Among biomedical journals, person-first language is considered preferable to identity person-first language. However, not all populations of people with certain medical diagnoses, such as deafness, prefer person-first language. Moreover, adherence to person-first language is poor among the literature on some neurological diagnoses. Therefore, it is most appropriate to consult with people with a given diagnosis regarding their preferred identifier. Here, we describe a consensus process undertaken by the REsearch objectives and COmmon Data Elements for Degenerative Cervical Myelopathy (RECODE-DCM, recode-dcm.com) steering committee members with cervical myelopathy to determine our preferred identifier.


Immediate implant and rehabilitation based on All-on-4 concept in patients with generalized aggressive periodontitis: A medium-term prospective study.
Aggressive periodontitis renders a great challenge to the conventional implant due to the risks of infection and ongoing marginal bone loss (MBL). A study about full-arch immediate implant and restoration in patients with advanced generalized aggressive periodontitis (GAP) was not read, even though the All-on-4 concept has been proven to be predictable for edentulous patients. This prospective study aimed to evaluate the feasibility and medium-term outcomes of immediate implant and rehabilitation based on the All-on-4 concept in patients with advanced GAP via clinical and radiographic analyses. Seventeen patients (mean age 39.4 years) with advanced GAP received immediate postextraction implant and rehabilitation based on the All-on-4 concept between January 2009 and January 2014. Eighty implants were inserted into 20 arches (7 maxillae and 13 mandibles). The average follow-up duration was 5 years (range 2-7). Complications, probing depth, and plaque, bleeding, and gingiva indices were evaluated. MBL was measured based on the panoramic radiographs taken immediately after surgery and annually thereafter. The cumulative survival rate (CSR) of the implants was 98.75% (79/80) after an average of 5 years. One tilted implant failed due to peri-implantitis. The average peri-implant MBL was 0.8 ± 0.4 and 1.2 ± 0.3 mm after 1 and 7 years, respectively. The CSR was 100% (20/20) for definite prostheses, while 85% (17/20) for provisional prostheses. The average probing depth, and plaque, bleeding, and gingiva indices at the last recall visit were 3.0 ± 0.5, 1.2 ± 0.4, 0.5 ± 0.5, and 0.4 ± 0.4 mm, respectively. Patient showed high satisfaction to the overall effects. Based on this study, the All-on-4 concept provided predictable outcomes in patients with GAP in 2- to 7-year follow-ups, and averted the severe bone defect area of aggressive periodontitis.


Liquid crystal contact thermography and lacrimal tract inflammation: a preliminary report.
Inflammation within the lacrimal drainage pathways can be assessed by conventional thermography. Liquid crystal contact thermography, a simpler technique, also provides objective evidence of lacrimal tract inflammation and may prove particularly useful in cases in which no inflammation can be detected clinically.


Kinetic isotope effects in the water forming reaction H2/D2 + OH from rigorous close-coupling quantum dynamics simulations.
Thermal rate constants and kinetic isotope effects (KIE) for the prototypical water-forming D2 + OH → D + DHO reaction are calculated for temperatures between 150 K and 1000 K using rigorous quantum dynamics simulations including all degrees of freedom. Very good comparison with experimental results is found for the thermal rate constants and overall good comparison with experimental KIE is obtained. Thermal rate constants and KIE for temperatures above 300 K are obtained with rigorous close-coupling calculations and employing the J-shifting approximation for overall rotational motion. Very good agreement is found validating the J-shifting approximation for this reaction. Thermal rate constants and KIE below 300 K are thus only obtained employing J-shifting. Good comparison with approximate methods for the calculation of thermal rate constants is found. The KIE for the title reaction increases notably below 250 K, which is found to be mostly due to an increased tunneling contribution in the H2 + OH reaction at these temperatures. Furthermore, microcanonical rates are obtained which can serve as benchmarks for the further development of approximate rate constant calculations.


Monocytes in Tumorigenesis and Tumor Immunotherapy.
Monocytes are highly plastic innate immune cells that display significant heterogeneity during homeostasis, inflammation, and tumorigenesis. Tumor-induced systemic and local microenvironmental changes influence the phenotype, differentiation, and distribution of monocytes. Meanwhile, monocytes and their related cell subsets perform an important regulatory role in the development of many cancers by affecting tumor growth or metastasis. Thanks to recent advances in single-cell technologies, the nature of monocyte heterogeneity and subset-specific functions have become increasingly clear, making it possible to systematically analyze subset-specific roles of monocytes in tumorigenesis. In this review, we discuss recent discoveries related to monocytes and tumorigenesis, and new strategies for tumor biomarker identification and anti-tumor immunotherapy.


Avian polyomavirus identified in a nestling Gouldian finch (Erythrura gouldiae) in New Zealand.
Abstract Four 4-5-month-old nestlings and one adult in a commercial aviary of 53 Gouldian finches (Erythrura gouldiae) died over a 2-week period in July 2000. One nestling was necropsied and showed bronze-tinged skeletal muscles, a swollen liver with haemorrhagic margins and numerous haemorrhages on serosal surfaces. The histological lesions included multifocal hepatic necrosis and haemorrhage associated with the presence of large clear or basophilic intranuclear inclusions in hepatocytes and Kupffer cells suggestive of avian polyomavirus infection. Similar inclusion bodies were present in splenic histiocytes. DNA was subsequently extracted from archived portions of liver, spleen, gizzard, heart, lung and kidney. A broad spectrum nested PCR was used to detect polyomavirus which sequence analysis confirmed as finch polyomavirus. Avian polyomavirus. Avian virus infections such as polyomavirus should be suspected in cases of sudden death in nestlings, particularly in susceptible species such as psittacine and passerine birds. The archiving of tissues from unconfirmed disease outbreaks provides a valuable resource for retrospective investigations.


Demonstration of a metabolically active glucose-6-phosphate pool in the lumen of liver microsomal vesicles.
Glucose-6-phosphate transport was investigated in rat or human liver microsomal vesicles using rapid filtration and light-scattering methods. Upon addition of glucose-6-phosphate, rat liver microsomes accumulated the radioactive tracer, reaching a steady-state level of uptake. In this phase, the majority of the accumulated tracer was glucose, but a significant intraluminal glucose-6-phosphate pool could also be observed. The extent of the intravesicular glucose pool was proportional with glucose-6-phosphatase activity. The relative size of the intravesicular glucose-6-phosphate pool (irrespective of the concentration of the extravesicular concentration of added glucose-6-phosphate) expressed as the apparent intravesicular space of the hexose phosphate was inversely dependent on glucose-6-phosphatase activity. The increase of hydrolysis by elevating the extravesicular glucose-6-phosphate concentration or temperature resulted in lower apparent intravesicular glucose-6-phosphate spaces and, thus, in a higher transmembrane gradient of glucose-6-phosphate concentrations. In contrast, inhibition of glucose-6-phosphate hydrolysis by vanadate, inactivation of glucose-6-phosphatase by acidic pH, or genetically determined low or absent glucose-6-phosphatase activity in human hepatic microsomes of patients suffering from glycogen storage disease type 1a led to relatively high intravesicular glucose-6-phosphate levels. Glucose-6-phosphate transport investigated by light-scattering technique resulted in similar traces in control and vanadate-treated rat microsomes as well as in microsomes from human patients with glycogen storage disease type 1a. It is concluded that liver microsomes take up glucose-6-phosphate, constituting a pool directly accessible to intraluminal glucose-6-phosphatase activity. In addition, normal glucose-6-phosphate uptake can take place in the absence of the glucose-6-phosphatase enzyme protein, confirming the existence of separate transport proteins.


Insular hypoperfusion correlates with the severity of delusions in individuals with Alzheimer's disease.
The aim of this study was to identify the brain regions associated with the severity of delusions in individuals with Alzheimer's disease (AD) using single-photon emission computed tomography. Thirty-five AD patients participated in this study. Multiple regression analysis was performed to identify a linear correlation between the delusion subscale score of the Neuropsychiatric Inventory (NPI) and regional cerebral blood flow (rCBF) in AD patients with delusions (n = 14). In addition, rCBF in patients with (n = 14) and without delusions (n = 21) was compared using a 2-sample t test. The delusion subscale score of the NPI was negatively correlated with rCBF in the right anterior insula (uncorrected p < 0.01 at the voxel level and corrected p < 0.05 at the cluster level). Compared to patients without delusions, however, rCBF in the right anterior insula was not significantly decreased in patients with delusions. We suggest that right anterior insular dysfunction may exacerbate delusions, although it is not responsible for the onset of delusions.


Nucleotide sequence of the xth gene of Escherichia coli K-12.
The xth gene of Escherichia coli K-12, which encodes exonuclease III, has been sequenced. Exonuclease III from a cloned copy of the E. coli K-12 gene has been purified and characterized. The molecular weight (30,921), the amino-terminal amino acid sequence, and the amino acid composition of the polypeptide predicted from the nucleotide sequence are in excellent agreement with those properties determined for the purified enzyme. The xth promoter was mapped by primer extension of in vivo transcripts. Inspection of the nucleotide sequence reveals that a region of dyad symmetry which could form a hairpin stem-loop structure in RNA characteristic of a rho-dependent terminator lies immediately downstream from the xth gene.


Associations between breast milk intake volume, macronutrient intake and infant growth in a longitudinal birth cohort: the Cambridge Baby Growth and Breastfeeding Study (CBGS-BF).
Growth patterns of breastfed infants show substantial inter-individual differences, partly influenced by breast milk (BM) nutritional composition. However, BM nutritional composition does not accurately indicate BM nutrient intakes. This study aimed to examine the associations between both BM intake volumes and macronutrient intakes with infant growth. Mother-infant dyads (n 94) were recruited into the Cambridge Baby Growth and Breastfeeding Study (CBGS-BF) from a single maternity hospital at birth; all infants received exclusive breast-feeding (EBF) for at least 6 weeks. Infant weight, length and skinfolds thicknesses (adiposity) were repeatedly measured from birth to 12 months. Post-feed BM samples were collected at 6 weeks to measure TAG (fat), lactose (carbohydrate) (both by 1H-NMR) and protein concentrations (Dumas method). BM intake volume was estimated from seventy infants between 4 and 6 weeks using dose-to-the-mother deuterium oxide (2H2O) turnover. In the full cohort and among sixty infants who received EBF for 3+ months, higher BM intake at 6 weeks was associated with initial faster growth between 0 and 6 weeks (β + se 3·58 + 0·47 for weight and 4·53 + 0·6 for adiposity gains, both P < 0·0001) but subsequent slower growth between 3 and 12 months (β + se - 2·27 + 0·7 for weight and -2·65 + 0·69 for adiposity gains, both P < 0·005). BM carbohydrate and protein intakes at 4-6 weeks were positively associated with early (0-6 weeks) but tended to be negatively related with later (3-12 months) adiposity gains, while BM fat intake showed no association, suggesting that carbohydrate and protein intakes may have more functional relevance to later infant growth and adiposity.


The fragmented neuronal Golgi apparatus in amyotrophic lateral sclerosis includes the trans-Golgi-network: functional implications.
The Golgi apparatus (GA) of spinal cord motor neurons is fragmented in sporadic amyotrophic lateral sclerosis (ALS), and in asymptomatic and symptomatic transgenic mice expressing the G93A mutation of the gene of the human Cu,Zn superoxide dismutase, found in certain cases of familial ALS (FALS) [Gonatas NK (1994) Am J Pathol 145:751-761; Mourelatos Z, et al. (1996) Proc Natl Acad Sci USA 93:5472-5477]. A similar fragmentation of the GA has been described in cells treated with microtubule-depolymerizing drugs, where the organelle is functional and contains both Golgi stacks and trans-Golgi network (TGN), the compartment of exit and targeting of proteins processed by the GA. To gain a better definition of the structure of the fragmented neuronal GA in ALS, four cases of sporadic ALS with numerous Bunina bodies in spinal cord motor neurons were stained with antibodies against human TGN and against the lumenal and cytoplasmic domains of MG160, a protein of the medial cisternae of the GA. The fragmented GA was stained with the three antibodies, indicating the presence of both Golgi stacks and TGN. Furthermore, the staining of the fragmented GA by the antiserum against the cytoplasmic domain of MG160 indicates that the fragmentation of the GA is not the result of a terminal and global cytoplasmic lytic event. The Bunina bodies were not stained by the anti-MG160 antibodies, suggesting that they are not derived from the GA. The perikarya of neurons with fragmented GA showed normal immunoreactivity with antibodies against the heavy neurofilament subunit and alpha-tubulin. However, because of the lack of appropriate antibodies the localization of proteins such as spectrin, ankyrin, centractin and others which link the microtubules with the GA were not done. The findings support the hypothesis that, in ALS, the fragmented neuronal GA is functional. Additional work with animal models of ALS may establish whether the fragmentation of the GA is a sign of early degeneration or a compensatory reaction of the injured motor neuron.


Enhanced D-lactic acid production from renewable resources using engineered Lactobacillus plantarum.
D-lactic acid is used as a monomer in the production of poly-D-lactic acid (PDLA), which is used to form heat-resistant stereocomplex poly-lactic acid. To produce cost-effective D-lactic acid by using all sugars derived from biomass efficiently, xylose-assimilating genes encoding xylose isomerase and xylulokinase were cloned into an L-lactate-deficient strain, Lactobacillus plantarum. The resulting recombinant strain, namely L. plantarum NCIMB 8826 ∆ldhL1-pLEM-xylAB, was able to produce D-lactic acid (at optical purity >99 %) from xylose at a yield of 0.53 g g(-1). Simultaneous utilization of glucose and xylose to produce D-lactic acid was also achieved by this strain, and 47.2 g L(-1) of D-lactic acid was produced from 37.5 g L(-1) glucose and 19.7 g L(-1) xylose. Corn stover and soybean meal extract (SBME) were evaluated as cost-effective medium components for D-lactic acid production. Optimization of medium composition using response surface methodology resulted in 30 % reduction in enzyme loading and 70 % reduction in peptone concentration. In addition, we successfully demonstrated D-lactic acid fermentation from corn stover and SBME in a fed-batch fermentation, which yielded 61.4 g L(-1) D-lactic acid with an overall yield of 0.77 g g(-1). All these approaches are geared to attaining high D-lactic acid production from biomass sugars to produce low-cost, highly thermostable biodegradable plastics.


One-year-old male with accelerated growth and development.
A 1-year-old male child with isosexual central (gonadotropin-dependent) precocious puberty caused by hypothalamic hamartoma is reported. Details of the diagnosis based solely on neuromaging characteristics, and satisfactory results of medical treatment with gonadotropin releasing hormone agonist analogues, are highlighted.


Glucose desensitization in INS-1 cells: evidence of impaired function caused by glucose metabolite(s) rather than by the glucose molecule per se.
Type 2 diabetes characteristically involves disturbances of the beta-cell function including reduced insulin secretion in response to elevated glucose. In experimental diabetes, beta cells are often "blind" to glucose, and clonal beta-cell lines chronically exposed to glucose show impaired glucose sensing. The present study focuses on the effect of long-term exposure to high-glucose concentrations on insulin secretion, insulin store, and insulin mRNA content in the beta-cell line INS-1. The cellular insulin mRNA content has been shown to be reduced by approximately 90% on such exposure for 4 days. This decrement could be partly counteracted by subsequent culture for 4 days at low glucose, while daily alternate culture in high and low glucose did not prevent the insulin mRNA content from being reduced. The insulin release from cells cultured at high glucose was simultaneously reduced by 50%. This change was, however, not reversed by subsequent culture at low glucose, a pattern also found for the intracellular insulin stores. The suppression of insulin mRNA, insulin release, and intracellular insulin stores induced by high glucose was completely neutralized by the metabolic glucokinase blocker, mannoheptulose, while 2-deoxyglucose, a phosphoglucose isomerase blocker, had no impact. This suggests that glucokinase activity may have a negative regulatory effect. Addition of D-glyceraldehyde (DG) induced an increase in insulin release, while insulin mRNA remained unaltered. It would therefore seem that at least one glucose metabolite is involved in the glucose desensitization in INS-1 cells, which opens the prospect of regulatory factor(s), which possess(es) negative, as well as positive, actions.


Characterizing Disease Burden and Progression of Geographic Atrophy Secondary to Age-Related Macular Degeneration.
To understand levels of disease burden and progression in a real-world setting among patients from the United Kingdom with bilateral geographic atrophy (GA) secondary to age-related macular degeneration (AMD). Retrospective cohort analysis of a multicenter electronic medical record (EMR) database. Patients who were aged ≥50 years with bilateral GA and no history of choroidal neovascularization (CNV) and who attended 1 of 10 clinical sites using the EMR. A deidentified data set was constructed from the records held at the 10 sites. An algorithm was used to extract cases with a GA diagnosis, of which 1901 had bilateral GA and form the basis of this report. A sample of records randomly selected from each center was used to validate disease definitions. Progression to blindness (visual acuity [VA] <20 letters or Snellen 3/60 in the better-seeing eye), driving ineligibility (VA ≤70 letters or Snellen 6/12 in the better-seeing eye), progression to CNV, loss of 10 or more letters, and mean change in VA over time. At first record of GA, 7.1% had a VA in the better-seeing eye equal to or lower than the cutoff for blindness registration and 71.1% had a VA that would have rendered them ineligible to drive. Over time, 16% became legally blind (median time to outcome, 6.2 years) and 66.7% became ineligible to drive (median time to outcome, 1.6 years). In the worse-seeing eye, 40.1% lost ≥10 letters in 2.4 years. Among patients with baseline and 24-month VA measurements, mean VA decline was 6.1 letters in the worse-seeing eye (n = 413) and 12.4 letters in the better-seeing eye (n = 414). The rate of progression to CNV in either eye was 7.4% per patient-year. At initial diagnosis, based on VA in the better-seeing eye, a high proportion of patients with bilateral GA were ineligible to drive and approximately 7% were eligible for UK blindness registration. The subsequent reduction in VA that occurred in the better-seeing eye would render a further two-thirds ineligible to drive. These findings emphasize the severity of the visual disability associated with GA secondary to AMD.


Fetal neck tumors - antenatal and intrapartum management.
In this review article we describe the ante- and perinatal management of fetal neck tumors. Although there are rare congenital anomalies, the clinical consequences for the fetus may be fatal and include airways obstruction, heart insufficiency, pulmonary hypoplasia and cosmetic effect. The right management allows to decrease the fetal and neonatal mortality and morbidity associated with the disease. It includes intrauterine therapy in some cases, mostly in a goitrus hypothyroidism of the fetus, but firstly, an assessment of the fetal airways patency with a subsequent, eventual Ex-Utero Intrapartum Treatment (EXIT).


Atractylenolide I improves behaviors in mice with depression-like phenotype by modulating neurotransmitter balance via 5-HT2A.
To explore the antidepressant effects and targets of atractylenolide I (ATR) through a network pharmacological approach. Relevant targets of ATR and depression analyzed by network pharmacology were scored (identifying 5-HT2A targets). Through elevated plus maze, open field, tail suspension, and forced swimming tests, the behavioral changes of mice with depression (chronic unpredictable mild stress [CUMS]) were examined, and the levels of neurotransmitters including serotonin, dopamine, and norepinephrine (5-HT, DA, and NE) were determined. The binding of ATR to 5-HT2A was verified by small molecular-protein docking. ATR improved the behaviors of CUMS mice, elevated their levels of neurotransmitters 5-HT, DA, and NE, and exerted a protective effect on their nerve cell injury. After 5-HT2A knockout, ATR failed to further improve the CUMS behaviors. According to the results of small molecular-protein docking and network pharmacological analysis, ATR acted as an inhibitor by binding to 5-HT2A. ATR can improve the behaviors and modulate the neurotransmitters of CUMS mice by targeting 5-HT2A.


Alkyl and Aromatic Amines as Digestive Ripening/Size Focusing Agents for Gold Nanoparticles.
Both long chain alkyl thiols and alkyl amines behave as size focusing agents for gold nanoparticles, a process that is under thermodynamic control. However, amines do not oxidize surface gold atoms while thiols do oxidize surface gold to gold(I) with evolution of hydrogen gas. Therefore, alkyl amines participate in digestive ripening by a different mechanism. The efficiency of alkyl amines for this process is described and compared, and ultimate gold particle size differences are discussed. Reported herein is a detailed investigation of alkyl chain lengths for alkyl amines, aromatic amines (aniline), and unusually reactive amines (2-phenylethyl amine). Also, two methods of preparation of the crude gold nanoparticles were employed: gold ion reduction/inverse micelle vs. metal vaporization (Solvated Metal Atom Dispersion-SMAD).


Phenotypic differentiation and diversifying selection in populations of Eruca sativa along an aridity gradient.
The aridity gradient in the eastern Mediterranean offers an opportunity to investigate intra-specific genetic differentiation and local adaptation in plant populations. Here we used genetic (FST) and quantitative trait (PST) differentiation to assess local adaptation among three natural populations of Eruca sativa (Brassicaceae) distributed along a climatic range representing desert, semi-arid and Mediterranean habitats. Amplified fragment length polymorphism (AFLP) analysis revealed high genetic diversity in each population, but low genetic differentiation between populations and relatively high gene flow. Further phenotypic evaluation in a common garden experiment (conduced in a Mediterranean habitat) showed clear differences in phenological traits among populations (day of flowering and duration of the reproductive stage), shoot and root biomass, as well as fitness-related traits (total number of fruits and total seed weight). FST-PST comparison showed that PST values of the phenological traits, as well as below- and above-ground biomass and fitness-related traits, were higher than the FST values. Overall, our results support the identification of genotypic and phenotypic differentiation among populations of E. sativa. Furthermore, the FST-PST comparison supports the hypothesis that these were subjected to past diversifying selection. Thus, the results clearly demonstrate adaptive divergence among populations along an aridity gradient, emphasize the ecological value of early flowering time in arid habitats, and contribute to our understanding of the possible impact of climate change on evolutionary processes in plant populations.


Nuclear and mitochondrial deoxyribonucleic acid replication during mitosis in Saccharomyces cerevisiae.
To study nuclear and mitochondrial deoxyribonucleic acid (DNA) synthesis during the cell cycle, a 15N-labeled log-phase population of Saccharomyces cervisiae was shifted to 14N medium. After one-half generation, the cells were centrifuged on a sorbitol gradient in a zonal rotor to fractionate the population according to cell size and age into fractions representing the yeast cell cycle. DNA samples isolated from the zonal rotor cell samples were centrifuged to equilibrium in CsC1 in an analytical ultracentrifuge to separate the nuclear and mitochondrial DNA components. The amount of 14N incorporated into each 15N-labeled DNA species was measured. The extent of nuclear DNA replication per sample was obtained by measuring the amount of hybrid DNA. The percentage of hybrid nuclear DNA increased from 6 to 68% and then decreased to 44% during the cell cycle. Upon ultracentrifugation, mitochondrial DNA banded as a unimodal peak in all zonal rotor samples. Mitochondrial DNA replication could be ascertained only by the 14N level in each mitochondrial peak and not, as with nuclear DNA, by hybrid DNA level. In contrast to the nuclear incorporation pattern, the 14N percentage in mitochondrial DNA remained effectively constant during the cell cycle. Comparison of the data to theoretical distributions showed that nuclear DNA was replicated discontinuously during the cell cycle, whereas mitochondrial DNA was replicated continuously throughout the entire mitotic cycle.


A prototype low-cost secondary standard calorimeter for reference dosimetry with ultra-high pulse dose rates.
Ultra-high pulse dose rate modalities present significant dosimetry challenges for ionisation chambers due to significant ion recombination. Conversely, calorimeters are ideally suited to measure high dose, short duration dose deliveries and this work describes a simple calorimeter as an alternative dosemeter for use in the clinic. Calorimeters were constructed featuring a disc-shaped core and single sensing thermistor encased in a 3D-printed body shaped like a Roos ionisation chamber. The thermistor forms one arm of a DC Wheatstone bridge, connected to a standard DMM. The bridge-out-of-balance voltage was calibrated in terms of temperature. A graphite-core calorimeter was calibrated in terms of absorbed dose to water (J/kg) in Co-60 and conventional 6, 10 and 15 MV X-rays. Similarly, an aluminium-core calorimeter was calibrated in a conventional 20 MeV electron beam and tested in a research high dose per pulse 6 MeV electron beam. Calorimeters were successfully calibrated in terms of absorbed dose to water in conventional radiotherapy beams at approximately 5 Gy/min with an estimated uncertainty of ±2-2.5% (k = 2), and performed similarly in a 6 MeV electron beam delivering approximately 180 Gy/s. A simple, low-cost calorimeter traceably calibrated to existing primary standards of absorbed dose could be used as a secondary standard for dosimetry for ultra-high pulse dose rates in the clinic. Secondary standard calorimeters for routine measurements are not available commercially; this work presents the basis of a simple, low-cost solution for reference dosimetry for ultra-high pulse dose rate beams.


Synthesis and Characterization of Bioinspired [Mo2 Fe2 ]-Hydride Cluster Complexes and Their Application in the Catalytic Silylation of N2.
The hydride-supported [Mo2 Fe2 ] cluster complex {Cp*Mo(PMe3 )}2 {FeN(SiMe3 )2 }2 (H)8 (2 a; Cp*=η5 -C5 Me5 ) and its [Mo2 Mn2 ] congener 2 b were synthesized from the reactions of Cp*Mo(PMe3 )(H)5 (1) with M{N(SiMe3 )2 }2 (M=Fe, Mn). The amide-to-thiolate ligand-exchange reactions of complex 2 a with bulky thiol reagents (HSR; R=2,4,6-iPr3 C6 H2 (Tip), 2,6-(SiMe3 )2 C6 H3 (Btp)) furnished the corresponding hydride-supported [Mo2 Fe2 ](SR)2 cluster complexes. The [Mo2 Fe2 ] clusters served as catalyst precursors for the reductive silylation of N2 and yielded ≈65-69 equivalents of N(SiMe3 )3 relative to the [Mo2 Fe2 ] clusters. Treatment of complexes 2 a and b with an excess of CNtBu resulted in the formation of dinuclear Mo-Fe and Mo-Mn complexes, which indicated that the [Mo2 M2 ] cores (M=Fe, Mn) split into two dinuclear species upon accommodation of substrates.


[Monoclonal antibodies in the treatment of thyroid eye disease].
Thyroid eye disease (TED) is an autoimmune condition affecting extraocular muscles and orbital fat that is most often a manifestation of the Graves' disease or Hashimoto thyroiditis. This disease significantly worsens the quality of life of patients, and therefore requires the use of effective treatment methods. Traditional therapy involves glucocorticosteroids and x-rays aimed at reducing the inflammatory process, rather than proptosis and diplopia, while targeted therapy is better able to influence the clinical course of the disease. The review presents a modern understanding of the pathogenesis of TED and analysis of clinical studies concerning the use of monoclonal antibodies for its treatment.


Novel, non-crinophagic, degradation of connecting peptide in transformed pancreatic beta cells.
Proinsulin is converted to equimolar amounts of insulin and connecting peptide (C-peptide) in pancreatic beta cell secretory granules. The fate of C-peptide, which remains soluble in granules while insulin is crystallized, was studied in a rat insulinoma cell line (INS cells). A pulse-chase approach demonstrated that insulin to C-peptide ratios (I/CP) began to rise in INS cells as soon as conversion took place, reaching 2.2:1 at 6 h of chase, and 5.3:1 by 24 h. During the first 4 h of chase, this ratio was inverted in the medium under basal conditions, possibly reflecting preferential secretion of C-peptide via the post granular constitutive-like pathway. However, the amount of C-peptide secreted to the medium (1.4%/h) was trivial compared with the net loss of C-peptide from the INS cells, estimated to be approximately 33% during the 1st h. As no extracellular degradation of insulin or C-peptide was observed, the data show progressive, extensive intracellular degradation of C-peptide. Stimulation of INS granule exocytosis revealed that the I/CP ratio in secretory granules is representative of that of cell extracts, indicating that C-peptide, but not insulin, is lost from functionally competent granules with time. This particular degradative route for C-peptide is thus distinct from crinophagy (fusion of granules and lysosomes) and was found to be much more active in INS cells than in primary islet beta cells.


CD8 functions as an inhibitory ligand in mediating the immunoregulatory activity of CD8+ cells.
Antisense and sense transfection technologies were employed in cellular coculture systems to investigate the physiologic requirements for CD8-dependent immunoregulation. Our data indicate that cells bearing genetically engineered CD8 molecules incorporating a glycoinositolphospholipid membrane anchor, as well as fixed cells bearing natural CD8 molecules, retain specific, CD8-dependent immunoregulatory activity. These findings together support the novel concept that CD8, a molecule traditionally thought of as a receptor, can function as an inhibitory ligand. CD8-dependent inhibition was shown to induce nonresponsiveness, persisting for at least 24 h, in Ag-specific responders. Moreover, only cells undergoing primary, but not secondary, antigenic stimulation through their TCR were found to be susceptible to CD8-dependent inhibition. Both CD4+ and CD8+ responding T cells were inhibited by CD8+ modulatory cells. These functional analyses of inhibitory and responder cells in CD8-dependent inhibition lay the groundwork for developing artificial CD8-based immunomodulatory peptides and deciphering CD8's role in natural immunoregulation.


Interleukin-18, interleukin-8, and CXCR2 and the risk of silicosis.
Molecular mechanisms in the pathogenesis of silicosis are not fully understood. Exposure to crystalline silica leads to the activation of signaling pathways controlling the production and secretion of inflammatory mediators. Inflammatory cytokines are noted as important candidate genes for fibrotic lung diseases. Cytokines, chemokines, and variations of their genes have been associated with upregulation or downregulation of chronic inflammatory mediators. Variations in the interleukin (IL)-18, IL-8 and chemokine receptor CXCR2 genes are believed to influence the risk of silicosis in stone-grinding factory workers in Iran. Allele-specific oligonucleotide polymerase chain reaction (PCR) procedure was carried out for IL-18 -137 and IL-18 -607, meanwhile touchdown PCR was performed for IL-8 -251 and CXCR2 +1208 genotyping. Variation in genotypic and allelic frequencies was not statistically different among cases versus controls (p > 0.05). These findings indicated for the first time that IL-18 -137, IL-18 -607, IL-8 -251, and CXCR2 +1208 are suggested not to influence the risk of silicosis in tested occupational group.


Long-term outcomes after critical illness: recent insights.
Intensive care survivors often experience post-intensive care sequelae, which are frequently gathered together under the term "post-intensive care syndrome" (PICS). The consequences of PICS on quality of life, health-related costs and hospital readmissions are real public health problems. In the present Viewpoint, we summarize current knowledge and gaps in our understanding of PICS and approaches to management.


KIR gene content in amerindians indicates influence of demographic factors.
Although the KIR gene content polymorphism has been studied worldwide, only a few isolated or Amerindian populations have been analyzed. This extremely diverse gene family codifies receptors that are expressed mainly in NK cells and bind HLA class I molecules. KIR-HLA combinations have been associated to several diseases and population studies are important to comprehend their evolution and their role in immunity. Here we analyzed, by PCR-SSP (specific sequencing priming), 327 individuals from four isolated groups of two of the most important Brazilian Amerindian populations: Kaingang and Guarani. The pattern of KIR diversity among these and other ten Amerindian populations disclosed a wide range of variation for both KIR haplotypes and gene frequencies, indicating that demographic factors, such as bottleneck and founder effects, were the most important evolutionary factors in shaping the KIR polymorphism in these populations.


Translation of the glenohumeral joint in patients with multidirectional and posterior instability: awake examination versus examination under anesthesia.
The purpose of this study was to assist in establishing guidelines to determine the degree of shoulder instability, the value of awake clinical examination, and the value of examination under anesthesia (EUA). Forty-three patients with clinical diagnosis of multidirectional shoulder instability (MDI) and 28 patients with posterior instability underwent bilateral shoulder translation testing, both awake and while under anesthesia. Two surgeons using guidelines and translation grades developed by the American Shoulder and Elbow Surgeons examined each patient and assigned a single grade for the anterior, posterior, and inferior directions. A comparison of translational grade was performed with the use of Pearson chi2 and McNemar symmetry to determine association. The patients with MDI showed increased translation in the anterior, inferior, and posterior directions when the affected limb was compared with the noninvolved side in both preoperative examination and EUA. Furthermore, the patients with MDI showed increased anterior translation on the affected side during EUA compared with the clinical examination. Patients with posterior instability demonstrated increased anterior translation for both affected and noninvolved limbs during EUA. However, the posterior translation obtained before surgery did not change during EUA for both the affected and noninvolved limbs, and there was no side-to-side difference in posterior translation.


[Nutrition in pregnancy].
Review about during pregnancy. Nutrition of a pregnant woman has to cover the maternal need of energy and additionally the stuff for the fetus as preconditions for its growth. In the trimester an extra intake of 15 grams protein is necessary, but no one of fat and carbohydrates. An increase of 400-500 mg Calcium, 10 mg fernum and vitamins A, B1, B2 and C is very important. Smoking, alcohol and drug abuse may be injurious to the fetus. Passive smoking of the mothers, therefore automatically of the fetus, too, is harmful for the fetus. Our recommendations for nutrition of pregnant women demonstrate, that feeding during pregnancy is simple.


Patterns and environmental drivers of greenhouse gas fluxes in the coastal wetlands of China: A systematic review and synthesis.
Coastal wetlands play an increasingly important role in regulating greenhouse gas (GHG) fluxes and thus affecting climate change. However, the overall magnitude, trend, and environmental drivers of GHG fluxes in these wetlands of China remain uncertain. Herein, we synthesized data from 70 publications involving 187 field observations to identify patterns and drivers of GHG fluxes across coastal wetlands in China. Average methane (CH4), nitrous oxide (N2O) fluxes, and carbon dioxide (CO2) emissions (ecosystem respiration) across coastal wetlands were estimated as 2.20±0.31 mg·m-2·h-1, 16.44±2.96 μg·m-2·h-1, and 388.76±42.28 mg·m-2·h-1, respectively. GHG emissions varied with tidal inundation, where CH4 and CO2 emissions during tidal inundation were lower than during ebbing. CH4 and CO2 emissions from wetlands decreased linearly with increasing latitude, while N2O did not. CH4 fluxes were positively related to air temperature and aboveground biomass, and CO2 emissions were positively related to soil organic carbon. N2O fluxes were lower with increasing soil pH, and CH4 and CO2 emissions were greater with increasing soil moisture. Based on the results of sustained-flux global warming potential and sustained-flux global cooling potential models, our paper indicate that the fluxes of CH4 and N2O in coastal wetlands have a positive feedback to global warming, which is mainly driven by the CH4 emission. Our synthesis improved understanding of the roles of coastal wetlands in the ecosystem C cycle under global change. We suggest that long-term field observations of GHG fluxes across a wider range of spatiotemporal scales are urgently required to improve the prediction accuracy in GHG fluxes and the assessment of net GHG balance and its contribution to the GWP of coastal wetlands.


Approaching the minimal metal ion binding peptide for structural and functional metalloenzyme mimicking.
The peptides Ac-His-Pro-His-Pro-His-NH(2) (L1) and Ac-Lys-His-Pro-His-Pro-His-Gln-NH(2) (L2) have been prepared and the equilibria of their proton, copper(II) and zinc(II) complexes in aqueous solution have been studied by the combination of pH-potentiometric titrations, UV/visible and circular dichroism (CD) spectroscopy. The latter methods also provided information on solution structure of the complexes formed under different conditions. Both ligands formed complexes with three imidazole nitrogens around the metal ion at pH ~7. In the L1 containing system precipitation of either copper(II) or zinc(II) complexes occurred upon slight increase of the pH. The re-titration of the filtered and acidified precipitates revealed that the insoluble materials were neutral complexes rather than metal-hydroxides. Indeed, by attaching amino acids with polar side-chains to the His-Pro-His-Pro-His template in L2 we could prevent any precipitation, and the soluble complexes around pH ~7 exerted three imidazole nitrogens and a (deprotonated) water molecule around the metal ions. To our knowledge L2 provides the first example of a short peptide preventing both the amide nitrogen coordination in copper(II) and the formation of copper(II) and zinc(II) hydroxides. The zinc(II) and copper(II) complexes at pH ~7 having similar structure to the natural hydrolytic and redox enzymes, respectively, showed considerable activity in hydrolytic cleavage assays with a model substrate (2-hydroxypropyl-4-nitrophenyl phosphate), as well as with native plasmid DNA, and in a superoxide dismutase-like reaction.


Conflict monitoring mechanism at the single-neuron level in the human ventral anterior cingulate cortex.
Life requires monitoring and adjusting behavior in the face of conflicts. The conflict monitoring theory implicates the anterior cingulate cortex (ACC) in these processes; its ventral aspect (vACC) specializes in emotional conflict. To elucidate the underpinning neural mechanism, we recorded vACC extracellular activity from 12 patients with mood disorders or epilepsy who performed the face-emotional Stroop task. Behaviorally, both conflict detection and adaptation to conflict were evident. The firing rate of neurons in the vACC represented current conflict, i.e., current-congruency. The late onset of the effect is compatible with a role in monitoring. Additionally, early responses of some neurons represented the immediate history of conflicts, i.e., previous-trial-congruency. Finally, in some neurons the response to the current-trial was modulated by previous-trial-congruency, laying the ground for adjusting-to-conflicts. Our results uncover a single neuron level mechanism in the vACC that encodes and integrates past and present emotional conflicts, allowing humans to accommodate their responses accordingly.


Onset Potential for Electrolyte Oxidation and Ni-Rich Cathode Degradation in Lithium-Ion Batteries.
High-capacity Ni-rich layered metal oxide cathodes are highly desirable to increase the energy density of lithium-ion batteries. However, these materials suffer from poor cycling performance, which is exacerbated by increased cell voltage. We demonstrate here the detrimental effect of ethylene carbonate (EC), a core component in conventional electrolytes, when NMC811 (LiNi0.8Mn0.1Co0.1O2) is charged above 4.4 V vs Li/Li+-the onset potential for lattice oxygen release. Oxygen loss is enhanced by EC-containing electrolytes-compared to EC-free-and correlates with more electrolyte oxidation/breakdown and cathode surface degradation, which increase concurrently above 4.4 V. In contrast, NMC111 (LiNi0.33Mn0.33Co0.33O2), which does not release oxygen up to 4.6 V, shows a similar extent of degradation irrespective of the electrolyte. This work highlights the incompatibility between conventional EC-based electrolytes and Ni-rich cathodes (more generally, cathodes that release lattice oxygen such as Li-/Mn-rich and disordered rocksalt cathodes) and motivates further work on wider classes of electrolytes and additives.


Punishment in the public goods game is evaluated negatively irrespective of non-cooperators' motivation.
The evolution of human cooperation toward strangers remains puzzling. While the punishment of non-cooperators is a possible explanation, whether punishments can help cooperation evolve depends on how people evaluate punishers. Thus, it is of vital importance to elucidate the perception of punishers. Previous studies have found that punishment is evaluated differently in different contexts; punishers are negatively and positively evaluated in the public goods game with punishment (PGG-P) and in the third-party punishment game (TPP), respectively. To disentangle the mixed evidence, our research focused on motivations for non-cooperation and examined whether different motivations for non-cooperation could explain the inconsistent findings. We hypothesized that people positively evaluate punishers when they punish those who non-cooperated to exploit others (greed), e.g., in a TPP situation. Contrastingly, people negatively evaluate punishers when they punish those who non-cooperated to avoid exploitation (fear), e.g., in a PGG-P situation. If so, in either game, punishers would be evaluated positively in situations where greed dominates, and negatively in situations where fear dominates the non-cooperation motivation. To test this, we conducted two online experiments with hypothetical scenarios in which the decision-making order was used to manipulate the motivations of non-cooperators. The results of Study 1 (N = 240) using only PGG-P and Study 2 (N = 602) using both PGG-P and TPP showed that the non-cooperation motivation did not have a significant effect on the evaluations of punishers and non-punishers. Punishers (vs non-punishers) were evaluated negatively in PGG-P and positively in TPP regardless of the decision-making order of non-cooperation. We discussed the role of higher-order information in the evaluation of punishment.


[Features of biochemical changes in blood serum according to the form of progression of idiopathic and dysplastic coxarthrosis].
Carried out biochemical studies of blood serum (collagenase activity, glycosamineglicans and hydroxyproline fractions) of 72 patients with hip idiopathic osteoarthrosis and 30 patients with dysplastic osteoarthrosis of the iv-th stage in.accordance with J. H. Kellgren and J. S. Lavrence depending on the form of pathologic process progression. It has been proved that both with idiopathic and dysplastic coxartrosis metabolism of basic protein of osteochonrous tissue was broken both in catabolic and in synthetic phase of this process. The most deep changes 1 of biochemical values (collagenase, free and proteinbinded hydroxyprolines, the content of glycosamines) have been observed with rapid form of course progression of idiopathic and dysplastic coxarthrosis. Definite appropriateness promotesin.patients better understanding of coxarthrosis pathogenese, development of diagnostic and medical measures for patients with this severe orthopedic pathology.


Human skeletal muscle type 1 fibre distribution and response of stress-sensing proteins along the titin molecule after submaximal exhaustive exercise.
Early responses of stress-sensing proteins, muscle LIM protein (MLP), ankyrin repeat proteins (Ankrd1/CARP and Ankrd2/Arpp) and muscle-specific RING finger proteins (MuRF1 and MuRF2), along the titin molecule were investigated in the present experiment after submaximal exhaustive exercise. Ten healthy men performed continuous drop jumping unilaterally on a sledge apparatus with a submaximal height until complete exhaustion. Five stress-sensing proteins were analysed by mRNA measurements from biopsies obtained immediately and 3 h after the exercise from exercised vastus lateralis muscle while control biopsies were obtained from non-exercised legs before the exercise. Decreased maximal jump height and increased serum creatine kinase activities as indirect markers for muscle damage and HSP27 immunostainings on muscle biopsies as a direct marker for muscle damage indicated that the current exercised protocol caused muscle damage. mRNA levels for four (MLP, Ankrd1/CARP, MuRF1 and MuRF2) out of the five studied stress sensors significantly (p < 0.05) increased 3 h after fatiguing exercise. The magnitude of MLP and Ankrd2 responses was related to the proportion of type 1 myofibres. Our data showed that the submaximal exhaustive exercise with subject's own physical fitness level activates titin-based stretch-sensing proteins. These results suggest that both degenerative and regenerative pathways are activated in very early phase after the exercise or probably already during the exercise. Activation of these proteins represents an initial step forward adaptive remodelling of the exercised muscle and may also be involved in the initiation of myofibre repair.


Maternal depression and anxiety disorders (MDAD) and child development: A Manitoba population-based study.
To examine the association between maternal depression and anxiety disorders (MDAD) and child development assessed during the kindergarten year. Administrative data from several health and social databases in Manitoba, Canada, were used to study 18,331 mother-child pairs. MDAD over the period from one year prior to the child's birth to the kindergarten year was defined using physician diagnoses and filled prescriptions. Child development was assessed during the kindergarten year using the Early Development Instrument (EDI) which measures vulnerability across five domains of development. Structural equation modeling was used to examine associations between timing, recurrence and severity of MDAD and child outcomes. Health at Birth (preterm, low birth weight, neonatal intensive care stay and long birth hospitalization), Family Context (teen mother, lone parent, socio-economic status (SES)), child age and child sex were covariates. MDAD had a modest negative association with child EDI scores across all models tested, particularly for social, emotional and physical development. Prenatal MDAD had a stronger negative association with outcomes than other time periods; however, recurrent MDAD had a stronger negative association with outcomes than any specific time period or MDAD severity. The influence of MDAD was mediated by Family Context, which had a strong, negative association with outcomes, particularly language and cognitive development. The number of time periods a child was exposed to MDAD in early childhood was more negatively associated with five areas of child development than timing or severity. Prenatal exposure may be more sensitive to MDAD than other time periods. The familial context (teen mother, lone parenthood and low SES) had a stronger influence on child outcomes than MDAD. Findings can be used to inform interventions which address maternal mental health from the prenatal period onward, and to support disadvantaged families to encourage healthy birth outcomes, early childhood development and school readiness.


Colloid-oil-water-interface interactions in the presence of multiple salts: charge regulation and dynamics.
We theoretically and experimentally investigate colloid-oil-water-interface interactions of charged, sterically stabilized, poly(methyl-methacrylate) colloidal particles dispersed in a low-polar oil (dielectric constant ε = 5-10) that is in contact with an adjacent water phase. In this model system, the colloidal particles cannot penetrate the oil-water interface due to repulsive van der Waals forces with the interface whereas the multiple salts that are dissolved in the oil are free to partition into the water phase. The sign and magnitude of the Donnan potential and/or the particle charge is affected by these salt concentrations such that the effective interaction potential can be highly tuned. Both the equilibrium effective colloid-interface interactions and the ion dynamics are explored within a Poisson-Nernst-Planck theory, and compared to experimental observations.


Molecular Mechanism of Mot1, a TATA-binding Protein (TBP)-DNA Dissociating Enzyme.
The essential Saccharomyces cerevisiae ATPase Mot1 globally regulates transcription by impacting the genomic distribution and activity of the TATA-binding protein (TBP). In vitro, Mot1 forms a ternary complex with TBP and DNA and can use ATP hydrolysis to dissociate the TBP-DNA complex. Prior work suggested an interaction between the ATPase domain and a functionally important segment of DNA flanking the TATA sequence. However, how ATP hydrolysis facilitates removal of TBP from DNA is not well understood, and several models have been proposed. To gain insight into the Mot1 mechanism, we dissected the role of the flanking DNA segment by biochemical analysis of complexes formed using DNAs with short single-stranded gaps. In parallel, we used a DNA tethered cleavage approach to map regions of Mot1 in proximity to the DNA under different conditions. Our results define non-equivalent roles for bases within a broad segment of flanking DNA required for Mot1 action. Moreover, we present biochemical evidence for two distinct conformations of the Mot1 ATPase, the detection of which can be modulated by ATP analogs as well as DNA sequence flanking the TATA sequence. We also show using purified complexes that Mot1 dissociation of a stable, high affinity TBP-DNA interaction is surprisingly inefficient, suggesting how other transcription factors that bind to TBP may compete with Mot1. Taken together, these results suggest that TBP-DNA affinity as well as other aspects of promoter sequence influence Mot1 function in vivo.


Regulation of CEACAM1 Protein Expression by the Transcription Factor ETS-1 in BRAF-Mutant Human Metastatic Melanoma Cells.
BRAF becomes constitutively activated in 50% to 70% of melanoma cases. CEACAM1 has a dual role in melanoma, including facilitation of cell proliferation and suppression of infiltrating lymphocytes, which are consistent with its value as a marker for poor prognosis in melanoma patients. Here we show that BRAFV600E melanoma cells treated with BRAF and MEK inhibitors (MAPKi) downregulate CEACAM1 mRNA and protein expression in a dose- and exposure time-dependent manners. Indeed, there is a significant correlation between the presence of BRAFV600E and CEACAM1 expression in melanoma specimens obtained from 45 patients. Vemurafenib-resistant cell systems reactivate the MAPK pathway and restore basal CEACAM1 mRNA and protein levels. These combined results suggest transcriptional regulation. Indeed, luciferase reporting assays show that CEACAM1 promoter (CEACAM1p) activity is significantly reduced by MAPKi. Importantly, we show that the MAPK-driven CEACAM1p activity is mediated by ETS1, a major transcription factor and downstream effector of the MAPK pathway. Phosphorylation mutant ETS1T38A shows a dominant negative effect over CEACAM1 expression. The data are consistent with independent RNAseq data from serial biopsies of melanoma patients treated with BRAF inhibitors, which demonstrate similar CEACAM1 downregulation. Finally, we show that CEACAM1 downregulation by MAPKi renders the cells more sensitive to T-cell activation. These results provide a new view on a potential immunological mechanism of action of MAPKi in melanoma, as well as on the aggressive phenotype observed in drug-resistant cells.


NFAT is a nerve activity sensor in skeletal muscle and controls activity-dependent myosin switching.
Calcineurin (Cn) signaling has been implicated in nerve activity-dependent fiber type specification in skeletal muscle, but the downstream effector pathway has not been established. We have investigated the role of the transcription factor nuclear factor of activated T cells (NFAT), a major target of Cn, by using an in vivo transfection approach in regenerating and adult rat muscles. NFAT transcriptional activity was monitored with two different NFAT-dependent reporters and was found to be higher in slow compared to fast muscles. NFAT activity is decreased by denervation in slow muscles and is increased by electrostimulation of denervated muscles with a tonic low-frequency impulse pattern, mimicking the firing pattern of slow motor neurons, but not with a phasic high-frequency pattern typical of fast motor neurons. To determine the role of NFAT, we transfected regenerating and adult rat muscles with a plasmid coding for VIVIT, a specific peptide inhibitor of Cn-mediated NFAT activation. VIVIT was found to block the expression of slow myosin heavy chain (MyHC-slow) induced by slow motor neuron activity in regenerating slow soleus muscle and to inhibit the expression of MyHC-slow transcripts and the activity of a MyHC-slow promoter in adult soleus. The role of NFAT was confirmed by the finding that a constitutively active NFATc1 mutant stimulates the MyHC-slow, inhibits the fast MyHC-2B promoter in adult fast muscles, and induces MyHC-slow expression in regenerating muscles. These results support the notion that Cn-NFAT signaling acts as a nerve activity sensor in skeletal muscle in vivo and controls nerve activity-dependent myosin switching.


Bioactive constituents from Turkish Pimpinella species.
A new 'phenylpropanoid', 4-(3-methyloxiran-2-yl)phenyl 2-methylbutanoate (1), a new trinorsesquiterpene, 4-(6-methylbicyclo[4.1.0]hept-2-en-7-yl)butan-2-one (2), and eight known compounds (3-10) were isolated from the essential oils of several Pimpinella species growing in Turkey. The structures of the new compounds were determined by 1D- and 2D-NMR analyses. The absolute configuration of 1 was established via comparison of its optical rotation with that of 'epoxypseudoisoeugenyl 2-methylbutyrate' (11), the absolute configuration of which was determined by chemical degradation and an appropriate Mosher ester formation. Direct bioautography revealed antifungal activity of 1 and 11 against Colletotrichum acutatum, C. fragariae, and C. gloesporioides. Subsequent evaluation of antifungal compounds in a 96-well microtiter assay showed that compounds 1 and 11 produced the most-significant growth inhibition in Phomopsis spp., Colletotrichum spp., and Botrytis cinerea. Compounds 1 and 6 displayed antimicrobial activities against Mycobacterium intracellulare, with IC50 values of 2.78 and 1.29 microM, respectively.