Evaluation of 2'-Fucosyllactose and Bifidobacterium longum Subspecies infantis on Growth, Organ Weights, and Intestinal Development of Piglets.
Human milk is rich in oligosaccharides that influence intestinal development and serve as prebiotics for the infant gut microbiota. Probiotics and 2'-fucosyllactose (2'-FL) added individually to infant formula have been shown to influence infant development, but less is known about the effects of their synbiotic administration. Herein, the impact of formula supplementation with 2'-fucosyllactose (2'-FL) and Bifidobacterium longum subsp. infantis Bi-26 (Bi-26), or 2'-FL + Bi-26 on weight gain, organ weights, and intestinal development in piglets was investigated. Two-day-old piglets (n = 53) were randomized in a 2 × 2 design to be fed a commercial milk replacer ad libitum without (CON) or with 1.0 g/L 2'-FL. Piglets in each diet were further randomized to receive either glycerol stock alone or Bi-26 (109 CFU) orally once daily. Body weights and food intake were monitored from postnatal day (PND) 2 to 33/34. On PND 34/35, animals were euthanized and intestine, liver and brain weights were assessed. Intestinal samples were collected for morphological analyses and measurement of disaccharidase activity. Dry matter of cecum and colon contents and Bifidobacterium longum subsp. infantis abundance by RT-PCR were also measured. All diets were well tolerated, and formula intake did not differ among the treatment groups. Daily body weights were affected by 2'-FL, Bi-26, and day, but no interaction was observed. There was a trend (p = 0.075) for greater total body weight gain in CON versus all other groups. Jejunal and ascending colon histomorphology were unaffected by treatment; however, there were main effects of 2'-FL to increase (p = 0.040) and Bi-26 to decrease (p = 0.001) ileal crypt depth. The addition of 2'-FL and/or Bi-26 to milk replacer supported piglet growth with no detrimental effects on body and organ weights, or intestinal structure and function.


Mycophenolate Mofetil Use in Pediatric Immune Thrombocytopenia Refractory to First-line Therapy: a Single-center Experience.
Most children treated for immune thrombocytopenia remit during the first year following diagnosis. For the ∼40% who develop persistent or chronic disease, second-line treatment options include immunomodulation and thrombomimetic agents. While immunomodulators target the underlying mechanism, prolonged immunosuppression may increase the risk of infection. We report the use of the reversible immunomodulating agent mycophenolate mofetil (MMF) in 16 pediatric patients with immune thrombocytopenia refractory to first-line treatment. Using escalating doses up to 2400 mg/m 2 /d, MMF treatment resulted in a 73% response rate. Adverse events were mostly mild and tolerable. Complete responders have been successfully tapered off MMF with sustained responses.


Diverse effects of ANXA7 and p53 on LNCaP prostate cancer cells are associated with regulation of SGK1 transcription and phosphorylation of the SGK1 target FOXO3A.
Tumor suppressor function of the calcium/phospholipid-binding Annexin-A7 (ANXA7) has been shown in Anxa7-deficient mice and validated in human cancers. In the androgen-resistant prostate cancer cells, ANXA7 and p53 showed similar cytotoxicity levels. However, in the androgen-sensitive LNCaP, ANXA7 greatly exceeded the p53-induced cytotoxicity. We hypothesized that the p53 underperformance in LNCaP could be due to the involvement of p53-responsive SGK1 and FOXO3A. In this study, we show that p53 failed to match programmed cell death (PCD) and G1-arrest that were induced by ANXA7 in LNCaP. WT-ANXA7 preserved total FOXO3A expression with no hyperphosphorylation that could enable FOXO3A nuclear translocation and proapoptotic transcription. In contrast, in the p53-transfected LNCaP cells with maintained cell proliferation, the phosphorylated (but not total) FOXO3A fraction was increased implying a predominantly cytoplasmic localization and, subsequently, a lack of FOXO3A proapoptotic transcription. In addition, p53 reduced the expression of aberrant SGK1 protein form in LNCaP. Using Ingenuity Pathway Analysis and p53-signature genes, we elucidated the role of distinct SGK1/FOXO3A-associated regulation in p53 versus ANXA7 responses and proposed that aberrant SGK1 could affect reciprocal SGK1-FOXO3A-Akt regulation. Thus, the failure of the cell growth regulator p53 versus the phospholipid-binding ANXA7 could be potentially attributed to its diverse effects on SGK1-FOXO3A-Akt pathway in the PTEN-deficient LNCaP.


[Planning in Brazilian Public Health System: the case of the Health Secretariat of State of Bahia].
The object of this article is the planning process of the Health Secretariat of State of Bahia from January 2007 to July 2009. It describes and analyses this process, discussing the difficulties and the advances to build the planning practice in this institution. It was used documental analyses and systematic registers of lived experience by the authors to do it. The process points for an approach between theory and practice articulating the Strategic-situational focus and the Communicative Action, including the principle of the Social Participation. The analyses explain that the process occurs in a pool of dialogic movements enlaced by the explicative, normative, strategic and tactic-operative moments that finished with the construction of the Health Secretariat of State of Bahia. There is a powerful planning revealed in plurality and multiplicity of approaches used to build a collective "vision of future" in organizations that adopted "plans" like instruments to qualify Government actions.


Anatomical study of the cavernous sinus emphasizing operative approaches.
The aim of this article is to describe the microsurgical anatomy of the cavernous sinus, the triangles, and the osseous relationships in the region with special attention to the relationships important in surgical approaches on the intracavernous structures. Fifty cavernous sinuses obtained from twenty-five cadaver heads were studied in detail using magnification. Stepwise dissections of the cavernous sinuses performed to demonstrate the intradural and extradural routes, anatomy of the triangles and osseous relationships in the region. The main branches of the intracavernous portion of the carotid artery were the meningohypophyseal, the inferior cavernous sinus, and McConnell;s capsular artery . The main branches of the meningohypophyseal trunk were the tentorial, the dorsal meningeal, and the inferior hypophyseal artery. There were variations of the main branches of the meningohypophyseal trunk. The sixth cranial may splite into rootlets as it passes lateral to the carotid artery. There were size and shape variation of the triangles. Aprecise understanding of the bony relationships, the anatomy of the triangels and neurovascular content of the cavernous sinus, together with the use of cranial base and microsurgical techniques are necessary for safer surgery. Asingle approach is not capable of providing exposure of all parts of the sinus.


Cranial bony decompressions in the management of head injuries: decompressive craniotomy or craniectomy?
Decompressive surgery is one of the available options in dealing with traumatic brain injury (TBI) when clinical and radiological evidence confirm that medical treatment may be insufficient. This can be achieved either by complete removal of the bone or by allowing it to float, but the indications and utility of these are yet to be resolved. This study examines the indications and outcome for both procedures. Review of all cases of bony decompression done at the Memfys Hospital for Neurosurgery, Enugu, Nigeria from August 2002 to May 2010. Prospectively recorded data of CT, MRI, operating room, clinics and wards were utilized. There were 38 patients out of whom 35 were males and 3 females. The mean age was 36 years (range 15-80). The causes of the predisposing TBI were road traffic accidents (RTA) (79%), gunshot (10.5%), and assault (7.9%). Decompressive surgery was unilateral in 36 and bi-frontal in 2. Decompressive craniectomy with bone stored in anterior abdominal wall pocket was done in 8 patients and decompressive craniotomy with bone left in situ in 30. Of the latter, bone was unsecured and allowed to float in 13 and the craniotomy was lightly anchored with sutures in 17 patients. Surgery was performed within 24 h in 68.4% of cases. Mortality was 21.1% overall but was up to 25% in the more severely injured patients who had craniectomy. Bony decompression is useful in the management of head trauma. Careful selection of cases and appropriate radiological assessment are important and will guide decision for either craniotomy or craniectomy.


Effect of Incorporation of Pretreatment Serum Carcinoembryonic Antigen Levels Into AJCC Staging for Colon Cancer on 5-Year Survival.
The American Joint Committee on Cancer (AJCC) has proposed the inclusion of pretreatment serum carcinoembryonic antigen (CEA) levels (C stage) into the conventional TNM staging system of colon cancer. The latter proposal has yet to be widely adopted because of the lack of long-term survival estimates of after C-stage incorporation into AJCC staging. To evaluate whether long-term overall and cancer-specific survival is affected by inclusion of C stage into the standard AJCC TNM staging and to study the implications on survival estimates. We performed a retrospective study of all patients diagnosed as having histologically proven colonic adenocarcinoma from January 1, 2004, through December 31, 2005, from the National Cancer Institute's Surveillance, Epidemiology, and End Results database. We stratified each AJCC stage as C0 (normal) or C1 (elevated) based on the pretreatment serum CEA level. Median follow-up was 71 months. Five-year estimates of overall and disease-specific survival and hazard ratios (HRs) for estimates of risk of overall and disease-specific mortality. A total of 16 619 patients were evaluated, and of these, 8878 patients had C0 disease and 7741 had C1 disease. C1 stage was independently associated with a 51% and 59% increased risk of overall (HR, 1.51; 95% CI, 1.44-1.59; P < .001) and disease-specific mortality (HR, 1.59; 95% CI, 1.49-1.69; P < . 001) at a median follow-up of 71 months. Analysis of survival of the AJCC stages subdivided as C0 or C1 revealed a significant worse prognosis of C1 AJCC stages compared with the respective C0 AJCC stages. The magnitude of change in survival was large enough to cause clustering of survival estimates of C1 vs C0 cancers across various AJCC stages. Analysis of patients with stage I, II, and III cancer revealed that node-negative C1 disease was associated with prognosis similar or worse than node-positive C0 disease. Inclusion of C stage into the AJCC TNM staging of colon cancer revealed significant differences dependent on C stage in terms of 5-year survival. C-stage inclusion resulted in substantial change in survival estimates, with C1 status portending a prognosis to certain stages similar to or worse than higher AJCC TNM stages with C0 status. We recommend routine pretreatment CEA testing as standard of care in colon cancer and use of C stage for multimodality treatment planning and risk stratification in prospective studies and randomized clinical trials.


[Ovarian development and analysis of mating effects on ovary maturation of Diaphorina citri Kuwayama (Hemiptera: Psyllidae)].
As many other insects with a restricted diet, the citrus psyllid Diaphorina citri Kuwayama, which vector the causing agent of the Huanglongbing, the bacteria Candidatus Liberibacter spp., is intimately associated with symbiotic microorganisms. These mutualist symbionts play a key role on their host nutritional ecology, and are vertically transmitted to the progeny. However, despite the role symbionts play on host development and reproduction, and the growing opportunities of exploitation of the association insect-symbiont to control insect vectored-pathogens, there are very few studies on the host reproductive biology and on the symbiont transovarial transmission. Therefore, we aimed at analyzing the ovary development during D. citri adulthood, and at verifying for the mating requirement as a trigger to initiate ovary development. Newly-emerged D. citri females were grouped as virgin or mated and ovary development was observed during adulthood. Newly-emerged females have immature ovaries, and ovaries remain without any mature eggs until females mate. Once female mates, the vitellogenesis synthesis and uptake are estimulated, and oocytes are quickly developed. Oocytes maturation in ovarioles is metachronic, with only one oocyte developing at a time in each oogenic cycle. Morphological observations of the reproductive system including the ovaries and spermatheca, after the first cluster of eggs is laid, indicated D. citri is polyandrous, and may require multiple mating to develop additional oogenic maturation cycles.


The measles virus hemagglutinin β-propeller head β4-β5 hydrophobic groove governs functional interactions with nectin-4 and CD46 but not those with the signaling lymphocytic activation molecule.
Wild-type measles virus (MV) strains use the signaling lymphocytic activation molecule (SLAM; CD150) and the adherens junction protein nectin-4 (poliovirus receptor-like 4 [PVRL4]) as receptors. Vaccine MV strains have adapted to use ubiquitous membrane cofactor protein (MCP; CD46) in addition. Recently solved cocrystal structures of the MV attachment protein (hemagglutinin [H]) with each receptor indicate that all three bind close to a hydrophobic groove located between blades 4 and 5 (β4-β5 groove) of the H protein β-propeller head. We used this structural information to focus our analysis of the functional footprints of the three receptors on vaccine MV H. We mutagenized this protein and tested the ability of individual mutants to support cell fusion through each receptor. The results highlighted a strong overlap between the functional footprints of nectin-4 and CD46 but not those of SLAM. A soluble form of nectin-4 abolished vaccine MV entry in nectin-4- and CD46-expressing cells but only reduced entry through SLAM. Analyses of the binding kinetics of an H mutant with the three receptors revealed that a single substitution in the β4-β5 groove drastically reduced nectin-4 and CD46 binding while minimally altering SLAM binding. We also generated recombinant viruses and analyzed their infections in cells expressing individual receptors. Introduction of a single substitution into the hydrophobic pocket affected entry through both nectin-4 and CD46 but not through SLAM. Thus, while nectin-4 and CD46 interact functionally with the H protein β4-β5 hydrophobic groove, SLAM merely covers it. This has implications for vaccine and antiviral strategies.


Semiautomated aseptic preparation of patient-individual antineoplastic intravenous solutions: first experiences in a German hospital pharmacy.
In 2013, the PharmaHelp device was introduced to daily patient care at the University Hospital of Heidelberg in order to improve process and staff safety and health. It is a semiautomated device located in a safety workbench for aseptic preparation with weight-and-visual-supported process check as well as radio-frequency identification of bags and vials. After qualification and a training period, several high-admixture-volume drugs were chosen stepwise to be prepared by the device. During a reporting period of 6 months, the number of processed production runs and produced bags were assessed as well as the productivity per hour of the device based on the processed admixing volume and time consumption for manual preprocessing and postprocessing steps. Weight conformity tests of 246 processed bags in 11 admixing-volume ranges showed 99.19% with a deviation of <5%. Validation of the automated preparation process with 40 media fills showed no microbiological growth. Daily patient care production in the first 6 months led to a nearly similar weight accuracy and an average productivity of 22.5 preparations per running hour of the device at an average admixing volume of 39.6 mL, excluding manual preprocessing and postprocessing tasks. Time consumption for all manual steps was about 76 min per run. The automated admixing process of the device showed remarkable effectivity and a satisfying accuracy, but the manual preprocessing and postprocessing steps are time consuming, which is associated with the local environment, and has to be reorganised.


Recent progress in the development of fluorescent, luminescent and colorimetric probes for detection of reactive oxygen and nitrogen species.
Reactive oxygen (ROS) and nitrogen (RNS) species cause oxidative and nitrosative stresses, respectively. These stresses are implicated not only in diverse physiological processes but also in various pathological processes, including cancer and neurodegenerative disorders. In addition, some ROS and RNS in the environment are pollutants that threaten human health. As a consequence of these effects, sensitive methods, which can be employed to selectively monitor ROS and RNS in live cells, tissues and organisms as well as in environmental samples, are needed so that their biological roles can be understood and their concentrations in environmental samples can be determined. In this review, fluorescent, luminescent and colorimetric ROS and RNS probes, which have been developed since 2011, are comprehensively discussed.


Improved procedure for recovery of Yersinia enterocolitica from meats.
A 1- to 3-day enrichment-KOH postenrichment procedure was evaluated and found to be as effective in recovering Yersinia enterocolitica from meats as a 14- to 21-day cold enrichment procedure, with or without KOH postenrichment. The shortened procedure consists of enriching 1.0- and 25-g samples of meat in phosphate-buffered saline (pH 7.2) at 25 degrees C. After incubation (48 and 72 h for 1.0-g samples and 24 and 48 h for 25-g samples); 0.5-ml portions of enrichment culture were treated with 4.5 ml of 0.25% KOH-0.5% NaCl for 2 min and 0.5% KOH-0.5% NaCl for 15 s, and 0.1-ml portions of treated culture were plated onto MacConkey or CIN agars or both. The procedure effectively recovered 2 to 12 cells of a number of both mouse-virulent and avirulent strains per g of ground beef with aerobic plate counts of approximately 10(6) to 10(7) CFU/g. Similarly, the procedure isolated both likely virulent and avirulent strains from porcine tongues (aerobic plate counts of 10(5) to 10(7) CFU/g) naturally contaminated with Y. enterocolitica. The organism was isolated from the tongues at similar rates by both shortened enrichment and cold enrichment procedures. Eight tongues were positive for serotype O:5,27 strains that agglutinate with WA-specific absorbed antiserum, an antiserum specific for mouse-virulent Y. enterocolitica (Doyle et al., Infect. Immun. 37:1234-1240, 1982), indicating that the oral cavity of swine is a reservoir of likely virulent serotype O:5,27 strains.


Intervening attenuation affects first-order statistical properties of ultrasound echo signals.
Previous studies show that first-order statistical properties of ultrasound echo signals are related to the effective number of scatterers in the "resolution cell" of a pulse-echo ultrasound system. When the effective number of scatterers is large (~10 or more) this results in echo signals whose amplitude follows a Rayleigh distribution, with the RF echo signal obeying Gaussian statistics; deviation from Rayleigh or Gaussian statistics yields information on scatterer number densities. In this paper, the influence of the medium's attenuation on non-Gaussian properties of the echo signal is considered. Preferential attenuation of higher frequency components of a pulsed ultrasound beam effectively broadens the beam and increases the resolution cell size. Thus, the resultant non-Gaussian parameter for broad bandwidth excitation of the transducer depends not only on the scatterer number density but also on the attenuation in the medium. These effects can be reduced or eliminated by using narrow-band experiments.


Analysis of autoantibodies to recombinant La (SS-B) peptides in systemic lupus erythematosus and primary Sjogren's syndrome.
Autoantibodies to a polymerase III transcription factor, La (SS-B), are frequently detected in the serum of patients with Sjogren's syndrome and systemic lupus erythematosus. To define the humoral immune response to this protein, we analyzed the patterns of antibody recognition toward 13 recombinant La peptides by immunoblotting and determined the heterogeneity of antibodies reactive with the immunodominant epitopes. The smallest epitopes that were strongly antigenic and recognized by greater than 70% of sera tested (immunodominant) were encoded by the subclones BgX and XA located in the 5' and 3' halves of the La cDNA, respectively. Conformation of the immunodominant La peptides played a major role in antibody recognition. Although greater diversity in antibody binding to carboxyl-terminal La peptides was observed, the overall pattern of peptide recognition by anti-La antibodies was similar in different diseases. The antibody responses to the immunodominant peptides were strongly correlated (r = 0.68, P less than 0.001). One- and two-dimensional isoelectric focusing of affinity purified IgG anti-La peptide antibodies revealed restricted heterogeneity and oligoclonal bands (kappa light chains). These observations suggest that anti-La antibodies are induced and/or maintained by the self antigen and that their diversity is constrained either by mechanisms related to tolerance or by affinity maturation of the humoral immune response.


Primary bone lymphoma of radius and tibia: A case report and review of literature.
Primary bone lymphoma (PBL) is a rare malignant entity. There is a better survival of PBL than any other malignant bone tumors and extranodal lymphomas. We report a rare case of PBL involving radius and tibia. The patient was a 14-year-old girl with left forearm pain and swelling after trauma. Six months later after the last chemotherapy and radiotherapy, pain and swelling of left knee was presented. Radiological imaging revealed a lytic destruction, periosteal reaction, and pathological fracture of radius and tibia with soft tissue mass. Surgical biopsy was performed, and the result of histopathological diagnosis was diffused large B-cell lymphoma (stage IV, group A). Chemotherapy combined with radiotherapy was applied before curation. Due to its uncommon presentation, PBL should be taken into consideration if differential diagnosis from other bone tumors is necessary in clinic.


TikTok in Plastic Surgery: A Systematic Review of Its Uses.
TikTok (San Jose, CA) is a popular and rapidly growing social media platform. With beauty and skincare among the top 5 most popular categories, TikTok represents an important platform for plastic surgery education and communication. However, given the vast array of content shared daily, regulating content for veracity is challenging. It may also be an important and potentially overlooked avenue for the dissemination of inaccurate information pertaining to plastic surgery. This systematic review evaluates TikTok's impact on plastic surgery. Following Preferred Reporting Items for Systematic Reviews and Meta-Analysis Guidelines, a systematic literature review was performed of the use of TikTok within the plastic surgery field. The following databases were queried: PubMed (National Institutes of Health; Bethesda, MD), EMBASE (Elsevier; Amsterdam, the Netherlands), and PsychInfo (American Psychological Association; Washington, DC). The search captured 31 studies of which 7 were included in the final analysis. The studies examined the following areas: gender-affirming surgery (n = 1), breast reconstruction (n = 1), aesthetic surgical procedures (n = 1), plastic surgeon profiles (n = 1), and profiles of videos relating to plastic surgery hashtags (n = 3). The videos' quality was assessed using the DISCERN scale. Physician videos scored notably higher than nonphysician videos. The mean DISCERN score across all the videos (n = 386) was 1.91 (range: 1.44-3.00), indicating poor quality. TikTok is a popular medium for sharing plastic surgery content. The existing literature has demonstrated overall poor-quality information on plastic surgery, and further study is needed to evaluate its impact in terms of perceptions of the specialty and healthcare behaviors. Future work should focus on promoting accurate, high-quality videos, potentially including a peer-review function for healthcare content. This can leverage TikTok's potential for disseminating content while upholding patient safety.


Vasopressin and angiotensin II receptors in rat aortic smooth muscle cells in culture.
Rat aortic smooth muscle cells were isolated and maintained in primary culture. After 2-3 days, cells recovered their contractile phenotype and could be induced to contract in response to vasopressin and angiotensin II. Vasopressin- and angiotensin-specific binding sites were detected on these cells, using tritiated Lys8-vasopressin, Asn1-Val5-angiotensin II, and Sarc1-Ile8-angiotensin II. Vasopressin binding sites had Kd values of 30 and 12 nM for Lys8-and Arg8-vasopressin, respectively, and a maximal binding capacity of 25,000 sites/cell. They displayed several of the expected characteristics of vasopressin receptors involved in the vasopressor response in vivo. A highly significant correlation was found between the relative agonistic or antagonistic vasopressor potencies of a series of vasopressin structural analogues and their relative abilities to inhibit [3H]vasopressin binding to aortic smooth muscle cells. Specific binding sites for Asn1-Val5-angiotensin II and Sarc1-Ile8-angiotensin II had the following characteristics: Kd = 2.3 and 1.3 nM, respectively; maximal capacity: 50,000 sites/cell. Vasopressin and angiotensin did not modify the intracellular cyclic AMP content of aortic smooth muscle cells.


Thrombotic risk and immobility in residents of long-term care facilities.
Hospitalized patients and residents of long-term care (LTC) facilities account for about 60% of all cases of venous thromboembolism (VTE), as incidence is correlated with increasing age, immobility, and underlying medical conditions. The primary aim of the study was to develop an evidence-based VTE risk stratification tool and definition of immobility for residents in LTC facilities. Using the Delphi process, a panel of vascular thrombotic and geriatric experts reviewed and ranked statements of VTE risk and immobility derived from randomized controlled trials, meta-analyses, cohort trials, case-control trials, and case series to arrive at consensus for the importance of each statement. Rating was conducted before and during an on-site meeting following discussion. Statements rated high and very high were used to develop a VTE risk stratification and immobility tool. A total of 1165 publications related to VTE risk were identified from which 137 (12%) pertained to subjects with a median age of 60 years or older; 42 (31%) met study criteria. Eight (29.6%) of 79 publications pertaining to immobility met study criteria. There were 4 studies related to VTE risk and 1 to immobility that were rated as high quality. VTE risk factors were age older than 60 years, active cancer, acute infectious disease, catheter in a central vein, chronic obstructive pulmonary disease, dehydration, history of VTE, having a first-degree relative with VTE, heart failure, hypercoagulable state, immobility, inflammatory bowel disease, obesity, rheumatoid arthritis and treatment with erythroid stimulating agents to a hemoglobin value greater than 12 g/dL, aromatase inhibitor, hormone replacement therapy, megestrol acetate, or selective estrogen receptor modulators. Immobility was defined as the presence of at least 1 of the following: being bedridden or bedridden except for bathroom privileges, unable to walk at least 10 feet, recent reduction in ability to walk at least 10 feet for at least 72 hours, and having a lower limb cast. A risk stratification tool for VTE and immobility was developed to assist clinicians in caring for residents of LTC facilities. A prospective trial is needed to validate the tool.


Isotopic Constraints on Sources and Transformations of Nitrate in the Mount Everest Proglacial Water.
Glacier melting exports a large amount of nitrate to downstream aquatic ecosystems. Glacial lakes and glacier-fed rivers in proglacial environments serve as primary recipients and distributors of glacier-derived nitrate (NO3-), yet little is known regarding the sources and cycling of nitrate in these water bodies. To address this knowledge gap, we conducted a comprehensive analysis of nitrate isotopes (δ15NNO3, δ18ONO3, and Δ17ONO3) in waters from the glacial lake and river of the Rongbuk Glacier-fed Basin (RGB) in the mountain Everest region. The concentrations of NO3- were low (0.43 ± 0.10 mg/L), similar to or even lower than those observed in glacial lakes and glacier-fed rivers in other high mountain regions, suggesting minimal anthropogenic influence. The NO3- concentration decreases upon entering the glacial lake due to sedimentation, and it increases gradually from upstream to downstream in the river as a soil source is introduced. The analysis of Δ17ONO3 revealed a substantial contribution of unprocessed atmospheric nitrate, ranging from 34.29 to 56.43%. Denitrification and nitrification processes were found to be insignificant in the proglacial water of RGB. Our study highlights the critical role of glacial lakes in capturing and redistributing glacier-derived NO3- and emphasizes the need for further investigations on NO3- transformation in the fast-changing proglacial environment over the Tibetan Plateau and other high mountain regions.


A multiplexed ion-exchange membrane-based miRNA (MIX·miR) detection platform for rapid diagnosis of myocardial infarction.
Micro RNAs (miRNAs) have shown great potential as rapid and discriminating biomarkers for acute myocardial infarction (AMI) diagnosis. We have developed a multiplexed ion-exchange membrane-based miRNA (MIX·miR) preconcentration/sensing amplification-free platform for quantifying in parallel a panel of miRNAs, including miR-1, miR-208b, and miR-499, from the same plasma samples from: 1) reference subjects with no evident coronary artery disease (NCAD); 2) subjects with stable coronary artery disease (CAD); and 3) subjects experiencing ST-elevation myocardial infarction (STEMI) prior to (STEMI-pre) and following (STEMI-PCI) percutaneous coronary intervention. The picomolar limit of detection from raw plasma and 3-decade dynamic range of MIX·miR permits detection of the miRNA panel in untreated samples from disease patients and its precise standard curve, provided by large 0.1 to 1 V signals and eliminates individual sensor calibration. The use of molecular concentration feature reduces the assay time to less than 30 minutes and increases the detection sensitivity by bringing all targets close to the sensors. miR-1 was low for NCAD patients but more than one order of magnitude above the normal value for all samples from three categories (CAD, STEMI-pre, and STEMI-PCI) of patients with CAD. In fact, miR-1 expression levels of stable CAD, STEMI-pre and STEMI-PCI are each more than 10-fold higher than the previous class, in that order, well above the 95% confidence level of MIX·miR. Its overexpression estimate is significantly higher than the PCR benchmark. This suggests that, in contrast to protein biomarkers of myocardial injury, miR-1 appears to differentiate ischemia from both reperfusion injury and non-AMI CAD patients. The battery-operated MIX·miR can be a portable and low-cost AMI diagnostic device, particularly useful in settings where cardiac catheterization is not readily available to determine the status of coronary reperfusion.


Starch Granule Re-Structuring by Starch Branching Enzyme and Glucan Water Dikinase Modulation Affects Caryopsis Physiology and Metabolism.
Starch is of fundamental importance for plant development and reproduction and its optimized molecular assembly is potentially necessary for correct starch metabolism. Re-structuring of starch granules in-planta can therefore potentially affect plant metabolism. Modulation of granule micro-structure was achieved by decreasing starch branching and increasing starch-bound phosphate content in the barley caryopsis starch by RNAi suppression of all three Starch Branching Enzyme (SBE) isoforms or overexpression of potato Glucan Water Dikinase (GWD). The resulting lines displayed Amylose-Only (AO) and Hyper-Phosphorylated (HP) starch chemotypes, respectively. We studied the influence of these alterations on primary metabolism, grain composition, starch structural features and starch granule morphology over caryopsis development at 10, 20 and 30 days after pollination (DAP) and at grain maturity. While HP showed relatively little effect, AO showed significant reduction in starch accumulation with re-direction to protein and β-glucan (BG) accumulation. Metabolite profiling indicated significantly higher sugar accumulation in AO, with re-partitioning of carbon to accumulate amino acids, and interestingly it also had high levels of some important stress-related metabolites and potentially protective metabolites, possibly to elude deleterious effects. Investigations on starch molecular structure revealed significant increase in starch phosphate and amylose content in HP and AO respectively with obvious differences in starch granule morphology at maturity. The results demonstrate that decreasing the storage starch branching resulted in metabolic adjustments and re-directions, tuning to evade deleterious effects on caryopsis physiology and plant performance while only little effect was evident by increasing starch-bound phosphate as a result of overexpressing GWD.


Complications of Charcot Reconstruction in Patients With Peripheral Arterial Disease.
The primary aim was to determine the rate of complications in patients with peripheral arterial disease and diabetic Charcot neuroarthropathy who underwent osseous reconstruction. Complications included delayed healing, dehiscence, and major lower extremity amputation. A review of patients with Charcot neuroarthropathy requiring reconstruction secondary to ulceration or acute infection was performed. Descriptive analysis compared outcomes between those with and without peripheral arterial disease. Bivariate analysis and multivariate logistic regression were analyzed for delayed healing, dehiscence, and major amputation. In a cohort of 284 patients with diabetic Charcot neuroarthropathy who underwent osseous reconstruction, the rate of peripheral arterial disease was 20.8% (59/284). Bivariate analysis for delayed healing found hypertension (p = .0352), peripheral arterial disease (p = .0051), and smoking history (p = .0276) to be statistically significant factors. Delayed healing was 2.012 times more likely in the presence of peripheral arterial disease [OR 2.012 (95% CI 1.088-3.720)]. Bivariate analysis for major lower extremity amputation found renal disease (0.0003) (renal disease: ESRD and CKD) and peripheral arterial disease (0.0001) to be statistically significant factors. Major amputation was 4.414 times more likely in the presence of peripheral arterial disease [OR 4.414 (95% CI 2.087-9.334)]. Peripheral arterial disease was identified in 20.8% (59/284) of diabetic patients who underwent Charcot osseous reconstruction. Peripheral arterial disease increased the risk of delayed healing by 2.012 fold, and increased the risk of major lower extremity amputation by 4.414 fold. The rates of complications in patients with peripheral arterial disease were significantly higher than those without peripheral arterial disease who underwent osseous reconstruction.


A robust and cost-effective approach to sequence and analyze complete genomes of small RNA viruses.
Next-generation sequencing (NGS) allows ultra-deep sequencing of nucleic acids. The use of sequence-independent amplification of viral nucleic acids without utilization of target-specific primers provides advantages over traditional sequencing methods and allows detection of unsuspected variants and co-infecting agents. However, NGS is not widely used for small RNA viruses because of incorrectly perceived cost estimates and inefficient utilization of freely available bioinformatics tools. In this study, we have utilized NGS-based random sequencing of total RNA combined with barcode multiplexing of libraries to quickly, effectively and simultaneously characterize the genomic sequences of multiple avian paramyxoviruses. Thirty libraries were prepared from diagnostic samples amplified in allantoic fluids and their total RNAs were sequenced in a single flow cell on an Illumina MiSeq instrument. After digital normalization, data were assembled using the MIRA assembler within a customized workflow on the Galaxy platform. Twenty-eight avian paramyxovirus 1 (APMV-1), one APMV-13, four avian influenza and two infectious bronchitis virus complete or nearly complete genome sequences were obtained from the single run. The 29 avian paramyxovirus genomes displayed 99.6% mean coverage based on bases with Phred quality scores of 30 or more. The lower and upper quartiles of sample median depth per position for those 29 samples were 2984 and 6894, respectively, indicating coverage across samples sufficient for deep variant analysis. Sample processing and library preparation took approximately 25-30 h, the sequencing run took 39 h, and processing through the Galaxy workflow took approximately 2-3 h. The cost of all steps, excluding labor, was estimated to be 106 USD per sample. This work describes an efficient multiplexing NGS approach, a detailed analysis workflow, and customized tools for the characterization of the genomes of RNA viruses. The combination of multiplexing NGS technology with the Galaxy workflow platform resulted in a fast, user-friendly, and cost-efficient protocol for the simultaneous characterization of multiple full-length viral genomes. Twenty-nine full-length or near-full-length APMV genomes with a high median depth were successfully sequenced out of 30 samples. The applied de novo assembly approach also allowed identification of mixed viral populations in some of the samples.


Adult cystic nephroma and mixed epithelial and stromal tumor of the kidney: clinical, radiographic, and pathologic characteristics.
Adult cystic nephroma (CN) and mixed epithelial and stromal tumor (MEST) are benign renal tumors readily distinguished from cystic renal cell carcinoma and other malignant variants based on histopathology. Clinical and radiographic data regarding these lesions are sparse, especially with respect to factors providing clinical suspicion for CN or MEST. Pathology of 22 CN (21 patients) and 10 MEST (9 patients) treated between 1987 and 2005 was re-reviewed according to 2004 WHO classification. Nineteen CN patients (90%) and 9 MEST patients (100%) were female and median ages were 55 (range: 39 to 79) and 52 (range: 39 to 67) years, respectively. Twenty-two patients (73%) presented symptomatically. CN were commonly Bosniak III lesions (77%), whereas 70% of MEST had solid enhancing components. An intrapelvic component was present in 5 CN (23%) and no MEST. Preoperative radiologic suspicion was documented in 36% of CN, but in only 1 patient with MEST. Nephron-sparing surgery was performed for 16 tumors (50%), including 5 suspected preoperatively and 3 that were 7 cm or greater. There was 1 case each of bilateral CN and MEST. Two patients with CN had concomitant renal cell carcinoma. One patient with MEST had sarcomatous component. At mean follow-up of 4.5 years (range: 1 to 18), no patient had novel or recurrent tumors develop. CN and MEST typically present symptomatically in perimenopausal women. Pedunculation of a multiloculated cystic lesion may allow for preoperative suspicion of these lesions in the proper clinical setting. Nephron-sparing surgery is the preferred treatment even for large and/or central lesions, if amenable.


Increased activation of GATA-3, IL-2 and IL-5 of cord blood mononuclear cells in infants with IgE sensitization.
Risk of allergic diseases has been linked to abnormal patterns of fetal immune development, suggesting that priming of the immune system may occur in utero. The aim of the study was to investigate whether the pattern of immune response in cord blood mononuclear cells (CBMC) shows association with allergic diseases and IgE sensitization at 2 yr of age, and to study the effect of maternal probiotic supplementation on CBMC immune responses. CBMC were isolated from 98 neonates in a randomized double-blinded intervention study. CBMC were stimulated with beta-lactoglobulin, and phytohemaglutinin (PHA). Secretion of interferon-gamma (IFN-gamma), interleukin-5 (IL-5), and IL-13 was measured by an ELISA; IL-2, IL-4, and IL-10 by a cytokine bead assay. T-cell polarization-associated IL-4 receptor and IL-12R expressions, and the respective transcription factors GATA-3 and T-bet were analyzed with RT-PCR. The above responses were compared with the development of allergic diseases and IgE sensitization at 2 yr of age, and with the maternal probiotic or placebo supplementation. PHA-stimulated GATA-3 expression and IL-2 secretion in CBMC were higher in IgE-sensitized children at an age of 2 yr than in the non-sensitized, non-allergic children (p = 0.03 and 0.026). PHA-induced expression of GATA-3 correlated with IL-5 (p = 0.003, r = 0.300) and IL-13 (p = 0.007, r = 0.278) secretion of CBMC, and IL-5 secretion of beta-lactoglobulin-stimulated CBMC was higher in IgE-sensitized children at 2 yr of age than in the non-sensitized, non-allergic children (p = 0.013). Probiotic bacteria had no effect on CBMC immune responses. In CBMC-enhanced induction of GATA-3, which activates several Th2 cytokines genes, was a risk factor for IgE sensitization. The immune deviation towards Th2-type immunity developed already in utero and seemed to modulate the pattern of immune response favoring an IgE response to environmental antigens.


Mononuclear and dinuclear manganese compounds stabilized by supramolecular interactions.
New manganese compounds [Mn(HphpzMe)(2)(H(2)phpzMe)(HCO(2))] (1), [Mn(2)(phpzMe)(2)(HphpzMe)(2)(OCH(3))]·2CH(3)OH (2), Na{[Mn(HphpzPh)(phpzPh)(MeOH)(2)](2)}(HCO(2)) (3), [Mn(HphpzPh)(2)(EtOH)(2)]ClO(4)·2EtOH (4) and [Mn(HphpzPh)(2)N(3)] (5) were synthesized and characterized with various techniques. 1, 4 and 5 are mononuclear manganese(III) compounds, 2 is a mixed-valence dinuclear manganese(III/IV) compound, and 3 is a trinuclear compound containing two manganese(III) ions and a sodium(I) ion. A remarkable feature is the spontaneous formation of the formate ion as a result of the methanol or methoxide oxidation in compounds 1 and 3. Using ethanol precludes the formation of the formate and compound 4 is obtained. The molecular structure of all compounds is stabilized by supramolecular interactions, including strong hydrogen bonding and π-π interactions.


Interdisciplinary Health Care Evaluation Instruments: A Review of Psychometric Evidence.
Teamwork among health care professionals has been found to improve patient outcomes and reduce burnout. Surveys from individual team members are often used to measure the effectiveness of teamwork performance, as they provide an efficient way to capture various constructs of teamwork. This allows evaluators to better understand team functioning, areas of strength, and to identify potential areas for improvement. However, the majority of published surveys are yet to be validated. We conducted a review of psychometric evidence to identify instruments frequently used in practice and identified in the literature. The databases searched included MEDLINE, EMBASE, CINAHL, and PsycINFO. After excluding duplicates and irrelevant articles, 15 articles met the inclusion criteria for full assessment. Seven surveys were validated and most frequently identified in the literature. This review aims to facilitate the selection of instruments that are most appropriate for research and clinical practice. More research is required to develop surveys that better reflect the current reality of teamwork in our evolving health system, including a greater consideration for patient as team members. Additionally, more research is needed to encompass an increasing development of team assessment tools.


Integrating CD4+ T cell help for therapeutic cancer vaccination in a preclinical head and neck cancer model.
Head and neck squamous cell carcinomas (HNSCC) are well suited for cancer vaccination strategies. In addition to tumor-associated antigens (TAAs) and endogenous retrovirus (ERV) encoded proteins, HNSCCs have a relatively high tumor mutational burden encoding potential neoepitopes. Peptide vaccine candidates are prioritized by predicted high-affinity to major histocompatibility complex (MHC) class I with MHC-II affinity largely not being considered. Herein, we extend previous studies to evaluate therapeutic vaccination in the mouse oral cancer (MOC) 22 model. Two distinct MOC22 derived SLPs were tested - a TSA-oriented mutant intercellular adhesion molecule 1 (mICAM1) and p15E, an ERV encoded antigen. In silico prediction revealed mICAM1 SLP bore strong MHC-I and MHC-II epitopes sharing a mutant residue with vaccination significantly increasing both antigen-specific IFN-γ producing CD4+ and CD8+ T cells. By contrast, p15E SLP had a predicted high-affinity MHC-I epitope but lacked an MHC-II epitope and vaccination induced antigen-specific CD8+ but not CD4+ T cell responses. Therapeutic mICAM1 vaccination attenuated tumor growth effectively with mICAM1-specific T cells displaying durable IFN-γ production compared with p15E SLP. Furthermore, mICAM1 SLPs carrying weakened MHC-II binding epitopes were unable to control tumor growth. These data underscore the potential value of therapeutic targeting of HNSCC epitopes and highlight the importance of studying distinct antigen classes in this setting. Moreover, they raise the possibility that, at least in part, CD4+ T cell help is critical for cancer vaccination in this preclinical HNSCC model and suggest in silico prediction approaches prioritize overlapping MHC-I and MHC-II epitopes to generate potent cancer vaccines.


Work response of mildly mentally retarded adults to self- versus external regulation as a function of motivational orientation.
The Picture Motivation Scale was administered to 144 mildly mentally retarded adults from a community-based intermediate care facility. Those in the top and bottom quartiles on intrinsic motivation were classified as either relatively task intrinsically motivated or task extrinsically motivated and were assigned to behavior regulation conditions: self-regulated reinforcement, externally imposed reinforcement, or control. All subjects were given a work task consisting of placing washers into successive compartments of a container, with subjects in the external-reinforcement condition yoked to those in the self-regulation condition with respect to work goal and number of tokens received for their work. Across conditions intrinsically motivated subjects worked harder than did extrinsically motivated subjects; all of them worked harder under conditions of regulation of reinforcement matched to their motivational orientation (i.e., intrinsically motivated subjects under self-regulation, extrinsically motivated subjects under externally imposed reinforcement) than under the contrary condition. When not rewarded, intrinsically motivated subjects showed more sustained performance than did extrinsically motivated subjects. The importance of matching self-regulation expectations to individual differences in motivational orientation of retarded persons being prepared for relatively independent living was discussed.


Structure and enzymatic mechanism of a moonlighting dUTPase.
Genome integrity requires well controlled cellular pools of nucleotides. dUTPases are responsible for regulating cellular dUTP levels and providing dUMP for dTTP biosynthesis. In Staphylococcus, phage dUTPases are also suggested to be involved in a moonlighting function regulating the expression of pathogenicity-island genes. Staphylococcal phage trimeric dUTPase sequences include a specific insertion that is not found in other organisms. Here, a 2.1 Å resolution three-dimensional structure of a ϕ11 phage dUTPase trimer with complete localization of the phage-specific insert, which folds into a small β-pleated mini-domain reaching out from the dUTPase core surface, is presented. The insert mini-domains jointly coordinate a single Mg2+ ion per trimer at the entrance to the threefold inner channel. Structural results provide an explanation for the role of Asp95, which is suggested to have functional significance in the moonlighting activity, as the metal-ion-coordinating moiety potentially involved in correct positioning of the insert. Enzyme-kinetics studies of wild-type and mutant constructs show that the insert has no major role in dUTP binding or cleavage and provide a description of the elementary steps (fast binding of substrate and release of products). In conclusion, the structural and kinetic data allow insights into both the phage-specific characteristics and the generally conserved traits of ϕ11 phage dUTPase.


Network analysis of patient flow in two UK acute care hospitals identifies key sub-networks for A&E performance.
The topology of the patient flow network in a hospital is complex, comprising hundreds of overlapping patient journeys, and is a determinant of operational efficiency. To understand the network architecture of patient flow, we performed a data-driven network analysis of patient flow through two acute hospital sites of King's College Hospital NHS Foundation Trust. Administration databases were queried for all intra-hospital patient transfers in an 18-month period and modelled as a dynamic weighted directed graph. A 'core' subnetwork containing only 13-17% of all edges channelled 83-90% of the patient flow, while an 'ephemeral' network constituted the remainder. Unsupervised cluster analysis and differential network analysis identified sub-networks where traffic is most associated with A&E performance. Increased flow to clinical decision units was associated with the best A&E performance in both sites. The component analysis also detected a weekend effect on patient transfers which was not associated with performance. We have performed the first data-driven hypothesis-free analysis of patient flow which can enhance understanding of whole healthcare systems. Such analysis can drive transformation in healthcare as it has in industries such as manufacturing.


X-ray fluorescence microscopy: A method of measuring ion concentrations in the ear.
This technical note describes synchrotron x-ray fluorescence microscopy (XFM) as a method for measuring the concentrations of different elements in cross-sections of the ear at extremely high resolution. This method could be of great importance for addressing many open questions in hearing research. XFM uses synchrotron radiation to evoke emissions from many biologically relevant elements in the tissue. The intensity and wavelength of the emitted radiation provide a fingerprint of the tissue composition that can be used to measure the concentration of the elements in the sampled location. Here, we focus on energies that target biologically-relevant elements of the periodic table between magnesium and zinc. Since a highly focused x-ray beam is used, the spot size is well below 1 μm and the samples can be scanned at a nanometer lateral resolution. This study shows that measurement of the concentrations of different elements is possible in a mid-modiolar cross-section of a mouse cochlea. Images are presented that indicate potassium and chloride "hot spots" in the spiral ligament and the spiral limbus, providing experimental evidence for the potassium recycling pathway and showing the cochlear structures involved. Scans of a section obtained from the incus, one of the middle ear ossicles, in a developing mouse have shown that zinc is not uniformly distributed This supports the hypothesis that zinc plays a special role in the process of ossification. Although limited by sophisticated sample preparation and sectioning, the method provides ample exciting opportunities, to understand the role of genetics and epigenetics on hearing mechanisms in ontogeny and phylogeny.


The action of the C-terminal octapeptide of cholecystokinin and related peptides on pancreatic exocrine secretion.
The effect of the C-terminal octapeptide of cholecystokinin-pancreozymin (CCK-PZ), caerulein, and the C-terminal tetrapeptide of gastrin on pancreatic secretion of fluid, electrolyte, amylase, and protein was studied in anaesthetized dogs prepared with pancreatic fistulae. Against background stimulation of fluid secretion with submaximal doses of secretin, all the polypeptides produced a qualitatively similar pancreatic response, causing a highly significant increase in amylase, protein, calcium, and zinc concentrations. Magnesium concentration was significantly increased only when the concentration preceding the administration of the peptide was below 100 mu-equiv/l. Octa-CCK-PZ was 13-35 times and 20-56 times more potent than tetragastrin on weight and molar bases, respectively, as a stimulant of amylase secretion. The threshold doses were largest for amylase, lower for calcium, and lowest for zinc. A significant linear correlation was observed between amylase and calcium concentration, zinc and protein concentration, and magnesium and calcium concentration. The peptides produced some increase in secretin-induced volume flow, whereas bicarbonate, chloride, sodium, and potassium concentrations remained unchanged. The direct relation between bicarbonate concentration and flow rate was limited to rates below 1.5 ml/5 minutes. At higher rates bicarbonate and chloride concentration reached a high and low plateau, respectively, although the first five-min sample of pancreatic juice after secretin stimulation exhibited a relatively low bicarbonate and high chloride concentration compared with its voluminous flow. Chloride concentration varied inversely with bicarbonate concentration, the sum of the two anions being constant.


Cellular interactions: lessons from the nitrogen-fixing cyanobacteria.
Marine nitrogen-fixing cyanobacteria play a central role in the open-ocean microbial community by providing fixed nitrogen (N) to the ocean from atmospheric dinitrogen (N2 ) gas. Once thought to be dominated by one genus of cyanobacteria, Trichodesmium, it is now clear that marine N2 -fixing cyanobacteria in the open ocean are more diverse, include several previously unknown symbionts, and are geographically more widespread than expected. The next challenge is to understand the ecological implications of this genetic and phenotypic diversity for global oceanic N cycling. One intriguing aspect of the cyanobacterial N2 fixers ecology is the range of cellular interactions they engage in, either with cells of their own species or with photosynthetic protists. From organelle-like integration with the host cell to a free-living existence, N2 -fixing cyanobacteria represent the range of types of interactions that occur among microbes in the open ocean. Here, we review what is known about the cellular interactions carried out by marine N2 -fixing cyanobacteria and where future work can help. Discoveries related to the functional roles of these specialized cells in food webs and the microbial community will improve how we interpret their distribution and abundance patterns and contributions to global N and carbon (C) cycles.


Cycloaddition of cyclohexa-2,4-dienones, ring-closing metathesis, and photochemical reactions: a common stereoselective approach to duprezianane, polyquinane and sterpurane frameworks.
A novel approach to three different types of carbocyclic frameworks belonging to dupreziananes, sterpuranes, and polyquinanes from simple aromatic precursors has been presented. Cycloaddition of appropriately appended cyclohexa-2,4-dienones with acyclic dienes gave bridged bicyclic octanes suitably disposed with olefinic chains, which upon ring-closing metathesis led to functionalized tricyclo[22.214.171.124(1,5)]undecanes related to dupreziananes. Photochemical sigmatropic 1,2- and 1,3-acyl shifts in tricyclo[188.8.131.52(1,5)] undecanes upon triplet and singlet excitation provided stereoselective routes to sterpurane and polyquinane frameworks.


Microglial stability and repopulation in the retina.
Parenchymal central nervous system microglia are repopulated by bone marrow derived monocytes more slowly than any other reticuloendothelial cells. The contribution of bone marrow derived monocytes to the uninflammed retina has not been studied. The present study sought to determine repopulation of retinal microglia in uniflammed retina by bone marrow derived monocytes in bone marrow chimeric rats. Chimeric (Y-->X) Lewis rats were constructed by transplanting 5 x 10(7) male bone marrow cells into lethally irradiated female recipient rats. The chimeras were sacrificed 8, 10, 12, 30, and 52 weeks after bone marrow transplant, and retina, brain, lung, and spleen samples were collected. DNA was extracted and quantified. Y positive infiltrating cells in the collected samples were detected by polymerase chain reaction amplification of a Y chromosome specific 104 bp fragment. There was a rapid repopulation of haematopoietic tissues in the spleen (at 8 weeks), confirming the establishment of chimerism, and to a lesser extent, of lung (at 30 weeks). This repopulation was absent in the brain parenchyma and retina until 52 weeks after transplantation. These data indicate that resident microglia in the retina, much like those in the brain, are stable in number in the retinal compartment (up to 1 year), and repopulation by bone marrow derived cells may be delayed for a year.


Neural crest-specific deletion of Ldb1 leads to cleft secondary palate with impaired palatal shelf elevation.
LIM domain binding protein 1 (LDB1) is a transcriptional co-factor, which interacts with multiple transcription factors and other proteins containing LIM domains. Complete inactivation of Ldb1 in mice resulted in early embryonic lethality with severe patterning defects during gastrulation. Tissue-specific deletions using a conditional knockout allele revealed additional roles of Ldb1 in the development of the central nervous system, hematopoietic system, and limbs. The goal of the current study was to determine the importance of Ldb1 function during craniofacial development in mouse embryos. We generated tissue-specific Ldb1 mutants using Wnt1-Cre, which causes deletion of a floxed allele in the neural crest; neural crest-derived cells contribute to most of the mesenchyme of the developing face. All examined Wnt1-Cre;Ldb1(fl/-) mutants suffered from cleft secondary palate. Therefore, we performed a series of experiments to investigate how Ldb1 regulated palate development. First, we examined the expression of Ldb1 during normal development, and found that Ldb1 was expressed broadly in the palatal mesenchyme during early stages of palate development. Second, we compared the morphology of the developing palate in control and Ldb1 mutant embryos using sections. We found that the mutant palatal shelves had abnormally blunt appearance, and failed to elevate above the tongue at the posterior domain. An in vitro head culture experiment indicated that the elevation defect was not due to interference by the tongue. Finally, in the Ldb1 mutant palatal shelves, cell proliferation was abnormal in the anterior, and the expression of Wnt5a, Pax9 and Osr2, which regulate palatal shelf elevation, was also altered. The function of Ldb1 in the neural crest-derived palatal mesenchyme is essential for normal morphogenesis of the secondary palate.


[State of binocular vision and comparative assessment of methods of its study in motor vehicle drivers].
Ophthalmologic study covered 100 truck drivers (categories C and E according to International Classification) aged 21 to 50 and with length of service from 2 to 25 years. Comparative evaluation of various methods studying deep and stereoscopic vision proved that professional drivers have high values of deep and stereoscopic vision. Those values appeared to correlate strongly and positively (r = 0.57). Making a decision on adequacy to driving the stated vehicles, experts should prefer simple and informative method studying stereoscopic vision through lens stereoscope with "three circles" table.


Early gas stripping as the origin of the darkest galaxies in the Universe.
The known galaxies most dominated by dark matter (Draco, Ursa Minor and Andromeda IX) are satellites of the Milky Way and the Andromeda galaxies. They are members of a class of faint galaxies, devoid of gas, known as dwarf spheroidals, and have by far the highest ratio of dark to luminous matter. None of the models proposed to unravel their origin can simultaneously explain their exceptional dark matter content and their proximity to a much larger galaxy. Here we report simulations showing that the progenitors of these galaxies were probably gas-dominated dwarf galaxies that became satellites of a larger galaxy earlier than the other dwarf spheroidals. We find that a combination of tidal shocks and ram pressure swept away the entire gas content of such progenitors about ten billion years ago because heating by the cosmic ultraviolet background kept the gas loosely bound: a tiny stellar component embedded in a relatively massive dark halo survived until today. All luminous galaxies should be surrounded by a few extremely dark-matter-dominated dwarf spheroidal satellites, and these should have the shortest orbital periods among dwarf spheroidals because they were accreted early.


[On the educational benefits of biological sciences in space].
Experimental results in Space Sciences are getting accumulated. It may be advisable for the educational activities of our society that we inform young people, not only of each experimental result, but also of a variety of novel views of life and the world introduced by the researches in Biological Sciences in Space. We are going to provide a data-base including elementary words and concepts in these fields.


How has alcohol advertising in traditional and online media in Australia changed? Trends in advertising expenditure 1997-2011.
The aim of this study was to determine changes in advertising expenditures across eight media channels for the four main alcohol beverage types and alcohol retailers in Australia. Yearly advertising expenditures between January 1997 and December 2011 obtained from a leading media-monitoring company. Media channels assessed were: free-to-air television, newspapers, magazines, radio, outdoors (billboards), cinema, direct mail (from 2005) and online (from 2008). Data were categorised into alcohol retailers (e.g. supermarkets, off-licences) or four alcoholic beverage types (beer, wine, spirits, premixed spirits/cider). Regression analyses examined associations between year and expenditure. Total alcohol advertising expenditure peaked in 2007, then declined to 2011 (P = 0.02). Television advertising expenditure declined between 2000 and 2011 (P < 0.001), while outdoor advertising expenditure increased between 1997 and 2007. Alcohol retailers' advertising expenditure increased over time (P < 0.001), and from the mid-2000s exceeded expenditure for any single beverage category. For both beer and spirits, television advertising expenditure declined over time (beer: P < 0.001; spirits: P < 0.001) while outdoor advertising expenditure increased (beer: P < 0.001; spirits: P = 0.02). However, the number of advertised beer (P < 0.001), spirits (P < 0.001) and wine (P = 0.01) products increased over time. Retailers are playing an increasing role in advertising alcohol. As our study excluded non-traditional advertising media (e.g. sponsorships, in-store) we cannot determine whether declines in television advertising have been offset by increases in advertising in newer media channels. However, our findings that media channels used for alcohol advertising have changed over time highlights the need for adequate controls on alcohol advertising in all media channels. [White V, Faulkner A, Coomber K, Azar D, Room R, Livingston M, Chikritzhs T, Wakefield M. How has alcohol advertising in traditional and online media in Australia changed? Trends in advertising expenditure 1997-2011. Drug Alcohol Rev 2015]2015;34:521-530.


Sequestration of CO2 by Phosphatrane Molecules.
The stationary points for the reaction between the CO2 and nine different phosphatranes molecules have been characterized by means of MP2 computational methods. Two minima structures have been located: a pnicogen bonded complex where one of the oxygen atoms of CO2 acts as electron donor and an adduct that presents a covalent P-C linkage. The corresponding transition state structure linking the two minima has also been characterized. In gas phase, the pnicogen bonded complex is more stable than the corresponding adduct except in one case. In contrast, the inclusion of the solvent effect (toluene and THF), reverts the stability, being in all cases the different adducts more stable than the pnicogen bonded complexes. The electronic properties of the systems have been analysed with the Quantum Theory of Atoms in Molecules (QTAIM) and Electron Density Shift (EDS) methods.


Experimental Mixed-Gas Permeability, Sorption and Diffusion of CO₂-CH₄ Mixtures in 6FDA-mPDA Polyimide Membrane: Unveiling the Effect of Competitive Sorption on Permeability Selectivity.
The nonideal behavior of polymeric membranes during separation of gas mixtures can be quantified via the solution-diffusion theory from experimental mixed-gas solubility and permeability coefficients. In this study, CO₂-CH₄ mixtures were sorbed at 35 °C in 4,4'-(hexafluoroisopropylidene)diphthalic dianhydride (6FDA)-m-phenylenediamine (mPDA)-a polyimide of remarkable performance. The existence of a linear trend for all data of mixed-gas CO₂ versus CH₄ solubility coefficients-regardless of mixture concentration-was observed for 6FDA-mPDA and other polymeric films; the slope of this trend was identified as the ratio of gas solubilities at infinite dilution. The CO₂/CH₄ mixed-gas solubility selectivity of 6FDA-mPDA and previously reported polymers was higher than the equimolar pure-gas value and increased with pressure from the infinite dilution value. The analysis of CO₂-CH₄ mixed-gas concentration-averaged effective diffusion coefficients of equimolar feeds showed that CO₂ diffusivity was not affected by CH₄. Our data indicate that the decrease of CO₂/CH₄ mixed-gas diffusion, and permeability selectivity from the pure-gas values, resulted from an increase in the methane diffusion coefficient in mixtures. This effect was the result of an alteration of the size sieving properties of 6FDA-mPDA as a consequence of CO₂ presence in the 6FDA-mPDA film matrix.


Driver distraction and its effects on partially automated driving performance: A driving simulator study among young-experienced drivers.
Drivers of partially automated vehicles (PAVs) are relieved from parts of the driving tasks allocated to the automated driver. Ironically, these drivers are obligated to continuously monitor the driving task at all times and keep their attention on the roadway. This reduction in the driving task's demands and cognitive workload may encourage drivers to engage with non-driving related tasks (NDRT), which may impair drivers' awareness of the road environment and, as a result, compromise safety. This study examined how engagement with a visual-manual NDRT affects the driving performance of PAV drivers. Thirty-seven participants were randomly assigned to one of two experimental conditions in a driving simulator. Each consisted of two simulated drives. The first experimental condition included one drive under manual driving conditions and another under partially automated driving conditions (i.e., L2). Both drives had no NDRT involved. The second experimental condition included one drive under L2 without an NDRT and one drive under L2, including engagement with an NDRT. Participants' eye movements and heart rate were recorded throughout the experiment. Across various measures, the findings showed that under L2 driving conditions, engagement with an NDRT impairs driving performance in two primary aspects: (1) drivers were less aware of road hazards, and (2) their mental workload was higher when they engaged with an NDRT. In addition, the findings reveal that for drivers engaged with an NDRT, the attentional time-sharing strategy between the NDRT and the roadway monitoring task affected the probability of identifying a hazard. This study shows the adverse effects of engagement with an NDRT under L2 driving conditions on driving performance. Future studies should examine different interventions to mitigate these effects, assuring that drivers are constantly aware of the roadway environment.


In vitro biofilm activity of non-Candida albicans Candida species.
Candidosis has been attributed to C. albicans; however, infections caused by non-Candida albicans Candida (NCAC) species are increasingly being recognised. The ability of Candida to grow as a biofilm is an important feature that promotes both infection and persistence in the host. The biofilms' activity is significant since high activity might be associated with enhanced expression of putative virulence factors, whilst in contrast low activity has previously been suggested as a mechanism for resistance of biofilm cells to antimicrobials. The aim of this study was to determine the metabolic activity of in vitro biofilms formed by different clinical isolates of NCAC species. The in situ total metabolic activity of C. parapsilosis, C. tropicalis and C. glabrata biofilms was determined using 2,3-(2-methoxy-4-nitro-5-sulphophenyl)-5-[(phenylamino) carbonyl]-2H-tetrazolium hydroxide (XTT) reduction assay, and the number of cultivable cells was also established by CFU (colony forming unit) counts. The biofilm structure was assessed by scanning electron microscopy (SEM). Results showed that total biofilm metabolic activity was species and strain dependent. C. glabrata exhibited the lowest biofilm metabolic activity despite having the highest number of biofilm cultivable cells. Similarly, the metabolic activity of resuspended C. glabrata biofilm and planktonic cells was lower than that of the other species. This study demonstrates the existence of intrinsic activity differences amongst NCAC species, which could have important implications in terms of species relative virulence. Furthermore, the absence of an obvious correlation, between cultivable cells number and total biofilm activity, raises the question about which parameter is the most appropriate for the in vitro assessment of biofilms and their potential clinical significance.


HIV-1 protease inhibitor mutations affect the development of HIV-1 resistance to the maturation inhibitor bevirimat.
Maturation inhibitors are an experimental class of antiretrovirals that inhibit Human Immunodeficiency Virus (HIV) particle maturation, the structural rearrangement required to form infectious virus particles. This rearrangement is triggered by the ordered cleavage of the precursor Gag polyproteins into their functional counterparts by the viral enzyme protease. In contrast to protease inhibitors, maturation inhibitors impede particle maturation by targeting the substrate of protease (Gag) instead of the protease enzyme itself. Direct cross-resistance between protease and maturation inhibitors may seem unlikely, but the co-evolution of protease and its substrate, Gag, during protease inhibitor therapy, could potentially affect future maturation inhibitor therapy. Previous studies showed that there might also be an effect of protease inhibitor resistance mutations on the development of maturation inhibitor resistance, but the exact mechanism remains unclear. We used wild-type and protease inhibitor resistant viruses to determine the impact of protease inhibitor resistance mutations on the development of maturation inhibitor resistance. Our resistance selection studies demonstrated that the resistance profiles for the maturation inhibitor bevirimat are more diverse for viruses with a mutated protease compared to viruses with a wild-type protease. Viral replication did not appear to be a major factor during emergence of bevirimat resistance. In all in vitro selections, one of four mutations was selected: Gag V362I, A364V, S368N or V370A. The impact of these mutations on maturation inhibitor resistance and viral replication was analyzed in different protease backgrounds. The data suggest that the protease background affects development of HIV-1 resistance to bevirimat and the replication profiles of bevirimat-selected HIV-1. The protease-dependent bevirimat resistance and replication levels can be explained by differences in CA/p2 cleavage processing by the different proteases. These findings highlight the complicated interactions between the viral protease and its substrate. By providing a better understanding of these interactions, we aim to help guide the development of second generation maturation inhibitors.


Cell-free DNA extraction from urine of lung cancer patients and healthy individuals: Evaluation of a simple method using sample volume up-scaling.
Urine holds promise as a source for cell-free DNA (cfDNA) analysis of cancer genetics due to its nature as a self-collectable biospecimen available in large quantities. However, pre-analytical variables such as preservation of cfDNA or efficiency of up-scaling specimen volume need to be better explored to increase analysis sensitivity. Initially effects of pH levels on cfDNA stability of urine preserved with EDTA were investigated in three healthy probands. Furthermore, the efficiency of urine volume up-scaling was examined using a simple DNA extraction method and cfDNA in urine of 32 individuals. Quantification was performed by PCR detection of three relevant targets used for EGFR and KRAS gene mutational analysis. Only samples preserved with EDTA at alkaline pH levels showed cfDNA stability of up to 10 days at room temperature. Moreover, it was found that increasing the volume up to 100 mL was highly efficient. A similar amount of copies was detected in three different gene sites in all specimens indicating both a good availability and a non-random distribution pattern across genes. Since the median cfDNA copy number was 1642 copies/mL, abundance of cfDNA in this type of liquid biopsy is low. Predictable sensitivities with different urine volumes on the ground of detectable cfDNA in our study population revealed that up-scaling (>5 mL) is mandatory if the mutation allele frequency is less than 1%.


The Ability of the Eating Assessment Tool-10 to Detect Aspiration in Patients With Neurological Disorders.
Dysphagia is common in patients with neurological disorders. There is a need to identify patients at risk early by a useful clinical tool to prevent its serious complications. The study aims to determine the ability of the Turkish version of Eating Assessment Tool-10 (T-EAT-10) to detect aspiration in patients with neurological disorders. Two hundred fifty-nine patients with neurological disorders who had complaints about swallowing difficulty and referred for a swallowing evaluation were included. Oropharyngeal dysphagia was evaluated with the T-EAT-10 and videofluoroscopic swallowing study in the same day. The penetration-aspiration scale (PAS) was used to document the penetration and aspiration severity. The mean age of the patients was 59.72 ± 17.24 years (minimum [min] = 18, maximum [max] = 96), of which 57.1% were male. The mean T-EAT-10 of patients who had aspiration (PAS > 5) was 25.91 ± 10.31 (min = 1, max = 40) and the mean T-EAT-10 of patients who did not have aspiration (PAS < 6) was 15.70 ± 10.54 (min = 0, max = 40) (P < 0.001). Patients with a T-EAT-10 score higher than 15 were 2.4 times more likely to aspirate. A linear correlation was found between T-EAT-10 and PAS scores of the patients (r = 0.416, P < 0.001). The sensitivity of a T-EAT-10 higher than 15 in detecting aspiration was 81.0% and the specificity was 58.0%. A T-EAT-10 score of higher than 15 has a positive predictive value of 72.0% and a negative predictive value of 69.0%. The T-EAT-10 can be used to detect unsafe airway protection in neurology clinics to identify and refer dysphagic patients for further evaluation.


The whey proteins of the milk of red deer (Cervus elaphus L.). A homologue of bovine beta-lactoglobulin.
1. The whey proteins from the milk of red deer are compared with those of cattle. Gel chromatography and electrophoresis showed a close similarity between the whey proteins of the two species in the size, mobility and relative amounts of the main constituents and in the changes in their relative amounts with time after parturition. 2. The major constituent of the deer whey was isolated. It appeared to be homologous with bovine beta-lactoglobulin and had the following properties: m=-5.2X10(-9)m2-s-1-V-1 at 4 degrees C and pH 8.6; pI=5.17; S020, w =2.89S; v=0.748 ml/g; E1g/dl 1cm= 9.12 at 278 nm; deltan/c=1.794 X 10(-3)dl/g at 579 nm (all at 20 degrees C except m). Its molecular weight was that of a dimer with a subunit weight of 18 000. 3. Amino acid analyses of this protein, adjusted to lysine = 15 residues showed that it contains one more residue of aspartic acid, alanine and methionine and one less glutamic acid residue and two less leucine residues than bovine beta-lactoglobulin A. 4. On starch-gel electrophoresis at pH 8.2, this protein migrated at the same rate as bovine beta-lactoglobulin B, although its isoelectric point is close to that of the bovine A variant. Milk from three out of 27 hinds examined showed a variant. This migrated in starch gel at the same rate as the bovine A variant but had a more acid pI = 5.02. 5. The two species whose milk whey proteins are compared represent two different families of ruminants. The similarities found support the view that the milk whey proteins of the bovids are probably typical of the suborder as a whole.


Anastomosis in minimally invasive Ivor Lewis esophagectomy via two ports provides equivalent perioperative outcomes to open.
Minimally invasive esophagectomy (MIE) is becoming a selective treatment of esophageal cancer; however, it's a complex and technically demanding surgical operation. MIE can be performed in high volume centers in a variety of ways using different techniques. Transthoracic staplers have traditionally been used in open transthoracic Ivor Lewis Esophagectomy (ILE) with good success. An investigation of the safety and utility of transthoracic stapler via two ports on thorax for esophageal anastomosis in minimally invasive ILE is reviewed. Patients of esophageal cancer were selected between November 2012 and July 2014. All the patients received minimally invasive (MIE) or open transthoracic ILE. Transthoracic stapler for MIE anastomosis was performed through the major port located at subaxillary region. Patients' demographics, indications for esophagectomy, perioperative treatments, intraoperative data, postoperative complications, hospital length of stay, 7 and in-hospital mortality were evaluated. Totally, 63 consecutive patients underwent MIE or ILE. All the patients were Han with a mean age of 60 years (52-74). The indication of surgery is esophageal cancer, and squamous cell carcinoma was defined by pathologist before operation. None of the patients had neoadjuvant chemotherapy or radiation. All the MIE patients were no conversions to open thoracotomy or laparotomy. Mean operative time was 4.5 h. One patient (3.03%) suffered postoperative pneumonia, no leak from the gastric conduit staple line or esophageal anastomoses, no postoperative complication required surgical intervention was observed. The median hospital length of stay was 13 days (range 7-18). There were no in-hospital mortalities. In our study, transthoracic stapler through the major port at subaxillary seems technically feasible and safe for minimally invasive ILE with comparable morbidity and oncologic data to open.


Evaluation of cobalt-labeled octreotide analogs for molecular imaging and auger electron-based radionuclide therapy.
The somatostatin receptor, which is overexpressed by many neuroendocrine tumors, is a well-known target for molecular imaging and peptide receptor radionuclide therapy. Recently, (57)Co-labeled DOTATOC, an octreotide analog, was shown to have the highest affinity yet found for somatostatin receptor subtype 2. The aim of this study was to evaluate the biologic effects of novel cobalt-labeled octreotide analogs targeting the somatostatin receptor to identify promising candidates for molecular imaging and Auger electron-based radionuclide therapy. Cobalt-labeled DOTATATE, DOTATOC, and DOTANOC were prepared with (57)Co or (58m)Co for SPECT or Auger electron-based therapy, respectively. The cellular uptake and intracellular distribution of the radioligands were characterized with the pancreatic tumor cell line AR42J in vitro, including assessment of the therapeutic effects of (58m)Co-DOTATATE via DNA double-strand break and proliferation assays. Comparisons with the therapeutic effects of (111)In- and (177)Lu-DOTATATE were also performed. Tumor uptake and normal tissue uptake were characterized in a subcutaneous pancreatic tumor mouse model. All 3 cobalt-conjugated peptides resulted in time-dependent and receptor-specific uptake, with a high level (≥88%) of cellular internalization in vitro of the total cell-associated radioactivity. The DNA double-strand break yield showed a dose-dependent increase with activity, whereas cell survival showed a dose-dependent decrease. (58m)Co-DOTATATE was significantly more efficient in cell killing per cumulated decay than (111)In- and (177)Lu-DOTATATE. The in vivo pharmacokinetic studies showed a high level of receptor-specific tumor uptake. All cobalt-labeled radioligands showed a high level of receptor-specific uptake both in vitro and in vivo in tumor-bearing mice. Furthermore, (58m)Co-DOTATATE showed considerable therapeutic effects in vitro and, thus, could be an effective agent for eradicating disseminated tumor cells and micrometastases.


Left hemispheric cortical watershed infarcts triggered by carotid sinus self-massage.
A 69-year-old man was presented to our emergency department with acute onset of hemianopsia, aphasia and dizziness. He reported that while he was sitting in front of his computer at home, he had performed a bilateral self-massage of his carotid arteries when suddenly the symptoms occurred. A neurological examination revealed a hemianopsia with a visual field loss on the right side. In addition, a mild aphasic syndrome with agraphia and a word-finding disorder (National Institutes of Health Stroke Scale (NIHSS): 3 points) was diagnosed. The initial brain CT scan with CT angiography showed neither an intracerebral haemorrhage nor a cerebral infarction. Also, no occlusion or any signs of artery dissection or a flow relevant stenosis of the brain supplying arteries were found. After excluding other contraindications, an intravenous thrombolysis with weight-adapted alteplase was performed. The symptoms of the patient significantly improved in the short-term follow-up. Three days after admission no neurological deficits remained. The MRI of the brain revealed multifocal, small, left hemispherical strokes in the middle cerebral artery territory. In general, watershed infarcts after carotid sinus self-massage follow a rare ischaemic stroke mechanism. This case emphasises the importance of a detailed anamnestic evaluation to determine the aetiological classification of ischaemic stroke as well as educating patients' (poststroke) behaviour.


Multiple regulatory effects of varicella-zoster virus (VZV) gL on trafficking patterns and fusogenic properties of VZV gH.
Varicella-zoster virus (VZV) is an extremely cell-associated alphaherpesvirus; VZV infection is spread almost exclusively via cell membrane fusion. The envelope glycoprotein H (gH) is highly conserved among the herpesviruses. A virus-encoded chaperone, glycoprotein L (gL), associates with gH, and the gH:gL complex is required for gH maturation and membrane expression. We recently demonstrated that in the VZV system, the gH:gL complex facilitated cell membrane fusion and extensive polykaryon formation in transfected cells (K. M. Duus, C. Hatfield, and C. Grose, Virology 210:429-440, 1995). To further define the functions of the unusual VZV gL chaperone protein, we have performed a series of mutagenesis experiments with both gH and gL and analyzed the mutants by laser scanning confocal microscopy in a transfection-based fusion assay. We established the fact that immature gH exited the endoplasmic reticulum (ER) when coexpressed with either gE or gI and appeared on the cell surface in a patch pattern. A similar effect was observed on the cell surface with gH with a cytoplasmic tail mutagenized to closely resemble the vaccinia virus hemagglutinin cytoplasmic tail. Site-directed mutagenesis of the five gL cysteine residues demonstrated that four of five cysteines participated in the gL chaperone function required for proper maturation of gH. On the other hand, the same gL mutants facilitated transport of immature gH to the cell surface, where patching occurred. Studies of gL processing demonstrated that maturation did not require transport beyond the medial-Golgi; furthermore, gL was not detected in the outer cell membrane, nor was it secreted into the medium. Colocalization studies with 3,3'-dihexyloxa-cabocyanine iodide and N-(e-7-nitrobenz-2-oxa-1,3-diazol-4-yl-aminocaproyl)-D-erythro-sphingosine confirmed that gL was found primarily in the ER and cis/medial-Golgi when expressed alone. When all of these data were considered, they suggested a posttranslational gH:gL regulation model whereby the gL chaperone modulated gH expression via retrograde flow from the Golgi to the ER. In this schema, mature gL returns to the ER, where it escorts immature gH from the ER to the Golgi; thereafter, mature gH is transported from the trans-Golgi to the outer cell membrane, where it acts as a major fusogen.


The Competing Endogenous RNAs Regulatory Genes Network Mediates Leaf Shape Variation and Main Effector Gene Function in Mulberry Plant (Morus alba).
Mulberry plants (Morus alba) have leaf shapes, ranging from unlobed to lobed, which are crucial for yield, growth, and adaptability, indicating their ability to adapt to their environment. Competing endogenous RNAs (ceRNAs) constitute a web of RNAs within the organism's transcriptional regulatory system, including protein-coding genes (mRNAs), microRNAs (miRNAs), long non-coding RNAs (lncRNAs), circular RNAs (circRNAs), and others. In this study, samples for ceRNA sequencing were categorized into two groups: whole leaves and lobed leaves, each group with three replicates. In addition, we isolated, cloned, and characterized the precursor miRNA (miR156x) from the leaves of M. alba. miR156x precursor had a length of 107 base pairs and a minimum folding free energy of 50.27 kcal/mol. We constructed a pCAMBIA-35S-GUS-miR156x dual overexpression vector and established a transient transformation system for mulberry. At an optimal transformation solution (OD600 = 0.7), the GUS gene showed a higher expression in the leaves of transiently transformed mulberry with miR156x overexpression, four days after transformation, while the target genes of miR156x had decreased expression in the same leaves. Investigations into the transgenic mulberry plants uncovered various modifications to physio-chemical parameters including POD, SOD, PRO, MDA, soluble proteins and sugars, and chlorophyl content. miRNAs in the plants were found to act as negative regulators of gene expression in response to changes in leaf shape regulation, which was confirmed in vitro using dual-luciferase reporter assays. Subsequently, we cloned Maspl3 in vitro and conducted GST-Pull down assays, obtaining multiple proteins that interacted with the Maspl3 gene. This indicates that the miR156x/Maspl3/MSTRG.25812.1 regulatory module contributes to the differences in mulberry leaf shape.


Brain ageing in the new millennium.
This paper examines the current literature pertaining to brain ageing. The objective of this review is to provide an overview of the effects of ageing on brain structure and function and to examine possible mediators of these changes. A MEDLINE search was conducted for each area of interest. A selective review was undertaken of relevant articles. Although fundamental changes in fluid intellectual abilities occur with age, global cognitive decline is not a hallmark of the ageing process. Decline in fluid intellectual ability is paralleled by regionally specific age related changes apparent from both structural and functional neuroimaging studies. The histopathological mediators of these changes do not appear to be reduction in neuronal number, which, with the exception of selected hippocampal regions, remain relatively stable across age. At the molecular level, several mechanisms of age related change have been postulated. Such theoretical models await refinement and may eventually provide a basis for therapy designed to reduce effects of the ageing process. The role of possible protective factors such as 'brain reserve', neuroprotective agents and hormonal factors in modifying individual vulnerability to the ageing process has been the focus of a limited number of studies. Our understanding of the functional and structural changes associated with both healthy and pathological ageing is rapidly gaining in sophistication and complexity. An awareness of the fundamental biological substrates underpinning the ageing process will allow improved insights into vulnerability to neuropsychiatric disease associated with advancing age.


The XPO1 Inhibitor KPT-8602 Synergizes with Dexamethasone in Acute Lymphoblastic Leukemia.
KPT-8602 (Eltanexor) is a second-generation exportin-1 (XPO1) inhibitor with potent activity against acute lymphoblastic leukemia (ALL) in preclinical models and with minimal effects on normal cells. In this study, we evaluated whether KPT-8602 would synergize with dexamethasone, vincristine, or doxorubicin, three drugs currently used for the treatment of ALL. First, we searched for the most synergistic combination of KPT-8602 with dexamethasone, vincristine, or doxorubicin in vitro in both B-ALL and T-ALL cell lines using proliferation and apoptosis as a readout. Next, we validated this synergistic effect by treatment of clinically relevant B- and T-ALL patient-derived xenograft models in vivo. Finally, we performed RNA-sequencing (RNA-seq) and chromatin immunoprecipitation sequencing (ChIP-seq) to determine the mechanism of synergy. KPT-8602 showed strong synergism with dexamethasone on human B-ALL and T-ALL cell lines as well as in vivo in three patient-derived ALL xenografts. Compared with single-drug treatment, the drug combination caused increased apoptosis and led to histone depletion. Mechanistically, integration of ChIP-seq and RNA-seq data revealed that addition of KPT-8602 to dexamethasone enhanced the activity of the glucocorticoid receptor (NR3C1) and led to increased inhibition of E2F-mediated transcription. We observed strong inhibition of E2F target genes related to cell cycle, DNA replication, and transcriptional regulation. Our preclinical study demonstrates that KPT-8602 enhances the effects of dexamethasone to inhibit B-ALL and T-ALL cells via NR3C1- and E2F-mediated transcriptional complexes, allowing to achieve increased dexamethasone effects for patients.


Individuals living in areas with high background radon: a GIS method to identify populations at risk.
to identify and link populations and individuals that live within high risk areas. census registers and disease registers which contain data on individuals can only give aggregate statistics relating to postal code districts, town, county or state boundaries. However environmental risk factors rarely, if ever, respect these man-made boundaries. What is needed is a method to rapidly identify individuals who may live within a described area or region and to further identify the disease(s) occurring among these individuals and/or in these areas. this paper describes a method for linking the standard registers available in Sweden, notably the residence-property addresses they contain and the geographical coordinate setting of these, to map the population as a point coverage. Using standard GIS methods this coverage could be linked, merged or intersected with any other map to create new subsets of population. Representation of populations down to the individual level by automatised spatialisation of available census data is in its simplicity a new informatics method which in the designated GIS medium adds a new power of resolution. We demonstrate this using the radon maps provided by the local communes. The Swedish annual population registration records of 1991 for the county of Ostergötland and the property register available at the Central Statistical Bureau of Sweden formed the main data sources. By coupling the address in the population register to the property register each individual was mapped to the centroid of a property. By intersecting the population coverage with the radon maps, the population living in high, normal or low risk areas was identified and then analysed and stratified by commune, sex and age. The resulting tables can be linked to other database registers, to visualise and analyse geographical and related patterns. The methodology can be adapted for use with any other environmental map or small area. It can also be expanded to the fourth dimension by linking likewise available migration information to generate immediately coordinate-set, accumulated exposition and similar data.


Continuous Dropout.
Dropout has been proven to be an effective algorithm for training robust deep networks because of its ability to prevent overfitting by avoiding the co-adaptation of feature detectors. Current explanations of dropout include bagging, naive Bayes, regularization, and sex in evolution. According to the activation patterns of neurons in the human brain, when faced with different situations, the firing rates of neurons are random and continuous, not binary as current dropout does. Inspired by this phenomenon, we extend the traditional binary dropout to continuous dropout. On the one hand, continuous dropout is considerably closer to the activation characteristics of neurons in the human brain than traditional binary dropout. On the other hand, we demonstrate that continuous dropout has the property of avoiding the co-adaptation of feature detectors, which suggests that we can extract more independent feature detectors for model averaging in the test stage. We introduce the proposed continuous dropout to a feedforward neural network and comprehensively compare it with binary dropout, adaptive dropout, and DropConnect on Modified National Institute of Standards and Technology, Canadian Institute for Advanced Research-10, Street View House Numbers, NORB, and ImageNet large scale visual recognition competition-12. Thorough experiments demonstrate that our method performs better in preventing the co-adaptation of feature detectors and improves test performance.


Outcome, observer reliability, and patient preferences if CTA, MRA, or Doppler ultrasound were used, individually or together, instead of digital subtraction angiography before carotid endarterectomy.
To evaluate the accuracy of routinely available non-invasive tests (spiral computed tomographic angiography (CTA), time of flight magnetic resonance angiography (MRA), and colour Doppler ultrasound (DUS)), individually and together, compared with intra-arterial digital subtraction angiography (DSA) in patients with symptomatic tight carotid stenosis; and to assess the effect of substituting non-invasive tests for DSA on outcome, interobserver variability, and patient preference. Patients referred from a neurovascular clinic were subjected prospectively to DUS imaging. The operator was blind to symptoms. Patients with a tight carotid stenosis on the symptomatic side were admitted for DSA. CTA and MRA were performed during the admission. The CTA, MRA, and DSA films were each read independently by two of six experienced radiologists, blind to all other data. 67 patients were included (34 had all four imaging procedures). DUS, CTA, and MRA all agreed with DSA in the diagnosis of operable v non-operable disease in about 80% of patients. CTA tended to underestimate (sensitivity 0.65, specificity 1.0), MRA to overestimate (sensitivity 1.0, specificity 0.57), and DUS to agree most closely with (sensitivity 0.85, specificity 0.71) the degree of stenosis as shown by DSA. When using any two of the three non-invasive tests in combination, adding the third if the first two disagreed would result in very few misdiagnoses (about 6%). MRA had similar interobserver variability to CTA (both worse than DSA). Patients preferred CTA over MRA and DSA. DUS, CTA, and MRA all show similar accuracy in the diagnosis of symptomatic carotid stenosis. No technique on its own is accurate enough to replace DSA. Two non-invasive techniques in combination, and adding a third if the first two disagree, appears more accurate, but may still result in diagnostic errors.