Establishment and characterization of a new human myxoid liposarcoma cell line (DL-221) with the FUS-DDIT3 translocation.
Myxoid liposarcoma has the pathognomonic fusion oncogene FUS-DDIT3 encoding a chimeric transcription factor. Metastatic risk is higher with an increased round cell component and has been linked to aberrations involving the IGFR/PI3K/AKT pathway. These molecular insights have yet to translate to targeted therapies, and the lack of experimental models is a major hindrance. We describe the initial in-depth characterization of a new cell line (DL-221) and establishment of a mouse xenograft model. The cell line DL-221 was derived from a metastatic pleural lesion showing myxoid and round cell histology. This newly established cell line was characterized for phenotypic properties and molecular cytogenetic profile, using PCR, COBRA-FISH, and western blot. Next-generation whole-exome sequencing was performed to further characterize the cell line and the parent tumor. NOD-SCID-IL2R gamma knockout mice were xenograft hosts. DL-221 cells grew an adhering monolayer and COBRA-FISH showed an aneuploid karyotype with t(12;16)(q13;p11) and several other rearrangements; RT-PCR demonstrated a FUS-DDIT3 fusion transcript type 1. Both the cell line and the original tumor harbored a TP53 compound heterozygous mutation in exon 4 and 7, and were wild-type for PIK3CA. Moreover, among the 1254 variants called by whole-exome sequencing, there was 77% concordance between the cell line and parent tumor. The recently described hotspot mutation in the TERT promoter region in myxoid liposarcomas was also found at C228T in DL-221. Xenografts suitable for additional preclinical studies were successfully established in mice after subcutaneous injection. The established DL-221 cell line is the only published available myxoid liposarcoma cell line that underwent spontaneous immortalization, without requiring SV40 transformation. The cell line and its xenograft model are unique and helpful tools to study the biology and novel potential-targeted treatment approaches for myxoid liposarcoma.


Interferon-alpha enhances artemisinin-induced differentiation of HL-60 leukemia cells via a PKC alpha/ERK pathway.
Interferon-alpha (IFN-alpha) has been used in the treatment of several cancers, including chronic myeloid leukemia. Artemisinin, a sesquiterpene lactone endoperoxide that exists in several medicinal plants, is a well known anti-malarial agent. We previously reported that artemisinin by itself caused a relatively low level of HL-60 cell differentiation. In this study, we investigated the effects of IFN-alpha in combination with artemisinin on cell growth and differentiation in HL-60 leukemia cells. Combination of IFN-alpha and artemisinin synergistically induced the levels of leukemia cell differentiation, although IFN-alpha by itself did not affect cell proliferation and differentiation. The increased cell differentiation by IFN-alpha and artemisinin was significantly suppressed by the inhibitors for protein kinase C (PKC), extracellular signal-regulated kinase (ERK) and jun N-terminal kinase (JNK), but not by the inhibitors for phosphatidylinositol 3-kinase (PI3-K) and p38 mitogen-activated protein kinase (MAPK). Furthermore, co-treatment with IFN-alpha increased levels of PKC alpha and phosphorylated ERK. Taken together, these results indicate the enhancement of artemisinin-induced HL-60 cell differentiation by IFN-alpha through the activation of a PKC alpha/ERK signaling pathway, and suggest a possible use of IFN-alpha and artemisinin in the treatment of leukemic diseases.


Accessory limb production by nerve-induced cell proliferation.
The deviation of large limb nerves to a more proximal skin wound yielded a high proportion of accessory limb responses in different age groups of Ambystoma mexicanum (axolotls). In some instances the deviated nerve was positioned on skin previously grafted from an animal of different age and pigmentation from that of the host. Grafts were found not to be a necessary prerequisite for accessory limb induction, but the presence of wound epithelium was required. The rule of distal morphogenesis was expressed in reference to the level at which the nerve was cut, not in reference to the wound site where the accessory actually developed. The upper arm proved to be a more favorable site for accessory limb production than the flank or the leg under the conditions of the present experiments, in which little or no damage was done to the underlying muscles. The orientation of the accessory limb was extremely varied despite the uniformity of the surgical procedure.


Determination of glycemic monitoring marker 1,5-anhydroglucitol in plasma by liquid chromatography-electrospray tandem mass spectrometry.
Previous studies have shown that plasma 1,5-anhydroglucitol (1,5-AG) is markedly reduced among diabetic patients and therefore serves as a sensitive marker for short-term glycemic control. The current study describes the development of the liquid chromatography negative ion electrospray tandem mass spectrometry (LC-MS/MS) method to measure 1,5-AG in human plasma. The samples were pre-treated with protein precipitation and an isotope-labeled internal standard was used. Chromatographic separation was achieved on amide column (150 mm x 2.0mm i.d., 5 microm) followed by detection with multiple reaction monitoring mode. Linearity, accuracy, precision, recovery, matrix effect, and stability were evaluated during method validation over the range of 1-50 microg/mL. The validated method has been clinically applied among 159 type 2 diabetic patients and 290 control subjects. A marked reduction in 1,5-AG levels among the diabetic patients and significant between-gender difference in nondiabetic subjects were observed.


The influence of action-outcome contingency on motivation from control.
The sense of agency is defined as one's sense of control over one's actions and their consequences. A recent theory, the control-based response selection framework (Karsh and Eitam, Motivation from control: a response selection framework. The sense of agency, Oxford University Press, New York, 2015a), suggests that actions associated with a high sense of agency are intrinsically rewarding and thus motivate response selection. Previous studies support this theory by demonstrating that factors impacting on sense of agency (e.g. probability of an outcome following an action) also motivate selection of actions. Here we report a novel test of the control-based response selection framework in the domain of action-outcome contingency. The contingency between actions and their outcome has previously been demonstrated to impact the sense of agency, but its impact on the motivation to perform actions has not yet been examined. Participants were asked to press one of four buttons as randomly as possible. Each of the buttons was assigned a different probability of causing an outcome when pressed. Additionally, a contingency manipulation was employed where the probability of an outcome occurring in the absence of a button press was also varied in blocks throughout the experiment. Results demonstrated a significant influence of contingency on response speed, and a significant effect of probability on response selection, consistent with predictions from the control-based response selection framework. Furthermore, some evidence was observed for a positive correlation between influence of contingency and autistic traits, with individuals with higher autistic traits showing a greater influence of contingency on reaction times. The current findings support the idea that actions associated with an increased sense of agency are intrinsically rewarding, and identify how individual differences may impact on this process.


Flow injection analysis of nitrate and nitrite in commercial baby foods.
Commercial baby foods are an easy alternative to home-made meals especially for working parents in a nuclear family therefore it is imperative to determine the nitrate and nitrite content in commercially available baby foods varieties marketed in Fiji. A total of 108 baby food samples were analyzed for nitrate and nitrite using our standardized flow injection analysis (FIA) technique with colorimetric detection technique employing sulfanilamide and N-(1-naphthyl)ethylenediamine dihydrochloride as color reagents where the samples throughput was 38 h(-1). The commercial baby food varieties chosen comprised of vegetables, cereals, fruits and milk. The study shows that the nitrate content of the baby foods studied ranges from 2.10 to 220.67 mg kg(-1) whereas the nitrite content ranges from 0.44 to 3.67 mg kg(-1). Typical recoveries of spiked nitrate residues ranged from 92% to 106%. The study shows that the average nitrate content of commercially available baby foods in Fiji descends below the maximum level proposed by the European Union Legislation.


Interferon beta treatment in relapsing-remitting multiple sclerosis: a post-marketing study in Lombardia, Italy. Multiple Sclerosis Centers of Lombardia, Italy.
The aim of the study was to evaluate the efficacy of interferon beta-1a (IFN-beta-1a) and beta-1b (IFN-beta-1b) in clinical practice for the treatment of relapsing-remitting multiple sclerosis (RR MS). Patients were selected and prospectively monitored according to a predefined protocol. An appropriate form was prepared to collect clinical data of multiple sclerosis patients attending the MS Centers of Lombardia, Italy. On 30 June 1998, 317 patients were treated with IFN-beta-1b and 156 with IFN-beta-1a. Basal expanded disability status scale (EDSS) and relapse frequency were similar in both groups of patients. The annual relapse rate consistently decreased from 1.76 to 0.63 at 1 year and to 0.51 at 2 years for the IFN-beta-1b group and from 1.6 to 1.0 at 1 year for the IFN-beta-1a group. Disability remained stable in most patients. Dropouts (20.5%) were affected by more active disease compared to patients who continued to be treated. This study confirms the efficacy of both treatments, showing a more marked effect than expected from the clinical trials' results, probably due to differences in selection criteria and exclusion of dropouts.


The parAB gene products of Pseudomonas putida exhibit partition activity in both P. putida and Escherichia coli.
The bacteria for which there is evidence that proteins of the ParAB family act in chromosome segregation also undergo developmental transitions that involve the ParAB homologues, raising the question of whether the partition activity is equivalent to that of plasmid partition systems. We have investigated the role in partition of the parAB locus of a free-living bacterium, Pseudomonas putida, not known to pass through developmental phases. A parAB deletion mutant, compared with wild type, showed slightly higher frequencies of anucleate cells in exponentially growing cultures but much higher frequencies in deceleration phase. This increase was growth medium dependent. Oversupply of ParA and ParB proteins also raised anucleate cell levels, specifically in the deceleration phase, in wild-type and mutant strains and regardless of medium, as well as generating abnormal cell morphologies. Absence or oversupply of ParAB function had either slight or considerable effects on growth rate, depending on temperature and medium. The need for the Par proteins in chromosome partition thus appears to be subject to the cell's physiological state. Three sequences similar to cis-acting stabilization sites of Bacillus subtilis are present in the P. putida oriC-parAB region. One was inserted into an unstable mini-F and shown to stabilize it in E. coli in a ParAB-dependent manner.


Yohimbine-induced withdrawal and anxiety symptoms in opioid-dependent patients.
Alteration in noradrenergic regulation as well as alteration in the hypothalamic-pituitary-adrenal (HPA) axis have been associated with opioid dependence and acute abstinence symptoms. This double-blind, placebo-controlled study evaluated subjective, physiologic, and biochemical responses to yohimbine (.4 mg/kg, IV) in eight patients receiving methadone and compared results to those from a pool of nine healthy volunteers. All subjects were compared for panic anxiety symptom scale (PASS) scores, systolic and diastolic blood pressure, heart rate, plasma 3-methoxy-4 hydroxyphenethyleneglycol (MHPG), and cortisol. Yohimbine elicited objective and subjective opioid withdrawal and elevated craving for opioid drugs in methadone patients. Significant yohimbine effects were seen across the combined subject group for PASS, physiologic measures, MHPG, and cortisol. Methadone patients had lower baseline MHPG levels. Methadone group interactions with yohimbine were seen for systolic blood pressure and cortisol levels. Methadone-maintained patients are sensitive to the postsynaptic effects of noradrenergic-facilitating medications, experiencing greater physiologic and psychological symptoms, including an increase in craving. The effect on cortisol supports the above conclusion and is consistent with HPA axis perturbation in opioid dependence as reported in other studies and extends these observations to stable methadone-maintained patients. Medications that increase synaptic noradrenaline should be used with care in opioid-dependent patients.


Selection of D-amino-acid peptides that bind to Alzheimer's disease amyloid peptide abeta1-42 by mirror image phage display.
A mirror image phage display approach was used to identify novel and highly specific ligands for Alzheimer's disease amyloid peptide Abeta(1-42). A randomized 12-mer peptide library presented on M13 phages was screened for peptides with binding affinity for the mirror image of Abeta(1-42). After four rounds of selection and amplification the peptides were enriched with a dominating consensus sequence. The mirror image of the most representative peptide (D-pep) was shown to bind Abeta(1-42) with a dissociation constant in the submicromolar range. Furthermore, in brain tissue sections derived from patients that suffered from Alzheimer's disease, amyloid plaques and leptomeningeal vessels containing Abeta amyloid were stained specifically with a fluorescence-labeled derivative of D-pep. Fibrillar deposits derived from other amyloidosis were not labeled by D-pep. Possible applications of this novel and highly specific Abeta ligand in diagnosis and therapy of Alzheimer's disease are discussed.


Effect of light filters on contrast sensitivity function in normal and retinal degeneration subjects.
Contrast sensitivity functions were measured with five light filters and without a filter, in the presence of a glare source, in 12 retinal degeneration subjects and 9 normal subjects. The light filters included yellow-tinted, CPF 527, NoIR 111, 0.6 neutral density and sunglasses with a 95% UV filter. Retinal degeneration subjects showed a mild improvement in contrast sensitivity at the higher spatial frequencies and maintained mean log contrast sensitivity with light filters which reduced photopic light transmission up to 75%. Normal subjects exhibited a systematic decrease of contrast sensitivity at higher spatial frequencies with all light filters and overall mean log contrast sensitivity was significantly correlated (r = 0.86, P < 0.025) with photopic light transmission of the filters. It is concluded that, when in the presence of a glare source, the benefits of certain light filters to retinal degeneration subjects is related to the reduction of overall photopic luminance.


Ticarcillin compared with carbenicillin in the treatment of exacerbations of bronchopulmonary infection in cystic fibrosis.
Sixteen adults with cystic fibrosis and chronic bronchopulmonary infection with Pseudomonas aeruginosa, admitted to hospital for chemotherapy, were randomized to treatment with either carbenicillin or ticarcillin. In addition all patients received gentamicin. Both antibiotic treatments produced clinical improvement and significant improvement in respiratory function, but there was no difference between them. Pseudomonas aeruginosa was not eradicated from sputum with either treatment regimen.


Widespread decay of vitamin-related pathways: coincidence or consequence?
The advent of modern genomics has provided an unparalleled opportunity to consider the gene complement of an organism, and scrutinize metabolic pathways that are no longer active. This approach has led to an increasing number of reports of vitamin-associated pathway deterioration, with many indicating that independent gene-loss events of one or a few key genes have led to vitamin auxotrophy. Nonfunctional unitary pseudogenes belonging to these pathways are found in several species, demonstrating that these are recent evolutionary processes. Here, we examine the commonalities in the cellular roles and metabolism of vitamins that might have led to these losses. The complex pattern of vitamin auxotrophy across the eukaryotic tree of life is intimately connected with the interdependence between organisms. The importance of this process in terms of shaping communities on the one hand, and facilitating symbioses between organisms on the other, is only just beginning to be recognized.


Telomerase-specific attenuated viruses, a definitive strategy or just one more in circulating tumor cells detection anthology?
The quantification and isolation of Circulating Tumor Cells (CTC) is being the battleground during last years. There are many groups that are investing economic resources in trying to solve this jigsaw. Technological platforms based on different proofs of concept have been developed achieving in some cases excellent results despite not having been able to detect the total compute of the patient's CTC population. The handicap of this matter has been the lack of universal markers. Several years have gone so that in detection of CTC is take into account a basic characteristic that possesses the most of tumor cells, the loss of inactivation of the enzyme telomerase. Gene therapy has been combined with telomerase activity concept for develop a molecular tool that makes it possible to identify CTC: Telomerase-specific replication-selective viruses. This review includes for the first time all the scientific studies that have been published to date with this advanced technology. Furthermore, it describes the role in the diagnosis and prognosis that Telomerase-specific attenuated viruses have been playing in cancer patients study during this last decade.


Glycosuria and insulitis in NOD mice expressing the HLA-DQw6 molecule.
To investigate HLA-linked genes controlling the susceptibility and resistance to insulin dependent diabetes mellitus (IDDM), HLA-DQ alleles of 45 Japanese patients with IDDM were analysed, using sequence specific oligonucleotide (SSO). DQA1*0301 and DQB1*04 were positively associated (R.R = 6.6, Pc less than 0.05 and R.R. = 4.7 Pc less than 0.01) and DQA1*0103 and DQB1*0104 were negatively associated (R.R. = 0.2, Pc less than 0.01) with IDDM. DQA1*0103 and DQB1*0104 were in strong linkage disequilibrium to encode for DQw6 molecule. Therefore, in a Japanese population, the DQw6 molecule seems to control the resistance to IDDM. To determine whether or not the DQw6 molecule itself can protect against glycosuria and insulitis in NOD mice, these animals were mated with HLA-DQw6 transgenic-C57BL/6 mice (DQw6-B6) and the F1 progeny expressing the DQw6 molecule were backcrossed with NOD mice. Eighty-five female backcross progenies were classified into four groups, according to the MHC classII phenotype; I-ANOD/I-ANOD DQw6(-), I-ANOD/I-ANOD DQw6(+), I-ANOD/I-Ab DQw6(-) and I-ANOD/I-Ab DQw6(+). At the age of 16 weeks, 9.1% of the DQw6(-) I-Ab(-) mice had a glycosuria whereas none of the DQw6(+) I-Ab(-) mice had a glycosuria. At the age of 30 weeks 13.6% of the DQw6(-) I-Ab(-) mice had a glycosuria and 7.7% of the DQw6(+) I-Ab(-) mice had a glycosuria. Histological examinations of the pancreas were performed in the 30 week old mice or after the development of glycosuria.(ABSTRACT TRUNCATED AT 250 WORDS)


Cation excretion and kallikrein secretion by the rat parotid gland in various hypertensive models.
The major salivary glands are distinguished by a high kallikrein content, localized particularly in regions with intense electrolyte transport (the apical border of striated duct cells and the luminal border of the main excretory ducts). For this reason we used the parotid gland as a test model for investigations on electrolyte (sodium and potassium) and kallikrein transport in various forms of experimental hypertension [spontaneously hypertensive rats, renovascular (two-kidney one-clip) hypertension, and deoxycorticosterone trimethylacetate (DOCA)-salt hypertension]. As compared with values for the controls, the sodium concentration in parotid saliva was decreased in all hypertensive models studied, indicating enhanced sodium reabsorption in the glandular duct system. In DOCA-salt hypertension the flow rate dependency of the sodium concentration was preserved. In contrast, in genetic and renovascular hypertension, the decreased sodium concentration was independent of the salivary flow rate. Salivary potassium concentration showed an inverse relationship to sodium level and was slightly increased in DOCA-salt and renovascular hypertension. In genetic hypertension potassium concentration in saliva was lowered. Salivary kallikrein activity was enhanced in all forms of hypertension studied, with the highest increase in genetic hypertension (factor 3.5), followed by DOCA-salt hypertension (factor 3.0) and finally by renovascular hypertension (factor 2.5). The pathogenesis of the exaggerated kallikrein secretion observed in hypertension is discussed with regard to a counterregulatory mechanism as well as sympathicoadrenergic activation.(ABSTRACT TRUNCATED AT 250 WORDS)


Meta-analysis of bipolar radiofrequency endometrial ablation versus thermal balloon endometrial ablation for the treatment of heavy menstrual bleeding.
Heavy menstrual bleeding is a common problem that can severely affect quality of life. To compare bipolar radiofrequency endometrial ablation and thermal balloon ablation for heavy menstrual bleeding in terms of efficacy and health-related quality of life (HRQoL). Online registries were systematically searched using relevant terms without language restriction from inception to November 24, 2016. Randomized control trials or cohort studies of women with heavy menstrual bleeding comparing the efficacy of two treatments were eligible. Data were extracted. Results were expressed as risk ratios (RRs) or weighted mean differences (WMDs) with 95% confidence intervals (CIs). Six studies involving 901 patients were included. Amenorrhea rate at 12 months was significantly higher after bipolar radiofrequency endometrial ablation than after thermal balloon ablation (RR 2.73, 95% CI 2.00-3.73). However, no difference at 12 months was noted for dysmenorrhea (RR 1.04, 95% CI 0.68-1.58) or treatment failure (RR 0.78, 95% CI 0.38-1.60). The only significant difference for HRQoL outcomes was for change in SAQ pleasure score (12 months: WMD -3.51, 95% CI -5.42 to -1.60). Bipolar radiofrequency endometrial ablation and thermal balloon ablation reduce menstrual loss and improve quality of life. However, bipolar radiofrequency endometrial ablation is more effective in terms of amenorrhea rate and SAQ pleasure.


Contribution of interaction between genetic variants of interleukin-11 and Helicobacter pylori infection to the susceptibility of gastric cancer.
Gastric cancer (GC) ranks the second leading cause of cancer-related mortality worldwide. We aimed to clarify the relevance of genetic variants of IL-11, a hub of various carcinogenic pathways, as well as their interactions with Helicobacter pylori (H. pylori) infection in the development of GC. A case-control study with 880 GC cases and 900 healthy controls was conducted in a Chinese population. Six tagSNPs were detected by Taqman Allelic Discrimination assay, while H. pylori status was detected by Typing Detection Kit for Antibody to H. pylori and serum IL-11 level was measured using ELISA method. We found that rs1126760 (C vs T: OR=1.39, 95% CIs=1.13-1.70, P=0.002) and rs1126757 (C vs T: OR=0.82, 95% CIs=0.72-0.93, P=0.002) were significantly associated with susceptibility of GC. Even adjusted for Bonferroni correction, the results were still significant (P=0.002×6=0.012). IL-11 rs1126760 was significantly associated with higher serum and expression level of IL-11, while rs1126757 was significantly associated with lower serum IL-11 level (P<0.001). Significant interaction with H. pylori infection was identified for rs1126760 (P for interaction =0.005). Higher expression of the IL-11 gene was significant with development and poor prognosis of GC. Our study provides strong evidence that genetic variants of the IL-11 gene may interact with H. pylori infection and contribute to the development of GC. Further studies with larger sample size and functional experiments are needed to validate our findings.


Cole-impedance parameters representing biceps tissue bioimpedance in healthy adults and their alterations following eccentric exercise.
The purpose of this study is to identify if participation in an eccentric exercise protocol altered the Cole-impedance model parameters that represent localized bicep tissue bioimpedance. This supports continued efforts to identify which features of tissue bioimpedance may be effective markers to non-invasively identify skeletal muscle damage. Here, the Cole-impedance model parameters that best fit the localized electrical impedance of exercised (using an eccentric stimulus) and unexercised biceps of 6 participants (collected before, immediately after and at 24 h, 48 h, 72 h and 96 h) are determined using a numerical optimization technique. Statistical tests comparing the pre-exercise and post-exercise model parameters report significant decreases in R ∞ and R 1 with significant increases in C at 72 h and 96 h post-exercise for exercised biceps (aligning with noted periods of peak swelling). These changes in R ∞ , R 1 , and C were not observed in the unexercised biceps. These results support that the C parameter of the Cole-impedance model fit to bioimpedance data may be a suitable marker for identifying skeletal muscle damage.


First steps in designing an all-in-one ICT-based device for persons with cognitive impairment: evaluation of the first mock-up.
This project Smart Assisted Living involving Informal careGivers++ (SALIG) intends to develop an ICT-based device for persons with cognitive impairment combined with remote support possibilities for significant others and formal caregivers. This paper presents the identification of the target groups' needs and requirements of such device and the evaluation of the first mock-up, demonstrated in a tablet. The inclusive design method that includes end-users in the design process was chosen. First, a scoping review was conducted in order to examine the target group's need of an ICT-based device, and to gather recommendations regarding its design and functionalities. In order to capture the users' requirements of the design and functionalities of the device three targeted focus groups were conducted. Based on the findings from the publications and the focus groups a user requirement specification was developed. After that a design concept and a first mock-up was developed in an iterative process. The mock-up was evaluated through interviews with persons with cognitive impairment, health care professionals and significant others. Data were analysed using content analysis. Several useful recommendations of the design and functionalities of the SALIG device for persons with cognitive impairment were identified. The main benefit of the mock-up was that it was a single device with a set of functionalities installed on a tablet and designed for persons with cognitive impairment. An additional benefit was that it could be used remotely by significant others and formal caregivers. The SALIG device has the potentials to facilitate everyday life for persons with cognitive impairment, their significant others and the work situation for formal caregivers. The results may provide guidance in the development of different types of technologies for the target population and for people with diverse disabilities. Further work will focus on developing a prototype to be empirically tested by persons with cognitive impairment, their significant others and formal caregivers.


Correction to: Multi-center, randomized, double-blind, placebocontrolled study of quetiapine extended-release formulation in Japanese patients with bipolar depression.
The original version of this article contained a mistake in Table 5. Correct version is presented here.


HL2 on chromosome 7D of wheat (Triticum aestivum L.) regulates both head length and spikelet number.
A major QTL QSpl.nau-7D, named HL2, was validated for its effects on head length and kernel number per spike using NIL, and mapped to a 0.2 cM interval using recombinants. Improvement in wheat inflorescence traits such as spike or head length and spikelet number provides an important avenue to increase grain yield potential. In a previous study, QSpl.nau-7D, the major QTL for head length on chromosome 7D, was identified in the recombinant inbred lines derived from Nanda2419 and Wangshuibai. To validate and precisely map this QTL, the Wangshuibai allele was transferred to elite cultivar Yangmai15 through marker-assisted selection. Compared with the recurrent parent, the resultant near-isogenic line (NIL) yielded not only 28% longer spikes on the average but also more spikelets and kernels per spike. Moreover, the NIL had a lower spikelet density and did not show significant kernel weight change. In the F2 population derived from the NIL, QSpl.nau-7D acted like a single semi-dominant gene controlling head length and was therefore designated as Head Length 2 (HL2). With this population, a high-density genetic map was constructed mainly using newly developed markers, and 100 homozygous recombinants including 17 genotypes were obtained. Field experiments showed that the recombinants carrying the 0.2-cM interval flanked by Xwgrb1588 and Xwgrb1902 from Wangshuibai produced longer spikes than those without this Wangshuibai allele. Comparative mapping of this interval revealed a conserved synteny among cereal grasses. HL2 is beneficial to wheat breeding for more kernels per spike at a lower spikelet density, which is a favored morphological trait for Fusarium head blight resistance.


Genetic susceptibility of increased intestinal permeability is associated with progressive liver disease and diabetes in patients with non-alcoholic fatty liver disease.
Increased intestinal permeability plays a key role in the pathogenesis of fat deposition in the liver. The aim of our study was to assess whether a single nucleotide polymorphism of protein tyrosine phosphatase non-receptor type 2 (PTPN2) (rs2542151 T→G), involved in intestinal permeability, may be associated with non-alcoholic fatty liver disease (NAFLD) and type 2 diabetes mellitus (T2DM). We recruited a prospective cohort of NAFLD subjects and matched controls. Clinical data, PTPN2 genotype and laboratory data were collected for each patient. Results were stratified according to liver histology and diabetes. We enrolled 566 cases and 377 controls. PTPN2 genotype distribution did not significantly differ between patients and controls. In the entire population, patients with PTPN2 rs2542151 T→G (dominant model) have a higher prevalence of diabetes; 345 patients (60.9%) underwent liver biopsy: 198 (57.4%) had steatohepatitis and 75 (21.7%) had advanced fibrosis. At multiple logistic regression analysis PTPN2 rs2542151 T→G was associated with T2DM (OR 2.14, 95% CI 1.04-4.40, P = 0.03). Patients who underwent liver biopsy, rs2542151 T→G of PTPN2 was independently associated with severe steatosis (OR 2.00, 95% CI 1.17-3.43, p = 0.01) and severe fibrosis (OR 2.23, 95% CI 1.06-4.72, P = 0.03). Our study shows that NAFLD patients with rs2542151 T→G of PTPN2 have a higher severity of fatty liver disease and a higher prevalence of T2DM. These results suggest that individual genetic susceptibility to intestinal permeability could play a role in liver disease progression.


Prolonged trastuzumab therapy in a patient with recurrent breast cancer and anthracycline-induced cardiac failure.
Current practice precludes patients with pre-existing cardiac dysfunction from trastuzumab therapy. A 57-year-old patient with HER2 positive metastatic breast cancer and anthracycline-induced cardiac failure was safely treated with trastuzumab. At 46 months, left ventricular ejection fraction (LVEF) did fall to 38.3%, but 8 months later has recovered to 47%. She remains disease free and asymptomatic from cardiac dysfunction more than 6 years following breast cancer recurrence. We review the evidence for the use of trastuzumab in patients with controlled cardiac dysfunction, and suggest this group of patients should be considered for treatment with trastuzumab if no other or only less efficacious therapeutic options are available.


Natural Formulation Based on Diatomaceous Earth and Botanicals against Stored Product Insects.
Diatomaceous earth (DE) has long been known as a potential protectant for stored cereals against various stored product insects. Despite favorable effect for the environment and human health, DE has some negative side effects on the treated commodity. In order to minimize negative response and to improve its efficacy, this paper represents a study of developed natural formulation based on DE SilicoSec® enhanced with botanicals (essential oil lavender, corn oil, and bay leaves dust) and silica gel. The activity of formulation (labeled as N Form) was tested against Sitophilus oryzae (L.), Rhyzopertha dominica (F.), and Tribolium castaneum (Herbst) in seed wheat and barley under controlled conditions. As a reference comparative value, DE SilicoSec® was used. N Form showed higher efficacy than DE, especially in barley at the lowest concentration, inducing higher mortality of all three insect species. The highest average progeny inhibition was recorded in R. dominica population both in seed wheat and barley with 94.9% and 96.3% of inhibition, respectively, followed with S. oryzae and T. castaneum inhibition of 90.6% and 86.1%, respectively, in wheat and 94.9% and 89.7%, respectively, in barley. Results indicate that the developed natural formulation N Form enhanced the activity of DE SilicoSec® using lower amount of DE dust and that it could be successfully implemented for storage of cereals as alternatives to chemical pesticides for stored product insect control.


Identification of Associated Proteins by Immunoprecipitation and Mass Spectrometry Analysis.
Protein-protein interactions play central roles in intercellular and intracellular signal transduction. Impairment of protein-protein interactions causes many diseases such as cancer, cardiomyopathies, diabetes, microbial infections, and genetic and neurodegenerative disorders. Immunoprecipitation is a technique in which a target protein of interest bound by an antibody is used to pull down the protein complex out of cell lysates, which can be identified by mass spectrometry. Here, we describe the protocol to immunoprecipitate and identify the components of the protein complexes of ElmoE in Dictyostelium discoideum cells.


Characterization of microRNAs expressed in the cystic legion of the liver of Mus musculus perorally infected with Echinococcus multilocularis Nemuro strain.
Alveolar echinococcosis (AE) is a zoonosis caused by the metacestode of Echinococcus multilocularis. The published genome of E. multilocularis showed that approximately 86% of its genome is non-coding. Micro RNAs (miRNAs) are small non-coding regulatory RNAs, and recent studies on parasitic helminths expect miRNAs as a promising target for drug development and diagnostic markers. Prior to this study, only a few studies reported the E. multilocularis miRNA profiles in the intermediate host. The primary objective of this study was to characterize miRNA profiles via small RNA-seq in E. multilocularis Nemuro strain, a laboratory strain of Asian genotype, using mice perorally infected with the parasite eggs. The data were then compared with two previously published small RNA-seq data. We identified 44 mature miRNAs as E. multilocularis origin out of the 68 mature miRNA sequences registered in the miRNA database miRbase. The highest quantities of miRNAs detected were miR-10-5p, followed by bantam-3p, let-7-5p, miR-61-3p, and miR-71-5p. The top two most abundant miRNAs (miR-10-5p and bantam-3p) accounted for approximately 80.9% of the total parasite miRNAs. The highly expressed miRNA repertoire is mostly comparable to that obtained from the previous experiment using secondary echinococcosis created by an intraperitoneal administration of metacestodes. A detailed characterization and functional annotations of these shared miRNAs will lead to a better understanding of parasitic dynamics, which could provide a basis for the development of novel diagnostic and treatment methods for AE.


The applause sign in Parkinson's disease patients is related to dysexecutive syndrome.
Recent publications report that a positive applause sign is not only present in patients with neurodegenerative diseases where the subcortical structures are affected but also in patients with cortical dementia. The nature of this sign remains unknown. This study aimed to determine the frequency of the applause sign and its correlation with cognitive impairment in patients with idiopathic Parkinson's disease. The study included 30 non-depressed patients diagnosed with idiopathic Parkinson's disease. Study patients underwent the Unified Parkinson Disease Rating Scale part III, Dementia Rating Scale (DRS), Raven's Colored Progressive Matrices, and Mill Hill Vocabulary tests. Statistical analysis was performed by use of the parametric Student's t-test, nonparametric Mann-Whitney U test and Fisher's exact test, with the level of significance set at p<0.05. Negative applause sign was recorded in 66.7% and positive applause sign in 33.3% of patients. There were no between-group differences according to age, disease duration, or severity of motor symptoms. The positive applause sign group had significantly lower scores on the initiation/perseveration subscale of the DRS and a significantly higher frequency of scores below the cut-off score on the conceptualization and construction subscales of the DRS. The applause sign appears to be part of a broader dysexecutive syndrome in idiopathic Parkinson's disease.


Regeneration of hair cell stereociliary bundles in the chick cochlea following severe acoustic trauma.
Examination of pure-tone acoustic damage in the chick cochlea revealed a significant amount of hair cell recovery over a 10 day period following the exposure. The recovery included both a regeneration of stereociliary bundles to replace those that were lost and a reshuffling of the mosaic pattern of the hair cell surfaces that survived. Ten-day-old chicks were exposed to a 1500 Hz pure tone at 120 dB SPL for 48 h and their cochleae were processed for scanning, transmission and light microscopy at 0 h, 24 h, 48 h, 4 d, 6 d and 10 d after exposure. Immediately after exposure the damaged region exhibited two types of hair cell trauma. The first was a defined area of complete hair cell loss and the second was an area where the hair cells survived but exhibited varying amounts of stereocilia injury. After 48 h of recovery, new hair cells were identifiable in the region of hair cell loss and with time they underwent a progressive maturation of their stereociliary bundles. The surviving hair cells showed a dramatic rearrangement and expansion of their surfaces but exhibited no repair of the damaged stereociliary bundles. These results suggest that the chick cochlea is capable of a significant amount of recovery and regeneration following acoustic trauma.


Coexistence of crowned and blackwinged plovers.
Coexisting Crowned Plovers Vanellus coronatus and Blackwinged Plovers V. melanopterus were studied to determine whether niche partitioning occurred or was necessary to facilitate their coexistence. The results of this study show that niche partitioning did not occur. We suggest that the association of these two species potentially enhances the ability of each to detect and avoid predation, in spite of any possible deleterious effect of interspecific competition (e.g. reduced per capita food intake).


Does the speciation clock tick more slowly in the absence of heteromorphic sex chromosomes?
Squamates may be an attractive group in which to study the influence of sex chromosomes on speciation rates because of the repeated evolution of heterogamety (both XY and ZW), as well as an apparently large number of taxa with environmental sex-determination.


Self-concept correlates between at-risk and not-at-risk ninth-grade students.
This study examined whether the Tennessee Self-concept Scales could be used to discriminate among 43 ninth-grade students who were designated as being at risk of dropping out of high school and 47 students who were thought to show probability of persisting. Scores on the scales were submitted to stepwise multivariate discriminant analysis. Scores on the Self-satisfaction Scale constituted a linear function that correctly classified 72.22% of the subjects. Further investigation confirmed a 13-item scale selected from the Tennessee Self-concept Scales correctly identified the classification of 76.67% of the students.


Fertility after intrauterine device discontinuation.
Infertility rates and outcomes of pregnancies were evaluated for 219 former IUD users who had either expelled their IUDs or had them removed for medical or personal reasons other than planned pregnancy. Six women (2.7%) were infertile. Infertility rates were highest for women who used IUDs for more than two years and for women who had expelled their IUDs. Five women (2.3%) had ectopic pregnancies. The type of IUD used (Cu-7, Dalkon Shield, Lippes Loop) was not associated with different infertility rates or outcomes of pregnancies. The study results show that prior IUD use did not appear to impair subsequent fertility or outcome of pregnancy for women who either expelled their IUDs or had them removed for reasons other than pregnancy or planned pregnancy.


Gait freezing and speech disturbance in Parkinson's disease.
Gait freezing and speech disturbance are disabling axial features of Parkinson's disease (PD). However, the pathogenesis of these features remains unclear. We investigated the relation between changes in gait freezing and speech disturbance using visual and auditory cues in PD. 18 PD patients, comprising of 9 patients with freezing (PDGF) and 9 without gait freezing were studied. Patients performed a 7-m back-and-forth walk in a baseline state and with visual and auditory cues. Gait velocity, stride length and cadence were evaluated using a three-dimensional gait analysis system. For speech evaluation, patients read ten sentences in a baseline state and with visual and auditory cues. The time delay of speech initiation, speech rate and the number of repetitions per sentence were quantified. In PDGF patients, the increase in gait velocity positively correlated with the decrease in the time delay of the speech initiation. Also, the increase in the gait velocity and cadence positively correlated with the decrease in the number of repetitions per sentence. The increase in the stride length positively correlated with the increase in speech rate. Lastly, the increase in stride length positively correlated with the decrease in the number of repetitions per sentence. These findings suggest that there is a common pathomechanism of gait freezing and speech disturbance in PD.


A robust autonomous method for blood demand forecasting.
Blood supply chain management requires estimates about the demand of blood products. The more accurate these estimates are, the less wastage and fewer shortages occur. While the current literature demonstrates tangible benefits from statistical forecasting approaches, it highlights issues that discourage their use in blood supply chain optimization: there is no single approach that works everywhere, and there are no guarantees that any favorable method performance continues into the future. We design a novel autonomous forecasting system to solve the aforementioned issues. We show how possible changes in blood demand could affect prediction performance using partly synthetic demand data. We use these data then to investigate the performances of different method selection heuristics. Finally, the performances of the heuristics and single method approaches were compared using historical demand data from Finland and the Netherlands. The development code is publicly accessible. We find that a shift in the demand signal behavior from stochastic to seasonal would affect the relative performances of the methods. Our autonomous system outperforms all examined individual methods when forecasting the synthetic demand series, exhibiting meaningful robustness. When forecasting with real data, the most accurate methods in Finland and in the Netherlands are the autonomous system and the method average, respectively. Optimal use of method selection heuristics, as with our autonomous system, may overcome the need to constantly supervise forecasts in anticipation of changes in demand while being sufficiently accurate in the absence of such changes.


Flow-assisted surgical technique in cerebrovascular surgery.
Cerebrovascular surgery is a technically challenging subspecialty of neurosurgery. A flow-based approach to cerebrovascular surgery can help to optimize neurosurgical interventions. Direct intraoperative flow measurements can be made using a quantitative ultrasonic microvascular flow probe. In this article, we review the applications and utility of the FAST in aneurysm and EC-IC bypass surgery. In aneurysm surgery, flow measurements provide a quick and repeatable method to assess vessel patency and avoid complications related to vascular compromise. In bypass for flow augmentation in the setting of ischemia, flow measurements assess bypass patency and can assess the physiologic success of the bypass strategy. In bypass for flow replacement in the setting of planned vessel sacrifice, flow measurements can measure the flow deficit resulting from the vessel occlusion, assess the adequacy of in situ donor vessels, and confirm the ultimate adequacy of the bypass at the completion of surgery. The FAST strategy provides a useful approach to optimizing decision making and outcomes during cerebrovascular surgery.


Hydrogen sulfide attenuates ferroptosis and stimulates autophagy by blocking mTOR signaling in sepsis-induced acute lung injury.
Sepsis often leads to multiple organ failure or even death and is a significant health problem that contributes to a heavy economic burden. The lung is the first organ to be affected by sepsis. Presently, there is no specific drug or method to treat sepsis and sepsis-induced acute lung injury (ALI). H2S, along with CO and NO, is a physiological gas that acts as a signaling molecule and plays an active role in fighting various lung infections. GYY4137 is a novel H2S donor that is stable in vivo and in vitro. However, particularly in the context of ferroptosis, GYY4137 affects cecal ligation and puncture (CLP)-induced ALI by a mechanism that is not understood. Ferroptosis is a new form of cell necrosis. The primary mechanism is the accumulation of cellular lipid ROS in an iron-dependent manner. The principal objective of this project was to investigate the effects of GYY4137 on ferroptosis and autophagy in a mouse model of sepsis-induced ALI. We divided the experimental mice randomly into 5 groups: (1) sham group; (2) CLP group; (3) CLP + DMSO group: (4) CLP + GYY4137 (25 mg/kg) group; and (5) CLP + GYY4137 (50 mg/kg) group. (6) CLP + Rapamycin (2.0 mg/Kg) group. (7) CLP + Chloroquine (80 mg/Kg) group. (8) the Chloroquine (80 mg/Kg) + GYY (50 mg/Kg) group. The findings showed that GYY4137 significantly protected against CLP-induced ALI by improving sepsis-induced lung histopathological changes, diminishing lung tissue damage, ameliorating oxidative stress, and attenuating the severity of lung injury in mice. In this study, we found that GYY4137 could alleviate septicemia-induced ferroptosis in ALI by increasing the expression of GPx4 and SLC7A11 in lung tissue after CLP. One unexpected finding was the extent to which the levels of ferritin and ferritin light chain increased after CLP, which may be a compensatory mechanism for storing abnormally increased iron. We also found that the expression of p-mTOR, P62, and Beclin1 was significantly increased and that LC3II/LC3I declined after LPS stimulation, but the effect was inhibited by treatment with GYY4137, indicating that GYY4137 could inhibit the activation of autophagy in sepsis-induced ALI by blocking mTOR signaling.


Idiopathic left ventricular tachycardia originating from the mitral annulus.
Radiofrequency catheter ablation (RFCA) can eliminate most idiopathic repetitive monomorphic ventricular tachycardias (RMVTs) originating from the right and left ventricular outflow tracts (RVOT, LVOT). Here, we describe the electrophysiological (EP) findings of a new variant of RMVT originating from the mitral annulus (MAVT). MAVT was identified in 35 patients out of 72 consecutive left ventricular RMVTs from May 2000 to June 2004. All patients underwent an EP study and RFCA. The sites of origin of the MAVT were grouped into four groups according to the successful ablation sites around the mitral annulus. Group I included the anterior sites (n = 11), group II the anterolateral sites (n = 9), group III the lateral sites (n = 6), and group IV the posterior sites (n = 9). The MAVTs were a wide QRS tachycardia with a delta wave-like beginning of the QRS complex. The transitional zone of the R wave occurred between V1-V2 in all cases. The 12-lead electrocardiogram (ECG) pattern might reflect the site of the origin of MAVTs around the mitral annulus. We proposed an algorithm for predicting the site of the focus and the tactics needed for successful RFCA of the MAVT. We described the EP findings of the new variant of RMVT, MAVT. Most MAVTs could be eliminated by RF applications to the endocardial mitral annulus using our proposed tactics.


Oviductal epithelial cells transcriptome and extracellular vesicles characterization during thermoneutral and heat stress conditions in dairy cows.
In this study, the transcriptome of oviductal epithelial cells and certain characteristics of their extracellular vesicles of dairy cows were described under thermoneutral and heat stress conditions. Twenty cows were compared in springtime at THI = 65.6 ± 0.90 and in summertime at THI = 78.36 ± 2.73. During each season, the estrous cycles of the cows were synchronized, and on day 3 of the ensuing cycle, a blood sample was collected for progesterone determination, while their oviducts were collected after slaughter. Epithelial cells and oviductal fluid were collected from the oviduct ipsilateral and contralateral to the corpus, respectively. For the gene expression study, a comparative transcriptomic approach, using RNASeq, was performed on cells collected from the ipsilateral and the contralateral oviducts. The size and the concentration of extracellular vesicles (EVs) at both seasons were analyzed using Transmission Electron Microscopy and Nanoparticle tracking analysis and specific proteins were detected by Western blotting. Progesterone concentration was higher during the thermoneutral period. Between seasons, divergent expression of genes related to immune system, contractility, gamete protection and lncRNAs was found. The size and the concentration of the EVs did not differ between seasons, however, the concentration in the ipsilateral oviduct tended to be lower (p = 0.09) from the contralateral one in the summer, but not in the spring. Our results show for the first time that HS could be involved with alterations in the oviductal cells' gene expression and in the changes in concentration of EVs in the oviductal lumen. Our results imply that the altered oviductal environment during HS could be associated with the suppressed summer fertility in dairy cows.


Three years experience with a very low nitrogen diet supplemented with essential amino acids and keto-analogues in the treatment of chronic uraemia.
A low nitrogen, low phosphorus diet, supplemented with essential amino acids and keto-analogues, was administered to 48 chronic uraemics for a maximum of 36 months. In 10 cases renal function deteriorated and dialysis was started; eight patients changed to the dialytic therapy having difficulties in complying to the diet and three died for reasons not directly related to renal failure. The remaining twenty-seven patients are still on dietetic treatment and their renal function has not changed significantly. Their serum inorganic phosphorus and their circulating iPTH decreased significantly. Their subjective and objective conditions are satisfactory and no manifestation of protein malnutrition or other unwanted effects are detectable.