Comparison of Targeted Mass Spectrometry Techniques with an Immunoassay: A Case Study for HSP90α.
The objective of this study is to better understand factors governing the variability and sensitivity in SRM and PRM, compared to immunoassay. A 2D-LC-MS/MS-based SRM and PRM assay is developed for quantitative measurements of HSP90α in serum. Forty-three control sera are compared by SRM, PRM, and ELISA following the manufacturer's instructions. Serum samples are trypsin-digested and fractionated by strong cation exchange chromatography prior to SRM and PRM measurements. Analytical parameters such as linearity, LOD, LOQ, repeatability, and reproducibility of the SRM, PRM, and ELISA are determined. PRM data obtained by high-resolution MS correlate better with ELISA measurements than SRM data measured on a triple quadrupole mass spectrometer. While all three methods (SRM, PRM, and ELISA) are able to quantify HSP90α in serum at the ng mL-1 level, the use of PRM on a high-resolution mass spectrometer reduces variation and shows comparable sensitivity to immunoassay. Using fractionation, it is possible to measure ng mL-1 levels of HSP90α in a reproducible, selective, and sensitive way using PRM in serum. This opens up the possibility to use PRM in a multiplexed way as an attractive alternative for immunoassays without the use of antibodies or comparable binders.


[The peculiarities of the microwave in the frequency range of 51-52 GHz spectrum effects on E. coli cells].
The effects of microwaves on conformation of nucleoids in E. coli cells were studied by the method of anomalous viscosity time dependence (AVTD) at various frequencies in the range of 51-52 GHz and the power flux density of 100 microW/cm(2) . Linearly polarized microwaves resulted in significant effects within specific frequency windows of resonance type. The distances between frequency windows were in the range of 55-180 MHz. Only one of two possible circular polarizations, left-handed or right-handed, was shown to be effective at each frequency window. The sign of effective circular polarization alternated between frequency windows. We show that the effects of microwaves on E. coli cells as measured by the AVTD technique are not caused by adhesion of cells. The half-width of the 51.575 GHz resonance was measured to be 120+/-20 MHz. This value is very close to the half-width of the 51.755 GHz resonance as it has previously been determined at the same power flux density. The obtained data suggest similar targets for effects of microwaves at these two resonance frequencies and provide evidence for non-thermal nature of observed microwave effects.


Detection and localization of peptidases in Lactococcus lactis with monoclonal antibodies.
Monoclonal antibodies against peptidases of Lactococcus lactis were isolated and characterized: PEPN1-4 against a lysyl aminopeptidase PepN, PEPT1-5 against a tripeptidase PepT and PEPD1-3 against a dipeptidase PepD. These monoclonal antibodies reacted specifically with their respective antigens in crude cell extracts of Lc. lactis subspp. cremoris and lactis. A number of monoclonal antibodies cross reacted with proteins of other (lactic acid) bacteria. PEPT1, 2, 4 and 5 cross reacted weakly with a 35 kDa protein in Lactobacillus delbrueckii, while PEPT1 and PEPT2 reacted with proteins in the cell-free extract of Streptococcus thermophilus and Clostridium fervidus. Of the four isolated monoclonal antibodies against PEPN, only PEPN3 cross reacted weakly with a 90 kDa protein in Escherichia coli cell-free extract, and the other three antibody species against PEPN3 cross reacted with 80 kDa proteins of Lb. casei, Lb. delbrueckii, and Str. bovis, but not of Esch. coli. Of the three monoclonal antibodies against PepD, only PEPD1 and PEPD2 cross reacted with 40 kDa proteins of Lb. casei, Lb. delbrueckii and Str. bovis. All PEPN, PEPD and PEPT antibodies reacted with components in cell-free extracts of eleven different Lc. lactis strains, indicating that the peptidases of these strains were very similar to those of Lc. lactis subsp. cremoris WG2. However, Lc. lactis subsp. hordniae appeared to differ from the other Lc. lactis subspecies since only PEPT1, 2 and 5 reacted with a protein in the cell-free extract. Immunogold labelling of Lc. lactis WG2 with the isolated monoclonal antibodies revealed that PepN, PepD and PepT were located intracellularly. The intracellular location of these peptidases is discussed in relation to the supply of essential amino acids and peptides.


Dentists' knowledge, attitudes, and professional behavior toward the COVID-19 pandemic: A multisite survey of dentists' perspectives.
COVID-19 continues to affect the dental community worldwide. The authors conducted a cross-sectional electronic study using a multisite survey to examine dentists' knowledge, attitudes, and professional behavior toward COVID-19 and the affect on their livelihood. A questionnaire was circulated via e-mail and social media platforms to dentists in North America, Europe, Eastern Mediterranean, and Western Pacific regions. It covered demographic characteristics and questions about the COVID-19 outbreak related to dentists' levels of comfort for preventive and safety measures, provision of treatment, affect on work, and financial implications. Responses were tabulated and analyzed by means of χ2 or Fisher exact test. Mann-Whitney and Kruskal-Wallis tests were used to compare means. P < .05 was considered significant. A total of 1,251 dentists responded. General dentists represented 63.9% of the respondents and 62.5% worked in private practice. The global score for level of comfort with the preventive measures and provisions of treatment during the COVID-19 pandemic was low (14 of 30), and differences between regions were significant (P < .01); dentists working in private practice and general dental practitioners were less comfortable (P < .01). The respondents reported a lack of preparedness to confront a highly infectious respiratory disease. A new level of protective armamentarium in the dental operatory and updated operational guidance and policies are required, necessitating educators and regulators to ensure the delivery of knowledge and skills to oral health care providers. Dentists need to address the sustainability of their practices and have a robust business plan.


Density of functional Ca2+ release-activated Ca2+ (CRAC) channels declines after T-cell activation.
CRAC channel-mediated Ca(2+) entry plays a crucial role in T lymphocyte activation. Activated T cells display enhanced Ca(2+) signaling compared with resting T cells; this is partially attributed to activation-induced upregulation of CRAC channel expression. Orai and Stim family genes encode CRAC channel structural elements and regulatory proteins, respectively, but studies of their expression in T cells have led to controversial results. We re-examined Orai and Stim gene expression in resting, activated, and Jurkat T cells. Levels of Orai1 transcripts, encoding the human T cell CRAC channel subunit, were not significantly different between resting T and activated T cells. The total amount of all Orai transcripts was 2-fold higher in activated T cells than in resting T cells. Orai1 and total Orai transcript levels were significantly higher in Jurkat T cells than those in resting T cells. Stim expression did not vary significantly among cell types. Maximal whole-cell CRAC current amplitudes were 1.4-fold and 2.3-fold higher in activated and Jurkat T cells, respectively, than in resting T cells. Due to the small size of resting T cells, the surface CRAC channel density was 2.5-fold and 1.6-fold higher in resting T cells than in activated and Jurkat T cells, respectively. Predicted the rates of cytosolic Ca(2+) elevation calculated using the average values of CRAC channel currents and cell volumes showed that < 2-fold increase in the functional CRAC channel expression level cannot account for the enhanced rate of store-operated Ca(2+) entry in activated T cells compared with resting T cells.


Low folate in seminal plasma is associated with increased sperm DNA damage.
To determine associations between vitamin B status, homocysteine (tHcy), semen parameters, and sperm DNA damage. Observational study. A tertiary referral fertility clinic. Two hundred fifty-one men of couples undergoing in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI) treatment, with subgroups of fertile (n = 70) and subfertile men (n = 63) defined according to semen concentration and proven fertility. None. The DNA fragmentation index (DFI) as marker of sperm DNA damage determined using the sperm chromatin structure assay (SCSA), and semen parameters assessed according to World Health Organization criteria; tHcy, folate, cobalamin, and pyridoxine concentrations determined in seminal plasma and blood. In the total group of fertile and subfertile men, all biomarkers in blood were statistically significantly correlated with those in seminal plasma. No correlation was found between the biomarkers in blood and the semen parameters. In seminal plasma, both tHcy and cobalamin positively correlated with sperm count. Folate, cobalamin, and pyridoxine were inversely correlated with ejaculate volume. In fertile men, seminal plasma folate showed an inverse correlation with the DNA fragmentation index. Low concentrations of folate in seminal plasma may be detrimental for sperm DNA stability.


Technical note: Vetiver can grow on coal fly ash without DNA damage.
Fly ash is a by-product of coal-fired electricity generation plants. The prevalent practice of disposal is as slurry of ash and water to open lands or ash ponds located near power plants and this has lain to waste thousands of hectares all over the world. Wind and leaching are often the causes of off-site contamination from fly ash dumpsites. Vetiver (Vetiveria zizanioides) grown on fly ash for three months showed massive, mesh-like growth of roots which could have a phytostabilizing effect. The plant achieved this without any damage to its nuclear DNA as shown by comet assay done on the root nuclei, which implies the long-term survival of the plant on the remediation site. Also, when Vetiver is used for phytoremediation of coal fly ash, its shoots can be safely grazed by animals as very little of heavy metals in fly ash were found to be translocated to the shoots. These features make planting of Vetiver a practical and environmentally compatible method for restoration of fly ash dumpsites. Lack of DNA damage in Vetiver has been compared to that in a sensitive plant i.e. Allium cepa. Our results suggested that apart from traditional end-points viz. growth parameters like root length, shoot length and dry weight, comet assay could also be included in a battery of tests for initial, rapid and effective selection of plants for restoration and phytoremediation of polluted sites.


Male-female relatedness at specific SNP-linkage groups influences cryptic female choice in Chinook salmon (Oncorhynchus tshawytscha).
In a range of taxa, the relatedness between mates influences both pre- and post-mating processes of sexual selection. However, relatively little is known about the genetic loci facilitating such a bias, with the exception of the major histocompatibility complex. Here, we performed tightly controlled replicated in vitro fertilization trials to explore the impact of relatedness on two possible mechanisms of cryptic female choice (CFC) in Chinook salmon (Oncorhynchus tshawytscha). We tested (i) whether relatedness of mates, assessed using 682 single nucleotide polymorphisms (SNPs) on 29 SNP-linkage groups (LGs), biases a male's sperm velocity in ovarian fluid (a parameter previously shown to predict male fertilization success), and (ii) whether relatedness of mates governs fertilization success via other mechanisms, probably via sperm-egg interactions. We found that relatedness on three LGs explained the variation in sperm velocity, and relatedness on two LGs explained fertilization success, which might indicate the presence of genes important in sperm-ovarian fluid and sperm-egg interactions in these genomic regions. Mapping of the SNPs on these LGs to the rainbow trout genome revealed two genes that affect fertility in humans and represent candidate genes for further studies. Our results thereby provide a novel contribution to the understanding of the mechanism of CFC.


Absolute distance measurement by dispersive interferometry using a femtosecond pulse laser.
We describe an interferometric method that enables to measure the optical path delay between two consecutive femtosecond laser pulses by way of dispersive interferometry. This method allows a femtosecond laser to be utilized as a source of performing absolute distance measurements to unprecedented precision over extensive ranges. Our test result demonstrates a non-ambiguity range of ~1.46 mm with a resolution of 7 nm over a maximum distance reaching ~0.89 m.


The Adaptation to Mandarin-Accented English by Older, Hearing-Impaired Listeners Following Brief Exposure to the Accent.
Purpose The language processing of Mandarin-accented English (MAE) by older hearing-impaired (OHI), older normally hearing (NH), and younger NH listeners was explored. We examined whether OHI adults have more difficulty than NH listeners in recognizing and adapting to MAE speech productions after receiving brief training with the accent. Method Talker-independent adaptation was evaluated in an exposure training study design. Listeners were trained either by four MAE talkers or four Australian English talkers (control group) before listening to sentences presented by a novel MAE talker. Speech recognition for both the training sentences and the experimental sentences were compared between listener groups and between the training accents. Results Listeners in all three groups (OHI, older NH, younger NH) who had been trained by the MAE talkers showed higher odds of speech recognition than listeners trained by the Australian English talkers. The OHI listeners adapted to MAE to the same degree as the NH groups despite returning lower overall odds of recognizing MAE speech. Conclusions Older listeners with mild-to-moderate hearing loss were able to benefit as much from brief exposure to MAE as did the NH groups. This encouraging result suggests that OHI listeners have access to and can exploit the information present in a relatively brief sample of accented speech and generalize their learning to a novel MAE talker.


Anti-diabetic and antioxidant effect of cinnamon in poorly controlled type-2 diabetic Iraqi patients: A randomized, placebo-controlled clinical trial.
To determine the effect of cinnamon on fasting blood glucose, hemoglobin (Hb) A1c, and oxidative stress markers in poorly controlled type 2 diabetes. A total of 25 type 2 diabetic patients of both sexes, aged 49.1 ± 6.0, treated only with hypoglycemic agent sulfonylurea (glibenclamide) were randomly assigned to receive either 1 g of cinnamon or placebo daily for 12 weeks. A highly significant (P ≤ 0.001) reduction (10.12%) of fasting blood glucose level after 6 and 12 weeks of treatment 10.12% and 17.4%, respectively, compared to baseline value and to placebo group at corresponding duration. Meanwhile, the value of glycosylated Hb reduced in cinnamon treated group by (2.625%) and (8.25%) after 6 and 12 weeks, respectively, although this reduction was non-significant compared to baseline value. Concerning the oxidative stress markers, the level of serum glutathione showed highly significant (P ≤ 0.001) elevation after 12 weeks as compared to baseline value and placebo group at corresponding duration, malondialdehyde serum level decreased after treatment of diabetic patients with cinnamon resulted in highly significant (P ≤ 0.001) reduction after 6 and 12 weeks compared to placebo group, but when compared to baseline value, there is a (15%) reduction only after 12 weeks of treatment which was considered highly significant (P ≤ 0.001) change, Finally, administration of cinnamon to diabetic patients for 12 weeks resulted in significant (P ≤ 0.05) elevation of superoxide dismutase level. Intake of 1 g of cinnamon for 12 weeks reduces fasting blood glucose and glycosylated Hb among poorly controlled type 2 diabetes patients, as well as, there is improvement in the oxidative stress markers, indicating the beneficial effect of adjuvant cinnamon as anti-diabetic and antioxidant along with conventional medications to treat poorly controlled type 2 diabetes mellitus.


Methylation of tumor suppressor genes in ovarian cancer.
Aberrant methylation of gene promoter regions is one of the mechanisms for inactivation of tumor suppressor genes in human malignancies. In this study, the methylation pattern of 24 tumor suppressor genes was analyzed in 75 samples of ovarian cancer using the methylation-specific multiplex ligation-dependent probe amplification (MS-MLPA) assay. Of the 24 tumor suppressor genes examined, aberrant methylation was observed in 17. The three most frequently methylated genes were CDKN2B, CDH13 and RASSF1, followed by ESR1 and MLH1. Methylation frequencies ranged from 1.3% for CDKN2A, RARβ, CASP8, VHL and TP73 to 24% for CDKN2B. The corresponding normal DNA from each patient was also investigated. Methylation was detected in tumors, although not in normal tissues, with the exception of two samples, indicating aberrant methylation in tumors. Clear cell carcinoma samples exhibited a higher frequency of CDKN2B promoter hypermethylation compared to those of other histological types (P=0.05). Our data indicate that methylation of the CDKN2B gene is a frequent event in ovarian carcinogenesis and that analysis of only three genes is sufficient to detect the presence of methylation in 35% of ovarian cancer cases. However, more studies using a much larger sample size are needed to define the potential role of DNA methylation as a marker for ovarian cancer.


Optimising the proportion of selection candidates measured for feed intake for a beef cattle breeding objective that includes methane emissions.
Reducing daily methane production (DMP) via selection for lower estimated daily (pasture) feed intake (DFI) has the potential to be more cost effective than direct selection for DMP. Daily feed intake has a high heritability and high genetic correlation to DMP and has a potential lower cost of measurement. This study's main aim was to determine for a breeding nucleus the optimal proportion of randomly selected young male and female cattle in which to estimate DFI. This optimum proportion was determined by modeling the measurement costs and response to selection of Angus cattle on a (standard industry) Angus breeding index (ABI) augmented with DFI and DMP in a combined breeding objective (BO), but without DMP being measured. For the assumed herd structure and considering a 20 yr planning horizon, the highest net present value (NPV) occurred when 64% of males and no females were measured for DFI. The highest breakeven DFI test cost (A$41.51/head) and highest returns on investment (ROI) occurred when 36% of males and no females had DFI estimates. Higher ROI were achieved when all males had DFI estimates before any females had DFI estimates. There was a diminishing increase in rate of genetic gain when moving from 36% to 64% of males with DFI estimates, thus ROI decreased from 29.7% to 23.1%. When 36% of males had DFI estimates (and no females), herd DMP genetic gain was slightly positive as the DMP reduction per generation from male selection (-0.086) was more than offset by the DMP increase per generation from female selection (+0.110). The selection response for DMP only became negative when at least 40% of males had DFI estimates. Having 64% of males with DFI estimates resulted in a predicted genetic decrease in DMP (-0.018 kgCOe/head per yr), compared to an increase of 0.052 kgCOe/head per yr when no animals had DFI estimates. The optimum proportion of males with DFI estimates (36 to 64%) depends on the breeders attitude toward ROI and the value of genetic change for DMP. Sensitivity analysis showed that the economic value (EV), heritability and genetic variance of DFI had a higher impact on the NPV and ROI outcomes than parameters related to ABI and DMP, so future work should focus on obtaining robust estimates for DFI parameters. Higher EV for feed intake and DMP would result in higher percentages of animals being profitably measured for DFI, leading to larger reductions in DMP.


Polymerization-Incompetent Uromodulin in the Pregnant Stroke-Prone Spontaneously Hypertensive Rat.
The kidney is centrally involved in blood pressure regulation and undergoes extensive changes during pregnancy. Hypertension during pregnancy may result in an altered urinary peptidome that could be used to indicate new targets of therapeutic or diagnostic interest. The stroke-prone spontaneously hypertensive rat (SHRSP) is a model of maternal chronic hypertension. Capillary electrophoresis-mass spectrometry was conducted to interrogate the urinary peptidome in SHRSP and the control Wistar-Kyoto strain at three time points: prepregnancy and gestational days 12 and 18. The comparison within and between the Wistar-Kyoto and SHRSP peptidome at all time points detected 123 differentially expressed peptides (fold change >1.5; P<0.05). Sequencing of these peptides identified fragments of collagen α-chains, albumin, prothrombin, actin, serpin A3K, proepidermal growth factor, and uromodulin. Uromodulin peptides showed a pregnancy-specific alteration in SHRSP with a 7.8-fold (P<0.01) and 8.8-fold (P<0.05) increase at gestational days 12 and 18, respectively, relative to the Wistar-Kyoto. Further investigation revealed that these peptides belonged to the polymerization-inhibitory region of uromodulin. Two forms of uromodulin (polymerization competent and polymerization incompetent) were found in urine from both Wistar-Kyoto and SHRSP, where the polymerization-incompetent form was increased in a pregnancy-specific manner in SHRSP. Nifedipine-treated pregnant SHRSP showed only polymerization-competent uromodulin, indicating that calcium may be mechanistically involved in uromodulin polymerization. This study highlights, for the first time, a potential role of uromodulin and its polymerization in hypertensive pregnancy.


[Research progress on transcription factors and signal pathways involved in congenital esophageal atresia].
Congenital esophageal atresia is one of the serious birth defects. Identifying the etiology and mechanism of esophageal development can provide clues for the effective prevention and treatment of congenital esophageal atresia. Recent studies have shown that a variety of transcription factors and signaling pathways (including Wnt signaling pathway, bone morphogenetic protein signaling pathway, SHH signaling pathway, vascular endothelial growth factor signaling pathway) are involved in esophageal proliferation, differentiation and other processes, promoting the normal development of esophagus. Understanding the regulatory mechanisms during the normal development of esophagus can give reference for the prevention and treatment of congenital esophageal atresia. This article reviews the research progress on related transcription factors and signaling pathways in esophageal development.


Insulin-stimulated glucose transport in circulating mononuclear cells from nondiabetic and IDDM subjects.
The objectives of this study were 1) to evaluate glucose transport and its regulation by insulin in easily accessible human cells, 2) to investigate the glucose transporter isoforms involved, and 3) to establish whether a defect in glucose transport is associated with peripheral insulin resistance, which is common in insulin-dependent diabetes mellitus (IDDM) patients. We measured 2-deoxyglucose (2-DG) uptake in circulating mononuclear cells from 23 nondiabetic adults, 16 adults with IDDM, and 10 children with IDDM. Circulating mononuclear cells were separated from whole blood by Ficoll gradients and incubated with +/- 1 nM insulin. 2-DG uptake was measured after incubation with [3H]2-DG and cell separation through corn oil-phthalate. Cytochalasin B-inhibitable 2-DG uptake (basal and insulin stimulated) was higher in control than in IDDM subjects (P less than 0.001). Insulin significantly increased 2-DG uptake or 3-O-methylglucose uptake in both groups. Basal and insulin-stimulated 2-DG uptake was similar for adults and children with IDDM and did not correlate with age or body mass index in any group or disease duration, insulin dosage, or HbA1c in IDDM. In separated monocytes and lymphocytes, 2-DG uptake increased in response to insulin only in the monocyte population. Insulin dose-response curves indicated maximal stimulation of hexose uptake at 1-2 nM insulin for both control and diabetic subjects and demonstrated a significant decrease in maximal insulin response in the latter. Immunoblotting with specific antibodies revealed that circulating mononuclear cells and separated monocytes express the GLUT1 but not the GLUT4 isoform of the glucose transporter.(ABSTRACT TRUNCATED AT 250 WORDS)


Results of pars plana vitrectomy in closed-globe injuries.
To assess surgical and visual outcomes with pars plana vitrectomy (PPV) in closed-globe injuries resulting in anterior and posterior segment pathologies and evaluate the factors influencing the prognosis. Retrospective observational study. Forty-seven eyes of 47 consecutive patients who underwent PPV because of closed-globe injury between January 1992 and August 2003 were reviewed. Surgical and visual outcomes and prognostic factors were analyzed according to the Ocular Trauma Classification System. Preoperative visual acuity was less than 4/200 in 49% of eyes, classified as grades IV to V. After surgery, this ratio was 23% (P < .05). Final visual acuity was statistically significantly better in grade I (P = .0001), grade II (P = .002), and relative afferent pupillary defect-negative (P = .0001) injuries. Maculopathy was the most common adverse outcome influencing final visual acuity (32%), followed by secondary glaucoma (13%) and proliferative vitreoretinopathy (8%). PPV is a favorable treatment modality in severe closed-globe injuries. Assessment of injuries with respect to the Ocular Trauma Classification System seemed to predict visual outcomes in this series.


Strategies for overcoming chemotherapy resistance in enterohepatic tumours.
When considered together, enterohepatic tumours, i.e., those affecting the liver, the biliary tree and gallbladder and the intestine, constitute the first cause of death due to cancer. Although in many cases surgery and radiotherapy are efficacious, these therapeutic strategies cannot always be implemented. Moreover, even when the removal of tumours is possible, pre- and post-operative pharmacological adjuvant regimens are often needed. However, one important limitation to the use of cytostatic drugs to treat enterohepatic tumours is that they generally exhibit marked refractivity to currently available pharmacological approaches. In addition, most of them increase their chemoresistance during treatment. In view of the high refractivity of these tumours to anti-cancer drugs and the existence of undesirable side effects, both of which are drawbacks in the available chemotherapy, several novel therapeutic approaches have been devised. The purpose of the present review is to offer some insight into the different types of strategies that have already been evaluated and incorporated into clinical practice, such as therapies based on the use of molecular targets, as well as into the approaches that are still under experimental development, such as the chemosensitization of cancer cells, genetic manipulation of tumour or host cells, and cell-specific enhancement of intracellular concentrations of the active agent by efficient targeting of pro-drugs or by using inhibitors of efflux pumps.


A predictive control-based approach to networked Hammerstein systems: design and stability analysis.
In this paper, a predictive control-based approach is proposed for a Hammerstein-type system which is closed through some form of network. The approach uses a two-step predictive controller to deal with the static input nonlinearity of the Hammerstein system and a delay and dropout compensation scheme to compensate for the communication constraints in a networked control environment. Theoretical results are presented for the closed-loop stability of the system. Simulation examples illustrating the validity of the approach are also presented.


Microdroplet-enabled highly parallel co-cultivation of microbial communities.
Microbial interactions in natural microbiota are, in many cases, crucial for the sustenance of the communities, but the precise nature of these interactions remain largely unknown because of the inherent complexity and difficulties in laboratory cultivation. Conventional pure culture-oriented cultivation does not account for these interactions mediated by small molecules, which severely limits its utility in cultivating and studying "unculturable" microorganisms from synergistic communities. In this study, we developed a simple microfluidic device for highly parallel co-cultivation of symbiotic microbial communities and demonstrated its effectiveness in discovering synergistic interactions among microbes. Using aqueous micro-droplets dispersed in a continuous oil phase, the device could readily encapsulate and co-cultivate subsets of a community. A large number of droplets, up to ∼1,400 in a 10 mm × 5 mm chamber, were generated with a frequency of 500 droplets/sec. A synthetic model system consisting of cross-feeding E. coli mutants was used to mimic compositions of symbionts and other microbes in natural microbial communities. Our device was able to detect a pair-wise symbiotic relationship when one partner accounted for as low as 1% of the total population or each symbiont was about 3% of the artificial community.


Preparation and evaluation of (89)Zr-Zevalin for monitoring of (90)Y-Zevalin biodistribution with positron emission tomography.
To evaluate whether (89)Zr can be used as a PET surrogate label for quantification of (90)Y-ibritumomab tiuxetan ((90)Y-Zevalin) biodistribution and dosimetry before myeloablative radioimmunotherapy. Zevalin was labelled with (89)Zr by introducing N-succinyldesferal (N-sucDf) as a second chelate. For comparison of the in vitro stability of (89)Zr-Zevalin and (88)Y-Zevalin (as a substitute for (90)Y), samples were incubated in human serum at 37 degrees C up to 6 days. Biodistribution of (89)Zr-Zevalin and (88)Y-Zevalin was assessed at 24, 48, 72 and 144 h p.i. by co-injection in nude mice bearing the non-Hodgkin's lymphoma (NHL) xenograft line Ramos. The clinical performance of (89)Zr-Zevalin-PET was evaluated via a pilot imaging study in a patient with NHL, who had undergone [(18)F]FDG-PET 2 weeks previously. Modification of Zevalin with N-sucDf resulted in an N-sucDf-to-antibody molar ratio of 0.83+/-0.04. After radiolabelling and purification, the radiochemical purity and immunoreactivity of (89)Zr-Zevalin always exceeded 95% and 80%, respectively. (89)Zr-Zevalin showed the same stability in serum as (88)Y-Zevalin, with a radiochemical purity >95% during a period of 6 days. The co-injected (89)Zr-Zevalin and (88)Y-Zevalin conjugates showed a very similar biodistribution, except for liver and bone accumulation at 72 and 144 h p.i., which was significantly higher for (89)Zr than for (88)Y. PET images obtained after injection of (89)Zr-Zevalin showed clear targeting of all known tumour lesions. (89)Zr-Zevalin and (88)Y-Zevalin showed a very similar biodistribution in mice, implying that (89)Zr-Zevalin-PET might be well suited for prediction of (90)Y-Zevalin biodistribution in a myeloablative setting.


The MatTek story - how the three Rs principles led to 3-D tissue success!
MatTek Corporation has been working diligently for over 15 years to replace traditional animal-based toxicity and efficacy tests with alternative test methods based on human-cell derived, three-dimensional (3-D) tissue models. First discussed in detail by W.M.S. Russell and R.L. Burch 50 years ago in their book, The Principles of Humane Experimental Technique, and now fully integrated into forward-looking publications such as Toxicity Testing in the 21st Century: A Vision and a Strategy, the concept of replacing animals in test procedures with human cells and/or human cell-derived in vitro 3-D tissues is being embraced by the world's research scientists and toxicologists at an ever-increasing rate. 3-D in vitro models are being utilised not only for humanitarian reasons, but also because human 3-D tissues, in particular, produce more-physiologically relevant scientific data. Early on in MatTek's efforts to develop this alternative test method, senior management sought the assistance of experts within the in vitro testing and animal rights communities, to help define the specific in vitro human 3-D tissue products needed and navigate the regulatory landscape, especially in Europe where the replacement of animal-based testing with non-animal alternative test methods was well underway. MatTek was fortunate to receive that expert assistance on both fronts from Professor Michael Balls, who at that time was the newly-elected first director of ECVAM. In 1997, with the guidance and support of Professor Balls and others in the animal rights community, MatTek began the effort to validate several of its human 3-D tissue-based alternative test methods. Today, two MatTek human cell-derived 3-D tissue-based test methods are validated as full replacements for existing animal-based tests, with more tests in the validation pipeline. In addition, MatTek in vitro tissue models are in use worldwide by chemical, pharmaceutical and consumer product companies, as evidenced by citations in hundreds of patents and scientific articles from these industries. This article concludes with MatTek's thoughts on the direction that human 3-D tissue-based in vitro testing will take in the future.


Functional morphology of proximal hindlimb muscles in the frog Rana pipiens.
Musculoskeletal models have become important tools in understanding motor control issues ranging from how muscles power movement to how sensory feedback supports movements. In the present study, we developed the initial musculotendon subsystem of a realistic model of the frog Rana pipiens. We measured the anatomical properties of 13 proximal muscles in the frog hindlimb and incorporated these measurements into a set of musculotendon actuators. We examined whether the interaction between this musculotendon subsystem and a previously developed skeleton/joint subsystem captured the passive behavior of the real frog's musculoskeletal system. To do this, we compared the moment arms of musculotendon complexes measured experimentally with moment arms predicted by the model. We also compared sarcomere lengths measured experimentally at the starting and take-off positions of a jump with sarcomere lengths predicted by the model at these same limb positions. On the basis of the good fit of the experimental data, we used the model to describe the multi-joint mechanical effects produced by contraction of each hindlimb muscle and to predict muscle trajectories during a range of limb behaviors (wiping, defensive kicking, swimming and jumping). Through these analyses, we show that all hindlimb muscles have multiple functions with respect to accelerating the limb in its three-dimensional workspace and that the balance of functions depends greatly on limb configuration. In addition, we show that muscles have multiple, task-specific functions with respect to the type of contraction performed. The results of this study provide important data regarding the multifunctional role of hindlimb muscles in the frog and form a foundation upon which additional model subsystems (e.g. neural) and more sophisticated muscle models can be appended.


Erratum to: An improved assembly of the loblolly pine mega-genome using long-read single-molecule sequencing.
The 22-gigabase genome of loblolly pine (Pinus taeda) is one of the largest ever sequenced. The draft assembly published in 2014 was built entirely from short Illumina reads, with lengths ranging from 100 to 250 base pairs (bp). The assembly was quite fragmented, containing over 11 million contigs whose weighted average (N50) size was 8206 bp. To improve this result, we generated approximately 12-fold coverage in long reads using the Single Molecule Real Time sequencing technology developed at Pacific Biosciences. We assembled the long and short reads together using the MaSuRCA mega-reads assembly algorithm, which produced a substantially better assembly, P. taeda version 2.0. The new assembly has an N50 contig size of 25 361, more than three times as large as achieved in the original assembly, and an N50 scaffold size of 107 821, 61% larger than the previous assembly.


In vivo evaluation of a new surfactant polymer coating mimicking the glycocalyx of endothelial cells.
The purpose of this study was to demonstrate that a proprietary surfactant polymer (SP) coating does not adversely affect the hemodynamic performance of cardiopulmonary bypass (CPB) or gas exchange in oxygenators. The new coating was applied to a CPB circuit including cannulae, reservoir, oxygenator, and blood pump implanted into 12 pigs, divided into groups with either coated or noncoated pumps. CPB flow was maintained at a fixed level of approximately 2.4 L/min for 6 hours with full heparinization. Hemodynamic data and pump performance were recorded every hour, and blood samples were taken every 2 hours. After sacrifice, the CPB circuit and major organs were macroscopically examined. There was no significant difference in the oxygen transfer rate between the two groups. The coating did not adversely affect oxygenator inlet or outlet pressures. There was no significant difference between the two groups in microthrombi seen in the oxygenators. No thromboemboli were noted in the major organs on gross or histologic examination. In conclusion, this new SP coating did not decrease gas exchange performance, and its biocompatibility evaluations revealed no differences between coated and noncoated groups under aggressive heparin use.


Exploring coherent electron excitation and migration dynamics by electron diffraction with ultrashort X-ray pulses.
Exploring ultrafast charge migration is of great importance in biological and chemical reactions. We present a scheme to monitor attosecond charge migration in molecules by electron diffraction with spatial and temporal resolutions from ab initio numerical simulations. An ultraviolet pulse creates a coherent superposition of electronic states, after which a time-delayed attosecond X-ray pulse is used to ionize the molecule. It is found that diffraction patterns in the X-ray photoelectron spectra show an asymmetric structure, which is dependent on the time delay between the pump-probe pulses, encoding the information of molecular orbital symmetry and chemical bonding. We describe these phenomena by developing an electronic time-dependent ultrafast molecular photoionization model of a coherent superposition state. The periodical distortion of electron diffraction patterns illustrates the evolution of the electronic coherence, providing a tool for attosecond imaging of ultrafast molecular reaction processes.


Creatinine generation from kinetic modeling with or without postdialysis serum creatinine measurement: results from the HEMO study.
A convenient method to estimate the creatinine generation rate and measures of creatinine clearance in hemodialysis patients using formal kinetic modeling and standard pre- and postdialysis blood samples has not been described. We used data from 366 dialysis sessions characterized during follow-up month 4 of the HEMO study, during which cross-dialyzer clearances for both urea and creatinine were available. Blood samples taken at 1 h into dialysis and 30 min and 60 min after dialysis were used to determine how well a two-pool kinetic model could predict creatinine concentrations and other kinetic parameters, including the creatinine generation rate. An extrarenal creatinine clearance of 0.038 l/kg/24 h was included in the model. Diffusive cross-dialyzer clearances of urea [230 (SD 37 mL/min] correlated well (R2 = 0.78) with creatinine clearances [164 (SD 30) mL/min]. When the effective diffusion volume flow rate was set at 0.791 times the blood flow rate for the cross-dialyzer clearance measurements at 1 h into dialysis, the mean calculated volume of creatinine distribution averaged 29.6 (SD 7.2) L], compared with 31.6 (SD 7.0) L for urea (P < 0.01). The modeled creatinine generation rate [1183 (SD 463) mg/day] averaged 100.1 % (SD 29; median 99.3) of that predicted in nondialysis patients by an anthropometric equation. A simplified method for modeling the creatinine generation rate using the urea distribution volume and urea dialyzer clearance without use of the postdialysis serum creatinine measurement gave results for creatinine generation rate [1187 (SD 475) mg/day; that closely matched the value calculated using the formally modeled value, R2 = 0.971]. Our analysis confirms previous findings of similar distribution volumes for creatinine and urea. After taking extra-renal clearance into consideration, the creatinine generation rate in dialysis patients is similar to that in nondialysis patients. A simplified method based on urea clearance and urea distribution volume not requiring a postdialysis serum creatinine measurement can be used to yield creatinine generation rates that closely match those determined from standard modeling.


Neural responses to facial expressions support the role of the amygdala in processing threat.
The amygdala is known to play an important role in the response to facial expressions that convey fear. However, it remains unclear whether the amygdala's response to fear reflects its role in the interpretation of danger and threat, or whether it is to some extent activated by all facial expressions of emotion. Previous attempts to address this issue using neuroimaging have been confounded by differences in the use of control stimuli across studies. Here, we address this issue using a block design functional magnetic resonance imaging paradigm, in which we compared the response to face images posing expressions of fear, anger, happiness, disgust and sadness with a range of control conditions. The responses in the amygdala to different facial expressions were compared with the responses to a non-face condition (buildings), to mildly happy faces and to neutral faces. Results showed that only fear and anger elicited significantly greater responses compared with the control conditions involving faces. Overall, these findings are consistent with the role of the amygdala in processing threat, rather than in the processing of all facial expressions of emotion, and demonstrate the critical importance of the choice of comparison condition to the pattern of results.


Caring for the dentate elderly.
The proportional increase of elderly persons in most communities and increasing tooth retention among them betoken considerable change in gerodontic needs. Servicing dentate older patients will occupy progressively more practising time. This committment requires the development of somewhat different skills, knowledge and philosophy from those customarily propounded in dental teaching to date which has concentrated mostly on matters relevant to young and middle-aged persons. Dentists must become proficient in managing clinical entities of prevalence in the elderly. Xerostomia, a frequent, but often neglected symptom can have distressing consequences. Practitioners have a responsibility to detect it, analyse its likely cause or causes and select appropriate therapy. Root caries is re-emerging as a common finding. Its aetiology must be understood in order to institute effective preventive measures and treatment. Dentate elderly persons typically have heavily restored teeth subject to various iatrogenic problems. Some of the restorative work required can be provided exactly as for younger patients. However, optimum care is not necessarily the most elaborate. It is often desirable, when health or financial considerations dictate, to adopt expedients which reduce treatment extent and/or expense. Adhesive materials are extremely useful. The 'sandwich technique' for bonding composite resin to dentine via glass ionomer cement often facilitates the provision of extensive restorations in a minimally invasive fashion. Adhesive procedures can also be applied to treat attrition, mend fatigue fractures, splint teeth deficient in periodontal support, and provide simplified cosmetic treatment for elderly patients.


Activation of a Classic Hunger Circuit Slows Luteinizing Hormone Pulsatility.
The central regulation of fertility is carefully coordinated with energy homeostasis, and infertility is frequently the outcome of energy imbalance. Neurons in the hypothalamus expressing neuropeptide Y and agouti-related peptide (NPY/AgRP neurons) are strongly implicated in linking metabolic cues with fertility regulation. We aimed here to determine the impact of selectively activating NPY/AgRP neurons, critical regulators of metabolism, on the activity of luteinizing hormone (LH) pulse generation. We employed a suite of in vivo optogenetic and chemogenetic approaches with serial measurements of LH to determine the impact of selectively activating NPY/AgRP neurons on dynamic LH secretion. In addition, electrophysiological studies in ex vivo brain slices were employed to ascertain the functional impact of activating NPY/AgRP neurons on gonadotropin-releasing hormone (GnRH) neurons. Selective activation of NPY/AgRP neurons significantly decreased post-castration LH secretion. This was observed in males and females, as well as in prenatally androgenized females that recapitulate the persistently elevated LH pulse frequency characteristic of polycystic ovary syndrome (PCOS). Reduced LH pulse frequency was also observed when optogenetic stimulation was restricted to NPY/AgRP fiber projections surrounding GnRH neuron cell bodies in the rostral preoptic area. However, electrophysiological studies in ex vivo brain slices indicated these effects were likely to be indirect. These data demonstrate the ability of NPY/AgRP neuronal signaling to modulate and, specifically, reduce GnRH/LH pulse generation. The findings suggest a mechanism by which increased activity of this hunger circuit, in response to negative energy balance, mediates impaired fertility in otherwise reproductively fit states, and highlight a potential mechanism to slow LH pulsatility in female infertility disorders, such as PCOS, that are associated with hyperactive LH secretion.


The effects of nano-materials on the behaviors of sludge mortar specimens.
In this research, nano-composites are added to sewage sludge ash to create a mixture, which then replaces part of cement. Nano-composites are manufactured from pure quartzose sand. The influences of different amounts of nano-composites and sludge ash on mortar are evaluated. Cement, sludge ash (0%, 10%, and 20%), and nano-composites (0%, 0.5%, 1%, 2%, and 3%), which defined as the percent weight of cement and sludge ash, are mixed together in batches to make mortar specimens. Results show that the flowability of sludge ash mortar reduces with increasing amount of cement replaced and of nano-composites added. The compressive strength of mortar lowers when more amounts of cement are replaced by sludge ash, but increases with more quantity of nano-composites added. Moreover, the study shows that nano-composites can fortify the compressive strength of mortar. With the help of efficiency of compressive strength, nano-composites benefit most to the mortar with replacement of 10% sludge ash, followed by the substitution of 20% and 0%.


Phosphotyrosine-modified proteins are concentrated at the membranes of epithelial and endothelial cells during tissue development in chick embryos.
We have used high affinity polyclonal antibodies specific for phosphotyrosine (PTyr) residues to examine the localization in various chick embryonic tissues in situ of PTyr-modified proteins by immunocytochemical methods. During the period from 9 to 21 d of development, most tissues exhibit elevated levels of PTyr-modified proteins as determined by immunoblotting experiments of tissue extracts with the anti-PTyr antibodies (Maher, P. A., and E. B. Pasquale. 1988. J. Cell Biol. 106:1747-1755). By immunofluorescence labeling of semithin frozen sections, the highest concentrations of PTyr immunolabeling in all of the embryonic tissues examined were localized to the membranes of the epithelial and endothelial cells with other cells showing no detectable labeling. These results were confirmed by immunoelectron microscopic labeling, which showed particularly high concentrations of PTyr-modified proteins close to the membranes at the apical junctions. The corresponding adult tissues showed no labeling. It is proposed that these results reflect the molecular basis for the functional plasticity of epithelial and endothelial cell junctions during embryonic development.


The quantitative buffy coat capillary tubes versus thin and thick blood films in the diagnosis of malaria in Saudi Arabia.
The Quantitative Buffy Coat (QBC) capillary tube was compared with both the thin and thick films in the diagnosis of malaria among three groups of hospitalized patients. Group (1) comprised subjects with no symptoms or signs, group (2) comprised patients with fever and rigors of unknown cause, and group (3) comprised treated malarial patients consulting the hospital for a follow up. The QBC test showed more sensitivity than the two conventional methods in the three groups as follows: in group (1) the QBC was positive in 14.2% compared to 9% only in either thin or thick blood films. In group (2) the positivity was 95.1% compared to 79.3% & 76.8% in the thin and thick blood films respectively. In group (3) the sensitivity was 22.9% compared to only 5.7% in both of the thin and thick blood films. The authors found that the QBC technique in addition to its high sensitivity is very practical, reliable, simple and easy to perform. No doubt, the thin blood film still the method of choice in species identification of malaria parasites.


First detection of TR46/Y121F/T289A and TR34/L98H alterations in Aspergillus fumigatus isolates from azole-naive patients in Denmark despite negative findings in the environment.
Azole-resistant Aspergillus fumigatus harboring the TR34/L98H or TR46/Y121F/T289A alterations is increasingly found in Europe and Asia. Here, we present the first clinical cases of TR46/Y121/T289A and three cases of TR34/L98H outside the cystic fibrosis (CF) population in Denmark and the results of environmental surveys. Four patients (2012 to 2014) with 11 A. fumigatus and 4 Rhizomucor pusillus isolates and 239 soil samples (spring 2010 and autumn 2013, respectively) with a total of 113 A. fumigatus isolates were examined. Aspergillus isolates were screened for azole resistance using azole-containing agar. Confirmatory susceptibility testing was done using the EUCAST microbroth dilution EDEF 9.1 reference method. For relevant A. fumigatus isolates, CYP51A sequencing and microsatellite genotyping were performed. Three patients harbored TR34/L98H isolates. Two were azole naive at the time of acquisition and two were coinfected with wild-type A. fumigatus or R. pusillus isolates, complicating and delaying diagnosis. The TR46/Y121F/T289A strain was isolated in 2014 from a lung transplant patient. Genotyping indicated that susceptible and resistant Aspergillus isolates were unrelated and that no transmission between patients occurred. Azole resistance was not detected in any of the 113 soil isolates. TR34/L98H and TR46/Y121F/T289A alterations appear to be emerging in the clinical setting in Denmark and now involve azole-naive patients. Two recent soil-sampling surveys in Denmark were unable to indicate any increased prevalence of azole-resistant A. fumigatus in the environment. These findings further support the demand for real-time susceptibility testing of all clinically relevant isolates and for studies investigating the seasonal variation and ecological niches for azole-resistant environmental A. fumigatus.


Urokinase activity in corneal fibroblasts may be modulated by DNA damage and secreted proteins.
Proteases like urokinase-type plasminogen activator (uPA) play an important role in tumor invasion. Cells derived from ultraviolet radiation (UVR)-induced corneal sarcomas of Monodelphis domestica produce relatively high levels of uPA compared to the untransformed keratocytes suggesting a mechanism for their invasiveness. Because UVR is known to stimulate uPA production in many cell types, UVR exposure may further increase uPA expression in corneal tumor cells, thus enhancing their ability to infiltrate. We investigated control of basal uPA levels and the induction of uPA by UVR in transformed and untransformed corneal keratocytes from Monodelphis. These studies took advantage of the fact that Monodelphis possesses an active photolyase that can be stimulated to remove UVR-induced pyrimidine dimers by exposure to long-wavelength visible photoreactivating light (PRL). Our studies showed that significant induction of uPA occurred in response to 200 J/m2 UVR. This induction was partially blocked by treatment with PRL, indicating that DNA damage, the pyrimidine dimer in particular, played a role in uPA induction. In untransformed cultured corneal fibroblasts, the heparin-binding protein inhibitor, suramin, reduced basal uPA levels, UVR-induced uPA production and cell proliferation. Basic fibroblast growth factor, a heparin-binding growth factor known to be UVR-inducible in mesenchymal cells, stimulated uPA production and cell proliferation; however, anti-bFGF antibodies did not significantly decrease proliferation or basal uPA production. These findings suggested that basal levels of uPA secretion were modulated in response to heparin-binding growth factors and that these growth factors may also have mediated the effect of UVR on uPA levels.


[Characteristics of the carcinogenic effectiveness of 645 MeV protons and 60Co gamma-irradiation].
The carcinogenic effect of 645 MeV protons and 60Co-gamma-rays (1, 2 and 4 Gy) on mongrel female rats was comparatively studied. The observations were made throughout the entire life span of the exposed animals. Revealed were a similar biological effectiveness of the two types of radiation and a common nature of the carcinogenic transformations.


Empirical models of the electron temperature and density in the nightside venus ionosphere.
Empirical models of the electron temperature and electron density of the late afternoon and nightside Venus ionosphere have been derived from Pioneer Venus measurements acquired between 10 December 1978 and 23 March 1979. The models describe the average ionosphere conditions near 18 degrees N latitude between 150 and 700 kilometers altitude for solar zenith angles of 80 degrees to 180 degrees . The average index of solar flux was 200. A major feature of the density model is the factor of 10 decrease beyond 90 degrees followed by a very gradual decrease between 120 degrees and 180 degrees . The density at 150 degrees is about five times greater than observed by Venera 9 and 10 at solar minimum (solar flux approximately 80), a difference that is probably related to the effects of increased solar activity on the processes that maintain the nightside ionosphere. The nightside electron density profile from the model (above 150 kilometers) can be reproduced theoretically either by transport of 0(+) ions from the dayside or by precipitation of low-energy electrons. The ion transport process would require a horizontal flow velocity of about 300 meters per second, a value that is consistent with other Pioneer Venus observations. Although currently available energetic electron data do not yet permit the role of precipitation to be evaluated quantitatively, this process is clearly involved to some extent in the formation of the nightside ionosphere. Perhaps the most surprising feature of the temperature model is that the electron temperature remains high throughout the nightside ionosphere. These high nocturnal temperatures and the existence of a well-defined nightside ionopause suggest that energetic processes occur across the top of the entire nightside ionosphere, maintaining elevated temperatures. A heat flux of 2 x 10(10) electron volts per square centimeter per second, introduced at the ionopause, is consistent with the average electron temperature profile on the nightside at a solar zenith angle of 140 degrees .


Practical guide to the use of chemotherapy in castration resistant prostate cancer.
Chemotherapy, once thought to be toxic and ineffective in men with castration resistant prostate cancer (CRPC), has a significant impact on survival and quality-of-life in these patients. This article summarizes recent studies performed with two Food and Drug Administration (FDA) approved agents which have improved survival in men with CRPC, docetaxel and cabazitaxel. The literature on cytotoxic chemotherapy for castration resistant prostate cancer was reviewed. The individual efficacy, mechanisms of chemotherapeutic action, and appropriate disease states of administration were identified. Recent clinical trial results of chemotherapy combined with targeted agents was also reviewed. Front line cytotoxic therapy consists of docetaxel combined with prednisone. In two randomized trials, docetaxel based therapy demonstrated a 20%-24% improvement in survival over the palliative standard of care, mitoxantrone combined with prednisone. Eight randomized trials combining docetaxel/prednisone with other antiangiogenic, bone targeted, vaccine or metabolic therapies failed to demonstrate an improvement in survival over docetaxel alone. Cabazitaxel, an analogue of docetaxel which has activity in taxane resistant cell lines, is approved by the FDA, for use in CRPC patients who have previous exposure to docetaxel. Docetaxel combined with prednisone remains the standard of care as first line cytotoxic therapy for CRPC. Cabazitaxel is an effective second line cytotoxic agent that improves survival; studies are underway comparing cabazitaxel to docetaxel as first line chemotherapy. Given its lack of survival benefit, as well as the emergence of new treatments for prostate cancer, mitoxantrone has a diminished role in the treatment of CRPC.


(meso-5,5,7,12,12,14-Hexamethyl-1,4,8,11-tetra-aza-cyclo-tetra-deca-ne)nickel(II) bis-[O,O'-bis-(4-methyl-phen-yl) thio-phosphate].
In the centrosymmetric title complex, [Ni(C(16)H(36)N(4))](C(14)H(14)O(3)PS)(2), the Ni(II) ion is coordinated by four N atoms and two O atoms within a slightly distorted NiN(4)O(2) octa-hedral geometry. The asymmetric unit consits of one Ni(II) ion that is located on a center of inversion, half of the macrocylic ligand and one anion occupying general positions. Intra-molecular N-H⋯O and N-H⋯S hydrogen bonding is found between the macrocyclic ligand and the monothio-phosphate anion.


The role of school psychologists in the support of grieving children.
School psychologists are in a unique position to both identify and work with grieving students; to provide guidance to schools, families, and peers on approaches to support such children and youth; and to offer training to classroom educators so that they are better prepared to support grieving students. Yet, there has been minimal research published on this topic in professional journals. This special section of School Psychology Quarterly is devoted to grief and loss with the hope of beginning to narrow this gap in the literature. The three articles in this special section add to an evolving evidence-base that grief in children is common and the impact can be profound, that school professionals can and should play a major role in supporting grieving children, and that school psychologists can play a key role in empowering classroom educators and other school professionals so that no child in the future has to grieve alone. (PsycINFO Database Record


[Effects of asymmetric dimethyl hydrazine on physical and chemical properties of blood plasma if experimental animals].
Asymmetric dimethyl hydrazine (ADH), a highly toxic propellant component, is known to cause metabolic disturbances in humans and laboratory animals. Focus of this investigation was placed on crystal-forming properties of blood serum, and types of plasma protein destructions in ADH-intoxicated rats. Object of investigation was blood plasma collected from rats exposed to 100 mg/kg of per oral ADH total (5 mg/kg per a day, 5 days a wk, 4 wks.). The crystal-forming ability of plasma was assessed with the teziographic technique. Severity of proteins destruction was determined by the content of carbonyl products of oxidative protein modification in plasma. Chronic exposure to low ADH doses caused oxidative destruction of proteins in blood plasma and, consequently, significant degradation of the plasma ability to form crystals reflected in an altered zonal phase structure and phase bursting on teziograms. Symmetry gave way to pathological structures, e.g. petals and loops. Amorphous areas grew in size, bursting lines thinned out. Concretions became less numerous. These data suggest a close relationship between the impairments in physical and chemical properties of blood plasma of experimental animals by highly toxic asymmetric dimethyl hydrazine.


In vitro replication of mouse hepatitis virus strain A59.
An in vitro replication system for mouse hepatitis virus (MHV) strain A59 was developed using lysolecithin to produce cell extracts. In extracts of MHV-infected cells, radiolabeled UMP was incorporated at a linear rate for up to 1 h into RNA, which hybridized to MHV-specific cDNA probes and migrated in denaturing formaldehyde-agarose gels to the same position as MHV genomic RNA. The incorporation of [32P]UMP into genome-sized RNA in vitro correlated with the observed increase of [3H]uridine incorporation in MHV-infected cells labeled in vivo. Incorporation of [32P]UMP into genome-sized RNA was inhibited when extracts were incubated with puromycin. The addition to the assay of antiserum to the MHV-A59 nucleocapsid protein N inhibited synthesis of genome-sized RNA by 90% compared with the addition of preimmune serum. In contrast, antiserum to the E1 or E2 glycoproteins did not significantly inhibit RNA replication. In vitro-synthesized RNA banded in cesium chloride gradients as a ribonucleoprotein complex with the characteristic density of MHV nucleocapsids isolated from virions. These experiments suggest that ongoing protein synthesis is necessary for replication of MHV genomic RNA and indicate that the N protein plays an important role in MHV replication.


Fragment library screening identifies hits that bind to the non-catalytic surface of Pseudomonas aeruginosa DsbA1.
At a time when the antibiotic drug discovery pipeline has stalled, antibiotic resistance is accelerating with catastrophic implications for our ability to treat bacterial infections. Globally we face the prospect of a future when common infections can once again kill. Anti-virulence approaches that target the capacity of the bacterium to cause disease rather than the growth or survival of the bacterium itself offer a tantalizing prospect of novel antimicrobials. They may also reduce the propensity to induce resistance by removing the strong selection pressure imparted by bactericidal or bacteriostatic agents. In the human pathogen Pseudomonas aeruginosa, disulfide bond protein A (PaDsbA1) plays a central role in the oxidative folding of virulence factors and is therefore an attractive target for the development of new anti-virulence antimicrobials. Using a fragment-based approach we have identified small molecules that bind to PaDsbA1. The fragment hits show selective binding to PaDsbA1 over the DsbA protein from Escherichia coli, suggesting that developing species-specific narrow-spectrum inhibitors of DsbA enzymes may be feasible. Structures of a co-complex of PaDsbA1 with the highest affinity fragment identified in the screen reveal that the fragment binds on the non-catalytic surface of the protein at a domain interface. This biophysical and structural data represent a starting point in the development of higher affinity compounds, which will be assessed for their potential as selective PaDsbA1 inhibitors.


Circulating Tumor Cells as a Biomarker to Assist Molecular Diagnosis for Early Stage Non-Small Cell Lung Cancer.
Compared with tissue biopsy, liquid biopsy is the most preferable non-invasive promising method in personalized medicine, although it has many limitations in isolating circulating tumor cells (CTC). Lung cancer associated mortality is drastically increased due to a shortfall of early-stage detection, which remains a challenge. Herein, we aimed to detect lung cancer at an early-stage using CellCollector device. 39,627 volunteers underwent low-dose computed tomography; 2508 cases with pulmonary nodules and 7080 with no pulmonary nodules were chosen. After follow-up, 24 patients were diagnosed with early-stage non-small cell lung cancer (NSCLC), and subjected to CTC detection using CellCollector, along with 72 healthy volunteers. Immunofluorescence staining for EpCAM/CKs and CD45 were performed for CTC validation. Fifteen out of twenty-four (stage I, n = 18; stage II, n = 6) early-stage lung cancer patients were found to be CTC-positive, whereas no CTC was found in the control group. Genetic mutation of TP53, ERBB2, PDGFRA, CFS1R and FGFR1 in the CTC revealed 71.6% of the mutation sites similar to the tumor tissues of 13 patients. Molecular characterization revealed higher expression of protein PD-LI in CTC (40%) as compared to tumor tissue (26.7%). Moreover, CTC clusters were detected in 40% of patients. CTC detection using the CellCollector in early-stage NSCLC had a relative high capture rate. Moreover, CTC analysis is a prospective setting for molecular diagnostic in cases when tumor tissue biopsy is not desirable.


AffyGCQC: a web-based interface to detect outlying genechips with extreme studentized deviate tests.
Affymetrix GeneChip oligonucleotide arrays are dedicated to analyzing gene expression differences across distinct experimental conditions. Data production for such arrays is an elaborate process with many potential sources of variability unrelated to biologically relevant gene expression variations. Therefore, rigorous data quality assessment is fundamental throughout the process for downstream biologically meaningful analyses. We have developed a program named AffyGCQC, which is the acronym for a bioinformatics tool designed to perform Affymetrix GeneChip Quality Control. This program implements a graphical representation of QC metrics recommended by Affymetrix for GeneChip oligonucleotide array technology. Most importantly, it performs extreme studentized deviate statistical tests for the set of arrays being compared in a given experiment, thus providing an objective measure for outlier detection. AffyGCQC has been designed as an easy-to-use Web-based interface (online supplementary information: http://www.transcriptome.ens.fr/AffyGCQC/; contact: email@example.com).


[Glucoamylase immobilization on silochrome using gossypol].
The paper is concerned with conditions of glucoamylase binding with silanized silochrome using gossypol, dialdehyde isolated from cotton-plant. Kinetic properties of the immobilized enzyme are studied. The enzyme pH optimum does not change with immobilization and the temperature optimum is shifted from 50 degrees to 60 degrees C; a certain increase of the seeming Km is also observed. A high yield of the enzyme activity in immobilization evidences for the possibility of using gossypol as a binding agent in glucoamylase immobilization.


IDOMEN: An Extension of Infectious Disease Ontology for MENingitis.
In sub-Saharan African countries the prevention and control of epidemic diseases requires the improvement of the surveillance system for these diseases. Biomedical ontologies are a growing field that can improve health information systems. Indeed biomedical ontologies allow semantic support, data integration, automated reasoning. We are building a meningitis ontology to assist filtering messages relevant to meningitis domain on social media in order to predict a possible epidemic. Indeed, the messages filtered are used for data and event extraction that serve as input for a meningitis surveillance system. In this paper we focused on the modeling and formalization of different perspectives of the meningitis disease such as biological perspective, clinical perspective, epidemiological and public health perspective. This paper presents the three modules in the global Infection Disease Ontology for Meningitis (IDOMEN) and at the end, we illustrate a case of reasoning with our ontology.


Molecular characterization of severe and mild cases of influenza A (H1N1) 2009 strain from Argentina.
While worldwide pandemic influenza A(H1N1) pdm case fatality rate (CFR) was 0.4%, Argentina's was 4.5%. A total of 34 strains from mild and severe cases were analyzed. A full genome sequencing was carried out on 26 of these, and a partial sequencing on the remaining eight. We observed no evidence that the high CFR can be attributed to direct virus changes. No evidence of re-assortment, mutations associated with resistance to antiviral drugs, or genetic drift that might contribute to virulence was observed. Although the mutation D225G associated with severity in the latest reports from the Ukraine and Norway is not observed among the Argentine strains, an amino acid change in the area (S206T) surrounding the HA receptor binding domain was observed, the same previously established worldwide.


Amelioration of Type 2 Diabetes Using Four Strains of Lactobacillus Probiotics: Effects on Gut Microbiota Reconstitution-Mediated Regulation of Glucose Homeostasis, Inflammation, and Oxidative Stress in Mice.
This study aims to explore the preventive effects and underlying mechanisms of Lactobacillus fermentum CKCC1858 (CKCC1), L. fermentum CKCC1369 (CKCC2), Lactobacillus plantarum CKCC1312 (CKCC3), and Lactobacillus gasseri CKCC1913 (CKCC4) on high-fat diet combined with streptozotocin (HFD/STZ)-stimulated type 2 diabetes (T2D) in mice. Generally, the results indicated that most of the four probiotics reduced weight loss and liver and pancreas damage, significantly (p < 0.05) improved glucose metabolism by regulating glucagon-like peptide-1 (GLP-1), fasting glucose and insulin levels, and increasing expression of glucose transporters. Probiotics improved hyperlipemia, inflammation, and oxidative stress by reducing the secretion of blood lipids and proinflammatory cytokines, increasing antioxidant enzymes. Metagenomic results revealed that probiotics restored gut microbiota via enhancing (reducing) the relative abundance of beneficial bacteria (harmful bacteria) and altered specific metabolic pathways in T2D mice. CKCC1, CKCC3, and CKCC4 showed excellent effects compared to CKCC2. These results indicated that probiotics potentially prevented T2D, which is strain-specific.


Neurogenesis and exercise: past and future directions.
Research in humans and animals has shown that exercise improves mood and cognition. Physical activity also causes a robust increase in neurogenesis in the dentate gyrus of the hippocampus, a brain area important for learning and memory. The positive correlation between running and neurogenesis has raised the hypothesis that the new hippocampal neurons may mediate, in part, improved learning associated with exercise. The present review gives an overview of research pertaining to exercise-induced cell genesis, its possible relevance to memory function and the cellular mechanisms that may be involved in this process.


[The study of the multifactorial anthropogenic effect on the ecosystems of the industrial reservoirs of "Maiak" industrial complex].
The analysis of the ecological situation of the Southern Urals industrial water reservoirs of the nuclear fuel cycle enterprise, "Mayak" PA is represented. The study was held in the 80s - early 90s. The subjects of the study were: a cooling water reservoir--Kysyl-Tash Lake (R-2) as well as a radioactive waste storage reservoir (R-10). Irtyash Lake, which is a drinking water reservoir for the city of Ozyorsk and Alabuga and Kazhakul Lakes, located on the boundary of the Eastern Urals Radioactive Trace (EURT), were taken as control ones. Such water reservoirs as Irtyash, Kysyl-Tash and the waste storage reservoir (R-10) are incorporated into the Techa River basin; while Alabuga and Kazhakul Lakes are related to the interfluve between the Techa River and the Sinara River. The complex effect from such man--caused factors as radiation, chemical and thermal to water reservoirs' ecosystems was studied. Radionuclide specific activities of the major reservoir components (water, bottom sediments, and biological objects), cumulative stock and radiation doses to the biota were determined. Assessment of the condition of biological structures of individual reservoirs was performed. It was found that the long-term complex influence of radiation, thermal and chemical factors resulted in the formation of the unique technology-induced ecosystems being a part of "Mayak" PA process cycle. Radiation doses to the fish of the cooling water reservoir and the radioactive waste storage reservoir were experimentally estimated. These doses from the incorporated beta-emitters were not less then 2-3 Gy/year. The long-term complex influence of radiation and chemical factors didn't cause any irreversible changes either in the fish population or in the ecosystem. Water purity indicators like crayfish (Astacus leptodactilus Esch) and mollusk (Anodonta cygnea L.) were found in the cooling water reservoir. The comparative analysis of the ecological situation of the reservoirs carried out on the basis of several qualitative indicators and with the help of the formalized scoring system allowed determining that the optimum ecological conditions can be observed in Irtyash Lake. The quality of the environment of Alabuga Lake is slightly lower. The ecological conditions in Kysyl-Tash Lake are up to the standard, while in Kazhakul Lake they are lower than the standard. This is the result of the natural salinization of the ecosystem. The lowest indicator was obtained for the radioactive waste storage reservoir.


CRISPR-Cas gene-editing reveals RsmA and RsmC act through FlhDC to repress the SdhE flavinylation factor and control motility and prodigiosin production in Serratia.
SdhE is required for the flavinylation and activation of succinate dehydrogenase and fumarate reductase (FRD). In addition, SdhE is conserved in proteobacteria (α, β and γ) and eukaryotes. Although the function of this recently characterized family of proteins has been determined, almost nothing is known about how their genes are regulated. Here, the RsmA (CsrA) and RsmC (HexY) post-transcriptional and post-translational regulators have been identified and shown to repress sdhEygfX expression in Serratia sp. ATCC 39006. Conversely, the flagella master regulator complex, FlhDC, activated sdhEygfX transcription. To investigate the hierarchy of control, we developed a novel approach that utilized endogenous CRISPR (clustered regularly interspaced short palindromic repeats)-Cas (CRISPR associated) genome-editing by a type I-F system to generate a chromosomal point mutation in flhC. Mutation of flhC alleviated the ability of RsmC to repress sdhEygfX expression, whereas RsmA acted in both an FlhDC-dependent and -independent manner to inhibit sdhEygfX. Mutation of rsmA or rsmC, or overexpression of FlhDC, led to increased prodigiosin, biosurfactant, swimming and swarming. Consistent with the modulation of sdhE by motility regulators, we have demonstrated that SdhE and FRD are required for maximal flagella-dependent swimming. Together, these results demonstrate that regulators of both metabolism and motility (RsmA, RsmC and FlhDC) control the transcription of the sdhEygfX operon.


Anomalous expression of epithelial differentiation-determining GATA factors in ovarian tumorigenesis.
Tumor cells often appear in a deviant differentiated stage, and dedifferentiation is a hallmark of malignancy; however, the causative mechanism of the global changes in dedifferentiation is not understood. The GATA transcription factors function in cell lineage specification during embryonic development and organ formation. The transcriptional targets of the GATA factors in early embryonic development include Disabled-2 and collagen IV, markers for epithelial lineages. GATA-4 and GATA-6 are expressed strongly and are localized in the nucleus in ovarian surface epithelial cells in tissues or primary cell cultures. By immunohistochemistry, we found that 82% of the 50 tumors analyzed had lost GATA-6 function, either by a complete absence of expression or by cytoplasmic mislocalization. The frequent loss of GATA-6 was also confirmed in a panel of ovarian surface epithelial and tumor cell lines. Although GATA-4 is absent only in a small percentage (14%) of ovarian tumors, it is lost in the majority of established cell lines in culture. The loss of GATA-6 correlates with the loss of Disabled-2, collagen IV, and laminin, markers for epithelial cell types. Loss of GATA factors was also found in an in vitro model for spontaneous transformation of rat ovarian epithelial cells. Repression of GATA-6 by small interfering (si)RNA approach in cultured cells leads to dedifferentiation as indicated by the loss of Disabled-2 and laminin expression. Restoration of GATA factors expression by ectopic transfection suppresses cell growth and is incompatible with the maintenance of the cells in culture. However, restoration of GATA-4 and GATA-6 expression is not able to induce expression of endogenous Disabled-2 in tumor cells, suggesting that the loss of GATA factors and dedifferentiation are irreversible processes. In conclusion, we observed the inappropriate expression and cellular localization of the GATA transcription factors in ovarian tumor tissues and cancer cell lines, and we have demonstrated that down-regulation of GATA factor expression leads to dedifferentiation. We propose that alterations of GATA transcription factor expression and aberrant nucleocytoplasmic localization may contribute to the anomalous epithelial dedifferentiation of the ovarian tumor cells.


Regional differences in presentation characteristics, use of treatments and outcome of patients with cardiogenic shock: Results from multicenter, international registry.
Concurrent evidence about cardiogenic shock (CS) characteristics, treatment and outcome does not represent a global spectrum of patients and is therefore limited. The aim of this study was to investigate these regional differences. To investigate regional differences in presentation characteristics, treatments and outcomes of patients treated with all types of cardiogenic shock (CS) in a single calendar year on a multi-national level. Consecutive patients from 19 tertiary care hospitals in 13 countries with CS who were treated between January 1, 2018 and December 31, 2018 were enrolled in this study. In total, 699 cardiogenic shock patients were included in this study. Of these patients, 440 patients (63%) were treated in European hospitals and 259 (37%) were treated in Non-European hospitals. Female patients (P<0.01) and patients with a previous myocardial infarction (P=0.02) were more likely to present at Non-European hospitals; whereas older patients (P=0.01) and patients with cardiogenic shock due to acute heart failure (P<0.01) were more likely to present at European hospitals. Vasopressor use was more likely in Non-European hospitals (P=0.04), whereas use of mechanical circulatory support (MCS) was more likely in European hospitals (P<0.01). Despite adjustment for relevant confounders, 30-day in-hospital mortality risk was comparably high in CS patients treated in European vs. Non-European hospitals (hazard ratio 1.08, 95% CI 0.84-1.39, P=0.56). Despite marked heterogeneity in characteristics and treatment of CS patients, including fewer use of MCS but more frequent use of vasopressors in Non-European hospitals, 30-day in-hospital mortality did not differ between regions.


Inactivation of bacterial pathogens following exposure to light from a 405-nanometer light-emitting diode array.
This study demonstrates the susceptibility of a variety of medically important bacteria to inactivation by 405-nm light from an array of light-emitting diodes (LEDs), without the application of exogenous photosensitizer molecules. Selected bacterial pathogens, all commonly associated with hospital-acquired infections, were exposed to the 405-nm LED array, and the results show that both gram-positive and gram-negative species were successfully inactivated, with the general trend showing gram-positive species to be more susceptible than gram-negative bacteria. Detailed investigation of the bactericidal effect of the blue-light treatment on Staphylococcus aureus suspensions, for a range of different population densities, demonstrated that 405-nm LED array illumination can cause complete inactivation at high population densities: inactivation levels corresponding to a 9-log(10) reduction were achieved. The results, which show the inactivation of a wide range of medically important bacteria including methicillin-resistant Staphylococcus aureus, demonstrate that, with further development, narrow-spectrum 405-nm visible-light illumination from an LED source has the potential to provide a novel decontamination method with a wide range of potential applications.


Targeted Isolation of Indole Alkaloids from Streptomyces sp. CT37.
Four compounds (1-4) were isolated from the extracts of Streptomyces sp. CT37 using bioassay in conjunction with mass spectrometric molecular networking (MN) driven isolation. Their complete structures were established by high-resolution electrospray ionization mass spectrometry (HR-ESIMS), and 1D and 2D nuclear magnetic resonance (NMR) data. Legonimide 1 was identified as a new alkaloid containing a rare linear imide motif in its structure, while compounds 2-4 were already known and their structures were elucidated as 1H-indole-3-carbaldehyde, actinopolymorphol B, (2R,3R)-1-phenylbutane-2,3-diol, respectively. The biosynthetic pathways of 1-4 were proposed based on the reported biogenesis of indole alkaloids in literature. Bioactivity tests for 1 and 2 revealed moderate growth inhibition activity against Candida albicans ATCC 10231 with MIC95 values of 21.54 µg/mL and 11.47 µg/mL, respectively.


Serum insulin-like growth factor I: tumor marker or etiologic factor? A prospective study of prostate cancer among Finnish men.
Recent epidemiological studies suggest an association between higher blood levels of insulin-like growth factor I (IGF-I) and increased risk of prostate cancer. We evaluated the association between prediagnostic levels of IGF-I and insulin-like growth factor binding protein 3 (IGFBP-3) and prostate cancer risk in a nested case-control study within the Alpha-Tocopherol, Beta-Carotene Cancer Prevention Study. Within the same cohort (using different cases and controls who had sequential serum samples available) we also examined changes in serum IGF-I and IGFBP-3 levels over time by case status. The risk association study included incident prostate cancer cases (n = 100) diagnosed at least 5 years after baseline blood draw (range, 5-12 years; median 9 years) and frequency-matched (4:1) controls. The sequential serum study included all of the prostate cancer cases (n = 21) with prediagnostic (2-3 years before diagnosis) and diagnostic serum available, and pair-matched controls (1:1). An ELISA was used to quantitate serum levels of IGF-I and IGFBP-3 for both studies. The association between IGF-I or IGFBP-3 and prostate cancer risk was assessed using conditional logistic regression, and paired t tests were used to evaluate case-control differences in change in serum analytes over time. We found no significant association between either IGF-I or IGFBP-3 and prostate cancer risk. In a multivariate analysis, we observed an odds ratio of 0.52 (95% confidence interval, 0.23-1.16) for the fourth versus the first quartile of serum IGF-I. Serum IGF-I, but not IGFBP-3, increased significantly over time in cases (18% increase) but not controls (4% decrease; P = 0.02). In contrast to previous reports, we found no evidence to support a causal association between serum IGF-I or IGFBP-3 and the risk of prostate cancer. It is possible that serum IGF-I may be serving as a tumor marker rather than an etiologic factor in prostate cancer.


Human, Animal and Plant Health Benefits of Glucosinolates and Strategies for Enhanced Bioactivity: A Systematic Review.
Glucosinolates (GSs) are common anionic plant secondary metabolites in the order Brassicales. Together with glucosinolate hydrolysis products (GSHPs), they have recently gained much attention due to their biological activities and mechanisms of action. We review herein the health benefits of GSs/GSHPs, approaches to improve the plant contents, their bioavailability and bioactivity. In this review, only literature published between 2010 and March 2020 was retrieved from various scientific databases. Findings indicate that these compounds (natural, pure, synthetic, and derivatives) play an important role in human/animal health (disease therapy and prevention), plant health (defense chemicals, biofumigants/biocides), and food industries (preservatives). Overall, much interest is focused on in vitro studies as anti-cancer and antimicrobial agents. GS/GSHP levels improvement in plants utilizes mostly biotic/abiotic stresses and short periods of phytohormone application. Their availability and bioactivity are directly proportional to their contents at the source, which is affected by methods of food preparation, processing, and extraction. This review concludes that, to a greater extent, there is a need to explore and improve GS-rich sources, which should be emphasized to obtain natural bioactive compounds/active ingredients that can be included among synthetic and commercial products for use in maintaining and promoting health. Furthermore, the development of advanced research on compounds pharmacokinetics, their molecular mode of action, genetics based on biosynthesis, their uses in promoting the health of living organisms is highlighted.


Early sleep duration trajectories and children's cognitive development: a prospective cohort study.
We aimed to investigate the association between sleep duration trajectories and cognitive performance in preschool-aged Chinese children. We included 2131 children from the Ma'anshan birth cohort (MABC) study. Sleep duration trajectories from 6 to 48 months of age were determined using the group-based trajectory modeling (GBTM). Children's intellectual development was assessed using the Wechsler Preschool and Primary Scale of Intelligence. Compared to those with a medium total sleep duration trajectory, children with a short total sleep duration trajectory had poorer cognitive performance on the Visual Spatial Index (VSI) (β = -3.65; 95% CI = -6.77 to -0.53), which was associated with an increased risk of a low full-scale intelligence quotient (FSIQ) (OR = 1.60; 95% CI = 1.02 to 2.51). The short total sleep duration trajectory was associated with a low VSI compared with both the medium total sleep duration trajectory and the long total sleep duration trajectory. Compared to children with normal nighttime sleep duration and normal daytime sleep duration trajectories, children with short nighttime sleep and long daytime sleep duration trajectories, normal nighttime sleep and long daytime sleep duration trajectories, and short nighttime sleep and normal daytime sleep duration trajectories all had lower cognitive performance. The restricted cubic spline (RCS) also showed that children with and appropriate total sleep duration, an adequate nighttime sleep duration, and a moderate daytime sleep duration had higher FSIQ. The results of this study emphasize that a medium total sleep duration, adequate sleep at nighttime, and appropriate sleep in the daytime appear to be more beneficial for children's cognitive development. • Sleep duration in infancy is strongly associated with neurocognitive development. • Medium and long total sleep duration trajectories are beneficial for children's cognitive performance compared to the short total sleep duration trajectory. • A medium total sleep duration, adequate sleep at nighttime and appropriate sleep in the daytime appear to be more beneficial for children's cognitive development.


Three, two, one! Revision of the long-bodied sphaerodorids (Sphaerodoridae, Annelida) and synonymization of Ephesiella, Ephesiopsis and Sphaerodorum.
Long-bodied sphaerodorids (Annelida, Sphaerodoridae) is the common name for members of the three closely and morphologically homogenous currently accepted genera of benthic marine bristle worms: Ephesiella, Ephesiopsis and Sphaerodorum. Members of this group share the presence of two dorsal and longitudinal rows of macrotubercles with terminal papillae, and two longitudinal rows of microtubercles, features that are unique among sphaerodorids. Genera are distinguished by the chaetae morphology. Members of Ephesiella are characterised by having compound chaetae (except, sometimes, simple chaetae in the first chaetigers), Sphaerodorum bear only simple chaetae, and Ephesiopsis have both compound and simple chaetae in all parapodia. Mitochondrial (partial COI and 16S rDNA) and nuclear (partial 18S rDNA and 28S rDNA) sequence data of long-bodied sphaerodorids with compound and simple chaetae, and an outgroup of additional seven sphaerodorid species were analysed separately and in combination using Bayesian inference (BA), and Maximum Likelihood (ML) methods. Long-bodied sphaerodorids from around the world (including type specimens) were examined under a range of optical equipment in order to evaluate putative generic and specific diagnostic features, in addition to intraspecific variability. Phylogenetic analyses of mitochondrial and nuclear DNA sequences of specimens identified as Ephesiella and Sphaerodorum, based on chaeta morphology, were performed. Sphaerodorum and Ephesiella were recovered as paraphyletic and nested within each other. Revision of current nominal species diagnostic features are performed and discussed. Results contradict current generic definitions. Recovery of paraphyletic compound and simple chaetae clades urge the synonymization of these two genera of long-bodied sphaerodorids. Morphological data also suggest the synonymization of Ephesiopsis.


Addition of carvedilol to University Wisconsin solution improves rat steatotic and nonsteatotic liver preservation.
Here we examine the effect of adding carvedilol (CVD) to University of Wisconsin (UW) solution on the preservation of steatotic and nonsteatotic livers during cold ischemia and after normothermic reperfusion. We used an isolated perfused rat liver model. The following protocols were evaluated. Protocol 1 concerned the effect of CVD after cold ischemia. Steatotic and nonsteatotic livers were preserved for 24 hours in UW solution alone or with CVD. Livers without cold ischemia were used as controls. Transaminases were evaluated in the flushing effluent. Protocol 2 involved the effect of CVD after reperfusion. Both liver types were preserved for 24 hours in UW solution alone or with CVD and then perfused ex vivo for 2 hours at 37 degrees C. Livers flushed and perfused without ischemia were used as controls. Hepatic injury and functionality [transaminases, bile production, and hepatic clearance of sulfobromophthalein (BSP)] were evaluated after reperfusion. In addition, factors potentially involved in hepatic ischemia-reperfusion injury, including oxidative stress (malondialdehyde and superoxide anion levels), mitochondrial damage (glutamate dehydrogenase activity), microcirculatory disorders (flow rate and vascular resistance), and adenosine triphosphate (ATP) depletion, were evaluated after reperfusion. After cold ischemia, steatotic livers preserved in UW solution showed higher transaminase levels than nonsteatotic livers. After reperfusion, steatotic livers preserved in UW solution showed higher transaminase levels and lower bile production and BSP clearance than nonsteatotic livers. Alterations in the perfusion flow rate and vascular resistance, mitochondrial damage, and reduced ATP content were more evident in steatotic livers preserved in UW solution. The addition of CVD to UW solution reduced hepatic injury, obstructed its mechanisms, and improved hepatic functionality in both liver types. We conclude that CVD is a useful additive for UW solution that improves the preservation of steatotic and nonsteatotic livers subjected to prolonged cold ischemia.


The p65 subunit is responsible for the strong transcription activating potential of NF-kappa B.
The nuclear form of the NF-kappa B transcription factor binds to DNA as a heterodimer of a 50 kDa (p50) and 65 kDa (p65) polypeptide. The two polypeptides are encoded by different genes but share a long region of homology, the NRD motif, encompassing domains required for DNA binding and dimerization. In this study we have analysed the contribution of the two subunits to the strong transactivating potential of NF-kappa B. Transient expression of the p65 subunit alone resulted in a potent transactivation of a CAT reporter construct under the control of two NF-kappa B binding sites in monkey COS and mouse L cells. The strongly DNA binding p50 subunit showed only very weak, if any, induction of gene expression. Co-expression of p50 suppressed the transactivation by p65 presumably by competitive DNA binding of transcriptionally inactive p50 dimers (KBF1). Fusion of p65 sequences to DNA binding domain of the yeast GAL4 transcription factor allowed detection of the principal transactivation domain of p65 (TA1) in the C-terminal 30 amino acid sequence. TA1 is likely to adopt an amphipathic alpha-helical structure which clusters serine residues on the hydrophilic surface, a structural feature conserved between human, mouse and Xenopus p65. The unique C-terminal third of p65 contained at least one more activation domain, TA2, within a 90 amino acid sequence directly adjacent to TA1. In two mammalian cell lines, TA1 and TA2 acted separately, while in an insect cell line, the two domains were inactive after their separation. Our study suggests that the p50 subunit in NF-kappa B might only serve a helper function in DNA binding whereas the p65 subunit is responsible for initiating transcription. Homodimers of p50 seem to have the potential of down-regulating kappa B-specific gene expression.


A review of factors influencing litter size in Irish sows.
Many factors influence litter size. These include genetics, gilt management, lactation length, parity distribution, disease, stress and boar fertility. In the past 20 years, litter size in Irish sows has increased by only one pig. Born alive figures now average at 11.2 pigs per litter. In this regard, Ireland is falling behind our European competitors who have made significant advances over this time. Denmark, for example, has an average figure of 12.7 pigs born alive per litter and France an average of 12.5. The single area that could be improved immediately is sow feeding. It is important that sows are fed correctly throughout pregnancy. If over-fed during pregnancy, sows will have depressed appetite during lactation. If underfed in pregnancy, sows will be too thin at farrowing. The correct way to feed a pregnant sow is to match her feed allocation to her requirement for maintenance, body growth and growth of her developing foetuses. During lactation, sows should be given as much feed as they can eat to prevent excessive loss of body condition. Liquid-feed curves should be such that lactating sows are provided with a minimum mean daily feed supply of 6.2 kg. A small proportion of sows will eat more and this could be given as supplementary dry feed. Where dry feeding is practised in the farrowing house, it is difficult to hand-feed sows to match their appetite. Ideally ad libitum wet/dry feeders should be used. From weaning to service, sows should once again be fed ad libitum. If liquid feeding, this means giving at least 60 MJ DE (digestible energy) per day during this period. If dry feeding, at least 4 kg of lactation diet should be fed daily. The effort spent perfecting sow feeding management on units should yield high dividends in the form of increased pigs born alive per litter.


Sites of Basal cell carcinomas and head and neck congenital clefts: topographic correlation.
The embryologic fusion planes might be related with the sites of onset of basal cell carcinoma (BCC), thus supporting an embryologic role for its pathogenesis. A study involving 495 patients with 627 BCCs of the head and neck was carried out over a period of 5 years by correlating the distribution of all BCCs with the sites of congenital clefts of the head and neck using (1) the original anatomic diagram of the Tessier classification of craniofacial clefts, (2) the anatomic diagram by Moore et al featuring the paths of the "hairline indicators" of craniofacial clefts that represent the cranial extensions of the Tessier classification, and (3) an anatomical diagram featuring the sites of congenital clefts of the neck. The proportion of BCCs localized within a cleft site was significantly higher than those in the noncleft sites. The age of patients with BCCs localized within the Tessier cleft number 3 was the lowest among all cleft regions. A topographic correspondence between the sites of BCCs and the sites of congenital clefts was demonstrated in the head and neck. This evidence would support the hypothesis of an embryologic role for the pathogenesis of BCC. The existence of clusters of embryological stem cells in the sites of fusion and/or merging of embryonic processes might therefore be proposed. There may be special biology/physiology along these cleft lines that predispose BCC formation.