Pseudomonas aeruginosa outer membrane permeability: isolation of a porin protein F-deficient mutant.
A mutant of Pseudomonas aeruginosa severely deficient in outer membrane protein F levels was isolated by screening heavily mutagenized strains for membrane protein alterations on sodium dodecyl sulphate-polyacrylamide gel electrophoresis. To provide a basis for phenotypic comparison, three independent spontaneous revertants with normal protein F levels were isolated. Neither the protein F-deficient mutant nor its revertants had gross surface alterations as judged by their sensitivities to 31 phages with diverse receptors and their low degrees of leakage of periplasmic beta-lactamase into the supernatant. Outer membrane permeability was measured in whole cells by examining the rates of hydrolysis of a chromogenic beta-lactam, nitrocefin, by periplasmic RP1-encoded beta-lactamase. It was found that the outer membrane permeabilities of wild-type and protein F revertant strains were similar, but low when compared with those of Escherichia coli and an antibiotic-supersusceptible mutant Z61 of P. aeruginosa. The loss of protein F caused a further significant decrease in outer membrane permeability. The results suggest that protein F is a pore-forming protein in vivo and that only a small proportion, as few as 1 in 400, of the protein F molecules form active functional channels in vivo.


The cardiovascular risk profile of middle age women previously diagnosed with premature ovarian insufficiency: A case-control study.
Cardiovascular disease (CVD) is the leading cause of death in women worldwide. The cardiovascular risk profile deteriorates after women enter menopause. By definition, women diagnosed with premature ovarian insufficiency (POI) experience menopause before 40 years of age, which may render these women even more susceptible to develop CVD later in life. However, prospective long-term follow up data of well phenotyped women with POI are scarce. In the current study we compare the CVD profile and risk of middle aged women previously diagnosed with POI, to a population based reference group matched for age and BMI. We compared 123 women (age 49.0 (± 4.3) years) and diagnosed with POI 8.1 (IQR: 6.8-9.6) years earlier, with 123 population controls (age 49.4 (± 3.9) years). All women underwent an extensive standardized cardiovascular screening. We assessed CVD risk factors including waist circumference, BMI, blood pressure, lipid profile, pulse wave velocity (PWV), and the prevalence of diabetes mellitus, metabolic syndrome (MetS) and carotid intima media thickness (cIMT), in both women with POI and controls. We calculated the 10-year CVD Framingham Risk Score (FRS) and the American Heart Association's suggested cardiovascular health score (CHS). Waist circumference (90.0 (IQR: 83.0-98.0) versus 80.7 (IQR: 75.1-86.8), p < 0.01), waist-to-hip ratio (0.90 (IQR: 0.85-0.93) versus 0.79 (IQR: 0.75-0.83), p < 0.01), systolic blood pressure (124 (IQR 112-135) versus 120 (IQR109-131), p < 0.04) and diastolic blood pressure (81 (IQR: 76-89) versus 78 (IQR: 71-86), p < 0.01), prevalence of hypertension (45 (37%) versus 21 (17%), p < 0.01) and MetS (19 (16%) versus 4 (3%), p < 0.01) were all significantly increased in women with POI compared to healthy controls. Other risk factors, however, such as lipids, glucose levels and prevalence of diabetes were similar comparing women with POI versus controls. The arterial stiffness assessed by PWV was also similar in both populations (8.1 (IQR: 7.1-9.4) versus 7.9 (IQR: 7.1-8.4), p = 0.21). In addition, cIMT was lower in women with POI compared to controls (550 μm (500-615) versus 684 μm (618-737), p < 0.01). The calculated 10-year CVD risk was 5.9% (IQR: 3.7-10.6) versus 6.0% (IQR: 3.9-9.0) (p = 0.31) and current CHS was 6.1 (1.9) versus 6.5 (1.6) (p = 0.07), respectively in POI versus controls. Middle age women with POI presented with more unfavorable cardiovascular risk factors (increased waist circumference and a higher prevalence of hypertension and MetS) compared to age and BMI matched population controls. In contrast, the current study reveals a lower cIMT and similar 10-year cardiovascular disease risk and cardiovascular health score. In summary, neither signs of premature atherosclerosis nor a worse cardiovascular disease risk or health score were observed among middle age women with POI compared to population controls. Longer-term follow-up studies of women of more advanced age are warranted to establish whether women with POI are truly at increased risk of developing CVD events later in life. ClinicalTrials.gov Identifier: NCT02616510.


Clinical disease upregulates expression of CD40 and CD40 ligand on peripheral blood mononuclear cells from cattle naturally infected with Mycobacterium avium subsp. paratuberculosis.
CD40 and CD40 ligand (CD40L) have costimulatory effects as part of a complex series of events in host immunity. In this study, the expression of CD40 and CD40L on peripheral blood mononuclear cells (PBMCs) isolated from cattle with Johne's disease were measured on freshly isolated PBMCs and on cells cultured for 8, 24, and 72 h in the presence or absence of live Mycobacterium avium subsp. paratuberculosis and exogenous gamma interferon, interleukin 10, and transforming growth factor β. Results demonstrated greater CD40 and CD40L expression on fresh PBMCs obtained from animals in the clinical stage of disease (symptomatic) than those from healthy control animals or cows in the subclinical stage of disease (asymptomatic). A similar expression profile with greater magnitude was noted for cultured PBMCs, with increased CD40 expression after 8 and 24 h of culture and increased CD40L expression between 24 and 72 h on PBMCs obtained from clinically infected animals. The addition of live M. avium subsp. paratuberculosis to cell cultures resulted in downregulation of CD40L expression in naturally infected cows, regardless of the disease stage. In contrast, the addition of live M. avium subsp. paratuberculosis to cultures resulted in upregulation of CD40 expression on cells obtained from clinically infected animals, while a decrease in expression was noted for healthy and subclinically infected cows. No effects of exogenous cytokines on CD40 or CD40L expression were observed. These results clearly point for the first time to a disparity in the expression of these costimulatory molecules on immune cells from cattle in different stages of Johne's disease and suggest further investigation into their roles in paratuberculosis pathogenesis.


Topical meclocycline sulfosalicylate, benzoyl peroxide, and a combination of the two in the treatment of acne vulgaris.
One hundred and six patients with acne vulgaris of the face were treated for 10 weeks with either topical meclocycline sulfosalicylate, topical benzoyl peroxide or both preparations. A randomized, double-blind parallel group study was used. Benzoyl peroxide proved more effective than meclocycline in reducing acne lesion counts, while local side effects were more common in the benzoyl peroxide-treated patients. The combined treatment was of intermediate efficacy with fewer local side effects.


Essential requirement of I-A region-identical host bone marrow or bone marrow-derived cells for tumor neutralization by primed L3T4+ T cells.
The antitumor activity of Meth A-hyperimmunized BALB/c mouse spleen cells (Meth A-Im-SPL) was assayed by the Winn test in H-2 incompatible bone marrow chimeras in closed colony CD-1 (nu/nu), inbred DDD/1(nu/nu) (H-2s), or inbred BALB/c(nu/nu) (H-2d) mice as recipients. We found that Meth A-Im-SPL suppressed Meth A growth in the chimera nude mice which were reconstituted with bone marrow cells of the H-2d haplotype (i.e., BALB/c, DBA/2 and B10.D2), but not in the chimeras which were reconstituted with bone marrow cells of the H-2a, H-2b, or H-2k haplotype (i.e., B10.A, B10, and B10.BR). These results suggested that H-2 restriction occurred between Meth A-Im-SPL and bone marrow or bone marrow-derived cells in tumor neutralization. Furthermore, Meth A-Im-SPL did not suppress Meth 1 tumors (antigenically distinct from Meth A tumors) in the presence or absence of mitomycin C-treated Meth A in a Winn assay. These results suggested that there is tumor specificity in the "effector phase" as well as in the "induction phase". The phenotype of the effectors in the Meth A-Im-SPL was Thy-1.2+ and L3T4+, because Meth A-Im-SPL lost their antitumor activity with pretreatment with anti-Thy-1.2 monoclonal antibody (mAb) and complement or anti-L3T4 mAb and complement, but not with anti-Lyt-2.2 mAb and complement or complement alone. Positively purified L3T4+ T cells from Meth A-Im-SPL (Meth A-Im-L3T4), obtained by the panning method, suppressed the tumor growth in the chimera nude mice which were reconstituted with bone marrow cells of B10.KEA2 mice (that were I-A region-identical with Meth A-Im-L3T4 cells but not others in H-2) as well as B10.D2 cells (that were fully identical with Meth A-Im-L3T4 cells in H-2). We conclude that Meth A-Im-SPL (L3T4+) neutralized the tumors in collaboration with I-A region-identical host bone marrow or bone marrow-derived cells, and the neutralization was not accompanied by the "bystander effect."


Potential Risk of Significant N2O Emission without Changing NOx Conversion on Commercial V2O5/TiO2 Catalyst under Working Conditions.
Vanadium-based catalysts play a pivotal role in the emission control of industrial NOx via selective catalytic reduction (SCR) technology. However, little attention has been paid to the potential emission of greenhouse gas N2O under complex working conditions. This work reports that a commercial V2O5/TiO2 catalyst may lead to significant N2O emission without greatly changing the outlet NOx concentration after chromium (Cr) deposition. With a Cr loading of 2 wt %, N2O concentration increased from 27.8 to 199.2 ppm at 350 °C with the value of outlet N2O/(N2O+N2) from 2.5% to 19.4%. Experimental results combined with DFT+U calculations suggest that nonselective catalytic reduction (NSCR) is the main route for N2O formation in a wide temperature range of 250 ∼ 400 °C. It is stemmed from the fact that the covalent interaction between Cr and V species on the V2O5/TiO2 surface accelerates the conversion of V4+ + Cr6+ → V5+ + Cr3+, leading to a larger proportion of surface V5+. More importantly, surface V5+ is highly related to the redox property of the V2O5/TiO2 catalyst, which is beneficial to NSCR reaction rather than the standard SCR process. The work suggests that to better inhibit the emission of greenhouse gases during the NH3-SCR process, monitoring N2O emission should be included along with the NOx concentrations, especially in complex flue gases.


Response of frugivorous primates to changes in fruit supply in a northern Amazonian forest.
Few attempts have been made to understand how spatiotemporal changes in fruit supply influence frugivores in tropical forests. The marked spatiotemporal variation in fruit supply can affect frugivore abundance and distribution, but studies addressing the effects of this variation on primates are scarce. The present study aimed to investigate how the spatiotemporal distribution of fruits influences the local distribution of three frugivorous primates in the eastern part of the Maracá Ecological Station, a highly seasonal Amazonian rainforest. Specifically, it was hypothesised that primate distribution will track changes in fruit supply, resulting that sites with high fruit availability should be heavily used by primates. During a 1-year study, fruit supply (ground fruit surveys) and primate density (line-transects) were monitored in twelve 2 km-long transects at monthly intervals. Fruit supply varied seasonally, being low during the dry season. The density of Ateles belzebuth was positively related to fruit supply during fruit shortage, but Cebus olivaceus and Alouatta macconnelli did not follow the same pattern. The supply of Sapotaceae fruit was an important component determining local distribution of A. belzebuth during the overall fruit shortage. Highly frugivorous primates such as A. belzebuth respond to seasonal decline in fruit supply by congregating at places with high fruit supply in this forest, particularly, those with many individuals of species of Sapotaceae. This study underscores the importance of small-scale spatiotemporal changes of fruit supply as a key component of frugivorous primate ecology in highly seasonal environments.


Latent inhibition and overswitching in schizophrenia.
Attention disorders in schizophrenia are manifested in two different ways. On the one hand, the schizophrenia patient tends to keep a learned response even after it ceases to be relevant (perseveration). On the other hand, the schizophrenia patient tends to replace an adaptive response without being given a reason to do so (overswitching). In the present study, overswitching was investigated in relation to latent inhibition (LI), which is the normal ability to ignore nonrelevant stimuli. A new tool--the Combined Attention Test--was used for this purpose in a group of 41 unmedicated schizophrenia patients, divided into subgroups of patients with predominantly positive and negative symptoms, and 24 normal controls. The results show that positive schizophrenia patients, who exhibited high levels of overswitching, also revealed impaired LI, while the negative schizophrenia group, as well as normal controls, exhibited intact LI. These findings suggest that overswitching is a specific attention deficit in positive schizophrenia. We discuss the possibility that impaired LI is a consequence of overswitching and comment on the putative neurophysiology.


Anterior development in the parthenogenetic and viviparous form of the pea aphid, Acyrthosiphon pisum: hunchback and orthodenticle expression.
In the dipteran Drosophila, the genes bicoid and hunchback work synergistically to pattern the anterior blastoderm during embryogenesis. bicoid, however, appears to be an innovation of the higher Diptera. Hence, in some non-dipteran insects, anterior specification instead relies on a synergistic interaction between maternally transcribed hunchback and orthodenticle. Here we describe how orthologues of hunchback and orthodenticle are expressed during oogenesis and embryogenesis in the parthenogenetic and viviparous form of the pea aphid, Acyrthosiphon pisum. A. pisum hunchback (Aphb) mRNA is localized to the anterior pole in developing oocytes and early embryos prior to blastoderm formation - a pattern strongly reminiscent of bicoid localization in Drosophila. A. pisum orthodenticle (Apotd), on the other hand, is not expressed prior to gastrulation, suggesting that it is the asymmetric localization of Aphb, rather than synergy between Aphb and Apotd, that regulates anterior specification in asexual pea aphids.


[Advance in glycolipid biosurfactants--mannosylerythritol lipids].
Mannosylerythritol lipids (MELs), mainly produced by Ustilago and Pseudozyma, are surface active compounds that belong to the glycolipid class of biosurfactants. MELs have potential application in food, pharmaceutical and cosmetics industries due to their excellent surface activities and other peculiar bioactivities. In recent years, the research field of MELs has regained much attention abroad. However, MELs are rarely studied in China. In this review, the producing microorganisms and production conditions, diverse structures, biochemical properties, structure-function relationship and biosynthetic pathways of MELs are described. Some research problems and prospects are summarized and discussed as well.


Microdosimetric investigations on collimated fast neutron beams for radiation therapy: II. The problem of radiation quality and RBE.
The correlation between radiation quality and cellular RBE of a mixed n-gamma field produced by a collimated fast-neutron beam produced by the bombardment of 14 MeV deuterons on Be in a water phantom is investigated. The microdosimetric parameters yF and YD are used for a physical description of the local distribution of radiation components. On the basis of the dose fractions of these components, the dependence of RBE on depth is calculated and is compared with experimental RBE data. Outside the beam centre, it is shown that the microdosimetric parameter y can be used for estimates of local changes of RBE, if particular conditions for the absorbed dose and for the range of neutron energies are met. The spatial distributions of y for the mixed radiation changes in RBE for equal absorbed doses of 1 Gy are at maximum 10% and thus perhaps negligible.


Changing paradigms in management of metastatic Castration Resistant Prostate Cancer (mCRPC).
Recently, the standard of care for metastatic Castration Resistant Prostate Cancer (mCRPC) has changed considerably. Persistent androgen receptor (AR) signaling has been identified as a target for novel therapies and reengages the fact that AR continues to be the primary target responsible for metastatic prostate cancer. Androgen receptor gene amplification and over expression have been found to result in a higher concentration of androgen receptors on tumor cells, making them extremely sensitive to low levels of circulating androgens. Additionally, prostate cancer cells are able to maintain dihydrotestosterone (DHT) concentration in excess of serum concentrations to support tumor growth. For many years ketoconazole was the only CYP17 inhibitor that was used to treat mCRPC. However, significant toxicities limit its use. Newly approved chemotherapeutic agents such as Abiraterone (an oral selective inhibitor of CYP17A), which blocks androgen biosynthesis both within and outside the prostate cancer cells), and enzalutamide (blocks AR signaling) have improved overall survival. There are also ongoing phase III trials for Orteronel (TAK- 700), ARN- 509 and Galeterone (TOK-001), which targets androgen signaling. In this review, we will present the rationale for the newly approved hormonal treatments, their indications and complications, and we will discuss ongoing trials that are being done to improve the efficacy of the approved agents. Finally, we will talk about the potential upcoming hormonal treatments for mCRPC.


Computer model for glucose-limited growth of a single cell of Escherichia coli B/r-A.
A computer model is described which is capable of predicting changes in cell composition, cell size, cell shape, and the timing of chromosome synthesis in response to changes in external glucose limitation. The model is constructed primarily from information on unrestricted growth in glucose minimal medium. The ability of the model to make reasonable quantitative predictions under glucose-limitation is a test of the plausibility of the basic biochemical mechanisms included in the model. Such a model should be of use in differentiating among competing hypotheses for biological mechanisms and in suggesting as yet unobserved phenomena. The last two points are illustrated with the testing of a mechanism for the control of the initiation of DNA synthesis and predictions on cellwidth variations during the division cycle.


Degradation and rearrangement of a lung surfactant lipid at the air-water interface during exposure to the pollutant gas ozone.
The presence of unsaturated lipids in lung surfactant is important for proper respiratory function. In this work, we have used neutron reflection and surface pressure measurements to study the reaction of the ubiquitous pollutant gas-phase ozone, O3, with pure and mixed phospholipid monolayers at the air-water interface. The results reveal that the reaction of the unsaturated lipid 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine, POPC, with ozone leads to the rapid loss of the terminal C9 portion of the oleoyl strand of POPC from the air-water interface. The loss of the C9 portion from the interface is accompanied by an increase in the surface pressure (decrease in surface tension) of the film at the air-water interface. The results suggest that the portion of the oxidized oleoyl strand that is still attached to the lipid headgroup rapidly reverses its orientation and penetrates the air-water interface alongside the original headgroup, thus increasing the surface pressure. The reaction of POPC with ozone also leads to a loss of material from the palmitoyl strand, but the loss of palmitoyl material occurs after the loss of the terminal C9 portion from the oleoyl strand of the molecule, suggesting that the palmitoyl material is lost in a secondary reaction step. Further experiments studying the reaction of mixed monolayers composed of unsaturated lipid POPC and saturated lipid dipalmitoyl-sn-glycero-3-phosphocholine, DPPC, revealed that no loss of DPPC from the air-water interface occurs, eliminating the possibility that a reactive species such as an OH radical is formed and is able to attack nearby lipid chains. The reaction of ozone with the mixed films does cause a significant change in the surface pressure of the air-water interface. Thus, the reaction of unsaturated lipids in lung surfactant changes and impairs the physical properties of the film at the air-water interface.


Swelling of the vesicle is prerequisite for PTH secretion.
Unlike most secretory cells, high extra cellular calcium inhibits rather than stimulates hormonal secretion in several cells such as parathyroid cells, Juxtaglomerular cells and osteoclast. To gain further insight into the common but unique stimulus-secretion coupling mechanism in these cells, bovine parathyroid slices were incubated in various conditions of Krebs-Ringer (KR) solution containing essential amino acids. Parathyroid cells showed the inverse dependency of secretion on extra cellular calcium concentration as we expected. Ammonium acetate overcame the inhibitory effect of 2.5 mM of calcium and the maximum effect was as much as the five times of the basal value, while there was a little additive effect under 0 mM CaCl2. PTH secretion was biphasic according to the change of extra cellular osmolarity and the lowest response was observed at 300 mOsm/l. In Na-rich KR solution, high concentration of nigericin (> 10(-4)M) completely overcame the inhibitory effect of 2.5 mM CaCl2 and the maximum stimulatory effect was 8 times greater whereas it was only 2 times greater without CaCl2. In K-rich KR solution that abolished the K-gradient between the extra cellular solution and the cytoplasm, the rate of PTH secretion increased, and furthermore the addition of nigericin increased the rate of secretion significantly. The results above suggested that the osmotic swelling of the secretory vesicle in parathyroid cells might promote exocytosis as in Juxtaglomerular cells. We propose that the swelling of the vesicle is also prerequisite for secretion in several cells inhibited paradoxically by Ca++, whatever the signal transduction pathway for swelling of the secretory granules induced by the lowering of Ca++ in cytoplasm are.


Internet-delivered applied relaxation for vasomotor symptoms in postmenopausal women: lessons from a failed trial.
Internet-delivered therapies have a short history and promising results have been shown for several health problems, particularly for psychiatric conditions. This study was a first attempt to evaluate whether Internet-delivered applied relaxation for hot flushes in postmenopausal women may be useful. Due to a high drop-out rate the study was prematurely terminated after inclusion of approximately two thirds of calculated women. The Internet-delivered applied relaxation must probably be modified for such populations and settings before it can be used further. This article will discuss the benefits and pitfalls to learn in order to meet the challenges of future studies. Clinical trial registration number: NCT01245907.


History of Keratinocyte Carcinoma and Risk of Melanoma: A Prospective Cohort Study.
The association between history of keratinocyte carcinoma (KC, also known as nonmelanoma skin cancer) and risk of developing invasive melanoma has not been assessed comprehensively using prospective data. We followed 91 846 women in the Nurses' Health Study (NHS; 1984-2010), 114 918 women in the NHSII (1989-2011), and 48 946 men in the Health Professionals Follow-up Study (1986-2010) for diagnoses of KC and melanoma biennially. Cox proportional hazards models were used to compute hazard ratios (HRs) and 95% confidence intervals (CIs) of melanoma associated with history of KC, basal cell carcinoma (BCC), and squamous cell carcinoma (SCC). All statistical tests were two-sided. We documented 1949 melanomas, 38 842 BCCs, and 7462 SCCs over 6.4 million person-years of follow-up. After adjustment for other risk factors, a personal history of KC was associated with an increased risk of melanoma (meta-analysis HR = 2.22, 95% CI = 1.73 to 2.85). The association was more apparent among participants with a history of both BCC and SCC (HR = 3.40, 95% CI = 1.60 to 7.19) than among participants with a history of BCC only (HR = 2.20, 95% CI = 1.80 to 2.70) or SCC only (HR = 1.56, 95% CI = 0.98 to 2.46), and there was a strong risk-increasing trend associated with a higher number of reported KCs removed by surgery ( P trend < .001). In women, KC history was more strongly associated with head/neck melanomas (HR = 4.17, 95% CI = 2.77 to 6.27) than with trunk or limb melanomas (both HRs < 2.50, P heterogeneity = .04). Our results provide novel insights for the relationship between KC history and risk of developing melanoma, which may be important for melanoma prevention.


Phloretin flavonoid exhibits selective antiproliferative activity in doxorubicin-resistant gastric cancer cells by inducing autophagy, inhibiting cell migration and invasion, cell cycle arrest and targeting ERK1/2 MAP pathway.
Studies have shown that Phloretin exerts anticancer effects on several types of cancer cells. Nonetheless, the anticancer effects of Phloretin have not been fully explored against the human gastric cancer cells. Therefore, this study was undertaken to evaluate the anticancer effects of Phloretin against the human gastric cancer cells. Cell proliferation was evaluated by WST-1 assay while cell cycle analysis was carried out by flow cytometry. The effects on cell migration and invasion were evaluated by wound healing assay and transwell assays, respectively. Electron microscopy and western blot methods were used to study effects on autophagy and ERK1/2/MAPK signalling pathway. The results showed that Phloretin inhibited the proliferation rate of the human SNU-1 gastric cancer cells and showed an IC50 of 18 µM. However, Phloretin showed very high IC50 (80 µM) against the normal GES-1 normal gastric cells. Electron microscopy showed that Phloretin triggered autophagy in the SNU-1 gastric cancer cells which was accompanied by enhancement in the expression of LC3B II and Beclin 1. Cell cycle analysis showed that Phloretin caused accumulation of the SNU-1 cells in the G0/G1 phase of the cell cycle triggering G0/G1 cell cycle arrest. The G0/G1 arrest of SNU-1 cells was also associated with depletion of cyclin D1 and D2 expression. Wound healing and transwell assays showed that Phloretin suppressed the migration of the SNU-1 gastric cancer cells, suggestive of the anti-metastatic potential of this molecule. Finally, this molecule also blocked the ERK1/2/MAPK signalling pathway in SNU-1 cells in a concentration-dependent manner. Phloretin may prove beneficial as a promising drug candidate for gastric cancer treatment provided further studies are carried out on it, especially toxicological studies.


A generic approach for the design of whole-genome oligoarrays, validated for genomotyping, deletion mapping and gene expression analysis on Staphylococcus aureus.
DNA microarray technology is widely used to determine the expression levels of thousands of genes in a single experiment, for a broad range of organisms. Optimal design of immobilized nucleic acids has a direct impact on the reliability of microarray results. However, despite small genome size and complexity, prokaryotic organisms are not frequently studied to validate selected bioinformatics approaches. Relying on parameters shown to affect the hybridization of nucleic acids, we designed freely available software and validated experimentally its performance on the bacterial pathogen Staphylococcus aureus. We describe an efficient procedure for selecting 40-60 mer oligonucleotide probes combining optimal thermodynamic properties with high target specificity, suitable for genomic studies of microbial species. The algorithm for filtering probes from extensive oligonucleotides libraries fitting standard thermodynamic criteria includes positional information of predicted target-probe binding regions. This algorithm efficiently selected probes recognizing homologous gene targets across three different sequenced genomes of Staphylococcus aureus. BLAST analysis of the final selection of 5,427 probes yielded >97%, 93%, and 81% of Staphylococcus aureus genome coverage in strains N315, Mu50, and COL, respectively. A manufactured oligoarray including a subset of control Escherichia coli probes was validated for applications in the fields of comparative genomics and molecular epidemiology, mapping of deletion mutations and transcription profiling. This generic chip-design process merging sequence information from several related genomes improves genome coverage even in conserved regions.


Variations in the origin of superior laryngeal artery.
The superior laryngeal artery is the principal artery supplying the laryngeal mucosa, musculature, and glands. Knowledge of variations in the origin of superior laryngeal artery could prove to be very useful during reconstructive surgeries of the larynx, partial laryngectomy, laryngeal transplantation, and also during procedures like super-selective intra-arterial chemotherapy for laryngeal and hypolaryngeal cancers. However, relatively few studies have been done on the superior laryngeal artery in comparison to its clinical importance. The present study was aimed at documenting the prevalence of variable origin of the superior laryngeal artery within the carotid triangle. Sixty hemi-necks obtained from 30 South Indian cadavers were dissected and studied for variations in the origin of superior laryngeal artery. It was observed that the superior laryngeal artery took origin from superior thyroid in 91.7% cases. Variable origin from the external carotid artery was noted in 5% cases. The superior laryngeal artery was found to arise from the lingual artery in one case alone (1.7%). In addition to the above findings, a very rare variation of superior laryngeal artery arising from the ascending pharyngeal (1.7%) was also observed in the hemi-neck of one cadaver. All the variations that were observed were unilateral and on the left side. These findings may help provide further insight to the anatomists, radiologists and surgeons and can help improve performances during surgical manipulations of the larynx.


Wood-boring bivalves, opportunistic species in the deep sea.
Wood exposed for 104 days at a depth of 1830 meters at the permanent station of the research submersible D.S.R.V. Alvin was completely riddled by two species of bivalve wood borers (subfamily Xylophagainae, family Pholadidae). Their high reproductive rate, high population density, rapid growth, early maturity, and utilization of a transient habitat classify them as opportunistic species, the first recorded from the deep sea. Xylophaga is shown to be the most important species involved in decomposing woody plant material in the deep sea.


Physico-chemical and Biological Properties of a New Portland Cement-based Root Repair Material.
To propose bismuth carbonate, a radiopacifying agent, as a new endodontic root repair material that was added to Portland cement (PC) at 2 wt%, 5 wt%, 10 wt% and 15 wt%, and physicochemical and biological properties of each formulation were evaluated in comparison to MTA-Angelus. Mixed and powder samples were analysed by scanning electron microscopy (SEM) and X-ray power diffraction (XRD), and the semiquantitative constitution of the powder was determined by energy-dispersive spectrometer (EDS). Setting time was evaluated by Vicat needle and radiopacity analysed with digital X-ray. The pH of all tested materials was observed after immersion in water for 3, 24, 48, 72 and 168 h (or 7 days). Solubility and calcium release were measured after immersion in water for 24 h. A multiparametric assay XTT-NR-CVDE was used to evaluate the cytotoxicity of the materials in human periodontal ligament (HPDL) fibroblasts. HPDL fibroblasts were exposed to PC 15% and mineral trioxide aggregate (MTA), and the expression of proinflammatory cytokines (IL1A, IL6, IL8, TNF) and bone formation genes (ALP, COL1, RUNX2) was evaluated by real-time PCR. Mineralisation of HPDL fibroblasts cocultured with PC, PC 15% and MTA was evaluated with Von Kossa staining. PC-based groups presented more irregular and larger particles than MTA. PC and MTA showed similarities as observed by XRD and EDS. Setting time of PC-based groups was increased with the addition of bismuth carbonate. All tested materials were alkaline, and pH tended to reduce over time. All cements had solubility lower than recommended, with no difference between them (P>0.05) and showed calcium release. PC 15% had similar radiopacity when compared with MTA (P>0.05). Cell viability was higher for the tested materials than the positive control (P<0.001), but there was no difference when they were compared with negative control (P>0.05). Gene expression levels were similar for all tested groups (P>0.05). Analysed cements had positive Von Kossa staining. Overall, the addition of 15% of bismuth carbonate did not result in significant changes to its physicochemical and biological properties when compared with MTA, except for the setting time, and may be considered a potential substitute for MTA.


A rapid one-step method for the isolation of bacteroids from root nodules of soybean plants, utilizing self-generating Percoll gradients.
Bacteroids were isolated from the nodules of soybean plants by means of self-generating Percoll density gradients. The entire procedure can be performed in less than 1 h using an ordinary refrigerated centrifuge and angle head rotor. All of the markers for cytosol and bacteroid fractions behaved in accord with other reports in the literature. Asparaginase, beta-hydroxybutyrate dehydrogenase, and alanine dehydrogenase were all localized in the bacteroid fraction. Invertase, phosphoenolpyruvate carboxylase, and leghaemoglobin were all found in the cytosol fraction. Very little (less than 7%) cross contamination between the fractions was observed. The isolated bacteroids were viable, and based on electron micrographs, were free from contaminating plant material. Since the entire procedure is performed isosmotically, very little damage to the bacteroids is likely to occur. No organic compounds, except Percoll, were added to the isolating media, thus aiding in the analysis of bacteroid and cytosol metabolites.


One- versus two-incision technique for distal biceps tendon repair.
There are several techniques that have been described for distal biceps tendon repair but there is still controversy regarding the optimal technique. Our hypothesis is that the single-incision technique will have a similar complication rate and functionally equivalent restoration of function compared with the two-incision approach. A retrospective review of consecutive biceps tendon repairs was performed at one institution over a 5-year period. Thirty-six patients met the inclusion criteria and 26 were available for follow-up including subjective assessment, physical examination, and strength testing. Patients were divided into two groups based on the surgical approach utilized: 12 patients underwent single-incision repair and 14 had a two-incision repair. The average follow-up was 33 months (minimum 13; maximum 75). There were no statistically significant differences in regards to flexion strength or endurance, supination strength or endurance, or complication rates between the two techniques. In conclusion, both surgical techniques led to adequate restoration of strength with a low complication rate. Both techniques are safe to perform and should be guided by surgeon comfort with the approach.


An automated segmentation methodology for quantifying immunoreactive puncta number and fluorescence intensity in tissue sections.
A number of human brain diseases have been associated with disturbances in the structure and function of cortical synapses. Answering fundamental questions about the synaptic machinery in these disease states requires the ability to image and quantify small synaptic structures in tissue sections and to evaluate protein levels at these major sites of function. We developed a new automated segmentation imaging method specifically to answer such fundamental questions. The method takes advantage of advances in spinning disk confocal microscopy, and combines information from multiple iterations of a fluorescence intensity/morphological segmentation protocol to construct three-dimensional object masks of immunoreactive (IR) puncta. This new methodology is unique in that high- and low-fluorescing IR puncta are equally masked, allowing for quantification of the number of fluorescently-labeled puncta in tissue sections. In addition, the shape of the final object masks highly represents their corresponding original data. Thus, the object masks can be used to extract information about the IR puncta (e.g., average fluorescence intensity of proteins of interest). Importantly, the segmentation method presented can be easily adapted for use with most existing microscopy analysis packages.


Active targeting behaviors of biotinylated pluronic/poly(lactic acid) nanoparticles in vitro through three-step biotin-avidin interaction.
In order to prepare targeted drug carriers, previously a biotin group has been attached by our group to the end of Pluronic F87/poly(lactic acid) and Pluronic P85/poly(lactic acid) block co-polymers to obtain B-F87-PLA and B-P85-PLA, respectively. In this paper, the active targeting properties of B-F87-PLA and B-P85-PLA nanoparticles in vitro were investigated through a three-step biotin-avidin interaction by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) tests and fluorescence microscopy (FM). Two kinds of human ovarian cancer cells (OVCAR-3 and SKOV-3) and paclitaxel were chosen for the cytotoxicity tests. CA-125 antigen is over-expressed on OVCAR-3 cells but not on SKOV-3 cells. The loading and release behavior of paclitaxel loaded in B-Pluronic-PLA nanoparticles were also studied. Paclitaxel loaded in both B-F87-PLA and B-P85-PLA nanoparticles shows an initial rapid release followed by a slow release period. Compared with SKOV-3 cells, the cytotoxicity results implied that paclitaxel-loaded B-Pluronic-PLA nanoparticles were delivered more effectively to OVCAR-3 cells due to the specific interaction between the biotin groups on the surface of B-Pluronic-PLA nanoparticles and the avidin/biotinylated MAb X306/CA-125 antigen complexes on the surface of OVCAR-3 cells. The active targeting properties of B-F87-PLA nanoparticles were further confirmed by FM.


Amino acid disposition by liver and gastrointestinal tract after protein and glucose ingestion.
Hepatic uptake and gut and splanchnic output of amino acids were determined after administration of protein and glucose loads in conscious dogs with indwelling catheters in the femoral artery and portal and hepatic veins. Oral or parenteral glucose given with a beef meal blunted the rise in arterial amino acids relative to that seen with ingestion of beef alone. Gut amino acid output was considerably delayed with oral hypertonic glucose, but was unchanged with parenteral glucose and with oral isotonic glucose. Ingestion of beef with oral hypertonic mannitol, a nonabsorbable sugar alcohol, delayed the rise in gut amino acid output in a manner similar to that seen with beef plus oral hypertonic glucose. We have evaluated the relationship between circulating amino acids, circulating hormone concentrations, and amino acid metabolism. Significant correlations with hepatic amino acid uptake were found for insulin, for arterial and portal amino acid levels, and for gut amino acid output. Partial correlation analysis suggests that gut amino acid output is the major determinant of hepatic amino acid uptake.


Hypercalcemia-Induced ST-Segment Elevation Mimicking Acute Myocardial Injury: A Case Report and Review of the Literature.
ST-segment elevation in absence of acute coronary syndrome can be seen in multiple conditions, including acute pericarditis and coronary vasospasm, but it is rarely seen with severe hypercalcemia. The authors present a case of an 81-year-old female with a history of stage 4 squamous cell cancer of the lung, who presented to the emergency room with profound fatigue, weakness, anorexia, and drowsiness two weeks after her first chemotherapy cycle. Additionally, she had complaints of right-sided chest pain associated with worsening shortness of breath, as well as right arm numbness. An EKG obtained on arrival to the hospital showed diffuse ST-segment elevation (leads V3-V6, I, II, III, and aVF). Basic lab work found a calcium level of 20.4 mg/dl with elevated parathyroid hormone-related protein (PTHrP) of 135 pg/ml. Troponin I remained within normal limits. Serial EKS obtained during the patient's hospitalization demonstrated resolution of the ST elevation as calcium level normalized. This case emphasizes the importance of hypercalcemia as a differential diagnosis for ST-segment elevation and QT shortening when acute coronary syndrome is not present. Awareness of these EKG changes is critical for early diagnosis, recognition, and appropriate treatment.


Simian virus 40-associated small RNA: mapping on the simian virus 40 genome and characterization of its synthesis.
The simian virus 40 (SV40)-associated small RNA (SAS-RNA) has previously been shown to arise late in SV40 lytic infection and to bear homology with the SV40 early mRNA's, or the SV40 genome, at map position 0.21. By using hybridization analysis, we determined that the SAS-RNA is between 62 and 65 nucleotides in length and its homology region lies between nucleotides 2760 and 2825 of the SV40 late(+) DNA strand. Viable deletion mutants which lacked part or all of this region made no SAS-RNA, strongly indicating that this is the coding region of the SAS-RNA. The expected sequence for the SAS-RNA, determined from the DNA sequence between nucleotides 2760 and 2825, appeared to be very pyrimidine rich (76% uridine and cytidine). Deletion or alteration of sequences immediately preceding the SAS-RNA coding region (approximately nucleotides 2716 to 2748) resulted in the loss of SAS-RNA production. These sequences may be part of a promotor for SAS-RNA synthesis or a processing site for its excision from long nuclear late transcripts. Under growth conditions where late transcription was not fully initiated (tsA58 at 41 degrees C; wild-type SV40 in the presence of 1-beta-D-arabinofuranosylcytosine), no SAS-RNA was produced, indicating that the expression of the SAS-RNA is regulated by a mechanism related to the control of late transcription.


Conditional Knockout of PKC-δ in Osteoclasts Favors Bone Mass Accrual in Males Due to Decreased Osteoclast Function.
Protein kinase C delta (PKC-δ) functions as an important regulator in bone metabolism. However, the precise involvement of PKC-δ in the regulation of osteoclasts remains elusive. We generated an osteoclast specific PKC-δ knockout mouse strain to investigate the function of PKC-δ in osteoclast biology. Bone phenotype was investigated using microcomputed tomography. Osteoclast and osteoblast parameters were assessed using bone histomorphometry, and analysis of osteoclast formation and function with osteoclastogensis and hydroxyapatite resorption assays. The molecular mechanisms by which PKC-δ regulated osteoclast function were dissected by Western Blotting, TUNEL assay, transfection and transcriptome sequencing. We found that ablation of PKC-δ in osteoclasts resulted in an increase in trabecular and cortical bone volume in male mice, however, the bone mass phenotype was not observed in female mice. This was accompanied by decreased osteoclast number and surface, and Cathepsin-K protein levels in vivo, as well as decreased osteoclast formation and resorption in vitro in a male-specific manner. PKC-δ regulated androgen receptor transcription by binding to its promoter, moreover, PKC-δ conditional knockout did not increase osteoclast apoptosis but increased MAPK signaling and enhanced androgen receptor transcription and expression, finally leding to significant alterations in gene expression and signaling changes related to extracellular matrix proteins specifically in male mice. In conclusion, PKC-δ plays an important role in osteoclast formation and function in a male-specific manner. Our work reveals a previously unknown target for treatment of gender-related bone diseases.


Mortality after chronic hepatitis B virus infection: a linkage study involving 2 million parous women from Taiwan.
Few studies have evaluated survival rates among women who have chronic hepatitis B virus infection. We investigated the overall and disease-specific mortality rates in a nationwide cohort of women after they were screened for hepatitis B surface antigen (HBsAg) during pregnancy. HBsAg prenatal screening data were available for 2,087,994 women in Taiwan between 1 January 1986 and 31 March 2000 in the National Hepatitis B Vaccination Registry. Their vital status and cause of death were ascertained by computerized linkage with the National Death Certification Registry. Cox proportional hazards models were used to estimate the association between HBsAg status and specific causes of death. Overall, 14,524 deaths were identified after a mean of 11.43 years of follow-up. The age-adjusted hazard ratio for mortality among HBsAg carriers compared with noncarriers was 1.24 (95% confidence interval [CI], 1.19-1.30), 6.59 (95% CI, 5.70-7.61), and 1.09 (95% CI, 1.04-1.14) for all-cause, liver-specific, and non-liver-related deaths, respectively. In addition to liver-specific causes, a significantly increased risk of mortality from non-Hodgkin lymphoma (P < .001) and gallbladder and extrahepatic bile duct cancer (P = .01) was observed. Our study found an excess risk of death due to both liver-specific and non-liver-related causes for HBsAg-positive women in Taiwan. Effective prevention and treatment of hepatitis B virus infection is an important public health priority.


Fungal mediator tail subunits contain classical transcriptional activation domains.
Classical activation domains within DNA-bound eukaryotic transcription factors make weak interactions with coactivator complexes, such as Mediator, to stimulate transcription. How these interactions stimulate transcription, however, is unknown. The activation of reporter genes by artificial fusion of Mediator subunits to DNA binding domains that bind to their promoters has been cited as evidence that the primary role of activators is simply to recruit Mediator. We have identified potent classical transcriptional activation domains in the C termini of several tail module subunits of Saccharomyces cerevisiae, Candida albicans, and Candida dubliniensis Mediator, while their N-terminal domains are necessary and sufficient for their incorporation into Mediator but do not possess the ability to activate transcription when fused to a DNA binding domain. This suggests that Mediator fusion proteins actually are functioning in a manner similar to that of a classical DNA-bound activator rather than just recruiting Mediator. Our finding that deletion of the activation domains of S. cerevisiae Med2 and Med3, as well as C. dubliniensis Tlo1 (a Med2 ortholog), impairs the induction of certain genes shows these domains function at native promoters. Activation domains within coactivators are likely an important feature of these complexes and one that may have been uniquely leveraged by a common fungal pathogen.


The use of panoramic images for identification of edentulous persons.
The aim of this study was to determine if edentulous persons could be identified using panoramic images by: I) investigating the possibility of matching two panoramic radiographs of the same person obtained on two different occasions, II) determining what anatomical features are used as the base for matching, III) investigating if oral and maxillofacial radiologists (OMR) and dentists who were not oral and maxillofacial radiologists (NOMR) differed in their ability to match the images, and IV) determining if the time elapsed between the images affected the results or the confidence of the match. Panoramic image pairs from 19 patients obtained on two different occasions were included, plus 10 images from other edentulous patients. The time elapsed between the image pairs varied between 4 months and 6 years. Four OMR and four NOMR were asked to match the image pairs depicting the same patient. The participants marked each match as "certain", "likely", or "possible" and what anatomical structure they used for matching. The OMR group correctly matched 100% of the images and the NOMR group correctly matched 96%. The anatomy of the mandible was most often used for matching. The OMR group was more certain in their decisions than the NOMR group. The time elapsed between the examinations did not affect the result. In conclusion, panoramic images can be used to identify edentulous patients. Both OMR and NOMR could identify edentulous individuals when only panoramic radiographic images were available and the OMR were especially confident in the identification process.


Distinct mechanistic responses to replication fork stalling induced by either nucleotide or protein deprivation.
Incidents that slow or stall replication fork progression, collectively known as replication stress, represent a major source of spontaneous genomic instability. Here, we determine the requirement for global protein biosynthesis on DNA replication and associated downstream signaling. We study this response side by side with dNTP deprivation; one of the most commonly used means to investigate replication arrest and replicative stress. Our in vitro interrogations reveal that inhibition of translation by cycloheximide (CHX) rapidly impairs replication fork progression without decoupling helicase and polymerase activities or inducing DNA damage. In line with this, protein deprivation stress does not activate checkpoint signaling. In contrast to the direct link between insufficient dNTP pools and genome instability, our findings suggest that replication forks remain stable during short-term protein deficiency. We find that replication forks initially endure fluctuations in protein supply in order to efficiently resume DNA synthesis upon reversal of the induced protein deprivation stress. These results reveal distinct cellular responses to replication arrest induced by deprivation of either nucleotides or proteins.


Single Nematode Transcriptomic Analysis, Using Long-Read Technology, Reveals Two Novel Virulence Gene Candidates in the Soybean Cyst Nematode, Heterodera glycines.
The soybean cyst nematode (Heterodera glycines, SCN), is the most damaging disease of soybean in North America. While management of this pest using resistant soybean is generally still effective, prolonged exposure to cultivars derived from the same source of resistance (PI 88788) has led to the emergence of virulence. Currently, the underlying mechanisms responsible for resistance breakdown remain unknown. In this study, we combined a single nematode transcriptomic profiling approach with long-read sequencing to reannotate the SCN genome. This resulted in the annotation of 1932 novel transcripts and 281 novel gene features. Using a transcript-level quantification approach, we identified eight novel effector candidates overexpressed in PI 88788 virulent nematodes in the late infection stage. Among these were the novel gene Hg-CPZ-1 and a pioneer effector transcript generated through the alternative splicing of the non-effector gene Hetgly21698. While our results demonstrate that alternative splicing in effectors does occur, we found limited evidence of direct involvement in the breakdown of resistance. However, our analysis highlighted a distinct pattern of effector upregulation in response to PI 88788 resistance indicative of a possible adaptation process by SCN to host resistance.


A novel endoplasmic reticulum stress-related lncRNA signature for prognosis prediction and immune response evaluation in Stomach adenocarcinoma.
Stomach adenocarcinoma (STAD) is a significant contributor to cancer-related mortality worldwide. Although previous research has identified endoplasmic reticulum stress (ERS) as a regulator of various tumor-promoting properties of cancer cells, the impact of ERS-related long non-coding RNAs (lncRNAs) on STAD prognosis has not yet been investigated. Therefore, our study aims to develop and validate an ERS-related lncRNA signature that can accurately predict the prognosis of STAD patients. We collected RNA expression profiles and clinical data of STAD patients from The Cancer Genome Atlas (TCGA) and identified ERS-related genes from the Molecular Signature Database (MSigDB). Co-expression analysis enabled us to identify ERS-related lncRNAs, and we applied univariate Cox, least absolute shrinkage, and selection operator (LASSO), and multivariate Cox regression analyses to construct a predictive signature comprising of 9 ERS-related lncRNAs. We assessed the prognostic accuracy of our signature using Kaplan-Meier survival analysis, and validated our predictive signature in an independent gene expression omnibus (GEO) cohort. We also performed tumor mutational burden (TMB) and tumor immune microenvironment (TIME) analyses. Enrichment analysis was used to investigate the functions and biological processes of the signature, and we identified two distinct STAD patient subgroups through consensus clustering. Finally, we performed drug sensitivity analysis and immunologic efficacy analysis to explore further insights. The 9 ERS related-lncRNAs signature demonstrated satisfactory predictive performance as an independent prognostic marker and was significantly associated with STAD clinicopathological characteristics. Furthermore, patients in the high-risk group displayed a worse STAD prognosis than those in the low-risk group. Notably, gene set enrichment analysis (GSEA) revealed significant enrichment of extracellular matrix pathways in the high-risk group, indicating their involvement in STAD progression. Additionally, the high-risk group exhibited significantly lower TMB expression levels than the low-risk group. Consensus clustering revealed two distinct STAD patient subgroups, with Cluster 1 exhibiting higher immune cell infiltration and more active immune functions. Drug sensitivity analysis suggested that the low-risk group was more responsive to oxaliplatin, epirubicinl, and other drugs. Our study highlights the crucial regulatory roles of ERS-related lncRNAs in STAD, with significant clinical implications. The 9-lncRNA signature we have constructed represents a reliable prognostic indicator that has the potential to inform more personalized treatment decisions for STAD patients. These findings shed new light on the pathogenesis of STAD and its underlying molecular mechanisms, offering opportunities for novel therapeutic strategies to be developed for STAD patients.


Low root reserve accumulation during drought may lead to winter mortality in poplar seedlings.
Climate models suggest that more frequent drought events of greater severity and length, associated with climate change, can be expected in the coming decades. Although drought-induced tree mortality has been recognized as an important factor modulating forest demography at the global scale, the mechanisms underlying drought-induced tree mortality remain contentious. Above- and below-ground growth, gas exchange, water relations and carbon reserve accumulation dynamics at the organ and whole-plant scale were quantified in Populus tremuloides and P. balsamifera seedlings in response to severe drought. Seedlings were maintained in drought conditions over one growing and one dormant winter season. Our experiment presents a detailed description of the effect of severe drought on growth and physiological variables, leading to seedling mortality after an extended period of drought and dormancy. After re-watering following the dormant period, drought-exposed seedlings did not re-flush, showing that the root system had died off. The results of this study suggest a complex series of physiological feedbacks between the measured variables in both Populus species. Further, they reveal that reduced reserve accumulation in the root system during drought decreases the conversion of starch to soluble sugars in roots, which may contribute to the root death of drought-exposed seedlings during the dormant season by compromising the frost tolerance of the root system.


Overall and subgroup prevalence of non-alcoholic fatty liver disease and prevalence of advanced fibrosis in the United States: An updated national estimate in National Health and Nutrition Examination Survey (NHANES) 2011-2018.
Data on the prevalence of non-alcoholic fatty liver disease (NAFLD) in subgroups of the United States (US) population are limited. This study was conducted to estimate NAFLD prevalence overall and by subgroup, and prevalence of NAFLD with advanced fibrosis. Using the National Health and Nutrition Examination Survey (NHANES) 2011-2018 data, a cross-sectional study was conducted. NAFLD was defined as having a US Fatty Liver Index (USFLI) ≥ 30 in the absence of other causes of liver disease, including excess alcohol intake, chronic hepatitis B, and chronic hepatitis C. Likelihood for having advanced fibrosis was determined by the calculated NAFLD fibrosis score (NFS; high ≥ 0.676; low < -1.445) and fibrosis-4 index (FIB-4; high ≥ 2.67; low < 1.30). The weighted national prevalence of NAFLD in US adults was 26.7% (95% confidence interval: 25.3%-28.1%). Prevalence was higher among those aged ≥ 65 years, males, Mexican Americans, with BMI ≥ 35 kg/m2 (class 2 and 3 obesity) and with type 2 diabetes (T2D). Of those meeting the USFLI criterion for NAFLD, 18.1% and 3.7% were determined as having a high probability of advanced fibrosis based on NFS ≥ 0.676 and FIB-4 ≥ 2.67 cut-off values, respectively. This study supports an increased prevalence of NAFLD in specific subpopulations (aged ≥ 65 years, males, Mexican Americans, obese population, and patients with T2D). The observed difference in the prevalence of advanced fibrosis as estimated by NFS and FIB-4 highlights the challenge of choosing optimal cut-off values.


Modulation of Corpus Striatal Neurochemistry by Astrocytes and Vasoactive Intestinal Peptide (VIP) in Parkinsonian Rats.
The neurotoxin 6-hydroxydopamine (6-OHDA) is widely used in animal models of Parkinson's disease. In various neurodegenerative diseases, astrocytes play direct, active, and critical roles in mediating neuronal survival and functions. Vasoactive intestinal peptide (VIP) has neurotrophic actions and modulates a number of astrocytic activities. In this study, the effects of VIP on the striatal neurochemistry were investigated in parkinsonian rats. Adult Sprague-Dawley rats were divided into sham-operated, unilaterally 6-OHDA-lesioned, and lesioned + VIP-administered (25 ng/kg i.p.) groups. VIP was first injected 1 h after the intrastriatal 6-OHDA microinjection and then every 2 days throughout 15 days. Extracellular striatal concentration of glutathione (GSH), gamma-aminobutyric acid (GABA), glutamate (GLU), and lactate were measured in microdialysates by high-performance liquid chromatography (HPLC). Quantification of GABA and activity dependent neuroprotective protein (ADNP)-expressing cells were determined by glutamic acid decarboxylase (GAD)/ADNP + glial fibrillary acidic protein (GFAP) double immunohistochemistry. Our results demonstrated that a 6-OHDA lesion significantly increased the density of astrocytes in the striatum and VIP treatment slightly reduced the gliosis. Extracellular concentration of GABA, GLU, and lactate levels did not change, but GSH level significantly increased in the striatum of parkinsonian rats. VIP treatment reduced GSH level comparable to sham-operated groups, but enhanced GABA and GLU levels. Our double labeling results showed that VIP primarily acts on neurons to increase ADNP and GAD expression for protection. These results suggest that, in the 6-OHDA-induced neurodegeneration model, astrocytes were possibly activated for forefront defensiveness by modulating striatal neurochemistry.


The effect of latanoprost, brimonidine, and a fixed combination of timolol and dorzolamide on circadian intraocular pressure in patients with glaucoma or ocular hypertension.
To compare the circadian intraocular pressure (IOP) reductions induced by latanoprost, brimonidine tartrate, and a fixed combination of timolol maleate and dorzolamide hydrochloride in patients with primary open-angle glaucoma (POAG) or ocular hypertension (OHT). In this crossover study, 10 patients with POAG and 10 with OHT were treated with latanoprost once a day, brimonidine twice a day, and a fixed combination of timolol and dorzolamide twice a day for 1 month. Four 24-hour tonometric curves were obtained for each patient. Intraocular pressure (IOP) was measured at 3, 6, and 9 AM, and at noon and at 3, 6, and 9 PM, and at midnight, using a handheld electronic tonometer with the patient in supine and sitting positions and a Goldmann applanation tonometer with the patient sitting at the slitlamp. Reduction of circadian IOP. All the drugs significantly reduced IOP compared with the baseline at all times, except for brimonidine at midnight, 3 AM, and 6 AM. Latanoprost was more effective than brimonidine in lowering IOP at 3 and 6 AM and at 3 PM (P=.03), and the combination of timolol and dorzolamide was more effective than brimonidine at 3 and 9 AM (P=.04) and at 3 and 6 PM (P =.05) and more effective than latanoprost at 9 AM (P=.05). Latanoprost and the fixed combination of timolol and dorzolamide led to similar circadian reductions in IOP, whereas brimonidine was less effective, particularly during the night.


Mechanical History Dependence in Carbon Black Suspensions for Flow Batteries: A Rheo-Impedance Study.
We studied the effects of shear and its history on suspensions of carbon black (CB) in lithium ion battery electrolyte via simultaneous rheometry and electrical impedance spectroscopy. Ketjen black (KB) suspensions showed shear thinning and rheopexy and exhibited a yield stress. Shear step experiments revealed a two time scale response. The immediate effect of decreasing the shear rate is an increase in both viscosity and electronic conductivity. In a much slower secondary response, both quantities change in the opposite direction, leading to a reversal of the initial change in the conductivity. Stepwise increases in the shear rate lead to similar responses in the opposite direction. This remarkable behavior is consistent with a picture in which agglomerating KB particles can stick directly on contact, forming open structures, and then slowly interpenetrate and densify. The fact that spherical CB particles show the opposite slow response suggests that the fractal structure of the KB primary units plays an important role. A theoretical scheme was used to analyze the shear and time-dependent viscosity and conductivity. Describing the agglomerates as effective hard spheres with a fractal architecture and using an effective medium approximation for the conductivity, we found the changes in the derived suspension structure to be in agreement with our qualitative mechanistic picture. This behavior of KB in flow has consequences for the properties of the gel network that is formed immediately after the cessation of shear: both the yield stress and the electronic conductivity increase with the previously applied shear rate. Our findings thus have clear implications for the operation and filling strategies of semisolid flow batteries.


Low-resolution models for ribosomal particles reconstructed from electron micrographs of tilted two-dimensional sheets.
Models of the whole ribosome (70S) and its large subunit (50S) were obtained at low resolution (47 and 28 A respectively) by three-dimensional image reconstruction using diffraction data collected from electron micrographs of two-dimensional ordered arrays. The comparison of the various reconstructed images, using interactive computer graphics, enabled the assessment of the reliability of the method, the derivation of the shape of the small subunit (30S) and the assignment of several functional features such as the probable path taken by the nascent protein chain, the presumed site for the process of biosynthesis of proteins, and a feasible mode for tRNA binding. The reconstructed models of the various ribosomal particles may be used for phasing of X-ray diffraction data at low resolution.


In vivo antistress and antioxidant effects of fermented and germinated mung bean.
Mung bean has been traditionally used to alleviate heat stress. This effect may be contributed by the presence of flavonoids and γ-aminobutyric acid (GABA). On the other hand, fermentation and germination have been practised to enhance the nutritional and antioxidant properties of certain food products. The main focus of current study was to compare the antistress effect of none-process, fermented and germinated mung bean extracts. Acute and chronic restraint stresses were observed to promote the elevation of serum biochemical markers including cholesterol, triglyceride, total protein, liver enzymes, and glucose. Chronic cold restraint stress was observed to increase the adrenal gland weight, brain 5-hydroxytryptamine (5-HT), and malondialdehyde (MDA) level while reducing brain antioxidant enzyme level. However, these parameters were found reverted in mice treated with diazepam, high concentration of fermented mung bean and high concentration of germinated mung bean. Moreover, enhanced level of antioxidant on the chronic stress mice was observed in fermented and germinated mung bean treated groups. In comparison between germinated and fermented mung bean, fermented mung bean always showed better antistress and antioxidant effects throughout this study.


YycH regulates the activity of the essential YycFG two-component system in Bacillus subtilis.
Of the numerous two-component signal transduction systems found in bacteria, only a very few have proven to be essential for cell viability. Among these is the YycF (response regulator)-YycG (histidine kinase) system, which is highly conserved in and specific to the low-G+C content gram-positive bacteria. Given the pathogenic nature of several members of this class of bacteria, the YycF-YycG system has been suggested as a prime antimicrobial target. In an attempt to identify genes involved in regulation of this two-component system, a transposon mutagenesis study was designed to identify suppressors of a temperature-sensitive YycF mutant in Bacillus subtilis. Suppressors could be identified, and the prime target was the yycH gene located adjacent to yycG and within the same operon. A lacZ reporter assay revealed that YycF-regulated gene expression was elevated in a yycH strain, whereas disruption of any of the three downstream genes within the operon, yycI, yycJ, and yycK, showed no such effect. The concentrations of both YycG and YycF, assayed immunologically, remained unchanged between the wild-type and the yycH strain as determined by immunoassay. Alkaline phosphatase fusion studies showed that YycH is located external to the cell membrane, suggesting that it acts in the regulation of the sensor domain of the YycG sensor histidine kinase. The yycH strain showed a characteristic cell wall defect consistent with the previously suggested notion that the YycF-YycG system is involved in regulating cell wall homeostasis and indicating that either up- or down-regulation of YycF activity affects this homeostatic mechanism.


Rectification of current responds to incorporation of fullerenes into mixed-monolayers of alkanethiolates in tunneling junctions.
This paper describes the rectification of current through molecular junctions comprising self-assembled monolayers of decanethiolate through the incorporation of C60 fullerene moieties bearing undecanethiol groups in junctions using eutectic Ga-In (EGaIn) and Au conducting probe AFM (CP-AFM) top-contacts. The degree of rectification increases with increasing exposure of the decanethiolate monolayers to the fullerene moieties, going through a maximum after 24 h. We ascribe this observation to the resulting mixed-monolayer achieving an optimal packing density of fullerene cages sitting above the alkane monolayer. Thus, the degree of rectification is controlled by the amount of fullerene present in the mixed-monolayer. The voltage dependence of R varies with the composition of the top-contact and the force applied to the junction and the energy of the lowest unoccupied π-state determined from photoelectron spectroscopy is consistent with the direction of rectification. The maximum value of rectification R = |J(+)/J(-)| = 940 at ±1 V or 617 at ±0.95 V is in agreement with previous studies on pure monolayers relating the degree of rectification to the volume of the head-group on which the frontier orbitals are localized.


Interaction between deglutition, tongue posture, and malocclusion: A comparison of intraoral compartment formation in subjects with neutral occlusion or different types of malocclusion.
To test the null hypothesis of no significant difference in terms of intraoral pressure curve characteristics assessed simultaneously at the subpalatal space (SPS) and the vestibular space (VS), during different oral postures, between four groups with either an Angle Class II/1 (II1), Angle Class II/2 (II2), anterior open bite (O) malocclusion, or a neutral occlusion control group (I). Intraoral pressure recordings were performed simultaneously in the VS and SPS of 69 consecutive subjects (nII1 = 15; nII2 = 17; nO = 17; nI = 20; mean age/standard deviation 18.43/6.60 years). Assessments included defined sections of open mouth posture (OMP, 30 seconds), anteriorly closed mouth condition (60 seconds), dynamics by a tongue-repositioning maneuver (TRM, 60 seconds), swallowing, and positive pressure generation (PP, 10 seconds). Interactions of malocclusion, compartment location, and posture on pressure curve characteristics were analyzed by Kruskal-Wallis and Mann-Whitney U-tests, adopting an α level of 5%. Globally significant group differences were detected at the VS (plateau duration and median peak heights during TRM; area under pressure curve [AUC] during PP) and SPS (AUC during TRM and PP). Subjects with anteriorly nonopen dental configurations (groups I and II2) were able to keep negative pressure levels at the VS for longer time periods during TRM, compared to groups O and II1. The null hypothesis was rejected for mean VS plateau durations and peak heights and for SPS AUC. Negative pressures at the VS may stabilize outer soft tissues passively and may explain the dental arch form shaping effect by mimic muscles.


Discrete effects in stellar feedback: Individual Supernovae, Hypernovae, and IMF Sampling in Dwarf Galaxies.
Using high-resolution simulations from the FIRE-2 (Feedback In Realistic Environments) project, we study the effects of discreteness in stellar feedback processes on the evolution of galaxies and the properties of the interstellar medium (ISM). We specifically consider the discretization of supernovae (SNe), including hypernovae (HNe), and sampling the initial mass function (IMF). We study these processes in cosmological simulations of dwarf galaxies with z = 0 stellar masses M * ~ 104-3 × 106 M⊙ (halo masses ~109-1010 M⊙). We show that the discrete nature of individual SNe (as opposed to a model in which their energy/momentum deposition is continuous overtime, similar to stellar winds) is crucial in generating a reasonable ISM structure and galactic winds and in regulating dwarf stellar masses. However, once SNe are discretized, accounting for the effects of IMF sampling on continuous mechanisms such as radiative feedback and stellar mass-loss (as opposed to adopting IMF-averaged rates) has weak effects on galaxy-scale properties. We also consider the effects of rare HNe events with energies ~1053 erg. The effects of HNe are similar to the effects of clustered explosions of SNe - which are already captured in our default simulation setup - and do not quench star formation (provided that the HNe do not dominate the total SNe energy budget), which suggests that HNe yield products should be observable in ultra-faint dwarfs today.


[Pulsed Doppler in children operated on for aortic coarctation: reliability in the evaluation of minor residual lesions].
The usual non invasive diagnostic methods may not be able to estimate a mild residual gradient in infants treated for Coarctation of the Aorta (CA) so we tried to determine whether Pulsed Doppler (PD) may be useful in these cases. We studied 16 children (Group A) treated for CA, clinically considered as good operative results (arm to leg gradient less than or equal to 10 mmHg) and admitted for cardiac catheterization to study associated cardiac defects. Ten children with congenital heart disease different from CA were studied as a control (Group B). Percutaneous left cardiac catheterization was performed in all children with pull-back recording of the aortic pressure across aortoplasty before angiography. PD study was performed with an ATL Mark 600 and a 3 MHz transducer positioned in the suprasternal notch. The sample volume was moved into the descending aorta in order to obtain peak flow velocity (PFV) and estimate systolic gradient (DEG). Cardiac catheterization gradients (CCG) ranged from 0 to 20 mmHg (3.9 +/- 6.3 mmHg) in Group A while no gradient was recorded at the Isthmus in Group B. In Group A the frequency shift changed just below the aortoplasty; PFV and DEG ranged respectively from 1 to 2.3 m/s (1.6 +/- 0.3 m/s) and from 4 to 22 mmHg (10 +/- 4.8 mmHg). In Group B the descending aorta PFV ranged from 0.8 +/- 1.3 m/s (1 +/- 0.2 m/s). PFV in Group A was higher than in Group B (p less than 0.001). CCG were plotted against DEG; the linear regression gave a satisfactory coefficient of correlation (R = 0.93 and p less than 0.001).(ABSTRACT TRUNCATED AT 250 WORDS)


Phagocytic and postphagocytic activities of bovine neutrophils for pure and mixed bacterial cultures.
A comparative study of phagocytosis and postphagocytic oxidative metabolic activity of bovine blood neutrophils incubated with pure and mixed cultures of Escherichia coli, Staphylococcus aureus, and Streptococcus agalactiae was preformed. Most neutrophils when incubated with mixed cultures showed preferential phagocytosis for one species and a smaller number phagocytized both species of microorganisms. Percent phagocytosis for E. coli in pure culture was similar to that of Strep. agalactiae in pure culture and higher than that for Staph. aureus in pure culture. Neutrophils incubated with mixed cultures of E. coli and Staph. aureus or E. coli and Strep. agalactiae showed greater than expected phagocytosis of each microorganisms alone and reduced phagocytosis of both microorganisms together. Postphagocytic oxidative metabolic activity of neutrophils, measured by percent nitroblue tetrazolium reduction, did not differ following phagocytosis of these three microorganisms in pure cultures. In comparison, a synergistic effect on nitroblue tetrazolium reductive activity was seen in mixed cultures as evidenced by higher percent nitroblue tetrazolium reduction following phagocytosis of both E. coli and Staph. aureus or E. coli and Strep. agalactiae. These observations indicate that the phagocytic and metabolic activities of neutrophils for bacteria in mixed cultures may not be identical to those in pure cultures.


Model of patterned self-assisted nanowire growth.
Vertically oriented and ordered GaAs nanowire arrays have been grown by the self-assisted mechanism using substrates prepared with nano-patterned oxide templates. Patterned Ga-assisted GaAs nanowire growth on (111) silicon by molecular beam epitaxy showed that the axial and radial growth rates increased with increasing interhole spacing. A model is described which accounts for the correlation of the final length and diameter with pattern pitch. The model considers that growth material is supplied by a secondary flux of both gallium and arsenic adatoms desorbing from the oxide surface between the nanowires which subsequently impinge on the liquid droplet and nanowire sidewalls. We show that shading of the incident and scattered flux by neighboring nanowires in the array can strongly affect the axial and radial growth rates, leading to significant differences in final nanowire morphologies.


[The problem of assessment of efficacy of antihypertensive therapy using 24-hour blood pressure monitoring].
Main advantages of the use of 24-hour blood pressure (BP) monitoring for assessment of effects of antihypertensive therapy consist in more precise selection of patients requiring drug treatment and more objective evaluation of results of therapy. However methodological limitations also exist. Among them are susceptibility of results to regression to the mean, insufficient understanding of many characteristics of 24-hour BP profile, interdependence (co-linearity) of various parameters. Lack of convincing data on prognosis of patients whose antihypertensive treatment has been guided by 24-hour BP monitoring also presents a serious problem. Promising direction of further research is investigation of reproducibility and reliability of different parameters of BP monitoring in relatively short-term studies including those with surrogate end points.


Overseas recruitment activities of NHS Trusts 2015-2018: Findings from FOI requests to 19 Acute NHS Trusts in England.
Migrant nurses form an increasing proportion of the nursing workforce, with the United Kingdom (UK) being the third most popular destination for overseas nurses in the world. The migrant nurse workforce is highly susceptible to policy changes at the macro or professional level of the donor and recipient countries. Freedom of information requests were issued to 19 National Health Service [NHS] Trusts in England to determine their involvement in overseas nurse recruitment activity from 1998 onwards. These indicate a notable shift away from active European Union (EU) recruitment and towards overseas countries particularly the Philippines and India. Reasons given were as follows: diminishing returns from EU sources, high attrition among EU nurses and the introduction of English language tests for EU nurses in July 2016. This led to Trusts revisiting their recruitment strategies by increasing more direct/less resource-intensive methods and expanding their focus outside of the EU. Trusts frequently utilised private recruitment companies for their recruitment drives, including consulting and influencing the Trusts' workforce strategies. Policy adjustments have numerous influences on the composition of the overseas nursing workforce. While the NHS continues its efforts in expanding its international nursing workforce, this should not be at the expense of ethical and sustainable recruitment practices, which may be compromised indirectly as a result.


MetaNetter 2: A Cytoscape plugin for ab initio network analysis and metabolite feature classification.
Metabolomics frequently relies on the use of high resolution mass spectrometry data. Classification and filtering of this data remain a challenging task due to the plethora of complex mass spectral artefacts, chemical noise, adducts and fragmentation that occur during ionisation and analysis. Additionally, the relationships between detected compounds can provide a wealth of information about the nature of the samples and the biochemistry that gave rise to them. We present a biochemical networking tool: MetaNetter 2 that is based on the original MetaNetter, a Cytoscape plugin that creates ab initio networks. The new version supports two major improvements: the generation of adduct networks and the creation of tables that map adduct or transformation patterns across multiple samples, providing a readout of compound relationships. We have applied this tool to the analysis of adduct patterns in the same sample separated under two different chromatographies, allowing inferences to be made about the effect of different buffer conditions on adduct detection, and the application of the chemical transformation analysis to both a single fragmentation analysis and an all-ions fragmentation dataset. Finally, we present an analysis of a dataset derived from anaerobic and aerobic growth of the organism Staphylococcus aureus demonstrating the utility of the tool for biological analysis.


Evaluation of nasal delivery systems of olanzapine by desorption electrospray ionization mass spectrometry imaging.
Nose-to-brain delivery presents an attractive administration route for neuroactive drugs that suffer from compromised bioavailability or fail to pass the blood-brain barrier. However, the conventional gauge of effectiveness for intranasal delivery platforms primarily involves detecting the presence of the administered drug within the brain, with little insight into its precise localization within brain structures. This may undermine the therapeutic efficacy of drugs and hinder the design of systems that target specific brain regions. In this study, we designed two intranasal delivery systems for the antipsychotic drug, olanzapine, and evaluated its distribution in the rat brain following intranasal administration. The first evaluated system was an olanzapine-loaded microemulsion and the second one was nanoparticulate aqueous dispersion of olanzapine. Both systems exhibited characteristics that render them compatible for intranasal administration, and successfully delivered olanzapine to the brain. We further employed an ambient mass spectrometry imaging method, called desorption electrospray ionization mass spectrometry imaging, to visualize the signal intensity of olanzapine in different brain regions following the intranasal administration of these two systems. Substantial variations in the distribution patterns of olanzapine across various brain structures were revealed, potentially highlighting the importance of mass spectrometry imaging in designing and evaluating intranasal drug delivery platforms.


Mean 3D Dispersion for Automatic General Movement Assessment of Preterm Infants.
The General Movement assessment (GMA) is a validated assessment of brain maturation primarily based on the qualitative analysis of the complexity and the variation of spontaneous motor activity. The GMA can identify preterm infants presenting an early abnormal developmental trajectory before term-equivalent age, which permits a personalized early developmental intervention. However, GMA is time-consuming and relies on a qualitative analysis; these limitations restrict the implementation of GMA in clinical practice. In this study based on a validated dataset of 183 videos from 92 premature infants (54 males, 38 females) born <33 weeks of gestational age (GA) and acquired between 32 and 40 weeks of GA, we introduce the mean 3D dispersion (M3D) for objective quantification and classification of normal and abnormal GMA. Moreover, we have created a new 3D representation of skeleton joints which allows an objective comparison of spontaneous movements of infants of different ages and sizes. Preterm infants with normal versus abnormal GMA had a distinct M3D distribution (p <0.001). The M3D has shown a good classification performance for GMA (AUC=0.7723) and presented an accuracy of 74.1%, a sensitivity of 75.8%, and a specificity of 70.1% when using an M3D of 0.29 as a classification threshold.Clinical relevance- Our study paves the way for the development of quantitative analysis of GMA within the Neonatal Unit.


[The causes of blindness in the 3rd region of Mali. Study of 101 clinical records].
A retrospective study of 101 blind patients seen in daily consultation has been done in Sikasso's Yeelen Center from January 1988 to October 1989. The results obtained show that the frequency of blindness is 1.3% and that more than 75% of the causes of losing the seeing are due to the pathology of the posterior segment. The optical atrophy represents the major cause of blindness (63% of the cases) with usually a glaucoma excavation. Corneal opacities and the chorioretinal scarrings take respectively the second and the third place with 13% and 10%.


Effect of dopamine beta-hydroxylase inhibition on systemic hemodynamics in conscious, spontaneously hypertensive rats.
The acute and chronic effects of a potent and selective dopamine beta-hydroxylase inhibitor, SK&F 102698 [1-(3,5-difluorobenzyl)imidazole-2-thiol], on systemic hemodynamic variables were assessed in chronically instrumented spontaneously hypertensive and normotensive Wistar rats. The changes observed in the hypertensive rats were compared with those obtained by vehicle in a different group of rats. Following intragastric administration of SK&F 102698 (75 mg/kg), blood pressure and cardiac output decreased gradually in both strains of rats, while total peripheral vascular resistance remained unchanged. The antihypertensive effect of repeated daily administration of SK&F 102698 in the hypertensive rats was sustained for the duration of treatment. During the acute phase, the decrease in cardiac output was due to bradycardia. In addition, as a result of hemodynamic changes, SK&F 102698 significantly reduced the minute work output of the heart. The results indicate that SK&F 102698 lowers blood pressure primarily by decreasing cardiac output.


Tinnitus, attendance at night-clubs and social drug taking in students.
A questionnaire was used to collect data from 545 students of the Manchester Metropolitan University. The aim of the study was to investigate associations between the attendance of university students at night-clubs (NCs) that play loud music and the incidence and duration of post exposure tinnitus (PET) and spontaneous tinnitus (ST). The possible effects of taking social drugs in NCs on tinnitus was also investigated. Descriptive analyses and Chi squared association analyses were carried out. The study showed that 87% of students attended noisy NCs and there was a significant association between the attendance at NCs and the duration of PET. There was no significant association between frequency of attendance and the incidence of tinnitus, however non attendees were significantly less likely to get ST. The amount of social drug-taking was not as high as expected, only 19% taking drugs more than rarely, although the incidence may be under-reported. Taking drugs while at night-clubs would appear to exacerbate the effects of noise exposure on tinnitus. Non drug taking males were less susceptible to PET lasting longer than 2 hours and ST than non drug taking females, whereas drug taking males were more susceptible than drug taking females. Whereas PET has yet to be proved to be related to auditory damage, this study would suggest the effect of night-club noise exposure impedes tinnitus recovery and could prove to be an early sign of permanent damage.


Trends in the effectiveness of endovascular recanalization for acute stroke: is a change taking place?
Despite recent technical advances in endovascular recanalization, there is skepticism regarding its clinical effectiveness compared with intravenous thrombolysis for treating acute ischemic stroke. We aimed to delineate its effectiveness and safety and their change over time. Using a prospective, multicenter stroke registry database, we identified 872 patients with ischemic stroke who underwent recanalization therapy with intravenous thrombolysis alone (IVT; n = 533) or endovascular recanalization with or without intravenous thrombolysis (EVT; n = 339) between April 2008 and January 2012. All subjects had National Institute of Health Stroke Scale score of 10 or more and arrived at the hospital within 4.5 hours of onset. Propensity score was used to address baseline imbalances between treatment groups, but balance adjustment was not performed for subgroup analyses. The primary outcome was modified Rankin Scale score of 0-2 at discharge. The year-by-year effectiveness and safety of EVT and IVT were compared. Before 2010, the primary outcome was not associated with the recanalization method. However, in 2011, EVT increased the odds of having a primary outcome compared with IVT (adjusted odds ratio [OR], 1.87; 95% confidence interval [CI], 1.08-3.23). In 2011, EVT was superior to IVT regarding the achievement of a favorable outcome at 3 months after stroke (OR, 1.99; 95% CI, 1.10-3.59). The odds of in-hospital mortality and 3-month mortality were not different over 4 years. There might have been a change in the effectiveness of endovascular recanalization compared with intravenous thrombolysis, but the results remain tentative until prospectively evaluated.


The action of human articular-cartilage metalloproteinase on proteoglycan and link protein. Similarities between products of degradation in situ and in vitro.
Interleukin 1 stimulation of human articular cartilage in organ culture produced the concomitant release of proteoglycan fragments and latent metalloproteinase. The released fragments ranged in size from that of almost intact proteoglycan subunits to the product of limiting digestion generated by the activated metalloproteinase. None of the fragments possessed the ability to interact with hyaluronic acid. Analysis of proteoglycan aggregate digested with the activated metalloproteinase showed that isolated hyaluronic acid-binding regions were produced from the proteoglycan subunits, and that the two higher-Mr link-protein components (Mr 48,000 and 44,000) were converted into the lowest-Mr component (Mr 41,000). Link protein extracted from cartilage under stimulation with interleukin 1 showed a similar conversion. These results suggest that interleukin 1 stimulates the release of latent metalloproteinase from chondrocytes and that a proportion of the enzyme is activated in situ in the cartilage matrix. The mode of action of the activated enzyme is compatible with a role in the changes in proteoglycan structure seen in aging.


Dose-dependent bone-sparing effects of dietary isoflavones in the ovariectomised rat.
The dose-dependent bone-sparing effects of dietary isoflavones (IF) were investigated in adult (7-month-old) Wistar rats. Forty animals were ovariectomised, allocated into four groups of ten rats each, and immediately treated orally with IF at 0 (OVX), 20 (IF20), 40 (IF40) or 80 (IF80) microg/g body weight per d for 91 d; ten sham-operated (SH) controls received the same diet without added IF. Animals were killed on day 91. Both femoral failure load and total femoral, diaphyseal or metaphyseal bone mineral densities (BMD) were lower in OVX animals than in SH animals. Urinary deoxypyridinoline (DPD) excretion, a marker of bone resorption, and plasma osteocalcin (OC) levels, a marker of osteoblast activity, were higher in OVX animals than in SH animals. Total femoral and diaphyseal BMD and femoral failure load were similar in IF-treated rats and SH rats. Although metaphyseal BMD in IF40 or IF80 rats was similar to that in SH rats, its value was lower in IF20 rats than in controls. The day 91 urinary DPD excretion in IF40 and IF80 rats, but not in IF20 rats, was similar to that in SH rats. Day 91 plasma OC concentrations in IF-treated rats were similar to day 45 values, but were decreased in OVX and SH rats. Thus, daily IF consumption prevented ovariectomy-induced bone loss, both by depressing bone resorption and stimulating osteoblast activity. Moreover, as only the highest IF level induced a weak uterotrophic activity, the optimal IF dose which preserves both cancellous and cortical bone, but exhibits no oestrogen-like effects on the uterus, was 40 microg/g body weight per d.


Repeatability and reproducibility of product ion abundances in electron capture dissociation mass spectrometry of peptides.
Site-specific reproducibility and repeatability of electron capture dissociation (ECD) in Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS) are of fundamental importance for product ion abundance (PIA)-based peptide and protein structure analysis. However, despite the growing interest in ECD PIA-based applications, these parameters have not yet been investigated in a consistent manner. Here, we first provide a detailed description of the experimental parameters for ECD-based tandem mass spectrometry performed on a hybrid linear ion trap (LTQ) FT-ICR MS. In the following, we describe the evaluation and comparison of ECD and infrared multiphoton dissociation (IRMPD) PIA methodologies upon variation of a number of experimental parameters, for example, cathode potential (electron energy), laser power, electron and photon irradiation periods and pre- irradiation delays, as well as precursor ion number. Ranges of experimental parameters that yielded an average PIA variation below 5% and 15% were determined for ECD and IRMPD, respectively. We report cleavage site-dependent ECD PIA variation below 20% and correlation coefficients between fragmentation patterns superior to 0.95 for experiments performed on three FT-ICR MS instruments. Overall, the encouraging results obtained for ECD PIA reproducibility and repeatability support the use of ECD PIA as a complementary source of information to m/z data in radical-induced dissociation applied for peptide and protein structure analysis.


ABCB1 gene polymorphisms, ABCB1 haplotypes and ABCG2 c.421c > A are determinants of inter-subject variability in rosuvastatin pharmacokinetics.
A randomized cross-over pharmacokinetic study of rosuvastatin calcium (single dose: 5 mg, 10 mg and 20 mg; multiple doses: 10mg once daily for 7 days) was conducted in 12 healthy Chinese volunteers. Plasma concentrations of rosuvastatin were determined by an LC-ESI-MS-MS method. Single-nucleotide polymorphisms (SNPs) in ABCB1, ABCG2, SLCOB1, CYP2C9 and CYP3A5 were determined by TaqMan (MGB) genotyping assay. An impact of the aforementioned SNPs on steady state pharmacokinetic parameters [average steady state concentration (Cav,ss) and area under the plasma concentration versus time curve during the dosing interval at steady state (AUCss)], dose-normalized (based on 5 mg) pharmacokinetic parameters of single-dose rosuvastatin were further analyzed. Rosuvastatin exhibited linear pharmacokinetics and great inter-subject variability. Cav,ss, AUCss and dose-normalized peak plasma concentration (Cmax) and AUC(0-infinity) of single-dose rosuvastatin were significantly related with ABCB1 C1236T, G2677T/A and C3435T polymorphisms and ABCB1 haplotypes. Compared to homozygous wild type and heterozygous mutation gene carriers, subjects carrying the variant ABCB1 1236TT, 2677 non-G or 3435TT genotype had higher Cav,ss, AUCss, Cmax and AUC(0-infinity) (p < 0.05). ABCB1 haplotype (1236TT-2677TT-3435TT) had significant influence on dose-normalized pharmacokinetics of single-dose rosuvastatin. ABCB1 haplotype (1236TT-2677TT-3435TT) carriers (n = 12) had obvious higher Cmax (11.16 +/- 3.10 microg x L(-1) vs 8.35 +/- 3.31 microg x L(-1), p < 0.05) and AUC(0-infinity) (86.61 +/- 24.32 microg x h x L(-1) vs 62.60 +/- 26.19 microg x h x L(-1), p < 0.05) compared to non-1236TT-2677TT-3435TT carriers (n = 24). ABCG2 c.421C > A had a significant impact on rosuvastatin pharmacokinetics. Homozygotes (AA) carriers had obvious higher Cmax (12.20 +/- 4.09 microg x L(-1) vs 8.70 +/- 3.09 microg x L(-1), p < 0.05) and AUC(0-infinity) (98.74 +/- 25.36 microg x h x L(-1) vs 64.97 +/- 24.90 microg x h x L(-1), p < 0.05) values compared to heterozygotes (CA) and homozygotes (CC) carriers. There were no significant effects on single-dose and steady-state pharmacokinetics of rosuvastatin by CYP2C9*3 (1075A > C), CYP3A5*3 g.6986A > G, ABCG2 c.34G > A, SLCO1B1 c.521 T > C, c.388 A > G, g.11187 G > A, c.571 T > C and c.597 C > T. In addition, no difference in rosuvastatin pharmacokinetics was observed among subjects of different genders. We conclude that ABCB1 C1236T, G2677T/A and C3435T polymorphism, ABCB1 haplotypes and ABCG2 c.421C > A are determinants of inter-subject variability in rosuvastatin pharmacokinetics in healthy Chinese volunteers, and potentially affect the efficacy and toxicity of statin therapy.


Detection of bird nests during mechanical weeding by incremental background modeling and visual saliency.
Mechanical weeding is an important tool in organic farming. However, the use of mechanical weeding in conventional agriculture is increasing, due to public demands to lower the use of pesticides and an increased number of pesticide-resistant weeds. Ground nesting birds are highly susceptible to farming operations, like mechanical weeding, which may destroy the nests and reduce the survival of chicks and incubating females. This problem has limited focus within agricultural engineering. However, when the number of machines increases, destruction of nests will have an impact on various species. It is therefore necessary to explore and develop new technology in order to avoid these negative ethical consequences. This paper presents a vision-based approach to automated ground nest detection. The algorithm is based on the fusion of visual saliency, which mimics human attention, and incremental background modeling, which enables foreground detection with moving cameras. The algorithm achieves a good detection rate, as it detects 28 of 30 nests at an average distance of 3.8 m, with a true positive rate of 0.75.