Comparison of the histopathological diagnoses of preoperative dilatation and curettage and Pipelle biopsy.
To evaluate the accuracy of dilatation and curettage (D&C) and Pipelle biopsy for the diagnosis of endometrial pathologies and determine whether the amount of endometrial tissue obtained using these techniques is sufficient for further histopathology of hysterectomy specimens. Patients undergoing hysterectomy for various indications were evaluated via Pipelle endometrial biopsy or D&C from 2009-2011. A total of 267 women were included with 78 women enrolled in the Pipelle group and 189 in the D&C group. Uterine findings were grouped as normal, hyperplasia, focal lesion, atypia, and atrophy. Histological sections from the Pipelle biopsy or D&C specimens were compared to each other and hysterectomy specimens. The concordance rate between Pipelle biopsy and hysterectomy was 62% and between D&C and hysterectomy was 67%. The sensitivity of Pipelle biopsy and D&C for detecting hyperplasia was 41.7% and 45%, respectively, and for detecting atypia was 71.4% for both techniques. The sensitivity of detecting atrophic endometrial tissue was significantly higher in the D&C group at 80% compared to 37.5% in the Pipelle biopsy group (p = 0.030). All other parameters were similar in both groups. Pipelle biopsy and D&C were equally successful for diagnosing endometrial pathologies. Neither Pipelle biopsy nor D&C was adequate for detecting focal endometrial pathologies and endometrial hyperplasia. In contrast, both techniques were sufficient for the diagnosis of atypia. The Pipelle biopsy technique is a reasonable pre-hysterectomy procedure that is more economical, less invasive, and can easily be performed in multiple clinics.


The activation of N-methyl-d-aspartate receptors downregulates transient outward potassium and L-type calcium currents in rat models of depression.
Major depression is an important clinical factor in ventricular arrhythmia. Patients diagnosed with major depression overexpress N-methyl-d-aspartate receptors (NMDARs). Previous studies found that chronic NMDAR activation increases susceptibility to ventricular arrhythmias. We aimed to explore the mechanisms by which NMDAR activation may increase susceptibility to ventricular arrhythmias. Male rats were randomly assigned to either normal environments as control (CTL) group or 4 wk of chronic mild stress (CMS) to produce a major depression disorder (MDD) model group. After 4 wk of CMS, depression-like behaviors were measured in both groups. Varying doses (1-100 μM) of NMDA and 10 μM NMDA antagonist (MK-801) were perfused through ventricular myocytes isolated from MDD rats to measure the L-type calcium current (ICa-L) and transient outward potassium current (Ito). Structural remodeling was assessed using serial histopathology including Masson's trichrome dye. Electrophysiological characteristics were evaluated using Langendorff perfusion. Depression-like behaviors were observed in MDD rats. MDD rats showed longer action potential durations at 90% repolarization and higher susceptibility to ventricular arrhythmias than CTL rats. MDD rats showed lower ICa-L and Ito current densities than CTL rats. Additionally, NMDA reduced both currents in a concentration-dependent manner, whereas there was no significant impact on the currents when perfused with MK-801. MDD rats exhibited significantly more fibrosis areas in heart tissue and reduced expression of Kv4.2, Kv4.3, and Cav1.2. We observed that acute NMDAR activation led to downregulation of potassium and L-type calcium currents in a rat model of depression, which may be the mechanism underlying ventricular arrhythmia promotion by depression.


SEC-10 and RAB-10 coordinate basolateral recycling of clathrin-independent cargo through endosomal tubules in Caenorhabditis elegans.
Despite the increasing number of regulatory proteins identified in clathrin-independent endocytic (CIE) pathways, our understanding of the exact functions of these proteins and the sequential manner in which they function remains limited. In this study, using the Caenorhabditis elegans intestine as a model, we observed a unique structure of interconnected endosomal tubules, which is required for the basolateral recycling of several CIE cargoes including hTAC, GLUT1, and DAF-4. SEC-10 is a subunit of the octameric protein complex exocyst. Depleting SEC-10 and several other exocyst components disrupted the endosomal tubules into various ring-like structures. An epistasis analysis further suggested that SEC-10 operates at the intermediate step between early endosomes and recycling endosomes. The endosomal tubules were also sensitive to inactivation of the Rab GTPase RAB-10 and disruption of microtubules. Taken together, our data suggest that SEC-10 coordinates with RAB-10 and microtubules to form the endosomal tubular network for efficient recycling of particular CIE cargoes.


Increased vitamin C content accompanied by an enhanced recycling pathway confers oxidative stress tolerance in Arabidopsis.
Vitamin C (L-ascorbic acid, AsA) has important antioxidant and metabolic functions in both plants and animals. Once used, ascorbic acid can be regenerated from its oxidized form in a reaction catalyzed by dehydroascorbate reductase (DHAR, EC 18.104.22.168). To analyze the physiological role of DHAR catalyzing the reduction of DHA to ascorbate in environmental stress adaptation, we examined whether increasing the level of AsA through enhanced AsA recycling would limit the deleterious effects of oxidative stress. A chimeric construct consisting of the double CaMV35S promoter fused to the Myc-dhar gene was introduced into Arabidopsis thaliana. Transgenic plants were biochemically characterized and tested for responses to oxidative stress. Western blot indicated that the dhar-transgene was successfully expressed. In homozygous T(4) transgenic seedlings, DHAR overexpression was increased up to 1.5 to 5.4 fold, which enhanced foliar ascorbic acid levels 2- to 4.25-fold and ratio of AsA/DHA about 3- to 16-fold relative to wild type. In addition, the level of glutathione, the reductant used by DHAR, also increased as did its redox state. When whole plants were treated with high light and high temperature stress or in vitro leaf discs were subjected to 10 muM paraquat, transgenic plants showed a larger AsA pool size, lower membrane damage, and a higher level of chlorophyll compared with controls. These data suggested that increasing the plant vitamin C content through enhanced ascorbate recycling could limit the deleterious effects of environmental oxidative stress.


Burnout among public doctors in Hong Kong: cross-sectional survey.
The stressful life of doctors makes them prone to burnout. We evaluated the prevalence of burnout among Hong Kong public hospital doctors and correlated burnout with job characteristics, working hours, stressors, and stress-relieving strategies. Cross-sectional survey. Hong Kong. One thousand doctors were randomly sampled from the Hong Kong Public Doctors' Association registry. Self-administered, anonymous questionnaires with postage-paid envelopes were mailed twice in early 2009. The Maslach Burnout Inventory-Human Services Survey was used for burnout assessment. According to this scale, burnout is defined as emotional exhaustion, depersonalisation, and a reduced sense of personal accomplishment. Correlation analysis, as well as univariate and multivariate analyses, were performed to assess factors associated with high degrees of burnout. RESULTS. A total of 226 questionnaires were analysed, of which 31.4% of the respondents satisfied the criteria for high burnout. They were younger and needed to work shifts, and their median year of practice was 8.5. High-burnout doctors worked similar hours per week to non-high-burnout doctors (mean ± standard deviation, 56.2 ± 12.7 vs 54.7 ± 10.9; P=0.413) and reported suicidal thoughts more often (9.9% vs 2.6%; P=0.033). Moreover, 52.2% of high-burnout doctors were dissatisfied or very dissatisfied with their jobs. 'Excessive stress due to global workload' and 'feeling that their own work was not valued by others' were the most significant stressors associated with high emotional exhaustion and depersonalisation, while 'feeling that their own work was not valued by others' and 'poor job security' correlated with low personal accomplishment. A high proportion of public doctors who responded to our survey endured high burnout. Trainees with some experience were at heightened risk. Stressors identified in this study should be addressed, so as to improve job satisfaction.


Immune thrombocytopenia in infants: a retrospective study with comparison to toddlers.
Immune thrombocytopenia (ITP) is the most common cause of acquired bleeding in childhood, but little is known about the clinical course and outcomes in infants with ITP. This is a retrospective study of all infants (1-12 months of age) and toddlers (13-47 months of age) diagnosed with ITP from a single centre during a 13-year period. The following data were compared between the two patients groups: demographics, severity of bleeding, platelet counts, duration of illness, development of chronic ITP, treatment and association with recent vaccination. Twenty-two infants and 30 toddlers were diagnosed and followed up for ITP during the study period. Infants with ITP generally had minor or mild bleeding (19, 86.4%) and seldom required treatment (7, 31.8%), and their thrombocytopenia resolved at a mean of 1.90 months after diagnosis. Besides age, the sex ratio, severity of bleeding, platelet counts and proportion that required treatment were comparable between infants and toddlers. Fewer infants developed chronic ITP (1/22 vs. 9/30, P = 0.032), but more infants had a history of vaccination in the preceding 6 weeks prior to diagnosis of ITP (13/22 vs. 1/30, P < 0.001). ITP in infants is almost always a self-limiting and transient illness, and the majority of cases do not require treatment.


Development of cowpea mosaic virus-based vectors for the production of vaccines in plants.
Plant viruses are emerging as an attractive alternative to stable genetic transformation for the expression of foreign proteins in plants. The main advantages of using this strategy are that viral genomes are small and easy to manipulate, infection of plants with modified viruses is simpler and quicker than the regeneration of stably transformed plants and the sequence inserted into a virus vector will be highly amplified. One use of these virus expression systems is for vaccine production. Among plant viruses, cowpea mosaic virus makes an ideal candidate for the production of such vaccines because it grows extremely well in host plants, is very stable, and the purification of virus particles, if required, is straightforward. In this article, the authors review the progress made in the development of cowpea mosaic virus-based vectors for vaccine production, making use of two main approaches: epitope presentation and polypeptide expression.


Research note: young children's resolution of distress through play.
It has been suggested that play is important to the young child as a means of resolving a distressing experience. Pre-school children were observed as they exhibited distress on the first day of school. High- and low-anxiety children were allowed either the opportunity for free play or participation in a story-reading session, each of which was performed in either the presence or absence of peers. The anxious children in the play condition were found to decrease in distress and their play was more solitary and imaginative in nature compared to other conditions.


Podophyllum hexandrum Offers Radioprotection by Modulating Free Radical Flux: Role of Aryl-Tetralin Lignans.
We have evaluated the effect of variation in aryl-tetralin lignans on the radioprotective properties of Podophyllum hexandrum. Two fractionated fractions of P. hexandrum [methanolic (S1) and chloroform fractions (S2)], with varying aryl-tetralin lignan content were utilized for the present study. The peroxyl ion scavenging potentials of S1 and S2 were found to be comparable [i.e. 45.88% (S1) and 41% (S2)] after a 48 h interval in a time-dependent study, whereas in a 2 h study, S2 exhibited significant (P < 0.05) antioxidant activity in different metal ion + flux states. In the aqueous phase, S2 exhibited non-site-specific reactive oxygen species scavenging activity, i.e. 73.12% inhibition at 500 mug ml(-1). S1 exhibited 58.40 +/- 0.8% inhibition (at 0.025 mug ml(-1)) of the formation of reactive nitrite radicals, comparable to S2 (52.45 +/- 0.825%), and also showed 45.01% site-specific activity (1000 mug ml(-1)), along with significant (P < 0.05) electron donation potential (50-2000 mug ml(-1)) compared to S2. Such activities of S1 could be attributed to the significantly (P < 0.05) higher levels of podophyllotoxin beta-d-glucopyranoside (16.5 times) and demethyl podophyllotoxin glucoside (2.9 times) compared with S2. Together, these findings clearly prove that aryl-tetralin lignan content influences the radiation protective potential of the Podophyllum fractions to a great extent.


Development of a niobium-doped titania inert anode for titanium electrowinning in molten chloride salts.
The direct electrochemical reduction of solid titanium dioxide in a chloride melt is an attractive method for the production of titanium metal. It has been estimated that this type of electrolytic approach may reduce the costs of producing titanium sponge by more than half, with the additional benefit of a smaller environmental footprint. The process utilises a consumable carbon anode which releases a mixture of CO2 and CO gas during electrolysis, but suffers from low current efficiency due to the occurrence of parasitic side reactions involving carbon. The replacement of the carbon anode with a cheap, robust inert anode offers numerous benefits that include: elimination of carbon dioxide emissions, more efficient cell operation, opportunity for three-dimensional electrode configurations and reduced electrode costs. This paper reports a study of Nb-doped titania anode materials for inert anodes in a titanium electrolytic reduction cell. The study examines the effect of niobium content and sintering conditions on the performance of Nb-doped TiO2 anodes in laboratory-scale electrolysis tests. Experimental findings, including performance in a 100 h laboratory electrolysis test, are described.


Deuterium labeled peptides give insights into the directionality of class III lantibiotic synthetase LabKC.
The biosynthesis of a considerable number of ribosomally synthesized peptide antibiotics involves the modification of Ser and Thr residues of a precursor peptide. This post-translational processing is performed by one or multiple modifying enzymes encoded in the biosynthetic gene cluster. We present a deuterium-label based enzyme assay, utilizing a series of peptide substrates with α-deuterated Ser, for the determination of the dehydration order during the biosynthesis of class III lantibiotic labyrinthopeptin A2. Remarkably, the data show that, in contrast to other modifying enzymes of class I and II lantibiotics, LabKC has a C- to N-terminal processing mode. This surprising finding, which we consider relevant for the biosyntheses of other class III lantibiotics, underlines significant differences of this class of modifying enzymes compared to other investigated systems.


Granule proteinases define mast cell heterogeneity in the serosa and the gastrointestinal mucosa of the mouse.
In order to define further mast cell heterogeneity in the mouse, affinity-purified antibodies against a 28,000 MW serine proteinase from mouse intestinal mast cells (IMCP) and against rat mast cell proteinase I (RMCPI) were used to characterize mast cell cytoplasmic granules immunohistochemically. On Western blot, anti-IMCP cross-reacted with RMCPI and with a 25,000 MW antigen from isolated mouse serosal mast cells (SMC). Anti-RMCPI did not react with IMCP, although it identified the same 25,000 MW antigen from SMC. Isolated SMC (85-90% pure) lacked the 28,000 MW IMCP on Western blot, even though, immunohistochemically, the cells were stained with both anti-RMCPI and anti-IMCP. Anti-IMCP stained the granules of more than 85% of all mast cells detected with toluidine blue in the tongue or gastrointestinal mucosa. The specificity of anti-RMCPI which, in the rat, detects very few mucosal mast cells was almost identical to that of anti-IMCP for murine tongue and gastric and large intestinal mucosae, but a significant proportion of cells in distal jejunal, ileal and caecal mucosae were not stained with this antibody. The immunohistochemistry of the large numbers of mast cells recruited to jejunum following infection 10 days previously with 300 Trichinella spiralis muscle larvae was similar to that of uninfected control mice. The results show that considerable mast cell heterogeneity exists within the gastrointestinal mucosa of the mouse and indicate that there are both similarities and differences between mouse and rat in the distribution of mast cells and of their granule proteinases.


Atorvastatin and celecoxib inhibit prostate PC-3 tumors in immunodeficient mice.
To investigate the effects and mechanisms of atorvastatin and celecoxib administered individually or in combination on human prostate cancer PC-3 cells cultured in vitro or grown as xenograft tumors in immunodeficient mice. Human prostate cancer PC-3 cells in culture were treated with atorvastatin and celecoxib alone or in combination. Severe combined immunodeficient (SCID) mice were injected s.c. with PC-3 cells. The mice received daily i.p injections starting 2 days before tumor cell inoculation and continuing during the course of treatment with atorvastatin (10 microg/g body weight/d), celecoxib (10 microg/g/d), a combination of atorvastatin (10 microg/g/d) and celecoxib (10 microg/g/d), or a combination of atorvastatin (5 microg/g/d) and celecoxib (5 microg/g/d). Atorvastatin in combination with celecoxib had stronger effects on growth inhibition and apoptosis of PC-3 cells than either agent used individually. Atorvastatin and celecoxib in combination also had a stronger inhibitory effect on activation of nuclear factor-kappaB and extracellular signal-regulated kinase 1/2 in PC-3 cells than either agent alone. Treatment of SCID mice with combinations of atorvastatin and celecoxib more effectively inhibited the formation and growth of PC-3 tumors in the mice than either agent administered alone. A combination of atorvastatin and celecoxib had a more potent inhibitory effect on the growth of PC-3 cells cultured in vitro or grown in SCID mice than either agent alone. A combination of atorvastatin and celecoxib may be an effective strategy for the prevention of prostate cancer.


Contrast media enhancement reduction predicts tumor response to presurgical molecular-targeting therapy in patients with advanced renal cell carcinoma.
A quantitative tumor response evaluation to molecular-targeting agents in advanced renal cell carcinoma (RCC) is debatable. We aimed to evaluate the relationship between radiologic tumor response and pathological response in patients with advanced RCC who underwent presurgical therapy. Of 34 patients, 31 underwent scheduled radical nephrectomy. Presurgical therapy agents included axitinib (n = 26), everolimus (n = 3), sunitinib (n = 1), and axitinib followed by temsirolimus (n = 1). The major presurgical treatment-related adverse event was grade 2 or 3 hypertension (44%). The median radiologic tumor response by RECIST, Choi, and CMER were -19%, -24%, and -49%, respectively. Among the radiologic tumor response tests, CMER showed a higher association with tumor necrosis in surgical specimens than others. Ki67/MIB1 status was significantly decreased in surgical specimens than in biopsy specimens. The magnitude of the slope of the regression line associated with the tumor necrosis percentage was greater in CMER than in Choi and RECIST. Between March 2012 and December 2016, we prospectively enrolled 34 locally advanced and/or metastatic RCC who underwent presurgical molecular-targeting therapy followed by radical nephrectomy. Primary endpoint was comparison of radiologic tumor response among Response Evaluation Criteria in Solid Tumors (RECIST), Choi, and contrast media enhancement reduction (CMER). Secondary endpoint included pathological downstaging, treatment related adverse events, postoperative complications, Ki67/MIB1 status, and tumor necrosis. CMER may predict tumor response after presurgical molecular-targeting therapy. Larger prospective studies are needed to develop an optimal tumor response evaluation for molecular-targeting therapy.


Wildfires and Older Adults: A Scoping Review of Impacts, Risks, and Interventions.
Climate change is leading to worsening disasters that disproportionately impact older adults. While research has begun to measure disparities, there is a gap in examining wildfire-specific disasters. To address this gap, this scoping review analyzed literature to explore the nexus of wildfires and older adults. We searched peer-reviewed literature using the following inclusion criteria: (1) published in a peer-reviewed journal; (2) available in English; (3) examines at least one topic related to wildfires; and (4) examines how criterion three relates to older adults in at least one way. Authors screened 261 titles and abstracts and 138 were reviewed in full, with 75 articles meeting inclusion criteria. Findings heavily focused on health impacts of wildfires on older adults, particularly of smoke exposure and air quality. While many articles mentioned a need for community-engaged responses that incorporate the needs of older adults, few addressed firsthand experiences of older adults. Other common topics included problems with evacuation, general health impacts, and Indigenous elders' fire knowledge. Further research is needed at the nexus of wildfires and older adults to highlight both vulnerabilities and needs as well as the unique experience and knowledge of older adults to inform wildfire response strategies and tactics.


Estimation of posterior corneal power for IOL calculation after myopic LASIK.
To estimate the posterior corneal power for intraocular lens (IOL) calculation after myopic LASIK. A retrospective study of 50 patients (92 eyes) who underwent uneventful myopic LASIK was conducted. Pre- and postoperative data (12 months) were collected including refraction and Orbscan II (Bausch & Lomb) for simulated keratometry (Sim K), central corneal thickness, and posterior corneal power. Statistical analysis was performed to determine the correlation between the postoperative posterior corneal power and other variables. An empirical formula was generated to predict the postoperative posterior corneal power. The net corneal power was calculated based on the new formula and Gaussian optics formula in an additional 31 eyes that underwent myopic LASIK and compared to the Orbscan total optical power map. The net corneal power was retrospectively applied to calculate the IOL power in an additional 10 patients who underwent cataract extraction after myopic LASIK. The back-calculation for emmetropic IOL power was performed and compared to the IOL power derived from the net corneal power. By using multiple linear regression, a formula to predict postoperative posterior corneal power can be computed from postoperative Sim K, central corneal thickness, and the amount of myopia treated with R2 of 0.63 (standard error =0.25) (P<.05). The net corneal power was highly correlated to the total optical power (R2=0.96); no statistically significant difference (paired t test) was noted (P>.05). The error of the IOL power derived from the net corneal power was -0.30+/-0.20 diopters. Corneal power after myopic LASIK can be estimated using postoperative Sim K, central corneal thickness, and the amount of myopia treated, which can be used for the assessment of IOL power after myopic LASIK.


Clinical deterioration following middle cerebral artery hemodynamic changes after intravenous thrombolysis for acute ischemic stroke.
Little information exists regarding what occurs in the affected artery in the days after acute ischemic stroke and its impact in the outcome. We sought to determine the hemodynamic evolution and correlated this evoution with clinical outcome in stroke patients treated with intravenous thrombolysis. Using serial transcranial Doppler ultrasound (TCD) on days 1 (TCD1), 3 to 6 (TCD2), and 7 to 10 (TCD3) after stroke, we determined the hemodynamics in the affected artery by means of the thrombolysis in brain ischemia (TIBI) score and compared this with clinical outcome (National Institutes of Health Stroke Scale [NIHSS] score) and functional outcome (modified Rankin Scale score) at discharge and at 3 months. Thirty-four patients were studied. There were 24 men with a mean (± SD) age of 72.9 ± 16.2 years. The mean time from stroke onset to the administration of intravenous tissue plasminogen activator was 181 ± 54.4 minutes, and the mean NIHSS score at admission was 16.9 ± 9. Hemodynamic changes were observed in 23 (68%) patients, including improvement in 17 (50%) patients and worsening in 6 (18%) patients within the first 10 days poststroke. Clinical deterioration (NIHSS ≥4 points) was timely associated with hemodynamic deterioration in 3 cases. Patients achieving full recanalization at TCD3 had better mRS scores at 3 months (4 v 3; P = .02). Hemodynamic changes in the affected artery occurred in about two-thirds of patients within the first 10 days after receiving intravenous thrombolysis; 18% had hemodynamic deterioration, which was associated with clinical worsening in half of these cases.


Effect of screening for type 2 diabetes on healthcare costs: a register-based study among 139,075 individuals diagnosed with diabetes in Denmark between 2001 and 2009.
Trials have not demonstrated benefits to the population of screening for type 2 diabetes. However, there may be cost savings for those found to have diabetes. We therefore aimed to compare healthcare costs among individuals with incident type 2 diabetes in a screened group with those in an unscreened group. In this register-based, non-randomised controlled trial, eligible individuals were men and women aged 40-69 years without known diabetes who were registered with a general practice in Denmark (n = 1,912,392). Between 2001 and 2006, 153,107 individuals registered with 181 practices participating in the Anglo-Danish-Dutch Study of Intensive Treatment in People with Screen Detected Diabetes in Primary Care (ADDITION)-Denmark study were sent a diabetes risk-score questionnaire. Individuals with a moderate-to-high risk were invited to visit their family doctor for assessment of diabetes status and cardiovascular risk (screening group). The 1,759,285 individuals registered with all other practices in Denmark constituted the retrospectively constructed no-screening (control) group. In this post hoc analysis, we identified individuals from the screening and no-screening groups who were diagnosed with diabetes between 2001 and 2009 (n = 139,075). Using national registry data, we quantified the cost of healthcare services in these two groups between 2001 and 2012. From a healthcare sector perspective, we estimated the potential healthcare cost savings for individuals with diabetes that were attributable to the screening programme. In the screening group, 27,177 of 153,107 individuals (18% of those sent a risk-score questionnaire) attended for screening, 1533 of whom were diagnosed with diabetes. Between 2001 and 2009, 13,992 people were newly diagnosed with diabetes in the screening group (including those diagnosed by screening) and 125,083 in the no-screening group. Healthcare costs were significantly lower in the screening group compared with the no-screening group (difference in mean total annual healthcare costs -€889 per individual with incident diabetes; 95% CI -€1196, -€581). The screening programme was associated with a cost saving per person with incident diabetes over a 5-year period of €2688 (95% CI €1421, €3995). Healthcare costs were lower among individuals with incident type 2 diabetes in the screened group compared with the unscreened group. The relatively modest cost of screening per person discovered to have developed diabetes was offset within 2 years by savings in the healthcare system.


Genetic analysis of iron uptake in the yeast Saccharomyces cerevisiae.
We used the methods of yeast genetics to identify genes involved in acquisition of iron by eukaryotic cells. Mutants were identified with defects in cellular iron uptake. These were organized into an upstream group and a downstream group. The upstream group was involved in the delivery of copper to the multicopper oxidase FET3. Mutants of this group were characterized by defective iron uptake that could be corrected by exposure of the cells to large amounts of copper. The downstream group was more directly involved in iron uptake. Mutant phenotypes from these genes could not be corrected by copper exposure. Genes in the upstream group encoded the regulator of copper transport, MAC1, and two copper transporters, CTR1 and CCC2. Genes in the downstream group encoded the multicopper oxidase FET3 and its partner the iron permease FTR1. In addition, the downstream genes encoded the surface reductases FRE1 and FRE2 and the iron regulatory protein AFT1. The iron and copper uptake processes in yeast intersect because the FET3 gene encodes a multicopper oxidase that is required for iron transport. In human beings, an analogous function may be served by ceruloplasmin, a multicopper oxidase with a role in iron homeostasis.


TiO2 Nanorods Decorated with Pd Nanoparticles for Enhanced Liquefied Petroleum Gas Sensing Performance.
Development of highly sensitive and selective semiconductor-based metal oxide sensor devices to detect toxic, explosive, flammable, and pollutant gases is still a challenging research topic. In the present work, we systematically enhanced the liquefied petroleum gas (LPG) sensing performance of chemical bath deposited TiO2 nanorods by decorating Pd nanoparticle catalyst. Surface morphology with elemental mapping, crystal structure, composition and oxidation states, and surface area measurements of pristine TiO2 and Pd:TiO2 nanorods was examined by high resolution transmission electron microscopy with energy-dispersive X-ray spectroscopy, X-ray diffraction, X-ray photoelectron spectroscopy, and nitrogen adsorption-desorption characterization techniques. LPG sensing performance of pristine TiO2 and Pd:TiO2 nanorods was investigated in different LPG concentration and operating temperature ranges. The LPG response of 21% for pristine TiO2 nanorods is enhanced to 49% after Pd catalyst decoration with reasonably fast response and recovery times. Further, the sensor exhibited long-term stability, which could be due to the strong metal support (Pd:TiO2) interaction and catalytic properties offered by the Pd nanoparticle catalyst. The work described herein demonstrates a general and scalable approach that provides a promising route for rational design of variety of sensor devices for LPG detection.


Activation of p56lck through mutation of a regulatory carboxy-terminal tyrosine residue requires intact sites of autophosphorylation and myristylation.
Mutation of the major site of in vivo tyrosine phosphorylation of p56lck (tyrosine 505) to a phenylalanine constitutively enhances the p56lck-associated tyrosine-specific protein kinase activity. The mutant polypeptide is extensively phosphorylated in vivo at the site of in vitro Lck autophosphorylation (tyrosine 394) and is capable of oncogenic transformation of rodent fibroblasts. These observations have suggested that phosphorylation at Tyr-505 down regulates the tyrosine protein kinase activity of p56lck. Herein we have attempted to examine whether other posttranslational modifications may be involved in regulation of the enzymatic function of p56lck. The results indicated that activation of p56lck by mutation of Tyr-505 was prevented by a tyrosine-to-phenylalanine substitution at position 394. Furthermore, activation of p56lck by mutation of the carboxy-terminal tyrosine residue was rendered less efficient by substituting an alanine residue for the amino-terminal glycine. This second mutation prevented p56lck myristylation and stable membrane association and was associated with decreased in vivo phosphorylation at Tyr-394. Taken together, these findings imply that lack of phosphorylation at Tyr-505 may be insufficient for enhancement of the p56lck-associated tyrosine protein kinase activity. Our data suggest that activation of p56lck may be dependent on phosphorylation at Tyr-394 and that this process may be facilitated by myristylation, membrane association, or both.


Elevated 3,5-diiodothyronine concentrations in the sera of patients with nonthyroidal illnesses and brain tumors.
This study reports the development of a highly sensitive and reproducible RIA for the measurement of 3,5-diiodothyronine (3,5-T2) in human serum and tissue. The RIA employs 3-bromo-5-[125I]iodo-L-thyronine (3-Br-5-[125I]T1) as tracer, which was synthesized carrier free by an interhalogen exchange from 3,5-dibromo-L-thyronine (3,5-Br2T0). The detection limits were 1.0 fmol/g and 0.8 pmol/L in human brain tissue and serum, respectively. T3, diiodothyroacetic acid, and 3-monoiodothyronine cross-reacted with a 3,5-T2 antibody to the extent of 0.06%, 0.13%, and 0.65%, respectively. Serum concentrations of 3,5-T2 were measured in 62 healthy controls and 4 groups of patients with nonthyroidal illness, i.e. patients with sepsis (n = 24), liver diseases (n = 23), head and/or brain injury n = 15), and brain tumors (n = 21). The mean serum level of 3,5-T2 in the healthy subjects was 16.2 +/- 6.4 pmol/L. Concentrations of 3,5-T2 were significantly elevated in patients with sepsis (46.7 +/- 48.8 pmol/L; P < 0.01), liver diseases (24.8 +/- 14.9 pmol/L; P < 0.01), head and/or brain injury (24.1 +/- 11.3 pmol/L; P < 0.05), and brain tumors (21.6 +/- 4.8 pmol/L; P < 0.01). In all 4 patient groups, serum levels of T3 were significantly reduced, confirming the existence of a low T3 syndrome in these diseases. Serum concentrations of 3,5-T2 were significantly elevated in patients with hyperthyroidism (n = 9) and were reduced in patients with hypothyroidism (n = 8). The levels of T4, T3, and 3,5-T2 were measured in normal human tissue samples from the pituitary gland and various brain regions and in brain tumors. In normal brain tissue, the concentrations of 3,5-T2 ranged between 70-150 fmol/g, and the ratio of T3 to 3,5-T2 was approximately 20:1. In brain tumors, however, T3 levels were markedly lower, resulting in a ratio of T3 to 3,5-T2 of approximately 1:1. Recent findings suggest a physiological, thyromimetic role of 3,5-T2, possibly stimulating mitochondrial respiratory chain activity. Should this prove to be correct, then the increased availability of 3,5-T2 in nonthyroidal illness may be one factor involved in maintaining clinical euthyroidism in patients with reduced serum levels of T3 during nonthyroidal illness.


Partially fluidized shear granular flows: continuum theory and molecular dynamics simulations.
The continuum theory of partially fluidized shear granular flows is tested and calibrated using two-dimensional soft particle molecular dynamics simulations. The theory is based on the relaxational dynamics of the order parameter that describes the transition between static and flowing regimes of granular material. We define the order parameter as a fraction of static contacts among all contacts between particles. We also propose and verify by direct simulations the constitutive relation based on the splitting of the shear stress tensor into a"fluid part" proportional to the strain rate tensor, and a remaining "solid part." The ratio of these two parts is a function of the order parameter. The rheology of the fluid component agrees well with the kinetic theory of granular fluids even in the dense regime. Based on the hysteretic bifurcation diagram for a thin shear granular layer obtained in simulations, we construct the "free energy" for the order parameter. The theory calibrated using numerical experiments with the thin granular layer is applied to the surface-driven stationary two-dimensional granular flows in a thick granular layer under gravity.


Different clinical implications for ST depression and T wave inversion in non-Q wave myocardial infarction.
The differences between ST depression and T wave inversion in non-Q wave myocardial infarction were investigated in 42 patients with initial non-Q wave infarction, 22 patients with ST depression (ST group), and 20 patients with T wave inversion (T group). The extent of ischemic area estimated by electrocardiography and two-dimensional echocardiography, characteristic features of electrocardiographic changes, and clinical findings on admission and outcome were estimated. ST elevation preceded T wave inversion in the same leads in 80% (16/20) of the T group, and transient Q waves developed in 55% (11/20). However, neither ST elevation nor transient Q waves were observed in the ST group. Two or three ischemic segments were present in 86% (19/22) of the ST group patients, but only one ischemic segment was present in 60% (12/20) of T group patients, predominantly the anterior segment. The short-axis view of the two-dimensional echocardiogram on the level of papillary muscle showed decreased contraction in two or three of the anterior, lateral, and inferior segments of the left ventricle in 78% (14/18) of ST group patients. Only one segment with decreased contraction was present in 100% (17/17) of T group patients. Cardiac status on admission was lower in the ST than the T group: Killip class II-IV, 59% (13/22) vs 20% (4/20), p < 0.05; mortality rate after 1 month, 41% (9/22) vs 0% (0/20), p < 0.05. Coronary angiograms, left ventriculograms, and autopsy findings also showed extensive myocardial lesions in accordance with multivessel disease in the ST group, but localized myocardial lesion suggesting one-vessel territory in the T group. T wave inversion in non-Q wave myocardial infarction indicates a recovery phase in transient transmural ischemia and localized subendocardial infarction within the presumed one-vessel territory, while ST depression suggests the presence of extensive ischemia in the subendocardium of multivessel territory, and infarction within that region.


Reducing gonad irradiation in pediatric diagnosis.
The greatest danger of carcinogenesis and of genetic damage through diagnostic radiologic procedures is in children, whose smaller bodies are more vulnerable and who have a longer life span in which this damage can be realized. Film badges placed on the gonad area during radiologic studies indicated widely varying degrees of gonad irradiation from similar procedures. These results emphasize the importance of technique in protecting children from unnecessary exposure. Such exposure can be reduced by greater beam filtration, use of higher tube potentials, careful beam collimation and centering, closer coning and shielding of the gonads. A new film tested reduced exposure time by 50 per cent. Further reduction was obtained by high-speed screens.A most important measure is avoidance of unnecessary, repetitious and undiagnostic studies. Fluoroscopy should be avoided if possible. If not, the operator must dark-adapt his eyes, use the smallest possible current, the narrowest beam, and the shortest exposure time. Image intensification promotes these aims. Modern equipment, properly shielded, assures against unsuspected exposure.


Corneal pachymetry by AS-OCT after Descemet's membrane endothelial keratoplasty.
Corneal thickness (pachymetry) maps can be used to monitor restoration of corneal endothelial function, for example after Descemet's membrane endothelial keratoplasty (DMEK). Automated delineation of the corneal interfaces in anterior segment optical coherence tomography (AS-OCT) can be challenging for corneas that are irregularly shaped due to pathology, or as a consequence of surgery, leading to incorrect thickness measurements. In this research, deep learning is used to automatically delineate the corneal interfaces and measure corneal thickness with high accuracy in post-DMEK AS-OCT B-scans. Three different deep learning strategies were developed based on 960 B-scans from 50 patients. On an independent test set of 320 B-scans, corneal thickness could be measured with an error of 13.98 to 15.50 μm for the central 9 mm range, which is less than 3% of the average corneal thickness. The accurate thickness measurements were used to construct detailed pachymetry maps. Moreover, follow-up scans could be registered based on anatomical landmarks to obtain differential pachymetry maps. These maps may enable a more comprehensive understanding of the restoration of the endothelial function after DMEK, where thickness often varies throughout different regions of the cornea, and subsequently contribute to a standardized postoperative regime.


Hepatitis after cardiosurgery. Frequency and causes. Results of a prospective study with use of autologous blood.
In a prospective study on the causes and frequency of hepatitis after operations on the open heart, two homogeneous groups of patients were formed. In one of them, the amount of homologous blood was reduced to half by means of preoperative donations of autologous blood, the total blood requirement remaining the same. Hepatitis occurred postoperatively in 10% of the cases. The factors which gave rise to the hepatitis were large-pool clotting preparations, transfusion with homologous blood and cross-infection in the hospital. Even after exclusion of other risk factors, the frequency of hepatitis among the recipients of coagulation preparations was around 60%. The frequency of the cases of hepatitis due to foreign blood was about 5 per 100 patients or about 1 per 100 units of blood. By use of autologous blood collected preoperatively, the risk due to transfusion could be lowered by 50%. In polytransfusion, a disproportionately high rise in the risk of hepatitis was observed. It is probably to be ascribed to unknown factors in the hospital environment ("nosocomial hepatitis").


Radiation-Associated Thyroid Cancer Surveillance and Management in a Cohort of Late Effects Patients.
Compared to the general population, the incidence of thyroid cancer in childhood and adolescent and young adult malignancy survivors is increased 14.0-18.0 times (CI 11.7-23.8). The cumulative incidence is variably reported as 0.5% by age 45 with 30-year incidence of 1.3% in women and 0.6% in men. This study aims to evaluate the incidence of radiation-associated thyroid cancer amongst patients treated with prior radiation to the thyroid followed up in a late effects service. A secondary aim was to assess screening compliance in this cohort. The medical records of all patients attending the late effects service from 1 January 2000 to 20 February 2013 were interrogated to identify patients exposed to thyroid irradiation. The screening compliance and incidence of thyroid cancer were assessed for the duration whilst under the guidance of the late effect service. Mode of diagnosis, all imaging and cytology were retrieved from the institutional electronic record. Cytology was categorized according to Bethesda. Four hundred and sixty-five patients were exposed to direct or scatter neck irradiation. Compliance with thyroid surveillance was observed in 76.9%. Ultrasound features of microcalcification and increased internal vascularity had a low sensitivity (62.5%) for predicting a malignant nodule, which improved when used in conjunction with a Bethesda IV-VI result (91.7%). However, cytological assessment was not performed in 45.6% of operative cases. Thirty-three patients had thyroid carcinoma of which 45.4% (n = 15) were incidental. The majority were papillary thyroid cancers (88.9%); of which 12.5% were node positive and 34.4% were multifocal. The incidence of thyroid cancer was elevated 57.6 times compared to the Australian general population (p < 0.001). Due to the high incidence of thyroid cancer, this study supports screening in this cohort. However, due to the risk of overtreatment, we endorse further investigation of thyroid nodules with ultrasound-guided fine-needle aspiration cytology based on sonographic criteria as for the general population and American Thyroid Association guidelines.


The effect of short-term labelling in (3H) thymidine on the viability of mouse blastomeres: alone and in combination with unlabelled blastomeres.
A method of labelling 8-cell-stage mouse blastomeres with [3H]thymidine is described, which allows them to be followed to the late blastocyst stage and is compatible with normal postimplantation development. However, the [3H]thymidine does affect the postimplantation vigour of the cells when placed in competition with unlabelled cells in blastomere aggregates. This suggests that caution should be used in interpreting results using [3H]thymidine as a label for early mammalian cells.


Integrins control epithelial stem cell proliferation in the Drosophila ovary by modulating the Notch pathway.
Cell proliferation and differentiation show a remarkable inverse relationship. The temporal coupling between cell cycle withdrawal and differentiation of stem cells (SCs) is crucial for epithelial tissue growth, homeostasis and regeneration. Proliferation vs. differentiation SC decisions are often controlled by the surrounding microenvironment, of which the basement membrane (BM; a specialized form of extracellular matrix surrounding cells and tissues), is one of its main constituents. Years of research have shown that integrin-mediated SC-BM interactions regulate many aspects of SC biology, including the proliferation-to-differentiation switch. However, these studies have also demonstrated that the SC responses to interactions with the BM are extremely diverse and depend on the cell type and state and on the repertoire of BM components and integrins involved. Here, we show that eliminating integrins from the follicle stem cells (FSCs) of the Drosophila ovary and their undifferentiated progeny increases their proliferation capacity. This results in an excess of various differentiated follicle cell types, demonstrating that cell fate determination can occur in the absence of integrins. Because these phenotypes are similar to those found in ovaries with decreased laminin levels, our results point to a role for the integrin-mediated cell-BM interactions in the control of epithelial cell division and subsequent differentiation. Finally, we show that integrins regulate proliferation by restraining the activity of the Notch/Delta pathway during early oogenesis. Our work increases our knowledge of the effects of cell-BM interactions in different SC types and should help improve our understanding of the biology of SCs and exploit their therapeutic potential.


Analyzing bone remodeling patterns after total hip arthroplasty using quantitative computed tomography and patient-specific 3D computational models.
Computational models in the form of finite element analysis technique that incorporates bone remodeling theories along with DEXA scans has been extensively used in predicting bone remodeling patterns around the implant. However, majority of such studies used generic models. Therefore, the aim of this study is to develop patient-specific finite element models of total hip replacement patients using their quantitative computed tomography (QCT) scans and accurately analyse bone remodelling patterns after total hip arthroplasty (THA). Patient-specific finite element models have been generated using the patients' QCT scans from a previous clinical follow-up study. The femur was divided into five regions in proximal-distal direction and then further divided into four quadrants for detailed analysis of bone remodeling patterns. Two types of analysis were performed-inter-patient and intra patient to compare them and then the resulting bone remodeling patterns were quantitatively analyzed. Our results show that cortical bone density decrease is higher in diaphyseal region over time and the cancellous bone density decreases significantly in metaphyseal region over time. In metaphyseal region, posterior-medial (P-M) quadrant showed high bone loss while diaphyseal regions show high bone loss in anterior-lateral (A-L) quadrant. Our study demonstrated that combining QCT with 3D patient-specific models has the ability of monitoring bone density change patterns after THA in much finer details. Future studies include using these findings for the development of a bone remodelling algorithm capable of predicting surgical outcomes for THA patients.


Teaching parasitology in a modular veterinary curriculum - The Vienna experience.
In a changing world with rapidly evolving new technologies, even in the "sheltered world" of teachers and lecturers at universities, we are challenged by new developments. As diversification is increasing in many aspects of our professional life, there is also a need to employ new ways of teaching, learning and assessments in veterinary curricula, taking into account the increasing numbers of students and limited teaching resources. The "classical" Austrian veterinary curriculum, with a series of consecutive lectures followed by practical courses in the different disciplines, separated from each other in the curricular time table and by annual examinations, has been a long-standing concept for teaching in the past. However, when veterinary practitioners and graduates where asked to judge the adequacy of undergraduate training for their professional skills and knowledge, several major shortcomings were revealed. The most commonly mentioned point was a lack of first-day competencies. As a consequence, the Vetmeduni Vienna developed a new veterinary curriculum implemented in 2014-2015. This curriculum covers 12 semesters and includes a "Diploma" (degree) thesis and several externships; graduates are eligible to practice in all branches of veterinary medicine. We abandoned the classical discipline-based teaching, established a modular, mostly organ-based system, and focus on student-centred and competency-based learning and teaching with defined learning outcomes and first-day skills. We also include training of scientific, managerial and communication skills in the curriculum. What does this mean for parasitology? We do not teach this subject in a closed lecture, but rather in a modular style, starting in the first year with the taxonomy and basic morphology of the animal kingdom, proceeding to parasite biology of the major groups in the second year, to the diagnostic and clinical training in practicals in the third year as well as joint lectures, seminars and conversatories in the organ- and species-based clinical training in years 3-6. Examinations are organised in different test formats, aligned to a list of skills and diseases. Quality assurance is a formal process and implemented at different levels. The ultimate aim of undergraduate teaching is to provide students with defined professional competencies and promoting life-long learning.


Numerical investigation of the mixed-mode failure of snow.
The failure of a weak snow layer underlying a cohesive slab is the primary step in the release process of a dry snow slab avalanche. The complex and heterogeneous microstructure of snow limits our understanding of failure initiation inside the weak layer, especially under mixed-mode shear-compression loading. Further complication arises from the dependence of snow strength on the loading rate induced by the balance between bond breaking and bond formation (sintering) during the failure process. Here, we use the discrete element method to investigate the influence of mixed-mode loading and fast sintering on the failure of a weak layer generated using cohesive ballistic deposition. Both fast and slow loading simulations resulted in a mixed-mode failure envelope in good agreement with laboratory experiments. We show that the number of broken bonds at failure and the weak layer strength significantly decreases with increasing loading angle, regardless of the loading rate. While the influence of loading rate appears negligible in shear-dominant loading (for loading angles above 30 ∘ ), simulations suggest a significant increase in the weak layer strength at low loading angles and low loading rates, characteristic of natural avalanches, due to the presence of an active sintering mechanism.


Synthesis and Characterization of a Thermoresponsive Copolymer with an LCST-UCST-like Behavior and Exhibiting Crystallization.
In this work, the diblock copolymer methoxy-poly(ethylene glycol)-block-poly(ε-caprolactone) (MPEG-b-PCL) was synthesized with a block composition that allows this polymer in aqueous media to possess both an upper critical solution temperature (UCST) and a lower critical solution temperature (LCST) over a limited temperature interval. The value of the UCST, associated with crystallization of the PCL-block, depended on heating (H) or cooling (C) of the sample and was found to be CPUCSTH = 32 °C and CPUCSTC = 23 °C, respectively. The LCST was not affected by the heating or cooling scans; assumed a value of 52 °C (CPLCSTH = CPLCSTC). At intermediate temperatures (e.g., 45 °C), dynamic light scattering (DLS), small-angle X-ray scattering (SAXS), and cryogenic transmission electron microscopy (cryo-TEM) showed that the solution consisted of a large population of spherical core-shell particles and some self-assembled rodlike objects. At low temperatures (below 32 °C), differential scanning calorimetry (DSC) and wide-angle X-ray scattering (WAXS) in combination with SAXS disclosed the formation of crystals with a cylindrical core-shell structure. Cryo-TEM supported a thread-like appearance of the self-assembled polymer chains. At temperatures above 52 °C, incipient phase separation took place and large aggregation complexes of amorphous morphology were formed. This work provides insight into the intricate interplay between UCST and LCST and the type of structures formed at these conditions in aqueous solutions of MPEG-b-PCL diblock copolymers.


A single-dose of intranasal vaccination with a live-attenuated SARS-CoV-2 vaccine candidate promotes protective mucosal and systemic immunity.
An attenuated SARS-CoV-2 virus with modified viral transcriptional regulatory sequences and deletion of open-reading frames 3, 6, 7 and 8 (∆3678) was previously reported to protect hamsters from SARS-CoV-2 infection and transmission. Here we report that a single-dose intranasal vaccination of ∆3678 protects K18-hACE2 mice from wild-type or variant SARS-CoV-2 challenge. Compared with wild-type virus infection, the ∆3678 vaccination induces equivalent or higher levels of lung and systemic T cell, B cell, IgA, and IgG responses. The results suggest ∆3678 as an attractive mucosal vaccine candidate to boost pulmonary immunity against SARS-CoV-2.


Degassing of 3H/3He, CFCs and SF6 by denitrification: measurements and two-phase transport simulations.
The production of N2 gas by denitrification may lead to the appearance of a gas phase below the water table prohibiting the conservative transport of tracer gases required for groundwater dating. We used a two-phase flow and transport model (STOMP) to study the reliability of 3H/3He, CFCs and SF6 as groundwater age tracers under agricultural land where denitrification causes degassing. We were able to reproduce the amount of degassing (R2=69%), as well as the 3H (R2=79%) and 3He (R2=76%) concentrations observed in a 3H/3He data set using simple 2D models. We found that the TDG correction of the 3H/3He age overestimated the control 3He/3He age by 2.1 years, due to the accumulation of 3He in the gas phase. The total uncertainty of degassed 3H/3He ages of 6 years (+/-2 sigma) is due to the correction of degassed 3He using the TDG method, but also due to the travel time in the unsaturated zone and the diffusion of bomb peak 3He. CFCs appear to be subject to significant degradation in anoxic groundwater and SF6 is highly susceptible to degassing. We conclude that 3H/3He is the most reliable method to date degassed groundwater and that two-phase flow models such as STOMP are useful tools to assist in the interpretation of degassed groundwater age tracer data.


Potential biomass yield per phosphorus and lipid accumulation property of seven microalgal species.
The potential biomass yield per phosphorus and lipid/triglyceride (TAG) accumulation properties of seven microalgal species: Scenedesmus sp. LX1, Chlorella ellipsoidea YJ1, Chlorella vuglaris, Chlorella sorokiniana, Chlorella pyrenoidosa, Dunaliella primolecta and Haematococcus pluvialis were investigated. Among the tested species, Scenedesmus sp. LX1 obtained the smallest minimal phosphorus content in cell (Q(0)) and the highest potential biomass yield of 6100kg-biomass/kg-P. After 12-day growth with intracellular phosphorus, Scenedesmus sp. LX1 accumulated about 30% lipid in biomass. Furthermore, the TAGs content per lipid of this strain (58.5%) as well as the lipid and TAGs yield per phosphorus (1800kg-lipid/kg-P and 680kg-TAGs/kg-P, respectively) were all significantly higher than that of any other species investigated in this study. Therefore, the phosphorus consumption to produce 1kg biodiesel using Scenedesmus sp. LX1 as feedstock was lowest among the tested species.


The Relationship Between "Protect People's Livelihood" and "Promote the Economy:" Provincial Evidence From China.
This study focused on medical care in a single country (China) and in regions with different economic backgrounds and different economic development levels to determine the effect of healthcare expenditure on short- and long-term economic growth. The study supported some interesting conclusions: (1) For most areas of China, increasing healthcare expenditure has a negative impact on economic growth in the short term but promotes growth in the long run; (2) Under different levels of economic development within China, there is significant heterogeneity in the interaction between healthcare expenditure and economic growth; (3) The negative effects of healthcare expenditure on short-term economic growth are greater during periods of economic turbulence than during times of stability; and (4) Healthcare expenditure has a negative effect on underdeveloped areas through the accumulation of material capital, while in economically developed areas, this channel has the opposite effect. To improve the quality of medical and health protection and the quality of life and welfare, China needs to consider the development characteristics of different economic zones and establish a multilevel, systematic and diversified medical and health protection system.


CYP2C9, CYP2C19, ABCB1 (MDR1) genetic polymorphisms and phenytoin metabolism in a Black Beninese population.
The genetically polymorphic cytochrome P450 2C9 (CYP2C9) metabolizes many important drugs. Among them, phenytoin has been used as a probe to determine CYP2C9 phenotype by measuring the urinary excretion of its major metabolite, S-enantiomer of 5-(4-hydroxyphenyl)-5-phenylhydantoin (p-HPPH). Phenytoin pharmacokinetic is also dependent on the activity of CYP2C19 and p-glycoprotein (ABCB1). To determine the influence of CYP2C9, CYP2C19 and ABCB1 genetic polymorphisms on phenytoin metabolism in a Black population, 109 healthy Beninese subjects received a single 300 mg oral dose of phenytoin. Blood was drawn 4 h after drug intake and urine was collected during the first 8 h. Plasma phenytoin and urine S- and R-enantiomers of p-HPPH were determined by high-performance liquid chromatography. Urinary excretion of (S)-p-HPPH [defined as urinary volumex(S)-p-HPPH urinary concentration] and PMR (defined as the ratio of p-HPPH in urine to 4 h phenytoin plasma concentration), both markers of CYP2C9 activity, were used to determine the functional relevance of new variants of CYP2C9 (*5, *6, *8, *9 and *11) in this population. Plasma phenytoin concentration was significantly associated with ABCB1 haplotype/genotype (P=0.05, Kruskal-Wallis test) and levels increased significantly in the genotype order: wild-type, T3421A and Block-2 genotypes (P=0.015, Jonckheere-Terpstra test). Urinary excretion of (S)-p-HPPH and PMR were significantly associated with the CYP2C9 genotype (P=0.001, analysis of variance (ANOVA) and P<0.0001, Kruskal-Wallis test, respectively) and decreased in the order: CYP2C9*1/*1, CYP2C9*1/*9, CYP2C9*9/*9, CYP2C9*1/*8, CYP2C9*8/*9, CYP2C9*9/*11, CYP2C9*1/*5, CYP2C9*6/*9, CYP2C9*1/*6, CYP2C9*8/*11, CYP2C9*5/*8 and CYP2C9*5/*6 (P<0.001, Jonckheere-Terpstra test). A combined analysis of CYP2C9, 2C19 and ABCB1 revealed that only ABCB1 predicted phenytoin concentration at 4 h and explained 8% of the variability (r=0.08, P=0.04). On the other hand, only CYP2C9 was predictive for the urinary excretion of (S)-p-HPPH and PMR (r=0.21, P=0.001 and r=0.25, P<0.001, respectively). Furthermore, significant relation was found between urinary excretion of (R)-p-HPPH and CYP2C9 genotype (P=0.035) and levels significantly increased in the genotype order: CYP2C9*1/*9, CYP2C9*1/*1, CYP2C9*9/*11, CYP2C9*1/*8 and CYP2C9*1/*5 (P<0.001, Jonckheere-Terpstra test). In summary, the present study demonstrates that, in a Black population, CYP2C9*5, *6, *8 and *11 variants, but not CYP2C9*9, are associated with a decreased phenytoin metabolism. The data also confirm the limited contribution of MDR1 gene to inter-individual phenytoin pharmacokinetic variation.


[Effects of arsenic trioxide and ATRA on PLZF-RARalpha-positive U937 leukemic cells].
To investigate effects of arsenic trioxide (As(2)O(3)) and alltrans retinoic acid (ATRA) on PLZF-RARalpha-positive cells. PLZF-RARalpha-positive U937 cells (U937/PLZF) were used as an in vitro model. The change of cell morphology was observed by Wright-Giemsa staining. Cell growth and proliferation were detected by methyl thiazolyl tetrazolium(MTT) assay. Cell cycle distribution and expression of cell membrane surface differentiation-related antigens (such as CD11b, CD64 and CD14) were determined by flow cytometry assay. Expression of PLZF was analyzed by immunofluorescence. Functional differentiation was reflected by nitroblue tetrazolium(NBT) reduction ability and cytochemistry staining. While U937/PLZF cells were incubated in tetracycline-withdrawn medium, the expression of PLZF-RARalpha; protein increased. After treated with As(2)O(3) (0.5 micromol/L) and ATRA (1 mumol/L), U937/PLZF cells presented some changes such as decreased nuclear/cytoplasm ratio, and partial disappearance of nucleoli, suggesting a certain degree of morphological differentiation. The cell growth and proliferation were inhibited in a dose- and time-dependent manner. The proportion of cells in S phage was decreased and CD11b level was increased. The expression of PLZF relocated in treated cells. However, no significant difference in NBT assay and cytochemistry staining was documented with the combination therapy. The combination of As(2)O(3) with ATRA can cause a slight tendency to morphological differentiation but is insufficient to induce functional differentiation of PLZF-RARalpha positive U937 leukemia cells.


The salivary microbiome is altered in the presence of a high salivary glucose concentration.
Type II diabetes (T2D) has been associated with changes in oral bacterial diversity and frequency. It is not known whether these changes are part of the etiology of T2D, or one of its effects. We measured the glucose concentration, bacterial counts, and relative frequencies of 42 bacterial species in whole saliva samples from 8,173 Kuwaiti adolescents (mean age 10.00 ± 0.67 years) using DNA probe analysis. In addition, clinical data related to obesity, dental caries, and gingivitis were collected. Data were compared between adolescents with high salivary glucose (HSG; glucose concentration ≥ 1.0 mg/d, n = 175) and those with low salivary glucose (LSG, glucose concentration < 0.1 mg/dL n = 2,537). HSG was associated with dental caries and gingivitis in the study population. The overall salivary bacterial load in saliva decreased with increasing salivary glucose concentration. Under HSG conditions, the bacterial count for 35 (83%) of 42 species was significantly reduced, and relative bacterial frequencies in 27 species (64%) were altered, as compared with LSG conditions. These alterations were stronger predictors of high salivary glucose than measures of oral disease, obesity, sleep or fitness. HSG was associated with a reduction in overall bacterial load and alterations to many relative bacterial frequencies in saliva when compared with LSG in samples from adolescents. We propose that hyperglycemia due to obesity and/or T2D results in HSG and subsequent acidification of the oral environment, leading to a generalized perturbation in the oral microbiome. This suggests a basis for the observation that hyperglycemia is associated with an increased risk of dental erosion, dental caries, and gingivitis. We conclude that HSG in adolescents may be predicted from salivary microbial diversity or frequency, and that the changes in the oral microbial composition seen in adolescents with developing metabolic disease may the consequence of hyperglycemia.


Studies on the O antigen of Salmonella typhosa. V. Enhancement of antibody response to protein antigens by the purified lipopolysaccharide.
Quantitative studies have demonstrated that a purified lipopolysaccharide (endotoxin) derived from the O-901 strain of Salmonella typhosa markedly enhanced the antibody response of rabbits when given separately or in conjunction with protein antigens. The augmentation of antibody levels varied from 2- to 40-fold with the number of injections, the dosage of antigen and endotoxin, and the route of administration. This antibody-enhancing property was found to be common to a broad group of endotoxins from Gram-negative bacilli and was not restricted to the lipopolysaccharide derived from S. typhosa. Factors affecting this enhancement were investigated, and data are presented which indicate that host susceptibility to endotoxin is a prerequisite for elevation of antibody levels; the intact lipopolysaccharide, as isolated, might not be essential for this activity; and the rate of clearance of antigen from the circulation of rabbits was accelerated when endotoxin was given in conjunction with protein. The data obtained are discussed in relation to postulated mechanisms on the antibody-enhancing action of endotoxin.


Transoral thyroidectomy and parathyroidectomy - A North American series of robotic and endoscopic transoral approaches to the central neck.
Most thyroid surgery in North America is completed via a cervical incision, which leaves a permanent scar. Approaches without cutaneous incisions offer aesthetic advantages. This series represents the largest series of transoral vestibular approaches to the central neck in North America, and the first published reports of robotic transoral vestibular thyroidectomy for thyroid carcinoma. Data was prospectively collected for patients that underwent transoral vestibular approach thyroidectomy and/or parathyroidectomy between April 2016 and February 2017. Fifteen patients underwent the procedure for removal of the thyroid (n=12), parathyroid (n=2) or both thyroid and parathyroid glands (n=1). The first case was converted to an open procedure. Fourteen were completed through these remote access incisions, including patients with a body mass index as high as 44. There were no permanent complications. The postoperative median Dermatology Life Quality Index score was 3, which indicates a small effect on quality of life. The transoral vestibular approach to the central neck is a promising technique for patients who desire to optimize aesthetics.


Capecitabine combined with weekly docetaxel in Chinese patients > 65 years with anthracycline-resistant metastatic breast cancer.
There are no data on more tolerable capecitabine doses in elderly patients in Chinese population. The aim of this study was to evaluate the activity and safety of capecitabine combined with weekly docetaxel for the treatment of anthracycline-resistant metastatic breast cancer (MBC) in older Chinese patients. MBC patients aged > 65 years pretreated with 1 - 5 prior chemotherapy regimens, including an anthracycline, received oral capecitabine 825 mg/m(2) twice daily, days 1 - 14, plus docetaxel 30 mg/m(2) on days 1 and 8 every 21 days. All 41 enrolled patients received at least 1 dose of treatment and were evaluable for safety; 38 received at least 2 cycles (median 4, range 2 - 8) and were evaluable for efficacy. The overall objective response rate was 47%, including complete responses in 8% of patients. Median time to progression was 8.9 months. Median overall survival was 17.6 months. The most common side effects were haematological and gastrointestinal toxicities and hand-foot syndrome. The only grade 3/4 adverse events were neutropenia (12%), alopecia (7%), grade 3 nausea and vomiting (2%) and grade 3 nail toxicity (2%). Capecitabine 825 mg/m(2) twice daily plus weekly docetaxel is active with an acceptable safety profile in Chinese women > 65 years with anthracycline-resistant MBC. Efficacy and tolerability compare favourably with previously reported trials evaluating higher capecitabine doses in combination with 3-weekly or weekly docetaxel.


New bisesquiterpenoid lactone from the wild rhizome of Atractylodes macrocephala Koidz grown in Qimen.
The rhizomes of Atractylodes macrocephala are used as both a food source and traditional Chinese medicine in China. A phytochemical investigation was carried out on wild A. macrocephala grown in Qimen County in eastern China, and yielded a novel bisesquiterpenoid lactone, namely, biatractylenolide II (1), along with two known compounds, atractylenolide II (2) and taraxeryl acetate (3). The structure and relative configuration of the new compound were elucidated mainly by 1D and 2D NMR spectroscopic methods in combination with HRESIMS experiments. This paper describes the isolation and structural elucidation of the new bisesquiterpenoid lactone (1).


Formation of amino acids from NH3 /NO2, CO2 and H2O: implications for the prebiotic origin of biomolecules.
The search for the conditions which must have prevailed in the long-distant past during the conversion of inanimate matter into animate matter is a fascinating area of research and it continues to draw the attention of the scientific community. The initiation of life on this planet must have been preceded by the development of biomolecules, amongst which amino acids have unique importance. Formation of amino acids under a certain set of conditions is shown in the present experiments. Solutions of ammonium carboxylates or the mixture of two such salts were prepared in 3:6.9:0.1 (v/v/v) acetonitrile/water/formic acid at a concentration of 50 μM. The studies were performed using a quadrupole time-of-flight (QqTOF) mass spectrometer. The formation of different amino acids was detected with high-resolution mass spectrometry. Here, we show the formation of amino acids when a solution of ammonium salts was injected into an electrospray ionization (ESI)-QqTOF-MS instrument. The ammonium salts were the source of NH3 and CO2 and H2 O was available in the medium. It seems that the combination of NH3 , CO2 , and H2 O leads to the formation of amino acids. Further to the literature reports of formation of amino acids under the reduced atmosphere represented by gases such as NH3 , CH4 , H2 and H2 O, here we demonstrate the formation of amino acids by the combination of NH3 /NO2 , CO2 and water vapours in the ESI source of the mass spectrometer.


On the limit cycle structure of threshold boolean networks over complete graphs.
In previous work, the limit structure of positive and negative finite threshold boolean networks without inputs (TBNs) over the complete digraph K(n) was analyzed and an algorithm was presented for computing this structure in polynomial time. Those results are generalized in this paper to cover the case of arbitrary TBNs over K(n). Although the limit structure is now more complicated, containing, not only fixed-points and cycles of length 2, but possibly also cycles of arbitrary length, a simple algorithm is still available for its determination in polynomial time. Finally, the algorithm is generalized to cover the case of symmetric finite boolean networks over K(n).


Plant-pathogen interaction, circadian rhythm, and hormone-related gene expression provide indicators of phytoplasma infection in Paulownia fortunei.
Phytoplasmas are mycoplasma-like pathogens of witches' broom disease, and are responsible for serious yield losses of Paulownia trees worldwide. The molecular mechanisms of disease development in Paulownia are of considerable interest, but still poorly understood. Here, we have applied transcriptome sequencing technology and a de novo assembly approach to analyze gene expression profiles in Paulownia fortunei infected by phytoplasmas. Our previous researches suggested that methyl methane sulfonated (MMS) could reverse the effects of the infection. In this study, leaf samples from healthy, infected, and both infected and methyl methane sulfonate treated plants were analyzed. The results showed that the gene expression profile of P. fortunei underwent dramatic changes after Paulownia witches' broom (PaWB) phytoplasma infection. Genes that encoded key enzymes in plant-pathogen interaction processes were significantly up-regulated in the PaWB-infected Paulownia. Genes involved in circadian rhythm and hormone-related genes were also altered in Paulownia after PaWB infection. However, after the PaWB-infected plants were treated with MMS, the expression profiles of these genes returned to the levels in the healthy controls. The data will help identify potential PaWB disease-resistance genes that could be targeted to inhibit the growth and reproduction of the pathogen and to increase plant resistance.


A global network for early warning and response to infectious diseases and bioterrorism: applied epidemiology and training programs, 2001.
In many ministries of health, applied epidemiology and training programs (AETPs) are responsible for detecting and responding to acute health events, including bioterrorism. In November 2001, we assessed the bioterrorism response capacity of 29 AETPs; 17 (59%) responded. Fifteen countries (88%) had bioterrorism response plans; in 6 (40%), AETPs took the lead in preparation and in 6 (40%) they assisted. Between September 11 and November 29, 2001, 12 AETPs (71%) responded to a total of 3024 bioterrorism-related phone calls. Six programs (35%) responded to suspected bioterrorism events. AETPs play an important role in bioterrorism surveillance and response. Support for this global network by various health agencies is beneficial for all developed and developing countries.


The Changing Safety Net for Low-Income Parents and Their Children: Structural or Cyclical Changes in Income Support Policy?
Refundable tax credits and food assistance are the largest transfer programs available to able-bodied working poor and near-poor families in the United States, and simultaneous participation in these programs has more than doubled since the early 2000s. To understand this growth, we construct a series of two-year panels from the 1981-2013 waves of the Current Population Survey Annual Social and Economic Supplement to estimate the effect of state labor-market conditions, federal and state transfer program policy choices, and household demographics governing joint participation in food and refundable tax credit programs. Overall, changing policy drives much of the increase in the simultaneous, biennial use of food assistance and refundable tax credits. This stands in stark contrast from the factors accounting for the growth in food assistance alone, where cyclical and structural labor market factors account for at least one-half of the growth, and demographics play a more prominent role. Moreover, since 2000, the business cycle factors as the leading determinant in biennial participation decisions in food programs and refundable tax credits, suggesting a recent strengthening in the relationship between economic conditions and transfer programs.


[Reinnervation of posterior cricoarytenoid muscle: a comparison of nerve implantation and neuromuscular pedicle transfer in an animal model].
Recently, methods of reinnervating posterior cricoarytenoid muscle (PCA) have been reported by many authors. This study was undertaken to compare direct implantation (NI) of a branch of the ansa cervicalis supplying the sternothyroid muscle with the transfer of a neuro-muscular pedicle of the ansa cervicalis innervating the sternothyroid muscle (NMP) in a denervated PCA dog model. Both were carried out immediately after denervation. The results showed that reinnervation with return of physiological function of PCA were got by both methods of nerve transfer. The NMP was apparently superior to the NI. The reason was that the NMP implanted more motor end-plates into the denervated PCA. So the NMP should be chosen first when possible. The NI is consistently effective when NMP is not available.


Decoy receptor-2 small interfering RNA (siRNA) strategy employing three different siRNA constructs in combination defeats adenovirus-transferred tumor necrosis factor-related apoptosis-inducing ligand resistance in lung cancer cells.
Tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) selectively induces apoptosis in cancer cells but not in normal cells. However, studies have indicated that more than half of human tumors exhibit TRAIL resistance. Although the mechanism of TRAIL resistance is not understood, it represents a barrier to any TRAIL-mediated gene therapy approach. In addition, no correlation between TRAIL receptor (TRAIL-R) expression profile and TRAIL resistance has been demonstrated in cancer cells. In this study, three different lung cancer cell lines and three different primary cell cultures established from patients with lung cancer (two patients with squamous cell lung carcinoma and one with adenocarcinoma) were screened for sensitivity to adenoviral delivery of TRAIL. Whereas TRAIL-resistant primary lung cell cultures and the A549 lung cancer cell line exhibited high levels of surface decoy receptor-2 (DcR2/TRAIL-R4) expression, TRAIL-sensitive lung cancer cell lines (HBE and H411) failed to express it. A DcR2 short interfering RNA (siRNA) approach involving three different siRNA constructs in combination downregulated DcR2/TRAIL-R4 expression and sensitized lung cancer cells to TRAIL-induced apoptosis. Immunohistochemical staining of samples from 10 patients with lung carcinoma suggested that high-level DcR2/TRAIL-R4 expression is a common phenotype observed in patients with non-small cell lung carcinoma.


Normal cognition and behavior in a Smith-Lemli-Opitz syndrome patient who presented with Hirschsprung disease.
Smith-Lemli-Opitz syndrome (SLOS) is an autosomal recessive disorder of cholesterol biosynthesis. It is caused by mutations in the gene encoding the enzyme 7-dehydrocholesterol Delta7-reductase (DHCR7), which catalyzes the final step in cholesterol biosynthesis, usually resulting in cholesterol deficiency. We report a 3.5-year-old girl who has cognition in the low average range and normal behavior, but in whom molecular studies identified two missense mutations in DHCR7: V326L and F284L. She was born at term following an uncomplicated pregnancy and delivery, and presented at 12 days of age with poor feeding, abdominal distention, and jaundice. Colonic biopsy was consistent with Hirschsprung disease. On physical examination she had mild ptosis, a long philtrum, mild micrognathia, a short, upturned nose, and subtle 2,3 syndactyly. Her 7-dehydrocholesterol (7-DHC) level was markedly elevated at 8.7 mg/dl (normal 0.10 +/- 0.05), and her cholesterol level was normal at 61 mg/dl (normal for newborn period 50-80 mg/dl). Karyotype analysis was normal, 46,XX. Breast milk feeding was initiated and continued for 18 months. Cholesterol supplementation was implemented at 100 mg/kg/day at 3 months, which resulted in increased cholesterol levels and reduced dehydrocholesterol levels. Neuropsychological testing has shown functioning in the low average range, between the 14th and 18th centiles when compared to peers. This is markedly higher than most children with SLOS. She has no behavioral problems. MRI and MRS testing of the brain revealed no structural abnormalities. This is in contrast to a recently reported case by Prasad et al. [2002: Am J Med Genet 108:64-68] with a mild phenotype, behavioral problems, and abnormal MRI, who is compound heterozygote for both a null and missense mutation. Our case suggests that patients with severe feeding disorders with or without Hirschprung disease and postnatal onset microcephaly may warrant screening for SLOS.


Overdiagnosis of gastric cancer by endoscopic screening.
Gastric cancer screening using endoscopy has recently spread in Eastern Asian countries showing increasing evidence of its effectiveness. However, despite the benefits of endoscopic screening for gastric cancer, its major harms include infection, complications, false-negative results, false-positive results, and overdiagnosis. The most serious harm of endoscopic screening is overdiagnosis and this can occur in any cancer screening programs. Overdiagnosis is defined as the detection of cancers that would never have been found if there is no cancer screening. Overdiagnosis has been estimated from randomized controlled trials, observational studies, and modeling. It can be calculated on the basis of a comparison of the incidence of cancer between screened and unscreened individuals after the follow-up. Although the estimation method for overdiagnosis has not yet been standardized, estimation of overdiagnosis is needed in endoscopic screening for gastric cancer. To minimize overdiagnosis, the target age group and screening interval should be appropriately defined. Moreover, the balance of benefits and harms must be carefully considered to effectively introduce endoscopic screening in communities. Further research regarding overdiagnosis is warranted when evaluating the effectiveness of endoscopic screening.


What is digital medicine?
Changes in health care are a fundamental part of social and intellectual evolution. The modern practice of scientific medicine depends on the existence of the written and printed word to store medical information. Because computers can transform information as well as store it, new digital tools cannot only record clinical data, they can also generate medical knowledge. In doing so, they make it possible to develop "digital medicine" that is potentially more precise, more effective, more experimental, more widely distributed, and more egalitarian than current medical practice. Critical steps in the creation of digital medicine are careful analysis of the impact of new technologies and coordinated efforts to direct technological development towards creating a new paradigm of medical care.


Demonstration of nitrogen coordination in metal--bleomycin complexes by electron spin--echo envelope spectroscopy.
We have studied the Cu(II), Co(II), and Fe(III) complexes of the antineoplastic drug bleomycin by using electron spin--echo envelope spectroscopy. For all three complexes, nitrogen coordination of the metal ions is demonstrated. For the Cu(II)-- and Co(II)--drug complexes, we have been able to identify imidazole as a metal ligand.


Convergent evolution of syringyl lignin biosynthesis via distinct pathways in the lycophyte Selaginella and flowering plants.
Phenotypic convergence in unrelated lineages arises when different organisms adapt similarly under comparable selective pressures. In an apparent example of this process, syringyl lignin, a fundamental building block of plant cell walls, occurs in two major plant lineages, lycophytes and angiosperms, which diverged from one another more than 400 million years ago. Here, we show that this convergence resulted from independent recruitment of lignin biosynthetic cytochrome P450-dependent monooxygenases that route cell wall monomers through related but distinct pathways in the two lineages. In contrast with angiosperms, in which syringyl lignin biosynthesis requires two phenylpropanoid meta-hydroxylases C3'H and F5H, the lycophyte Selaginella employs one phenylpropanoid dual meta-hydroxylase to bypass several steps of the canonical lignin biosynthetic pathway. Transgenic expression of the Selaginella hydroxylase in Arabidopsis thaliana dramatically reroutes its endogenous lignin biosynthetic pathway, yielding a novel lignin composition not previously identified in nature. Our findings demonstrate a unique case of convergent evolution via distinct biochemical strategies and suggest a new way to genetically reconstruct lignin biosynthesis in higher plants.


A survey of clinical and laboratory characteristics of the dengue fever epidemic from 2017 to 2019 in Zhejiang, China.
To explore the epidemic, clinical, and laboratory characteristics of dengue patients in Zhejiang and the possible mechanism. Epidemic, clinical and laboratory data of 231 dengue patients admitted to the Second Affiliated Hospital of Zhejiang Traditional Chinese Medicine University between August 2017 and December 2019 were collected. GSE43777 dataset was downloaded from the Gene Expression Omnibus database and was used for the immune cell infiltration analysis, logistic regression analysis, and nomogram construction. Gene set enrichment analysis (GSEA) was performed to explore the possible regulatory pathways in dengue infection. Further, the receiver operating characteristic curve analysis and decision curve analysis were conducted to evaluate the value of related immune cells in predicting dengue severity. Among the 231 patients, the gender ratio was 1:1.1 (male/female). The patients in the <60 years age group, 60 to 80 years age group, and >80 years age group were 47.2%, 45.5%, and 7.3%, respectively. The major symptoms were fever (100%), weak (98.3%), anorexia (76.6%), muscle and joint pain (62.3%), and nausea (46.8%). In dengue patients, 98.7% of serum samples had decreased platelet levels, 96.5% of them had decreased white blood cell (WBC) levels, 97.8% had elevated aspartate aminotransferase levels, 82.3% had elevated lactate dehydrogenase levels, 49.4% had increased creatinine levels, and 35.5% had increased creatine kinase levels. Pneumonia, pleural effusion, and bilateral pleural reaction were observed in 16.5%, 8.2%, and 4.8%, respectively of dengue patients. Gallbladder wall roughness and splenomegaly accounted for 6.1% and 4.3% of all cases. Moreover, the levels of T cell, B cell, and dendritic cells were significantly higher in the convalescent group and they were involved in immune- and metabolism-related pathways. Of note, low levels of these 3 immune cells correlated with high dengue infection risk, while only dendritic cells exhibited satisfactory performance in predicting dengue severity. Dengue fever patients often onset with fever, accompanied by mild abnormalities of the blood system and other organ functions. Moreover, T cells, B cells, and dendritic cells might be involved in dengue infection and development.


Computational Reproducibility of Molecular Phylogenies.
Repeated runs of the same program can generate different molecular phylogenies from identical data sets under the same analytical conditions. This lack of reproducibility of inferred phylogenies casts a long shadow on downstream research employing these phylogenies in areas such as comparative genomics, systematics, and functional biology. We have assessed the relative accuracies and log-likelihoods of alternative phylogenies generated for computer-simulated and empirical data sets. Our findings indicate that these alternative phylogenies reconstruct evolutionary relationships with comparable accuracy. They also have similar log-likelihoods that are not inferior to the log-likelihoods of the true tree. We determined that the direct relationship between irreproducibility and inaccuracy is due to their common dependence on the amount of phylogenetic information in the data. While computational reproducibility can be enhanced through more extensive heuristic searches for the maximum likelihood tree, this does not lead to higher accuracy. We conclude that computational irreproducibility plays a minor role in molecular phylogenetics.


Activation of protease-constitutive recA proteins of Escherichia coli by all of the common nucleoside triphosphates.
To understand why the RecA proteins of the protease-constitutive recA1202 and recA1211 mutants show very high protease activities in vivo without the usual need for DNA damage (E. S. Tessman and P. Peterson, J. Bacteriol. 163:677-687, 1985), we examined the activation of the mutant proteins by nucleoside triphosphates (NTPs) in vitro. In vivo, the mutant protease activities are resistant to inhibition by cytidine plus guanosine (C + G) in the growth medium, in contrast to the activities of weaker mutants, such as recA441, which are sensitive to C + G inhibition. We found that RecA1202 and RecA1211 proteins, in contrast to RecA+, can use natural NTPs other than ATP and dATP as cofactors in the cleavage of LexA repressor. The effectiveness of NTPs in promoting LexA cleavage by RecA1202 and RecA1211 proteins decreased in roughly the following order: dATP greater than ATP greater than UTP greater than ATP-gamma S greater than dCTP greater than CTP greater than dGTP greater than GTP greater than TTP. These mutant proteins showed higher affinities for ATP and single-stranded DNA and higher repressor cleavage activities than RecA+ protein. With the various effectors (single-stranded DNA or NTPs), the RecA1202 protein always showed more activity than RecA1211 in the cleavage of LexA repressor in vitro, which is consistent with the greater activity of the recA1202 mutant in vivo. The results explain, in part, why some recA mutants have unusually high constitutive RecA protease activity and why that activity is more or less resistant to C + G inhibition.


De Novo High-Titer Production of Delta-Tocotrienol in Recombinant Saccharomyces cerevisiae.
Delta-tocotrienol as a vitamin E isomer has received much attention because of its diverse biomedical applications. Microbial biosynthesis of delta-tocotrienol is a promising strategy for its economic and environmental advantages. Here, we accomplished complete biosynthesis of delta-tocotrienol in Saccharomyces cerevisiae from glucose. We first constructed and incorporated a heterologous pathway into the genome of S. cerevisiae by incorporating the genes hpd (from Pseudomonas putida KT2440), hpt (from Synechocystis sp. PCC 6803), and vte1 (from Arabidopsis thaliana) for the biosynthesis of delta-tocotrienol. We further enhanced the biosynthesis of the precursor geranylgeranyl diphosphate by overexpressing the thmg1 and ggppssa (from Sulfolobus acidocaldarius) genes, leading to a production titer of delta-tocotrienol of 1.39 ± 0.01 mg/L. Finally, we optimized the fermentation medium using the response surface methodology, enabling a high-titer production of delta-tocotrienol (3.56 ± 0.25 mg/L), ∼2.6-fold of that of the initial culture medium. Fed-batch fermentation in a 2 L fermenter was further used to enhance the production titer of delta-tocotrienol (4.10 ± 0.10 mg/L). To the best of our knowledge, this is the first report on the de novo biosynthesis of delta-tocotrienol in S. cerevisiae, and the highest titer obtained for microbial production of delta-tocotrienol.


[The structure of microtubules].
Cytoplasmic microtubules, plane monolayers of tubulin filaments and single filaments have been investigated by electron microscopy. The methods of optical diffraction, filtering and three-dimensional reconstruction have been used to analyse the micrographs and to construct a model of the dimeric molecule of tubulin. When viewed in the direction perpendicular to the longitudinal axes the molecule bear resemblance to the figure "8". When rotated by 90 degrees it looks as if composed of two globules. Two variants of the quoternary structure are considered: 1) each of two subunits is a deformed ring; 2) each rings of the "8"-like particles consists of two halves of different subunits. The second protein layer which is formed on microtubules in the presence of protamine consists of tubulin and is characterized by the crystal lattice similar to that of the microtubule itself, but rotated through the angle of about 95 degrees. In the presence of protamine we also detected flat spirals and stacks of such spirals forming short tubules.


Young Children's Moral Judgments, Justifications, and Emotion Attributions in Peer Relationship Contexts.
Children (n = 160, 4- to 9-year-olds; Mage = 6.23 years, SD = 1.46) judged, justified, attributed emotions, and rated intent for hypothetical physical harm, psychological harm, and resource distribution transgressions against close friends, acquaintances, disliked peers, or bullies. Transgressions against bullies were judged more acceptable than against friends and disliked peers and less deserving of punishment than against acquaintances and disliked peers. Transgressions against friends were judged least intended and resulting in more negative emotions for transgressors; actors transgressing against disliked peers, as compared to bullies or acquaintances, were happy victimizers. Across relationships, children viewed moral transgressions as wrong independent of rules and authority, based primarily on welfare and fairness justifications. Peer context colors but does not fundamentally change moral evaluations.


Genetic variation in IL28B (IFNL3) and response to interferon-alpha treatment in myeloproliferative neoplasms.
In myeloproliferative neoplasms (MPN), interferon-alpha (IFN-α) is an effective treatment with disease-modifying properties but currently with no clear predictors of treatment outcome. Recent genomewide association studies in chronic hepatitis C have found a strong influence of genetic polymorphism near the IL28B (IFNL3) gene in response to IFN-α treatment. In this study, we sought to evaluate the prognostic impact of IL28B rs12979860, rs8099917, and rs12980275 on IFN-α treatment response in myeloproliferative neoplasms. We retrospectively evaluated 100 patients with MPN treated with IFN-α. The hematologic treatment response on IFN-α was compared between patients and correlated with host genetic variations in IL28B. The genotypes of IL28B were determined by allelic discrimination assays. The CC genotype of rs12979860 was found significantly associated with hematologic response in polycythemia vera (PV) with a complete response (CR) in 79% (CC) compared to 48% (non-CC), (P = .036). No association between the genotypes and treatment response on hydroxyurea was found. These results imply an effect of IL28B genotype on the outcome of IFN-α treatment in MPN.


Latent synthesis of electrically conductive surface-silvered polyimide films.
A facile ambient temperature route to the fabrication of surface silver-metallized polyimide films is described. Silver(I) trifluoromethanesulfonate or silver(I) nitrate and a polyimide, derived from 2,2-bis(3,4-dicarboxyphenyl)hexafluoropropane dianhydride and an equimolar amount of 4,4'-oxydianiline and 3,5-diaminobenzoic acid, were dissolved together in dimethylacetamide. Silver(I)-doped films were prepared at thicknesses of 25-40 microm and depleted of solvent by evaporation at ambient temperature and low humidity. The silver(I)-ion-containing films were then treated with aqueous solutions of the reducing agents hydrazine hydrate and hydroxylamine, which brought forth surface-silvered films exhibiting conductivity on the order of bulk polycrystalline silver accompanied by modest-to-high specular reflectivity.


A study of aryl radical cyclization in enaminone esters.
Aryl radical cyclization in N-phenyl, N-benzyl, and N-phenethyl enaminone esters 1a-f was studied. N-Benzyl and N-phenethyl enaminones afforded 5-exo and 6-exo cyclization products, respectively, but radical cyclization did not occur in N-phenyl enaminones. The rate constants for the 5-exo and 6-exo cyclization processes in secondary enaminones were estimated as being on the order of 10(7) s(-1) at 353 K; since DNMR experiments showed the rate constant for rotation around the enaminone C3-N bond to be on the order of 10(4) s(-1) at this temperature, the initial enaminone configuration is maintained throughout the cyclization process. PM3 calculations suggested that the nonoccurrence of endo and 4-exo cyclizations is due to the corresponding transition structures involving significant distortion of the conjugated enaminone system.


Effect of changes in cruciate ligaments pretensions on knee joint laxity and ligament forces.
The knee joint cruciate ligaments are reconstructed with the rationale to avoid joint instability, recurrent injury, damage to soft tissues and osteoarthritis. Wide range of procedures with different stiffness, pretension, orientation and insertion locations have been proposed with the primary goal to restore the joint laxity. Apart from the general lack of success in preservation of force in the reconstructed ligament itself, the concern, not yet addressed, arises as to the effect of such perturbation on the other intact cruciate ligament. The interaction between cruciate ligament forces is hypothesized in this work. Using a 3-D nonlinear finite element model of the tibiofemoral joint, we examined this hypothesis by quantifying the extent of coupling between cruciate ligaments while varying the prestrain in each ligament under flexion with and without anterior-posterior (A-P) loads. A remarkable coupling was predicted between cruciate ligament forces in flexion thus confirming the hypothesis; forces in both cruciate ligaments increased as initial strain or pretension in one of them increased whereas they both diminished as one of them became slack. Moreover, changes in laxity and in ligament forces as a cruciate ligament prestrained or pretensioned varied with flexion angle and external loads. These findings have important consequences in joint functional biomechanics following a ligament injury or replacement surgery and in selection of laxity matched or ligament force matched pretensioning protocols.