Inhibition of Friend virus replication by a compound that reacts with the nucleocapsid zinc finger: anti-retroviral effect demonstrated in vivo.
The zinc finger structure that is found in the nucleocapsid protein of nearly all retroviruses has been proposed as a target for antiviral therapy. Since compounds that chemically attack the cysteines of the finger have been shown to inactivate both human immunodeficiency virus type 1 (HIV-1) and murine leukemia virus (MuLV) in vitro, 14 of these compounds were tested in an MuLV-induced Friend disease model to assess their ability to inhibit retroviral replication in vivo. Of the 14 compounds tested, only Aldrithiol-2 clearly exhibited anti-retroviral activity as measured indirectly by the delay of Friend disease onset (P < 0.05). These results were confirmed by quantitative competitive polymerase chain reaction studies which monitored viral spread by measuring the level of viral DNA in the peripheral blood mononuclear cells of treated mice. Comparison of treated mice with untreated mice revealed that Aldrithiol-2 produced a greater than 2-log reduction in virus levels. These results functionally demonstrate that a zinc finger-attacking compound can inhibit viral replication in vivo. Since only 1 of the 14 compounds studied was effective, this study also shows the importance of in vivo testing of these types of antiviral compounds in an animal model. Given the strict conservation of the metal-coordinating cysteine structure within HIV-1 and MuLV zinc fingers, our results support the proposal that anti-retroviral drugs which target the nucleocapsid zinc finger may be clinically useful against HIV-1.


Blueprints for green biotech: development and application of standards for plant synthetic biology.
Synthetic biology aims to apply engineering principles to the design and modification of biological systems and to the construction of biological parts and devices. The ability to programme cells by providing new instructions written in DNA is a foundational technology of the field. Large-scale de novo DNA synthesis has accelerated synthetic biology by offering custom-made molecules at ever decreasing costs. However, for large fragments and for experiments in which libraries of DNA sequences are assembled in different combinations, assembly in the laboratory is still desirable. Biological assembly standards allow DNA parts, even those from multiple laboratories and experiments, to be assembled together using the same reagents and protocols. The adoption of such standards for plant synthetic biology has been cohesive for the plant science community, facilitating the application of genome editing technologies to plant systems and streamlining progress in large-scale, multi-laboratory bioengineering projects.


[Risk of nerve damage by surgery of the lateral malleolus].
Fracture of the lateral malleolus is a common injury, which often requires surgery. The lateral approach is often used with a "cut straight to bone" method. In this 59-year-old woman with an ankle fracture we found a variant of the superficial peroneal nerve (SPN) crossing the operative field with great danger of laceration. A review of the literature reveals that in more than 10% of the cases significant nerve branches may be found in the operative field. This report emphasises the need for neurological exam and recommends that great care for nerve variants should be taken during surgery on and around the lateral malleolus.


Combined Laser Photocoagulation and anti-VEGF Injection Treatment in Radiation Retinopathy.
Background Radiation retinopathy can lead to a considerable reduction in visual acuity. We report 2 cases of radiation retinopathy in which a therapy with combined intravitreal anti-VEGF injection and laser photocoagulation (LPC) was used. Patient History Two 55-year-old women were referred to our clinic with radiation retinopathy, in the first case unilaterally after radiation due to endocrine orbitopathy (P1), in the second case in both eyes after palliative cerebral radiation for metastatic breast cancer (P2). Both cases were treated with combined intravitreal anti-VEGF injection and focal LPC. Therapy After the initiation therapy macular edema decreased considerably. One year after the beginning of therapy visual acuity increased in the first case from 0,05 to 0,16 p, in the second case from 0,5/0,4 to 0,6/0,5. Despite the positive response there is still continuing need for therapy in all three eyes. Conclusions Combined anti-VEGF injection and LT can reduce macular edema due to radiation retinopathy. The follow-up observation shows the necessity of long-term care.


Primary oat cell carcinoma of the larynx.
The aggressiveness of small (oat) cell carcinoma of the larynx presents a therapeutic challenge to the oncologist. Since the first description of this type of carcinoma in 1972, 52 patients have been reported in the literature and a variety of treatment regimens have been used. The purpose of this study was to report two new cases and review all previous reports to determine the disease's biological behavior, clinical manifestations, and optimum treatment. Thirty-five percent of the tumors were transglottic, and 27% were supraglottic. Fifty-four percent of patients had regional metastases at initial presentation and 17.6% had distant metastases. The median survival was 10 months for all patients. Patients who were treated with chemotherapy with or without other modalities had the best 2-year survival rates (52.2%). Forty-one percent of patients had regional recurrence only, 12.5% had regional recurrence and distant metastases, and 2% developed distant metastases only. We conclude that patients with oat cell carcinoma of the larynx should be treated with combination chemotherapy and radiation therapy. Surgery is best reserved for persistent and recurrent disease at the primary site and neck.


Identification of human hepatic cytochrome p450 enzymes involved in the biotransformation of cholic and chenodeoxycholic acid.
3alpha,7alpha,12alpha-trihydroxy-5beta-cholan-24-oic (cholic) and 3alpha,7alpha-dihydroxy-5beta-cholan-24-oic (chenodeoxycholic) acids are the predominant hepatic and biliary bile acids of most mammalian species including humans. Cholic and chenodeoxycholic acids are synthesized from cholesterol and accumulate in the liver during cholestasis. Biotransformation by hepatic cytochrome P450 (P450) enzymes represents a potentially effective pathway for elimination of these lipid-soluble bile acids. We developed a liquid chromatography/mass spectrometry-based assay to identify and quantify the human hepatic microsomal metabolites of cholic acid and chenodeoxycholic acid, and using a panel of human recombinant P450 enzymes, we determined the P450 enzymes involved. Incubation of cholic acid with human hepatic microsomes and NADPH produced a single metabolite, 7alpha,12alpha-dihydroxy-3-oxo-5beta-cholan-24-oic (3-dehydrocholic) acid. Of the recombinant P450 enzymes tested, only CYP3A4 catalyzed 3-dehydrocholic acid formation. Similar experiments with chenodeoxycholic acid revealed the formation of 7alpha-hydroxy-3-oxo-5beta-cholan-24-oic acid and 3alpha,6alpha,7alpha-trihydroxy-5beta-cholan-24-oic (gamma-muricholic) acid as major metabolites and 3alpha-hydroxy-7-oxo-5beta-cholan-24-oic (7-ketolithocholic) acid and cholic acid as minor metabolites. Among the human recombinant P450 enzymes examined, CYP3A4 exhibited the highest rates of formation for 7alpha-hydroxy-3-oxo-5beta-cholan-24-oic acid and gamma-muricholic acid from chenodeoxycholic acid. Formation of 7-ketolithocholic acid and cholic acid from chenodeoxycholic acid has not been reported previously and could not be attributed to any of the recombinant P450 enzymes tested. In conclusion, the predominant pathway for the biotransformation of both cholic and chenodeoxycholic acids in human hepatic microsomes was oxidation at the third carbon of the cholestane ring. This study highlights a major role for CYP3A4 and suggests a possible route for the elimination of these two bile acids.


Atopic dermatitis severity during exposure to air pollutants and weather changes with an Artificial Neural Network (ANN) analysis.
Epidemiological studies have shown an association between global warming, air pollution, and allergic diseases. Several air pollutants, including volatile organic compounds, formaldehyde, toluene, nitrogen dioxide (NO2 ), and particulate matter, act as risk factors for the development or aggravation of atopic dermatitis (AD). We evaluated the impact of air pollutants and weather changes on AD patients. Sixty AD patients ≥5 years of age (mean age: 23.5 ± 12.5 years), living in the Campania Region (Southern Italy), were followed for 18 months. The primary outcome was the effect of atmospheric and climatic factors on signs and symptoms of AD, assessed using the SCORAD (SCORing Atopic Dermatitis) index. We measured mean daily temperature (TOD), outdoor relative humidity (RH), diurnal temperature range (DTR), precipitation, particulate with aerodynamic diameter ≤ 10 μm (PM10 ), NO2 , tropospheric ozone (O3 ), and total pollen count (TPC). A multivariate logistic regression analysis was used to examine the associations of AD signs and symptoms with these factors. An artificial neural network (ANN) analysis investigated the relationships between weather changes, environmental pollutants, and AD severity. The severity of AD symptoms was positively correlated with outdoor temperatures (TOD, DTR), RH, precipitation, PM10 , NO2 , O3 , and TPC. The ANN analysis also showed a good discrimination performance (75.46%) in predicting disease severity based on environmental pollution data, but weather-related factors were less predictive. The results of the present study provide evidence that weather changes and air pollutions have a significant impact on skin reactivity and symptoms in AD patients, increasing the severity of the dermatitis. The knowledge of the single variables proportion on AD severity symptoms is important to propose alerts for exacerbations in patients with AD of each age. This finding represents a good starting point for further future research in an area of increasingly growing interest.


Bortezomib Treatment to Potentiate the Anti-tumor Immunity of Ex-vivo Expanded Adoptively Infused Autologous Natural Killer Cells.
Our results show that using an EBV-LCL feeder cell line, large numbers of pure populations of highly activated NK cells can be expanded ex vivo under GMP conditions. With the exception of thyroiditis and constitutional symptoms related to IL-2 therapy, infusions of up to 1 x 10(8) cells/kg of ex vivo expanded NK have been well tolerated and have provided preliminary clinical evidence for mediating anti-tumor immunity in patients with advanced cancer.


The effect of protein supplementation on quality of life, physical function, and muscle strength in patients with chronic obstructive pulmonary disease.
The combination of protein supplementation with exercise is successful in increasing weight and energy intake, as well as exercise capacity and health-related quality of life in sarcopenic patients diagnosed with chronic obstructive disease (COPD). However, the potential benefit of protein supplementation for non-sarcopenic patients with COPD has yet not previously been examined. The aim of this trial was to evaluate the effect of protein supplementation on quality of life, physical function, muscle strength and biochemical blood markers in patients diagnosed with COPD undergoing nine weeks of pulmonary rehabilitation. A prospective, parallel group randomised clinical trial. Patients referred from their general practitioners to the COPD rehabilitation outpatient programme at the local community rehabilitation centre. Patients (N.=53) with stable moderate to severe COPD diagnosed with COPD, 40 years or older and with a BMI<30. The participants were assigned to one of two groups to receive either twice daily protein supplementation (9.3 g of protein/566.4 KJ) plus exercise or exercise only. Before and after nine weeks of rehabilitation, mental state was measured by means of St George Respiratory Questionnaire, physical performance was evaluated by shuttle walking test and maximal muscle strength test, and fasting blood samples were analyzed. Supplementing exercise with protein had no additional effect on any of the outcome measures. However, shuttle walk time, St George total score and subscore for impact improved as effect of time. This trial was unable to provide evidence for the effect of protein supplementation on quality of life, physical function, and muscle strength in non-sarcopenic patients with moderate to severe COPD. The role of protein supplementation in COPD-rehabilitation should focus on identifying patients to receive supplement with protein and from those who will not benefit.


Suppression of autophagy by mycophenolic acid contributes to inhibition of HCV replication in human hepatoma cells.
Previous studies have shown that mycophenolic acid (MPA) has an anti-HCV activity. However, the mechanism of MPA-mediated inhibition of HCV replication remains to be determined. This study investigated whether MPA has an effect on autophagy, a cellular machinery required for HCV replication, thereby, inhibits HCV replication in Huh7 cells. MPA treatment of Huh7 cells could suppress autophagy, evidenced by decreased LC3B-II level and conversion of LC3B-I to LC3B-II, decreased autophagosome formation, and increased p62 level compared to MPA-untreated cells. Tunicamycin treatment or HCV infection could induce cellular autophagy, however, MPA also exhibited its inhibitory effect on tunicamycin- or HCV infection-induced autophagy. The expression of three autophagy-related genes, Atg3, Atg5, and Atg7 were identified to be inhibited by MPA treatment. Over-expression of these genes could partly recover HCV replication inhibited by MPA; however, silencing their expression by siRNAs could enhance the inhibitory effect of MPA on HCV. Collectively, these results reveal that suppression of autophagy by MPA plays a role in its anti-HCV activity. Down-regulating the expression of three autophagy-related genes by MPA involves in its antiviral mechanism.


A Bayesian Adaptive Design for Combination of Three Drugs in Cancer Phase I Clinical Trials.
We describe a Bayesian adaptive design for early phase cancer trials of a combination of three agents. This is an extension of an earlier work by the authors by allowing all three agents to vary during the trial and by assigning different drug combinations to cohorts of three patients. The primary objective is to estimate the Maximum Tolerated Dose (MTD) surface in the three-dimensional Cartesian space. A class of linear models on the logit of the probability of Dose Limiting Toxicity (DLT) are used to describe the relationship between doses of the three drugs and the probability of DLT. Trial design proceeds using conditional escalation with overdose control, where at each stage of the trial, we seek a dose of one agent using the current posterior distribution of the MTD of this agent given the current doses of the other two agents. The MTD surface is estimated at the end of the trial as a function of Bayes estimates of the model parameters. Operating characteristics are evaluated with respect to trial safety and percent of dose recommendation at dose combination neighborhoods around the true MTD surface.


Increased prevalence of antithyroid antibodies and thyroid diseases in pustulosis palmoplantaris.
A statistically significant increased prevalence of antithyroid antibodies was found by an indirect immunofluorescence technique in 37 patients with pustulosis palmoplantaris, as compared with an approximatley age- and sex-matched group of healthy controls. A history of thyroid disease was also more frequent in patients with PPP than in the population at large. The increased risk of thyroid dysfunction should be kept in mind in the long-term care of PPP cases. The prevalence of non-cutaneous autoantibodies was not increased in psoriatic subjects. The results accord with the concept that PPP and psoriasis may be separate entities.


Molecular characterization of the complete genome of a novel partitivirus hosted by the saprobic mushroom Leucocybe candicans.
Virus communities of uncultivated fungi stay largely unknown. In the current study, we characterized a new partitivirus species detected in the basidiomycetous, saprobic mushroom Leucocybe candicans, named "Leucocybe candicans partitivirus 1" (LcPV1). The full-length genome of LcPV1, determined using deep sequencing and RLM-RACE approaches, consists of two dsRNA segments with each having the same size of 1984 bp. Both dsRNA genome segments comprise a single open reading frame (ORF), encoding an RNA-dependent RNA polymerase (RdRp), and a capsid protein (CP), respectively. Based on BLASTp search, the sequences of the RdRp and CP show the highest identity (50.09% and 35.71% similarity, respectively) to those of partitiviruses reported from an oomycetous, plant pathogenic, stramenopile algae Plasmopara viticola and basidiomycetous, plant pathogenic fungus Ceratobasidium sp., respectively. Phylogenetic analyses performed based on the RdRp and CP sequences revealed that LcPV1 falls within a cluster that includes different alphapartitivirus species from the family Partitiviridae. In this study, we propose that LcPV1 is a new member of a species belonging to the genus Alphapartitivirus. To our knowledge, this is the first study reporting on a new fungal virus (mycovirus) identified in the basidiomycetous, saprobic mushroom Leucocybe candicans.


Social media use by orthodontic patients.
Internet-based social media sites have recently surged in popularity and are often used to share thoughts and seek support about health issues. The aim of this study was to investigate how orthodontic patients use Internet-based social media sites to share their treatment-related experiences and attitudes towards braces. A secondary objective was to investigate whether an online or mobile application would be considered helpful in improving co-operation with the use of orthodontic appliances. Patients visiting the orthodontic clinic at the University of Otago were asked to participate in a cross-sectional survey that sought details of their Internet-based social media use and their thoughts about the development of a reminder application. The sample comprised 130 orthodontic patients, with a mean age of 17.2 (SD 6.9) and a nearly equal sex distribution (52.3% were female). Internet-based social media sites were used by 80.8%, with Facebook being the most popular. Some 13.3% of the sample had posted comments about braces on these social media sites. Only 6.7% had considered obtaining information about orthodontic treatment from Internet-based social media sites, with the majority (81%) preferring to seek this information directly from their orthodontist. Nearly two-thirds of those who had difficulty remembering to wear their orthodontic appliances reported that a reminder application on their phone would be beneficial. A large proportion of orthodontic patients use Internet-based social media sites, although only a few currently use them to post about treatment-related topics. Social media sites may provide a useful channel of communication for patients seeking support. Further research is needed to evaluate the use of phone reminder applications in orthodontics.


Detection of transient interchain interactions in the intrinsically disordered protein alpha-synuclein by NMR paramagnetic relaxation enhancement.
NMR paramagnetic relaxation enhancement experiments were applied to the intrinsically disordered protein alpha-synuclein, the primary protein in Parkinson's disease, to directly characterize transient intermolecular complexes at neutral and low pH. At neutral pH, we observed weak N- to C-terminal interchain contacts driven by electrostatic interactions, while at low pH, the C- to C-terminal interchain interactions are significantly stronger and driven by hydrophobic contacts. Characterization of these first interchain interactions will provide fundamental insight into the mechanism of amyloid formation.


Targeted agents for adjuvant therapy of colon cancer.
Adjuvant therapy for colorectal cancer consists primarily of combinations of 5-fluorouracil/leucovorin (5-FU/LV) (with infusional or bolus 5-FU) with oxaliplatin or oral capecitabine. The angiogenesis inhibitor bevacizumab and the epidermal growth factor receptor inhibitor cetuximab have shown activity when combined with 5-FU/LV-based regimens as first-line treatment of advanced disease and are currently being evaluated as part of adjuvant therapy in colon cancer. Bevacizumab is being evaluated in combination with FOLFOX4 (5-FU/LV/oxaliplatin), FOLFOX6, or XELOX (capecitabine/oxaliplatin) in the National Surgical Adjuvant Breast and Bowel Project C08 trial, the AVANT (AVastin adjuvANT) trial, and the Intergroup Rectal Adjuvant trial. Cetuximab is being evaluated in combination with FOLFOX4 and FOLFOX6 in the North Central Cancer Treatment Group (NCCTG) N0147 trial and the Pan European Trials in Adjuvant Colon Cancer (PETTAC) 8 trial.


Prevalence and seasonal dynamics of blaCTX-M antibiotic resistance genes and fecal indicator organisms in the lower Lahn River, Germany.
Antibiotic-resistant bacteria represent an emerging global health problem and are frequently detected in riverine environments. Analyzing the occurrence of corresponding antibiotic-resistant genes in rivers is of public interest as it contributes towards understanding the origin and dissemination of these emerging microbial contaminants via surface water. This is critical for devising strategies to mitigate the spread of resistances in the environment. Concentrations of blaCTX-M antibiotic resistance genes were quantified weekly over a 12-month period in Lahn River surface water at two sampling sites using quantitative real-time PCR. Gene abundances were statistically assessed with regard to previously determined concentrations of fecal indicator organisms Escherichia coli, intestinal enterococci and somatic coliphages, as well as influential environmental factors. Similar seasonal patterns and strong positive correlations between fecal indicators and blaCTX-M genes indicated identical sources. Accordingly, linear regression analyses showed that blaCTX-M concentrations could largely be explained by fecal pollution. E. coli provided the best estimates (75% explained variance) at the upstream site, where proportions of blaCTX-M genes in relation to fecal indicator organisms were highest. At this site, rainfall proved to be more influential, hinting at surface runoff as an emission source. The level of agricultural impact increased from downstream to upstream, linking increasing blaCTX-M concentrations after rainfall events to the degree of agricultural land use. Exposure assessment revealed that even participants in non-swimming recreational activities were at risk of incidentally ingesting blaCTX-M genes and thus potentially antibiotic resistant bacteria. Considering that blaCTX-M genes are ubiquitous in Lahn River and participants in bathing and non-bathing water sports are at risk of exposure, results highlight the importance of microbial water quality monitoring with an emphasis on antibiotic resistance not only in designated bathing waters. Moreover, E. coli might serve as a suitable estimate for the presence of respective antibiotic resistant strains.


Increased mobility and stem-cell proliferation rate in Dugesia tigrina induced by 880nm light emitting diode.
The therapeutic effects elicited by photobiostimulation in the near infrared range may be associated with increased proliferation rate of particular cell-types. The present study utilized commercial light emitting diodes to investigate the effects of low-level near-infrared radiation on the proliferation rate of stem cells in amputated planarian. Whole and amputated animals were exposed to either ambient diurnal lighting, darkness, white light, red light, or near-infrared (880 nm) light. Irradiation was consistent for the duration of the experiments and was provided using commercial 5mm light emitting diodes (∼1.0 mW/m(2) in power density and ∼0.01 J/cm(2) in radiant exposure). Compared to other groups amputated planarian exposed to near-infrared displayed increased mobility by the 3rd day of exposure (F((4,26))=4.31, p<0.04, η(2)=41%). Higher densities of stem cells were measured in these worms 84 h post injury (F((4,72))=4.78, p<0.01, η(2)=21%). These findings suggest that non-coherent light sources with power-densities about 1000 times lower than contemporary low-power laser settings remain effective in generating photobiostimulation effects and warrants further investigation on stem-cell proliferation induced by near-infrared light emitting diodes.


Evaluation of equine oocyte developmental competence using polarized light microscopy.
The purpose of this study was to observe in vitro-matured equine oocytes with an objective computerized technique that involves the use of a polarized light microscope (PLM) in addition to the subjective morphological evaluation obtained using a classic light microscope (LM). Equine cumulus-oocyte complexes (COCs, n = 922) were subjected to different in vitro maturation times (24, 36 or 45 h), however, only 36-h matured oocytes were analyzed using CLM. The 36-h matured oocytes that reached maturity were parthenogenetically activated to evaluate the quality and meiotic competence. Average maturation percentages per session in groups 1, 2 and 3 (24-, 36- and 45-h matured oocytes respectively) were 29.31 ± 13.85, 47.01 ± 9.90 and 36.62 ± 5.28%, whereas the average percentages of immature oocytes per session were 28.78 ± 20.17, 7.83 ± 5.51 and 22.36 ± 8.39% respectively. The zona pellucida (ZP) birefringent properties were estimated and correlated with activation outcome. ZP thickness and retardance of the inner layer of the zona pellucida (IL-ZP) were significantly increased in immature oocytes compared with mature oocytes (P < 0.001 and P < 0.01 respectively). The comparison between parthenogenetically activated and non-activated oocytes showed a significant increase in the area and thickness of the IL-ZP in parthenogenetically activated oocytes (P < 0.01). These results show that the 36-h in vitro maturation (IVM) protocol allowed equine oocytes to reach maturity, and PLM observation of ZP can be used to distinguish mature and immature oocytes as well as activated and non-activated oocytes.


Insulin therapy waste produced in the households of people with diabetes monitored in Primary Care.
To analyze the insulin therapy waste disposal produced in the households of people with diabetes mellitus (DM). Cross-sectional study with 105 Primary Care patients. Socio-demographic and clinical variables and insulin therapy practice were analyzed through the absolute and relative frequencies, Fisher's exact test and prevalence ratio (PR). The association between types of insulin (60.0%), administered with a disposable syringe attached to a needle (80.9%), and a high percentage of reuse and disposal in normal household waste (57.1%) stood out. The professionals who most frequently provided guidance to people with diabetes were the nurses. Patients who had received instructions about disposal were 21 times more likely to dispose of waste properly than those who had not (PR 21.5; P < 0.0001). Age, gender, skin color, education, marital status, occupational status and type of DM did not interfere in the disposal (P > 0.05). People with diabetes served in Primary Health Care require guidance and material resources to carry out the appropriate disposal of insulin therapy waste.


SPATIAL POPULATION STRUCTURE IN THE WHIRLIGIG BEETLE DINEUTUS ASSIMILIS: EVOLUTIONARY INFERENCES BASED ON MITOCHONDRIAL DNA AND FIELD DATA.
The spatial population structure of the pond-living water beetle Dineutus assimilis (Coleoptera: Gyrinidae) was investigated through a field study of population dynamics and dispersal, with a concurrent assessment of the spatial distribution of mitochondrial DNA (mtDNA) restriction-fragment-length polymorphism (RFLP). A comprehensive 2-yr survey within a 60-km2 study area revealed pronounced fluctuations in local abundances, including extinctions and colonizations. The recapture of marked individuals showed that dispersal among ponds is frequent in both males and females and connects populations on a large geographic scale (maximum observed flight distance: 20 km). The population structure of D. assimilis is thus characterized by both pronounced genetic drift and frequent gene flow. Together, these two forces generate a pattern of very local and transient genetic differentiation. Mitochondrial DNA samples collected within a few kilometers indicate highly significant spatial structure, if newly founded demes or those that experienced recent bottlenecks are included. These results based on four demes within the study area were placed into a regional context by further samples collected at distances of 100 km and 200 km. Fst estimates computed on increasing spatial scales were variable but showed no increasing trend. Thus, gene flow exerts a strong homogenizing force over a wide geographic range but is counteracted locally by genetic drift. These findings highlight the need to supplement estimates of Fst with additional data to arrive at valid interpretations of the genetic information. More generally, this study raises questions about how to capture the relevant features of dynamic, subdivided populations to understand their evolutionary dynamics.


[Laparoscopic substitution of the right ureter by appendix onlay].
In highly-selected patients with long narrowing of the middle and upper third of the right ureter, ureteroplasty with an appendix can be performed. In the literature, publications on the possibility of ureteroplasty onlay with a detubularized appendix have been appeared. To study the results of laparoscopic ureteroplasty of the right ureter using appendix as onlay flap. In the Center of Urology of Mariinsky hospital, 5 patients (2 men and 3 women) with a long stricture of the middle or upper third of the right ureter underwent right-side ureteroplasty using appendix as onlay flap. The mean age of the patients was 42.5 years. In 4 cases, the stricture developed after ureteroscopy with lithotripsy, while in 1 patient, laparoscopic ureterolysis was previously performed. Three patients were admitted with nephrostomy tube, two with a ureteral stent. The average length of the stricture was 3.5 (2.8-5.2) cm. The ureteral stricture was visualized by laparoscopic access, dissected along its length and replaced with a detubularized appendix as onlay flap. The average procedure time was 225 (180-260) min, the volume of blood loss was 160 (70-245) ml, and the hospital stay was 3.6 (3-7) days. There were no intraoperative complications. In the postoperative period, fever was observed in one patient, and ileus, which resolved spontaneously, developed in another patient. With an average follow-up period of 13.5 (6.2-24.4) months, the procedure was effective in all cases. The urodynamics of the upper urinary tract recovered completely in 3 patients and improved in 2 cases. Pain disappeared in all patients. It was possible to remove external and internal drainage in all cases. In patients with long strictures of the middle and proximal parts of the right ureter, laparoscopic ureteroplasty using the appendix as onlay flap may be the method of choice. This procedure is less traumatic and has a low complication rate.


Fouling one's own nest revisited.
Transport of hazards from the workplace to the home by workers is a previously recognized but poorly addressed public health problem. Particularly disturbing are the childhood intoxications which may result from such paraoccupational exposure. Work clothes and shoes appear to be a common vehicle for hazard transport in recently reported cases as they were in the past, although other "markers" for potential paraoccupational exposure may be derived from case reviews. These include: poor workplace hygiene, occupational intoxications in adult workers, and the "cottage industry" phenomenon. These markers are reviewed in the context of recently reported cases and strategies are suggested to mitigate these easily preventable exposures.


Novel lanthanide(II) complexes supported by carbon-bridged biphenolate ligands: synthesis, structure and catalytic activity.
[Ln[N(SiMe3)2]2(THF)2](Ln = Sm, Yb) reacts with 1 equiv. of carbon-bridged biphenols, 2,2'-methylene-bis(6-tert-butyl-4-methylphenol)(L1H2) or 2,2'-ethylidene-bis(4,6-di-tert-butylphenol)(L2H2), in toluene to give the novel aryloxide lanthanide(II) complexes [[LnL1(THF)n]2](Ln = Sm, n = 3 (1); Ln = Yb, n = 2 (2)) and [[LnL2(THF)3]2](Ln = Sm (5); Ln = Yb (6)) in quantitative yield, respectively. Addition of 2 equiv. of hexamethylphosphoric triamide (HMPA) to a tetrahydrofuran (THF) solution of 1, 2 and 5 affords the corresponding HMPA-coordinated complexes, [[LnL1(THF)m(HMPA)n]2(THF)y](Ln = Sm, n = 2, m = 0, y = 2 (3); Ln = Yb, m = 1, n = 1, y = 6 (4)) and [[SmL2(HMPA)2]2](7) in excellent yields. The single-crystal structural analyses of 3, 4 and 7 revealed that these aryloxide lanthanide(II) complexes are dimeric with two Ln-O bridges. The coordination geometry of each lanthanide metal can be best described as a distorted trigonal bipyramid. Complexes 1-3, 5 and 7 can catalyze the ring-opening polymerization of epsilon-caprolactone (epsilon-CL), and 1-3, along with 5 show moderate activity for the ring-opening polymerization of 2,2-dimethyltrimethylene carbonate (DTC) and the copolymerization of epsilon-CL and DTC to give random copolymers with high molecular weights and relatively narrow molecular weight distributions..


[Using of 4-thiouridine and 4-thiothymidine for pyrimidine nucleoside phosphorylase studing].
4-Thiouridine and 4-thiothymidine were developed as efficient substrates for spectrophotometric determination of uridine phosphorylase and thymidine phosphorylase activity. 4-Thiouridine has maximum absorbance at 330 nm (pH 7.5). The change in extinction coefficient for 4-thiouridine/4-thiouracil, deltaepsilon is 3000 M(-1) x cm(-1). It appeared that 4-thiouridine is a good substrate for uridine phosphorylase with Michaelis-Menten constant 130 microM and kcat 49 s(-1). In the case of 4-thiothymidine/4-thiothymine deltaepsilon is even larger: 5000 M(-1) x cm(-1) at 336 nm.


Cutaneous findings in patients with chronic venous insufficiency.
Venous hypertension causes many different cutaneous findings such as varicosities, telangiectasia, edema, and pigmentation, dermatitis, and venous ulcers on the skin. This study aims to investigate the cutaneous signs and symptoms of chronic venous insufficiency (CVI) and to examine their contribution to early diagnosis. A total of 150 patients were included in the study who applied to the dermatology outpatient clinic and were diagnosed with skin disease related to CVI or had skin findings. Patient's age, gender, complaints, occupation, additional diseases, drug usage, history of prolonged standing and travel, smoking habit, number of pregnancies, history of varicose veins in the family, dermatological diagnosis/findings, and venous Doppler ultrasonography reports were examined retrospectively. 56% of patients were women. Mean age was 56.69 ± 13.6 years. Overall, 82.7% of total patients had at least one skin finding. The most frequent skin findings except varicose veins were insufficiency dermatitis accounted for 32.7% of patients, telangiectasia, and pigmentation which were comprised 25.3%, 19.3% of the total number of patients respectively. In addition to this, 48.7% of patients had itching problems and 32.7% had pain. Moreover, 46% of patients presented superficial vein insufficiency, while 8.7% had deep vein insufficiency. For 47.3% of patients, vein diameter dilation was observed and 11.3% suffered from perforating vein insufficiency. In terms of Clinical-Etiologic-Anatomic-Pathophysiologic (CEAP) classification, scores of 52% of the patients were C3 and lower, while scores of 48% of total patients were C4a and higher. Early diagnosis and treatment of chronic venous insufficiency could prevent further chronic processes such as venous ulceration which is an advanced CVI finding. Thus, assessing the early skin findings might be important to identify the underlying venous insufficiency disease.


Establishment of a novel experimental model for muscle-invasive bladder cancer using a dog bladder cancer organoid culture.
In human and dogs, bladder cancer (BC) is the most common neoplasm affecting the urinary tract. Dog BC resembles human muscle-invasive BC in histopathological characteristics and gene expression profiles, and could be an important research model for this disease. Cancer patient-derived organoid culture can recapitulate organ structures and maintains the gene expression profiles of original tumor tissues. In a previous study, we generated dog prostate cancer organoids using urine samples, however dog BC organoids had never been produced. Therefore we aimed to generate dog BC organoids using urine samples and check their histopathological characteristics, drug sensitivity, and gene expression profiles. Organoids from individual BC dogs were successfully generated, expressed urothelial cell markers (CK7, CK20, and UPK3A) and exhibited tumorigenesis in vivo. In a cell viability assay, the response to combined treatment with a range of anticancer drugs (cisplatin, vinblastine, gemcitabine or piroxicam) was markedly different in each BC organoid. In RNA-sequencing analysis, expression levels of basal cell markers (CK5 and DSG3) and several novel genes (MMP28, CTSE, CNN3, TFPI2, COL17A1, and AGPAT4) were upregulated in BC organoids compared with normal bladder tissues or two-dimensional (2D) BC cell lines. These established dog BC organoids might be a useful tool, not only to determine suitable chemotherapy for BC diseased dogs but also to identify novel biomarkers in human muscle-invasive BC. In the present study, for the 1st time, dog BC organoids were generated and several specifically upregulated organoid genes were identified. Our data suggest that dog BC organoids might become a new tool to provide fresh insights into both dog BC therapy and diagnostic biomarkers.


Changes in equity of maternal, newborn, and child health care practices in 115 districts of rural Ethiopia: implications for the health extension program.
Reducing within-country inequities in the coverage of maternal, newborn, and child health (MNCH) interventions is essential to improving a country's maternal and child health and survival rates. The community-based health extension program (HEP) of Ethiopia, launched in 2003, aims to provide equitable primary health care services. Since 2008 the Last Ten Kilometers Project (L10K) has been supporting the HEP in promoting equitable MNCH interventions in 115 districts covering about 14 million people. We report the inequities in MNCH programmatic indicators in 2008 and in 2010 in the L10K areas, along with changes in equity between the two survey periods, and the implications of these results for the national program. The study used cross-sectional surveys of 3932 and 3867 women from 129 representative kebeles (communities) conducted in December 2008 and December 2010, respectively. Nineteen HEP outreach activity coverage and MNCH care practice indicators were calculated for each survey period, stratified by the inequity factors considered (i.e. age, education, wealth and distance from the nearest health facility). We calculated relative inequities using concentration indices for each of the indicators and inequity factors. Ninety-five percent confidence intervals and survey design adjusted Wald's statistics were used to assess differentials in equity. Education and age related inequities in the MNCH indicators were the most prominent (observed for 13 of the 19 outcomes analyzed), followed in order by wealth inequity (observed for eight indicators), and inequity due to distance from the nearest health facility (observed for seven indicators). Age inequities in six of the indicators increased between 2008 and 2010; nevertheless, there was no consistent pattern of changes in inequities during that period. Some related issues such as inequities due to wealth in household visits by the health extension workers and prevalence of modern family household; and inequities due to education in household visits by community health promoters showed improvement. Addressing these inequities in MNCH interventions by age, education and wealth will contribute significantly toward achieving Ethiopia's maternal health targets for the Millennium Development Goals and beyond. HEP will require more innovative strategies to achieve equitable MNCH services and outcomes and to routinely monitor the effectiveness of those strategies.


Parallel synthesis of cell-penetrating peptide conjugates of PMO toward exon skipping enhancement in Duchenne muscular dystrophy.
We describe two new methods of parallel chemical synthesis of libraries of peptide conjugates of phosphorodiamidate morpholino oligonucleotide (PMO) cargoes on a scale suitable for cell screening prior to in vivo analysis for therapeutic development. The methods represent an extension of the SELection of PEPtide CONjugates (SELPEPCON) approach previously developed for parallel peptide-peptide nucleic acid (PNA) synthesis. However, these new methods allow for the utilization of commercial PMO as cargo with both C- and N-termini unfunctionalized. The synthetic methods involve conjugation in solution phase, followed by rapid purification via biotin-streptavidin immobilization and subsequent reductive release into solution, avoiding the need for painstaking high-performance liquid chromatography purifications. The synthesis methods were applied for screening of PMO conjugates of a 16-member library of variants of a 10-residue ApoE peptide, which was suggested for blood-brain barrier crossing. In this work the conjugate library was tested in an exon skipping assay using skeletal mouse mdx cells, a model of Duchene's muscular dystrophy where higher activity peptide-PMO conjugates were identified compared with the starting peptide-PMO. The results demonstrate the power of the parallel synthesis methods for increasing the speed of optimization of peptide sequences in conjugates of PMO for therapeutic screening.


Part II. rigour in qualitative research: complexities and solutions.
Anthony G Tuckett outlines the strategies and operational techniques he used to attain rigour in a qualitative research study through relying on Guba and Lincoln's trustworthiness criterion. Research strategies such as use of personal journals, audio recording and transcript auditing, and operational techniques including triangulation strategies and peer review, are examined.


IgH chain class switch recombination: mechanism and regulation.
IgH class switching occurs rapidly after activation of mature naive B cells, resulting in a switch from expression of IgM and IgD to expression of IgG, IgE, or IgA; this switch improves the ability of Abs to remove the pathogen that induces the humoral immune response. Class switching occurs by a deletional recombination between two switch regions, each of which is associated with a H chain constant region gene. Class switch recombination (CSR) is instigated by activation-induced cytidine deaminase, which converts cytosines in switch regions to uracils. The uracils are subsequently removed by two DNA-repair pathways, resulting in mutations, single-strand DNA breaks, and the double-strand breaks required for CSR. We discuss several aspects of CSR, including how CSR is induced, CSR in B cell progenitors, the roles of transcription and chromosomal looping in CSR, and the roles of certain DNA-repair enzymes in CSR.


Discovery of Selective, Substrate-Competitive, and Passive Membrane Permeable Glycogen Synthase Kinase-3β Inhibitors: Synthesis, Biological Evaluation, and Molecular Modeling of New C-Glycosylflavones.
Glycogen synthase kinase-3β (GSK-3β) is a key enzyme responsible for tau hyperphosphorylation and is a viable therapeutic target of Alzheimer's disease (AD). We developed a new class of GSK-3β inhibitors based on the 6- C-glycosylflavone isoorientin (1). The new inhibitors are passive membrane permeable and constitutively attenuate GSK-3β mediated tau hyperphosphorylation and amyloid neurotoxicity in an AD cellular model. Enzymatic assays and kinetic studies demonstrated that compound 30 is a GSK-3β substrate-competitive inhibitor with distinct kinase selectivity, isoform-selectivity and over 310-fold increased potency as compared to 1. Structure-activity relationship analyses and in silico modeling suggest the mechanism of actions by which the hydrophobic, π-cation, and orthogonal multipolar interactions of 30 with the substrate site are critical for the GSK-3β inhibition and selectivity. The results provide new insights into GSK-3β drug discovery. The new inhibitors are valuable chemical probes and drug leads with therapeutic potential to tackle AD and other GSK-3β relevant diseases.


Eye injury in migrant farm workers and suggested hazard controls.
The eyes are a common site of injury in agricultural operations. Identification of the cause of injury is important to inform preventive interventions. The objective of this study was to describe the hazards and mechanisms of acute traumatic injury to the eyes of agricultural workers who are hired in farming operations on a seasonal basis. A review of the literature was performed to summarize the mechanisms of eye injuries in agriculture. Field observations and informal interviews were performed to verify the literature and to determine whether there are eye hazards for farm workers that have not been reported in the literature. Additional mechanisms of injury were elicited, and suggested methods of injury prevention are presented here.


[Experimental study and clinical application of end-to-side (transverse incision) anastomosis between small arteries].
The results of end to side anastomoses with longitudinal or transverse incision on the transverse limb were investigated. The anastomosis of the superficial epigastric artery to the femoral artery in rabbits served as the experimental model. The results demonstrated that the microartery end to side anastomosis with transverse arteriotomy is a more preferable anastomosis style with higher patency, easier arteriotomy and suturing, and it effectively prevents anastomotic stoma spasm or stenosis, and in accord with vascular physiological function. It is worthy to be recommended. The clinical successful cases are also reported.


Cardiac structure and function in response to a multi-stage marathon over 4486 km.
To investigate whether participation in the Trans Europe Foot Race 2009 (TEFR), an ultramarathon race held over 64 consecutive days and 4486 km, led to changes in cardiac structure and function. Cardiac magnetic resonance imaging was performed in 20 of 67 participating runners (two women; mean ± SD age 47.8 ± 10.4 years) at three time points (baseline scan at 294 ± 135 km (B), scan two at 1735 ± 86 km (T1) and scan three at 3370 ± 90 km (T2)) during the TEFR. Imaging included an assessment of left ventricular structure (mass) and function (strain). In parallel, cardiac troponin I, NT-pro-BNP, myostatin and GDF11 were determined in venous blood samples. A subsample of ten runners returned for a follow-up scan eight months after the race. Left ventricular mass increased significantly (B, 158.5 ± 23.8 g; T1, 165.1 ± 23.2 g; T2, 167 ± 24.6 g; p < 0.001) over the course of the race, although no significant change was seen in the remaining structural and functional parameters. Serum concentrations of cardiac troponin I and NT-proBNP significantly increased 1.5 - and 3.5-fold, respectively, during the first measurement interval, with no further increase thereafter (cardiac troponin I, 6.8 ± 3.1 (B), 16.9 ± 10.4 (T1) and 17.1 ± 9.7 (T2); NT-proBNP, 30.3 ± 22.8 (B), 135.9 ± 177.5 (T1) and 111.2 ± 87.3 (T2)), whereas the growth markers myostatin and GDF11 did not change. No association was observed with functional parameters, including the ejection fraction and the volume of both ventricles. The follow-up scans showed a reduction to baseline values (left ventricular mass 157 ± 19.3 g). High exercise-induced cardiac volume load for >2 months in ultra-endurance runners results in a physiological structural adaptation with no sign of adverse cardiovascular remodelling.


Perceived Barriers and Benefits of COVID-19 Testing among Pacific Islanders on Guam.
Pacific Islanders residing in the U.S. Affiliated Pacific Islands have had among the highest COVID-19-associated morbidity and mortality rates in the U.S. To reduce this disparity, we conducted a study to increase the reach and uptake of COVID-19 testing in Guam. Participants, who completed a pre-survey on demographics, health status, history of COVID-19 testing and vaccination, access to COVID-19 testing, sources of COVID-19 information, and knowledge and attitudes towards COVID-19 test results and transmission, were invited to attend an online educational session about COVID-19 testing and transmission and to complete a post-survey. There were significant positive changes between pre- and post-survey in knowledge and perceptions about COVID-19 testing and transmission, but changes were not necessarily due to exposure to the educational session. Compared to CHamoru participants (n = 380), Other Micronesians (n = 90) were significantly less knowledgeable about COVID-19 transmission and testing, were significantly more likely to not want to know if they had COVID-19, were more likely to believe if they did have COVID-19 there was not much that could be done for them, and that they would have difficulty in getting the needed healthcare. This study is another example of disparities in health knowledge and perceptions of certain Pacific Islander groups.


Hyperfocus or flow? Attentional strengths in autism spectrum disorder.
The comorbidity of autism spectrum disorder (ASD) and attention-deficit/hyperactivity disorder (ADHD) diagnoses is well established. An ASD diagnosis is associated with elevated ADHD traits and symptoms, as well as strengths in attention. In the ASD literature, attentional strengths have been described as maladaptive (e.g., hyperfocus), in contrast with positive portrayals in the typically developing population (e.g., flow). The objective of this study was to (1) compare profiles of attentional strengths and weaknesses in ASD and ADHD and (2) determine whether attentional strengths in ASD are associated with impairment, poorer cognitive flexibility, and perseveration/perfectionism. In a community sample of 5,744 children and youth, 131 children were reported as having a diagnosis of ASD (mean age 10.3 years) and 346 children were reported as having a diagnosis of ADHD (mean age 10.7 years). We used the Strengths and Weaknesses of Attention-Deficit/Hyperactivity-symptoms and Normal-behaviors (SWAN) rating scale to calculate attentional and hyperactive/impulse control strength and weakness counts and scores. The Autism-Spectrum Quotient Switching factor served as a measure of cognitive flexibility. Impairment was assessed with the Columbia Impairment Scale. We used the symmetry/ordering factor on the Toronto Obsessive-Compulsive Scale as a measure of perseveration/perfectionism. No differences were found between the ADHD and ASD groups in SWAN weakness scores, symptoms, or hyperactive/impulse control strengths; however, autistic children had higher rates of attentional strengths [odds ratio: 5.7, 95% CI (2.8, 11.6), p < 0.0001]. Post-hoc pairwise testing identified four attentional strengths with significantly higher rates in ASD than in ADHD. Attentional strength scores were not associated with impairment or poor cognitive flexibility, but predicted levels of perseveration/perfectionism. The effect of attentional strengths on impairment and cognitive flexibility did not differ between autistic and Control children, but the higher perseveration/perfectionism scores seen in ASD were not found in Control children. ASD is associated with a pattern of attentional strengths that is not found in ADHD Characterizing the full range of attentional abilities in autistic children may explain variability in outcomes such as quality-of-life indicators and identify protective factors, providing targets for strength-based behavioral interventions. The clinical and etiological implications of the subgroup of autistic children with attentional strengths require further investigation.


Leisure-Time Physical Activity in Pregnancy and Maternal-Child Health: A Systematic Review and Meta-Analysis of Randomized Controlled Trials and Cohort Studies.
Evidence suggests that leisure-time physical activity (LTPA) during pregnancy is associated with a reduced risk of preeclampsia, gestational diabetes mellitus (GDM), and preterm birth. However, these results are inconsistent when comparing cohort studies and randomized controlled trials (RCTs). The purpose of our study was to compare the associations between LTPA in pregnancy and maternal (GDM, preeclampsia, and weight gain during pregnancy) and child health outcomes (preterm birth, birthweight, and fetal growth) between RCTs and cohort studies. We performed a systematic search in PubMed, Web of Science, and EBSCO up to 31 August 2015. Inclusion criteria for experimental studies required randomized trials with a control group and exposure to a physical activity structured program. The inclusion criteria for cohort studies required information on LTPA during pregnancy as an exposure and at least one maternal-child health outcome. We assessed the methodological quality of all studies and performed a meta-analysis to produce summary estimates of the effects using random models. We included 30 RCTs and 51 cohort studies. The meta-analysis of RCTs indicated that participation in LTPA was associated with lower weight gain during pregnancy, lower likelihood of GDM, and lower likelihood of delivering a large-for-gestational-age infant. Cohort studies indicated that participation in LTPA was associated with lower weight gain during pregnancy, lower likelihood of GDM, and lower risk of preterm delivery. Our findings support the promotion of LTPA in pregnancy as a strategy to improve maternal and child health.


The impact of cycle proficiency training on cycle-related behaviours and accidents in adolescence: findings from ALSPAC, a UK longitudinal cohort.
Cycle accidents are a common cause of physical injury in children and adolescents. Education is one strategy to reduce cycle-related injuries. In the UK, some children undertake National Cycle Proficiency Scheme [NCPS] training (now known as Bikeability) in their final years of primary school. It aims to promote cycling and safe cycling behaviours but there has been little scientific evaluation of its effectiveness. The sample (n = 5415) were participants in the Avon Longitudinal Study of Parents and Children who reported whether or not they had received NCPS training. Outcomes were self-reported at 14 and 16 years: cycling to school, ownership of cycle helmet, use of cycle helmet and high-visibility clothing on last cycle, and involvement in a cycle accident. An additional outcome, hospital admittance due to a cycle accident from 11 to 16 years, was also included for a subsample (n = 2222) who have been linked to Hospital Episode Statistics (HES) data. Approximately 40 % of the sample had received NCPS training. Trained children were more likely to cycle to school and to own a cycle helmet at both 14 and 16 years, to have worn a helmet on their last cycle at age 14, and to have worn high-visibility clothing at age 16, than those who had not attended a course. NCPS training was not associated with self-reported involvement in a cycle accident, and only six of those with HES data had been admitted to hospital due to a cycle accident. Irrespective of training, results indicate very low use of high-visibility clothing, very few girls cycling as part of their school commute, and less than half of helmet owners wearing one on their last cycle. Our results suggest cycle training courses for children can have benefits that persist into adolescence. However, the low use of cycle helmets, very low use of high-visibility clothing, and low levels of cycling to school for girls, indicate the further potential for interventions to encourage cycling, and safe cycling behaviours, in young people.


Development of social functioning in preschizophrenia children and adolescents: a systematic review.
Schizophrenia is associated with severe deficits in social functioning. Similar deficits may be present prior to psychosis onset, in childhood and adolescence. If so, then prepsychosis social deficits could provide clues to the development of pathological processes in preschizophrenia children and could potentially improve early identification of the disorder and suggest targets for intervention. Evidence is reviewed from birth cohort, case- control, and familial high-risk studies within distinct periods of development to clarify the nature, timing, and specificity of social deficits in preschizophrenia children and adolescents. The results indicate that poor social functioning does differentiate preschizophrenia children and adolescents from their peers and can be a sensitive and potentially specific predictor of schizophrenia, not just psychopathology in general. Furthermore, age (but not sex) appears to be an important moderator of the strength and specificity of the association between particular social deficits (e.g., externalizing, internalizing) and later schizophrenia. Results are discussed in the context of current developmental theories of timing and pathophysiology of schizophrenia involving hypothalamic- pituitary-adrenal dysregulation. Implications for the early identification and treatment of preschizophrenia individuals are also considered.


Catalytic synthesis of amides via aldoximes rearrangement.
Amide bond formation reactions are among the most important transformations in organic chemistry because of the widespread occurrence of amides in pharmaceuticals, natural products and biologically active compounds. The Beckmann rearrangement is a well-known method to generate secondary amides from ketoximes. However, under the acidic conditions commonly employed, aldoximes RHC=NOH rarely rearrange into the corresponding primary amides RC(=O)NH2. In recent years, it was demonstrated that this atom-economical transformation can be carried out efficiently and selectively with the help of metal catalysts. Several homogeneous and heterogenous systems have been described. In addition, protocols offering the option to generate the aldoximes in situ from the corresponding aldehydes and hydroxylamine, or even from alcohols, have also been developed, as well as a series of tandem processes allowing the access to N-substituted amide products. In this Feature article a comprehensive overview of the advances achieved in this particular research area is presented.


Myocardial Infarct Size by CMR in Clinical Cardioprotection Studies: Insights From Randomized Controlled Trials.
The aim of this study was to review randomized controlled trials (RCTs) using cardiac magnetic resonance (CMR) to assess myocardial infarct (MI) size in reperfused patients with ST-segment elevation myocardial infarction (STEMI). There is limited guidance on the use of CMR in clinical cardioprotection RCTs in patients with STEMI treated by primary percutaneous coronary intervention. All RCTs in which CMR was used to quantify MI size in patients with STEMI treated with primary percutaneous coronary intervention were identified and reviewed. Sixty-two RCTs (10,570 patients, January 2006 to November 2016) were included. One-third did not report CMR vendor or scanner strength, the contrast agent and dose used, and the MI size quantification technique. Gadopentetate dimeglumine was most commonly used, followed by gadoterate meglumine and gadobutrol at 0.20 mmol/kg each, with late gadolinium enhancement acquired at 10 min; in most RCTs, MI size was quantified manually, followed by the 5 standard deviation threshold; dropout rates were 9% for acute CMR only and 16% for paired acute and follow-up scans. Weighted mean acute and chronic MI sizes (≤12 h, initial TIMI [Thrombolysis in Myocardial Infarction] flow grade 0 to 3) from the control arms were 21 ± 14% and 15 ± 11% of the left ventricle, respectively, and could be used for future sample-size calculations. Pre-selecting patients most likely to benefit from the cardioprotective therapy (≤6 h, initial TIMI flow grade 0 or 1) reduced sample size by one-third. Other suggested recommendations for standardizing CMR in future RCTs included gadobutrol at 0.15 mmol/kg with late gadolinium enhancement at 15 min, manual or 6-SD threshold for MI quantification, performing acute CMR at 3 to 5 days and follow-up CMR at 6 months, and adequate reporting of the acquisition and analysis of CMR. There is significant heterogeneity in RCT design using CMR in patients with STEMI. The authors provide recommendations for standardizing the assessment of MI size using CMR in future clinical cardioprotection RCTs.


American trypanosomiasis in dogs from an urban and rural area of Yucatan, Mexico.
American trypanosomiasis in dogs is reported from South of the United States to Argentina and Chile. It is transmitted through the contact of dogs with reduviid insects when they feed; reduviid insects are well established in the southern state of Yucatan, Mexico. However, there are no reports available about trypanosomiasis prevalence in dogs of urban and rural areas. A cross-sectional study was performed in 345 stray dogs, 102 from a rural community of Yucatan (Tunkas) and 243 from Merida, capital city of the Yucatan state. Serum samples were obtained for detection of immunoglobulin G antibodies against Trypanosoma cruzi by indirect immunofluorescence assay and Western blot. DNA was extracted from whole blood of urban dogs for polymerase chain reaction (PCR) assay. Prevalence was determined and the risk to become infected in both urban and rural dogs was evaluated using a 2 x 2 contingency table. In addition to the effect of body condition score (BCS), the age and sex of dogs were also evaluated to determine the risk of infection. Antibody prevalence against T. cruzi in rural areas was 9.8%, whereas in urban dogs was 14.4%. When PCR results were included, prevalence in dogs from the urban area increased to 17.3%. PCR allowed the detection of active asymptomatic acute-phase disease. The risk to become infected was not different between urban and rural areas, suggesting that the vector is well adapted both to rural areas and urban sprawling. Dogs with poor BCS tended to have a higher probability of seroreactivity to T. cruzi proteins than dogs with regular or good BCS (p = 0.05).


Toward a Better Understanding of Hemiwicking: A Simple Model to Comprehensive Prediction.
The hemiwicking state has attracted much interest because of numerous important potential applications in inking, printing, boiling heat transfer, and condensation. However, the mechanism of the emergence of hemiwicking has not been well understood, especially the effects of geometry of patterned surfaces on the hemiwicking state has not been systematically investigated. Here, we presented a new method to study the critical conditions for hemiwicking on patterned surfaces. By minimizing the variation of the free energy, we obtain the corresponding stable height of the hemiwicking film and find that it is easier for a droplet to be in the hemiwicking state if the pillar surface has small spacing, large radius and height, and a small intrinsic contact angle. Our established model is applied to a flat-topped cylindrical pillar-patterned surface, and the modeling results are in well agreement with experiments and other existing theories. Besides, our model is also applied to other kinds of patterned surfaces including hemispherical-topped cylindrical and conical pillars, about which the other existing theories are deficient. Our theoretical results not only are in well agreement with the experimental observations but also provide some important predictions, which implies that the established model could be applicable to understanding the basic physical mechanism of the hemiwicking state and be useful in guiding the design and fabrication of hemiwicking surfaces.


Phosphatidylserine-specific phospholipase A1 involved in hepatitis C virus assembly through NS2 complex formation.
Several members of the phospholipase family have been reported to be involved in hepatitis C virus (HCV) replication. Here, we identified another phospholipase, phosphatidylserine-specific phospholipase A1 (PLA1A), as a host factor involved in HCV assembly. PLA1A was upregulated by HCV infection, and PLA1A knockdown significantly reduced J399EM (genotype 2a) HCV propagation at the assembly step but not the entry, RNA replication, and protein translation steps of the life cycle. Protein localization and interaction analysis further revealed a role of PLA1A in the interaction of NS2-E2 and NS2-NS5A, as the formation of the NS2-E2 and NS2-NS5A complexes was weakened in the absence of PLA1A. In addition, PLA1A stabilized the NS2/NS5A dotted structure during infection. These data suggest that PLA1A plays an important role in bridging the membrane-associated NS2-E2 complex and the NS5A-associated replication complex via its interaction with E2, NS2, and NS5A, which leads to a coordinating interaction between the structural and nonstructural proteins and facilitates viral assembly. Hepatitis C virus (HCV) genomic replication is driven by the replication complex and occurs at the membranous web, while the lipid droplet is the organelle in which virion assembly is initiated. In this study, we identified phosphatidylserine-specific phospholipase A1 (PLA1A), a member of phospholipase A 1 family, as a novel host factor involved in the assembly process of HCV. PLA1A, which is induced by HCV infection at a late infection stage, interacts with HCV E2, NS2, and NS5A proteins and enhances and stabilizes the NS2-E2 and NS2-NS5A complex formation, which is essential for viral assembly. Thus, PLA1A is an important host factor which is involved in the initiation of the viral assembly in close proximity to Core-decorated lipid droplets through bringing together the HCV replication complex and envelope complex.


Identification of variety-specific metabolites of basil by high performance thin layer chromatography-assisted metabolic profiling techniques.
The technological advances of analytical instrumentation and techniques has laid the ground for the rapid expansion of metabolomics or in a wider sense, untargeted analysis applied to life sciences themes. However, the objective of identifying all existing metabolites within organisms remains a daunting challenge. All analytical techniques exhibit varying degrees of sensitivity and versatility for the detection of metabolites and none of the existing analytical platforms can be expected to be ideal for exhaustive chemical profiling. Planar liquid chromatography, and in particular, high performance thin layer chromatography (HPTLC), has been used for chemical profiling of natural products in conjunction with metabolomics. HPTLC has specific advantages which include its ability to generate reliable chemical fingerprinting data and facilitate preparative work for metabolite isolation during later stages of metabolomics analysis. In this study, we investigated the chemical profiles of four commercially available basil cultivars, namely Dolly, Emily, Keira, and Rosie. We used HPTLC as the primary analytical tool for the separation of basil cultivars based on detected metabolites, and then compared the results with those obtained with other analytical platforms. We identified the characteristic marker compounds of each basil cultivar from the HPTLC plates and validated their potential using LC-MS and GC-MS analyses as a metabolomics tool. Firstly, we compared the HPTLC data of the four cultivars, obtained with two systems that used silica gel 60 and two mobile phases composed of toluene-EtOAc (8:2, v/v) and EtOAc-formic acid-acetic acid-water (100:11:11:27, v/v), with 1H NMR data to evaluate their separation power. Despite providing lower resolution and detecting fewer compounds, the HPTLC separation power was comparable, and in some cases even better than that of 1H NMR. Additionally, we investigated the potential of HPTLC as a tool for chemical fingerprinting and demonstrated its suitability for preparative purposes that are essential for identifying metabolites in mixture analysis. Metabolites were easily isolated from sample mixtures, and identified with the assistance of GC-MS, LC-MS, and TLC-densitometry.. Several marker compounds were thus identified, including 2,4 di-tertbutylphenol, palmitic acid, hexadecanamide, 9-octadecenamide, squalene, hentriacontane, methyl 3-(3,5-ditert‑butyl‑4-hydroxyphenyl)propanoic acid, sagerinic acid, and cyanidin-3-O-sophoroside.


A case of mixed adenoneuroendocrine carcinoma of the pancreas: Immunohistochemical analysis for histogenesis.
Tumors with multiple histological features, such as adenocarcinomas and neuroendocrine carcinomas, were included as a new category of neuroendocrine carcinomas in the 2010 World Health Organization classification. We recently experienced a rare case of a pancreatic carcinoma with both adenocarcinoma and neuroendocrine carcinoma components, a so-called mixed adenoneuroendocrine carcinoma. A 66-year-old man was referred to our hospital with obstructive jaundice. Contrast-enhanced computed tomography images indicated a tumor located at the pancreatic head measuring 3.0 × 2.5 cm in diameter and invading the common bile duct. Cytological examination of the bile juice obtained by endoscopic retrograde cholangiopancreatography revealed adenocarcinoma cells. Pancreaticoduodenectomy was performed safely as radical resection. Microscopically, the resected tumor consisted of tightly intermingled adenocarcinoma and neuroendocrine carcinoma components. On the immunohistochemical examination, p53 was ubiquitously positive in both components, whereas chromogranin A, synaptophysin and neuron-specific enolase, neuroendocrine markers, were limited to the neuroendocrine carcinoma component. Thus, such features of both adenocarcinoma and neuroendocrine carcinoma observed microscopically and immunohistochemically seemed to indicate a composite tumor. The findings of this case suggest that adenocarcinoma and neuroendocrine carcinoma may be derived from a single cancer stem cell.


Angiotensin Converting Enzyme Inhibitor Has a Protective Effect on Decompression Sickness in Rats.
Introduction: Commercial divers, high altitude pilots, and astronauts are exposed to some inherent risk of decompression sickness (DCS), though the mechanisms that trigger are still unclear. It has been previously showed that diving may induce increased levels of serum angiotensin converting enzyme. The renin angiotensin aldosterone system (RAAS) is one of the most important regulators of blood pressure and fluid volume. The purpose of the present study was to control the influence of angiotensin II on the appearance of DCS. Methods: Sprague Dawley rats have been pre-treated with inhibitor of angiotensin II receptor type 1 (losartan; 10 mg/kg), angiotensin-converting enzyme (ACE) inhibitor (enalapril; 10 mg/kg), and calcium-entry blocker (nifedipine; 20 mg/kg). The experimental groups were treated for 4 weeks before exposure to hyperbaric pressure while controls were not treated. Seventy-five rats were subjected to a simulated dive at 1000 kPa absolute pressure for 45 min before starting decompression. Clinical assessment took place over a period of 60 min after surfacing. Blood samples were collected for measurements of TBARS, interleukin 6 (IL-6), angiotensin II (ANG II) and ACE. Results: The diving protocol induced 60% DCS in non-treated animals. This ratio was significantly decreased after treatment with enalapril, but not other vasoactive drugs. Enalapril did not change ANG II or ACE concentration, while losartant decreased post dive level of ACE but not ANG II. None of the treatment modified the effect of diving on TBARS and IL-6 values. Conclusion: Results suggests that the rennin angiotensin system is involved in a process of triggering DCS but this has to be further investigated. However, a vasorelaxation mediated process, which potentially could increase the load of inert gas during hyperbaric exposure, and antioxidant properties were excluded by our results.


The key role of aminoglycosides in antibacterial therapy and prophylaxis.
The discovery that aminosugars found in nature kill common bacteria initiated use of a new class of antibiotics that have played a key role in the management of several major medical problems over the past 40 years. The rapid bactericidal effect of the aminoglycosides on aerobic Gram-negative bacilli has expanded their use to approximately 6% of all our general hospital patients. These agents have many desirable properties, but also have the potential for producing ototoxicity and nephrotoxicity that imposes pharmacological limitations on their optimum use. The experience of earlier investigations demonstrated the need for quantitative measurements and individual patient dosing which has reduced the risk of toxicity many fold. Using sensitive measurements, toxic manifestations can be classified as detectable by laboratory means only, some that have clinical expression and rarely a lasting handicap. Semisynthetic modification of kanamycin (amikacin) and sissomicin (netilmicin) protects these compounds against inactivation by most bacterial enzymes and preserves their effectiveness. Netilmicin seems to have less potential for causing toxicity, especially ototoxicity that is irreversible, and offers the possibility of further gains through pharmacological flexibility without crippling toxicity.


Test-retest reliability and predictive utility of a macroscale principal functional connectivity gradient.
Mapping individual differences in brain function has been hampered by poor reliability as well as limited interpretability. Leveraging patterns of brain-wide functional connectivity (FC) offers some promise in this endeavor. In particular, a macroscale principal FC gradient that recapitulates a hierarchical organization spanning molecular, cellular, and circuit level features along a sensory-to-association cortical axis has emerged as both a parsimonious and interpretable measure of individual differences in behavior. However, the measurement reliabilities of this FC gradient have not been fully evaluated. Here, we assess the reliabilities of both global and regional principal FC gradient measures using test-retest data from the young adult Human Connectome Project (HCP-YA) and the Dunedin Study. Analyses revealed that the reliabilities of principal FC gradient measures were (1) consistently higher than those for traditional edge-wise FC measures, (2) higher for FC measures derived from general FC (GFC) in comparison with resting-state FC, and (3) higher for longer scan lengths. We additionally examined the relative utility of these principal FC gradient measures in predicting cognition and aging in both datasets as well as the HCP-aging dataset. These analyses revealed that regional FC gradient measures and global gradient range were significantly associated with aging in all three datasets, and moderately associated with cognition in the HCP-YA and Dunedin Study datasets, reflecting contractions and expansions of the cortical hierarchy, respectively. Collectively, these results demonstrate that measures of the principal FC gradient, especially derived using GFC, effectively capture a reliable feature of the human brain subject to interpretable and biologically meaningful individual variation, offering some advantages over traditional edge-wise FC measures in the search for brain-behavior associations.


Atomistic simulations of the load dependant friction force between silicon tip and diamond substrate.
In this paper, the load dependence on the interfacial friction between a cubic silicon tip and diamond substrate was investigated using molecular dynamics simulations. With the increase of the applied load, the sliding process experiences the states of superlubricity, single slip instability, double slip instability and plastic stage. The transitions from one state to the next one occur at the contact pressure 5.3 GPa, 8.0 GPa and 10.8 GPa, sequentially. In the superlubricity state, both friction and dissipated energy approach zero, independent of the load. However, in the single slip state the friction has a linear relationship with load, while the double slip mode induces decreased frictional force and lower damping. The coupling of the structure and forces in the plastic regime leads to the reduction of friction. These behaviors show good agreement with the recent experimental observations and theoretical predictions.


Two mass-spectrometric techniques for quantifying serine enantiomers and glycine in cerebrospinal fluid: potential confounders and age-dependent ranges.
The recent discovery and specific functions of D-amino acids in humans are bound to lead to the revelation of D-amino acid abnormalities in human disorders. Therefore, high-throughput analysis techniques are warranted to determine D-amino acids in biological fluids in a routine laboratory setting. We developed 2 chromatographic techniques, a nonchiral derivatization with chiral (chirasil-L-val column) separation in a GC-MS system and a chiral derivatization with Marfey's reagent and LC- MS analysis. We validated the techniques for D-serine, L-serine, and glycine determination in cerebrospinal fluid (CSF), evaluated several confounders, and determined age-dependent human concentration ranges. Quantification limits for D-serine, L-serine, and glycine in cerebrospinal fluid were 0.14, 0.44, and 0.14 micromol/L, respectively, for GC-MS and 0.20, 0.41, and 0.14 micromol/L for LC-MS. Within-run imprecision was <3% for both methods, and between-run imprecision was <13%. Comparison of both techniques with Deming regression yielded coefficients of 0.90 (D-serine), 0.92 (L-serine), and 0.96 (glycine). Sample collection, handling, and transport is uncomplicated-there is no rostrocaudal CSF gradient, no effect of storage at 4 degrees C for 1 week before storage at -80 degrees C, and no effect of up to 3 freeze/thaw cycles. Conversely, contamination with erythrocytes increased D-serine, L-serine, and glycine concentrations. CSF concentrations for 145 apparently healthy controls demonstrated markedly and specifically increased (5 to 9 times) D-serine concentrations during early central nervous system development. These 2 clinically applicable analysis techniques will help to unravel pathophysiologic, diagnostic, and therapeutic issues for disorders associated with central nervous system abnormalities, NMDA-receptor dysfunction, and other pathology associated with D-amino acids.


Susceptibility of cells from patients with primary immunodeficiency diseases to transformation by simian virus 40.
Skin fibroblasts were cultured from 15 patients with primary immunodeficiency diseases associated with a high cancer risk, including sex-linked agammaglobulinemia, IgA deficiency, variable immunodeficiency, ataxia-telangiectasia (cerebellar malfunction and abnormalities of blood vessels and immune response), Wiskott-Aldrich syndrome (low platelet count, eczema, and abnormal immune mechanism), and severe combined system (cellular and humoral) immunodeficiency. Fourteen of 15 cell strains were found to have low or regular susceptibility to transformation with the tumor virus, simian virus 40. The data are consistent with the view that the frequent occurrence of malignancy in patients with primary immunodeficiency is due to abnormalities of the immunologic surveillance mechanism.


Ultralow ruthenium modification of cobalt metal-organic frameworks for enhanced efficient bifunctional water splitting.
Hydrogen economy has emerged as a promising alternative to the current hydrocarbon economy. It involves harvesting renewable energy to split water into hydrogen and oxygen and then further utilising clean hydrogen fuel for various applications. The rational exploration of advanced non-precious metal bifunctional electrocatalysts for the oxygen evolution reaction (OER) and hydrogen evolution reaction (HER) is critical for efficient water splitting. Herein, an ultralow Ru-modified cobalt metal-organic framework (CoRu0.06-MOF/NF) two-dimensional nanosheet array bifunctional catalyst was fabricated through a strategy under mild experimental conditions. The obtained CoRu0.06-MOF/NF exhibited excellent bifunctional electrocatalytic activity and stability in alkaline media, with low overpotentials of 37 and 181 mV and significant durability for more than 95 and 110 h toward the HER and OER at 10 mA cm-2, respectively. The experimental results showed that the two-dimensional nanoarray structure had a large specific surface area and abundant exposed active sites. Additionally, ultralow Ru modification optimized the electronic structure and improved the conductivity of the cobalt metal-organic frameworks, thereby reducing the energy barrier of the rate-limiting step and accelerating the water splitting reaction.


Beliefs of Palestinian Women From Israel About the Responsibility and Punishment of Violent Husbands and About Helping Battered Women.
This article presents a study that examined beliefs about violent husbands and about helping battered women among Palestinian women living in Israel from the perspective of patriarchal ideology. A convenience sample of 701 married women was obtained, and a self-report questionnaire was administered. The findings reveal that the majority of participants held violent husbands accountable for their behavior; however, the majority of them did not support punishing violent husbands through formal agencies (i.e., the police) or through informal social institutions (i.e., the family). In addition, contrary to expectations, the majority of women perceived wife beating as a social problem rather than as a private one that should be dealt with within the family. Regression and multiple regression analysis revealed that women's endorsement of patriarchal ideology was found to influence all three above-mentioned beliefs about violent husbands and battered women, over and above the amount of variance in each of these beliefs that could be attributed to the women's sociodemographic characteristics. The limitations of the study and its implications for future research are discussed.


Circadian rhythm of insulin-induced hypoglycemia in man.
Four healthy men were given an insulin tolerance test (0.1 IU/kg BW) six times in different days at the following clock hours: 0400 h, 0800h, 1200 h, 1600 h, 2000 h, and 2400 h, in random order. Four days separated two consecutive tests. The glucose disappearance rate (K value) was calculated for each test between 5 min and 25 min after iv injection of insulin. The statistical evaluation by Cosinor test showed a statistically significant circadian rhythm for the hypoglycemic action of exogenous insulin with a peak at 1030 h (95% condifence limits: 0830 to 1430 h).


Heparanase-1 activity and the early postnatal prostate development.
Ventral prostate (VP) morphogenesis starts during embryonic development and continues for the first three postnatal weeks. Heparan sulfate (HS) affects paracrine signaling. Heparanase-1 (HPSE) is the only enzyme capable of cleaving HS. HPSE releases the HS bioactive fragment and mobilizes growth factors. Little is known, however, about HS turnover and HPSE function during VP morphogenesis. In this study, we measured HSPG expression and analyzed the expression and distribution of HPSE in the rat VP. HPSE was predominantly expressed by the VP epithelium. The VP was treated with heparin in ex vivo cultures to interfere with HS and resulted in delayed epithelial growth. Hpse knockdown using siRNA delayed epithelial growth in the first postnatal week ex vivo, which was similar to treating with the lower concentration of heparin. Hpse silencing was related to changes in HS chain length (as determined by size-exclusion chromatography, up-regulation of Mmp9, and down-regulation of Mmp2 expression). It also down-modulated ERK1/2 phosphorylation, suggesting a reduction in signaling, likely due to decreased HS cleavage and growth factor bioavailability. Our results showed that HPSE played a role in early epithelial growth during the first week of VP postnatal development. Developmental Dynamics 248:211-220, 2019. © 2019 Wiley Periodicals, Inc.


Hierarchical V4C3TX@NiO-reduced graphene oxide heterostructure hydrogels and defective reduced graphene oxide hydrogels as free-standing anodes and cathodes for high-performance asymmetric supercapacitors.
Asymmetric supercapacitors (ASCs) based on a battery-type anode and a capacitive-type cathode have been attracting extensive interest because of their high energy density. Herein, NiO nanosheets are hydrothermally deposited onto a V4C3TX substrate, which are then assembled into a 3D porous heterostructure hydrogel through a graphene oxide-assisted self-convergence hydrothermal process at low temperatures. The resultant hierarchical V4C3TX@NiO-RGO heterostructure hydrogel exhibits an ultrahigh specific capacitance of up to 1014.5 F g-1 at 1 A g-1. In addition, a defective reduced graphene oxide (DRGO) hydrogel is prepared using a cost-effective hydrothermal procedure followed by cobalt-catalyzed gasification, which shows a higher specific capacitance (258 F g-1 at 1 A g-1) than the untreated RGO hydrogel (176 F g-1). These two electrodes are then assembled into an ASC; the device features a stable operating voltage of 1.8 V, a maximum energy density of 86.22 W h kg-1 at 900 W kg-1, and excellent cycling stability at 96.4% capacitance retention after 10 000 cycles at 10 A g-1. The results from this work highlight the unique potential of MXene-based materials for the construction of high-performance ASCs.


Clinical ethics and the suffering Christian.
Contrary to the ecumenical spirit of our time, the differences among the Christian religions bring into question what one can say or do in common with fellow Christians. This issue, echoing the program of this journal, accentuates those differences, specifically when we focus on the Christian who is ill and suffering. At the bedside, it is the specifics of a religion, including not only its doctrines, but its informing and sustaining narratives, that must particularly be brought into play for the sake of the patient. Given this, our focus in this article regards what such a view implies for the clinician who is caring for a Christian patient whose religion he may or may not share, in general or in its specifics. Our basic conclusion is that the tendency for the clinician to act as both the patient's spiritual counselor, as well as his clinician, is generally neither prudent nor appropriate. Both hats should not be worn concurrently. This view is advanced not only because of concerns regarding patient vulnerability and the possible abuse of power, but also because the two roles may collide with or undermine each other.


The BILAG2004-Pregnancy Index is a valid disease activity outcome measure for pregnant SLE patients.
This study was to determine whether the BILAG2004-Pregnancy Index (BILAG2004-P) has construct/criterion validity and is sensitive to change. This was an observational multicentre study that recruited pregnant SLE patients. Data were collected on disease activity [using the BILAG2004-P and Physician Global Assessment (PGA)], investigations and therapy at each assessment. The overall BILAG2004-P score as determined by the highest score achieved by any system was used in the analysis. Cross-sectional analysis was used for construct and criterion validity. The comparison was with C3, C4 and anti-dsDNA for construct validity, while it was with change in therapy and PGA in criterion validity. Sensitivity to change was assessed by determining the relationship between the change in BILAG2004-P and the change in therapy between two consecutive visits. A total of 97 patients with 112 pregnancies were recruited. There were 610 assessments available for construct/criterion validity analysis (98.2% of pregnancies had more than one assessment) and 497 observations for sensitivity to change analysis. Increasing BILAG2004-P scores were associated with low C3. The active BILAG2004-P score (grade A or B) was associated with an increase in therapy and the PGA of active disease. There was an increasing likelihood of higher overall scores with an increase in therapy and the PGA of active disease. In the sensitivity to change analysis, an increase in the BILAG2004-P score was associated with an increase in therapy and inversely associated with a decrease in therapy. A decrease in the BILAG2004-P score was associated with a decrease in therapy and inversely associated with an increase in therapy. The BILAG2004-P has criterion validity and is sensitive to change.


The increased activity of a transcription factor inhibits autophagy in diabetic embryopathy.
Maternal diabetes induces neural tube defects and stimulates the activity of the forkhead box O3 (Fox)O3a in the embryonic neuroepithelium. We previously demonstrated that deleting the FOXO3a gene ameliorates maternal diabetes-induced neural tube defects. Macroautophagy (hereafter referred to as "autophagy") is essential for neurulation. Rescuing autophagy suppressed by maternal diabetes in the developing neuroepithelium inhibits neural tube defect formation in diabetic pregnancy. This evidence suggests a possible link between FoxO3a and impaired autophagy in diabetic embryopathy. We aimed to determine whether maternal diabetes suppresses autophagy through FoxO3a, and if the transcriptional activity of FoxO3a is required for the induction of diabetic embryopathy. We used a well-established type 1 diabetic embryopathy mouse model, in which diabetes was induced by streptozotocin, for our in vivo studies. To determine if FoxO3a mediates the inhibitory effect of maternal diabetes on autophagy in the developing neuroepithelium, we induced diabetic embryopathy in FOXO3a gene knockout mice and FoxO3a dominant negative transgenic mice. Embryos were harvested at embryonic day 8.5 to determine FoxO3a and autophagy activity and at embryonic day 10.5 for the presence of neural tube defects. We also examined the expression of autophagy-related genes. C17.2 neural stem cells were used for in vitro examination of the potential effects of FoxO3a on autophagy. Deletion of the FOXO3a gene restored the autophagy markers, lipidation of microtubule-associated protein 1A/1B-light chain 3I to light chain 3II, in neurulation stage embryos. Maternal diabetes decreased light chain 3I-positive puncta number in the neuroepithelium, which was restored by deleting FoxO3a. Maternal diabetes also decreased the expression of positive regulators of autophagy (Unc-51 like autophagy activating kinase 1, Coiled-coil myosin-like BCL2-interacting protein, and autophagy-related gene 5) and the negative regulator of autophagy, p62. FOXO3a gene deletion abrogated the dysregulation of autophagy genes. In vitro data showed that the constitutively active form of FoxO3a mimicked high glucose in repressing autophagy. In cells cultured under high-glucose conditions, overexpression of the dominant negative FoxO3a mutant blocked autophagy impairment. Dominant negative FoxO3a overexpression in the developing neuroepithelium restored autophagy and significantly reduced maternal diabetes-induced apoptosis and neural tube defects. Our study revealed that diabetes-induced FoxO3a activation inhibited autophagy in the embryonic neuroepithelium. We also observed that FoxO3a transcriptional activity mediated the teratogenic effect of maternal diabetes because dominant negative FoxO3a prevents maternal diabetes-induced autophagy impairment and neural tube defect formation. Our findings suggest that autophagy activators could be therapeutically effective in treating maternal diabetes-induced neural tube defects.


Fidelity of structure representation in electron micrographs of negatively stained protein molecules.
We have investigated the fidelity of structure representation in electron micrographs of negatively stained proteins by conducting a systematic evaluation of such micrographs in terms of a known molecular structure, solved by x-ray crystallography. Microcrystals of immunoglobulin G Dob were used as specimens in this comparison between micrograph images, optimized by computer image processing, and reference images derived computationally from the crystal structure. To an effective resolution of 2 nm, we observed a remarkably good correlation between the experimental images and their idealized counterparts, which are unaffected by those factors--electron irradiation and dehydration--that are thought to be primarily responsible for perturbation of protein structure during electron microscopy. Separate structural features resolved in these micrographs do not, in general, correspond to specific components of individual molecules but arise instead from complex superpositions involving several overlapping molecules.


The accumulation and persistence of somatic mutations in vivo.
Mutant frequencies at the native Dlb-1 and the lacI transgene reach a plateau within 1 week of mutagenesis in the small intestine of mice. This indicates that these mutations are selectively neutral and should accumulate additively. In a test with three mutagens, one potent (ethylnitrosourea) and two weak (1,2-dimethylhydrazine and methyl methanesulphonate), ten weekly treatments induced ten times the frequency of Dlb-1 mutations induced by a single treatment in each case, a result that is consistent with perfect additivity. These observations indicate that the mutation frequency at neutral loci will be an integrated measure of the effects of aging, endogenous mutagens, environmental mutagens and antimutagens. They suggest a simple protocol for transgenic mutation assays which would increase sensitivity, require fewer animals, and conform to standard toxicological practice.


A natively paired antibody library yields drug leads with higher sensitivity and specificity than a randomly paired antibody library.
Deep sequencing and single-chain variable fragment (scFv) yeast display methods are becoming more popular for discovery of therapeutic antibody candidates in mouse B cell repertoires. In this study, we compare a deep sequencing and scFv display method that retains native heavy and light chain pairing with a related method that randomly pairs heavy and light chain. We performed the studies in a humanized mouse, using interleukin 21 receptor (IL-21R) as a test immunogen. We identified 44 high-affinity binder scFv with the native pairing method and 100 high-affinity binder scFv with the random pairing method. 30% of the natively paired scFv binders were also discovered with the randomly paired method, and 13% of the randomly paired binders were also discovered with the natively paired method. Additionally, 33% of the scFv binders discovered only in the randomly paired library were initially present in the natively paired pre-sort library. Thus, a significant proportion of "randomly paired" scFv were actually natively paired. We synthesized and produced 46 of the candidates as full-length antibodies and subjected them to a panel of binding assays to characterize their therapeutic potential. 87% of the antibodies were verified as binding IL-21R by at least one assay. We found that antibodies with native light chains were more likely to bind IL-21R than antibodies with non-native light chains, suggesting a higher false positive rate for antibodies from the randomly paired library. Additionally, the randomly paired method failed to identify nearly half of the true natively paired binders, suggesting a higher false negative rate. We conclude that natively paired libraries have critical advantages in sensitivity and specificity for antibody discovery programs.


Long noncoding miRNA gene represses wheat β-diketone waxes.
The cuticle of terrestrial plants functions as a protective barrier against many biotic and abiotic stresses. In wheat and other Triticeae, β-diketone waxes are major components of the epicuticular layer leading to the bluish-white glaucous trait in reproductive-age plants. Glaucousness in durum wheat is controlled by a metabolic gene cluster at the WAX1 (W1) locus and a dominant suppressor INHIBITOR of WAX1 (Iw1) on chromosome 2B. The wheat D subgenome from progenitor Aegilops tauschii contains W2 and Iw2 paralogs on chromosome 2D. Here we identify the Iw1 gene from durum wheat and demonstrate the unique regulatory mechanism by which Iw1 acts to suppress a carboxylesterase-like protein gene, W1-COE, within the W1 multigene locus. Iw1 is a long noncoding RNA (lncRNA) containing an inverted repeat (IR) with >80% identity to W1-COE The Iw1 transcript forms a miRNA precursor-like long hairpin producing a 21-nt predominant miRNA, miRW1, and smaller numbers of related sRNAs associated with the nonglaucous phenotype. When Iw1 was introduced into glaucous bread wheat, miRW1 accumulated, W1-COE and its paralog W2-COE were down-regulated, and the phenotype was nonglaucous and β-diketone-depleted. The IR region of Iw1 has >94% identity to an IR region on chromosome 2 in Ae. tauschii that also produces miRW1 and lies within the marker-based location of Iw2 We propose the Iw loci arose from an inverted duplication of W1-COE and/or W2-COE in ancestral wheat to form evolutionarily young miRNA genes that act to repress the glaucous trait.


[Animals' clever adaptation strategy for seasonal changes in environment].
Organisms living outside of tropical zones experience seasonal changes in environment. Organisms are using day length as a calendar to change their physiology and behavior such as seasonal breeding, hibernation, migration, and molting. A comparative biology approach revealed underlying mechanisms of vertebrate seasonal reproduction. Here we review the current understanding of vertebrate seasonal reproduction. We Aso describe the involvement of tissue-specific post-translational modification in functional diversification of a hormone.