L-amino acid sensing by the extracellular Ca2+-sensing receptor.
The extracellular calcium (Ca(2+)(o))-sensing receptor (CaR) recognizes and responds to (i.e., "senses") Ca(2+)(o) as its principal physiological ligand. In the present studies, we document that the CaR is activated not only by extracellular calcium ions but also by amino acids, establishing its capacity to sense nutrients of two totally different classes. l-Amino acids, especially aromatic amino acids, including l-phenylalanine and l-tryptophan, stereoselectively mobilized Ca(2+) ions in the presence of the CaR agonists, Ca(2+)(o), gadolinium (Gd(3+)(o)), and spermine in fura-2-loaded human embryonic kidney (HEK-293) cells stably transfected with the human CaR. l-amino acid-dependent effects were observed above, but not below, a threshold level of Ca(2+)(o) of approximately 1.0 mM. l-Amino acids, particularly aromatic amino acids, also stereoselectively enhanced the sensitivity of the CaR to its agonists, Ca(2+)(o) and spermine. Branched-chain amino acids were almost inactive, and charged amino acids, including arginine and lysine, were much less effective than aromatic and other amino acids. l-amino acid mixtures emulating the amino acid composition of fasting human plasma reproduced the effects of high concentrations of individual l-amino acids on Ca(2+) mobilization and enhanced the sensitivity of the CaR to Ca(2+)(o). The data presented herein identify the CaR as a molecular target for aromatic and other l-amino acids. Thus, the CaR can integrate signals arising from distinct classes of nutrients: mineral ions and amino acids. The actions of l-amino acids on the CaR may provide explanations for several long recognized but poorly understood actions of dietary protein on calcium metabolism.


Targeting TMPRSS2 and Cathepsin B/L together may be synergistic against SARS-CoV-2 infection.
The entry of SARS-CoV-2 into target cells requires the activation of its surface spike protein, S, by host proteases. The host serine protease TMPRSS2 and cysteine proteases Cathepsin B/L can activate S, making two independent entry pathways accessible to SARS-CoV-2. Blocking the proteases prevents SARS-CoV-2 entry in vitro. This blockade may be achieved in vivo through 'repurposing' drugs, a potential treatment option for COVID-19 that is now in clinical trials. Here, we found, surprisingly, that drugs targeting the two pathways, although independent, could display strong synergy in blocking virus entry. We predicted this synergy first using a mathematical model of SARS-CoV-2 entry and dynamics in vitro. The model considered the two pathways explicitly, let the entry efficiency through a pathway depend on the corresponding protease expression level, which varied across cells, and let inhibitors compromise the efficiency in a dose-dependent manner. The synergy predicted was novel and arose from effects of the drugs at both the single cell and the cell population levels. Validating our predictions, available in vitro data on SARS-CoV-2 and SARS-CoV entry displayed this synergy. Further, analysing the data using our model, we estimated the relative usage of the two pathways and found it to vary widely across cell lines, suggesting that targeting both pathways in vivo may be important and synergistic given the broad tissue tropism of SARS-CoV-2. Our findings provide insights into SARS-CoV-2 entry into target cells and may help improve the deployability of drug combinations targeting host proteases required for the entry.


[Costs and benefits of heart failure treatment].
Congestive heart failure constitutes an important public health problem, because of its prevalence and mortality, and also its morbidity and significant resource utilization. It is the main cause of hospitalization for patients aged 65 or over, and it absorbs 2% of the health care budget in industrialized countries. With the aging of the population and the advancing age of the baby boom generation, the situation will worsen: the prevalence of congestive heart failure in Europe may increase by 70% by the year 2010. To favourably alter these economic consequences, treatment must not only prolong life but also slow down or even prevent disease progression towards more severe stages and decrease resource utilization, mainly in reducing the need for hospital admissions. To date, the most convincing results of economic analyses concern vasodilators: the hydralazine-isosorbide dinitrate combination and angiotensin converting enzyme inhibitors (ACE). Even if all these studies do not come to the same conclusions in terms of numbers, they agree in showing that costs per year of life saved by vasodilators, and particularly by ACE, are very favourable (< $10,000 per year of life saved) and much lower than those of most other well accepted medical strategies.


Elevated Plasma Viral Loads in Romidepsin-Treated Simian Immunodeficiency Virus-Infected Rhesus Macaques on Suppressive Combination Antiretroviral Therapy.
Replication-competent human immunodeficiency virus (HIV) persists in infected people despite suppressive combination antiretroviral therapy (cART), and it represents a major obstacle to HIV functional cure or eradication. We have developed a model of cART-mediated viral suppression in simian human immunodeficiency virus (SIV) mac239-infected Indian rhesus macaques and evaluated the impact of the histone deacetylase inhibitor (HDACi) romidepsin (RMD) on viremia in vivo. Eight macaques virologically suppressed to clinically relevant levels (<30 viral RNA copies/ml of plasma), using a three-class five-drug cART regimen, received multiple intravenous infusions of either RMD (n = 5) or saline (n = 3) starting 31 to 54 weeks after cART initiation. In vivo RMD treatment resulted in significant transient increases in acetylated histone levels in CD4(+) T cells. RMD-treated animals demonstrated plasma viral load measurements for each 2-week treatment cycle that were significantly higher than those in saline control-treated animals during periods of treatment, suggestive of RMD-induced viral reactivation. However, plasma virus rebound was indistinguishable between RMD-treated and control-treated animals for a subset of animals released from cART. These findings suggest that HDACi drugs, such as RMD, can reactivate residual virus in the presence of suppressive antiviral therapy and may be a valuable component of a comprehensive HIV functional cure/eradication strategy.


The effect of asbestos-alternatives on the accuracy of cast veneer crowns.
The effect of three different lining materials on the investment expansion during the casting procedure, was compared. The accuracy of the castings was assessed by measuring slits at the gingival shoulder of a cast veneer crown. The liners were made of asbestosos, paper and a glassfiber material. They were used with a gypsum bonded and a phosphate bonded investment material using a regular type III and a high-fusing alloy. The glass-fiber material seemed to give a good and reproducible accuracy with both investments. The effect of this liner seemed to be comparable or better than that of the conventional asbest material. The paper liner sometimes allowed a sufficient expansion, but with varying results. Additional problems were connected to this material as it was burnt to ash during the mould heating procedure.


Characteristics of reporting diabetes mellitus research results in Japanese newspapers.
This study aims to characterize the coverage of research on diabetes mellitus by Japanese newspapers. Newspaper articles with the key words diabetes mellitus, diabetes disease, or blood sugar in the headline were selected from four major Japanese newspapers for the period 1988-2007 using the ELNET database and coded by two researchers. Of 152 newspaper articles examined, 92 (60.5%) were based on journal articles, and published in English. The remaining 60 (39.5 %) were based on academic meetings, of which 51 (85.0%) were conducted in Japan. Seventy-two articles covered non-human studies (47.4%), but only 26 used a word such as animal, mouse, or cell in the headline to describe the study subjects. Publications in countries where the native language is not English may have language and geographical barriers that affect the reporting of research results.


YKL-40 promotes chemokine expression following drug-induced liver injury via TF-PAR1 pathway in mice.
Background: The inflammatory factor YKL-40 is associated with various inflammatory diseases and is key to remodeling inflammatory cells and tissues. YKL-40 (Chi3l1) promotes the activation of tissue factor (TF), leading to intrahepatic vascular coagulation (IAOC) and liver injury. TF is a key promoter of the exogenous coagulation cascade and is also involved in several signaling involving cell proliferation, apoptosis, charring, migration and inflammatory diseases pathways. However, the effect of YKL-40-induced TF-PAR1 pathway on the expression of downstream chemokines remains unknown. Methods: We established a liver injury model using Concanavalin A (ConA) in C57 BL/6 mice. By adopting various experimental techniques, the effect of YKL-40 induced TF-PAR1 pathway on the expression of downstream chemokine ligand 2 (CCL2) and IP-10 was verified. Results: We found that overexpression of YKL-40 increased the expression of TF, protease-activated receptor 1 (PAR1), CCL2 and IP-10 in mice and exacerbated the severity of liver injury. However, blocking the expression of TF significantly reversed the extent of liver injury. Conclusion: We found that YKL-40 promotes the expression of downstream chemokines ligand 2 (CCL2) and IP-10 by activating the TF-PAR1 pathway, leading to increased recruitment of inflammatory cells and exacerbating the progression of liver injury. This provides a new approach for the clinical treatment of drug-induced liver injury.


Paraquat-induced phosphatidylserine oxidation and apoptosis are independent of activation of PLA2.
Paraquat is a pneumotoxin that causes lung injury by enhancing oxidative stress; however, the cellular responses to these redox events are undefined. We previously showed that paraquat produced selective peroxidation of phosphatidylserine that preceded apoptosis in 32D cells. We now report that the phospholipase A2 (PLA2) inhibitor quinacrine can attenuate phosphatidylserine oxidation and also block paraquat-induced apoptosis. Therefore, we investigated the potential for PLA2 to mediate apoptosis after paraquat. We found that, in contrast to quinacrine, the PLA2 inhibitors manoalide, aristolochic acid, and arachidonyl trifluoromethylketone failed to prevent paraquat-induced apoptosis. Moreover, no evidence of PLA2 activation was observed within 7 h after paraquat exposure. Finally, quinacrine failed to inhibit basal and 4-bromo-A-23187-induced release of [3H]arachidonic acid at concentrations that protected paraquat-induced apoptosis. We conclude that paraquat-induced phosphatidylserine oxidation and apoptosis occurred in the absence of PLA2 activation and that quinacrine protected phosphatidylserine and cell viability after paraquat in a PLA2-independent manner.


Regulatory aspects of glycosaminoglycan biosynthesis.
The control of glycosaminoglycan biosynthesis was investigated by studying the kinetic and regulatory properties of some enzymes involved in the formation of UDP-sugar precursors: UDP-N-acetylglucosamine 4'-epimerase, catalyzing the interconversion of hexosamine precursors and UDP-glucose dehydrogenase and UDP-glucose 4'-epimerase, utilizing UDP-glucose for the formation of uronic acid and galactose precursors. The study was carried out in tissues with different glycosaminoglycan production: bovine cornea, producing both chondroitin sulfate and keratan sulfate, and newborn-pig epiphysial-plate cartilage, producing mostly chondroitin sulfate. The biosynthesis of hexosamine precursors appeared to be regulated by the value of the NAD/NADH ratio. This control mechanism regulated also the activities of both UDP-glucose dehydrogenase and UDP-glucose 4'-epimerase and, therefore, it could correlate the biosynthesis of glycosaminoglycan precursors with the redox activity of the cell. At the level of UDP-glucose utilization two other control mechanisms were demonstrated: the different affinities of UDP-glucose dehydrogenase and UDP-glucose 4'-epimerase for UDP-glucose in tissues with different glycosaminoglycan production and the cellular concentration of UDP-xylose. This sugar-nucleotide inhibited UDP-glucose dehydrogenase, but did not affect the UDP-glucose 4'-epimerase activity; therefore, and increase of its cellular concentration may result in a decreased chondroitin sulfate synthesis and in an increased keratan sulfate formation.


Development of a von Willebrand-like syndrome after prolonged use of hydroxyethyl starch.
A 29-year-old female with no prior history of bleeding tendencies had gingival bleeding develop after 10 days of treatment with hydroxyethyl starch (HES), 1,000 mL/day. Laboratory studies revealed a marked reduction in Factor VIII coagulant activity (VIII:C), von Willebrand's factor antigen (vWF:Agn), and (VIII R:RCo), resembling severe type I von Willebrand's disease. The HES was stopped and the bleeding subsided. The levels of Factor VIII:C, vWF:Agn, and Factor VIII R:RCo gradually increased to normal over the following 17 days.


Effects of excitotoxic lesions in the ventral striatopallidal--thalamocortical pathway on odor reversal learning: inability to extinguish an incorrect response.
The role of the ventral striatopallidal pathway and related cortical areas in stimulus-reward association reversal behavior was studied by inducing bilateral lesions with the excitotoxin, N-methyl-D-aspartate (NMDA) at restricted sites in the system. The areas lesioned were the ventral pallidum (VP), the ventral striatum (VS), the medial prefrontal cortex (mPFC) [i.e., the prelimbic (PL) and infralimbic (IL) cortexes], and the orbital cortex [i.e., the dorsolateral orbital (DLO), ventral lateral orbital (VLO), and lateral orbital (LO) cortexes]. Rats with lesions of the dorsal caudate nucleus and putamen (CPu) served as a positive control in this study. Water-deprived rats were trained on a go, no-go two-odor olfactory discrimination task to respond to one odor (S+) with water as a reward and to suppress responding to the other odor (S-). The rats were then tested for their ability to reverse the associated stimuli. The number of errors made before successfully learning the stimulus-reward association were measured in relation to a sham lesion control group which did not receive injections of NMDA. In experimental rats, the lesions did not affect their ability to learn stimulus-reward associations for novel odors, but did result in an increase in the number of false alarms after the significance of the associated stimuli were reversed. That is, the lesioned animals persisted in responding to the formerly rewarded but now unrewarded stimulus. Rats with damage to the CPu did not show a significant effect when compared with the controls during reversal problems. The results support the hypothesis that the ventral striatopallidal system, together with related thalamic and frontal cortical structures, functions in reversal learning by suppressing inappropriate responses to stimuli that are no longer rewarded.


Long-term outcome after additional catheter-directed thrombolysis versus standard treatment for acute iliofemoral deep vein thrombosis (the CaVenT study): a randomised controlled trial.
Conventional anticoagulant treatment for acute deep vein thrombosis (DVT) effectively prevents thrombus extension and recurrence, but does not dissolve the clot, and many patients develop post-thrombotic syndrome (PTS). We aimed to examine whether additional treatment with catheter-directed thrombolysis (CDT) using alteplase reduced development of PTS. Participants in this open-label, randomised controlled trial were recruited from 20 hospitals in the Norwegian southeastern health region. Patients aged 18-75 years with a first-time iliofemoral DVT were included within 21 days from symptom onset. Patients were randomly assigned (1:1) by picking lowest number of sealed envelopes to conventional treatment alone or additional CDT. Randomisation was stratified for involvement of the pelvic veins with blocks of six. We assessed two co-primary outcomes: frequency of PTS as assessed by Villalta score at 24 months, and iliofemoral patency after 6 months. Analyses were by intention to treat. This trial is registered at ClinicalTrials.gov, NCT00251771. 209 patients were randomly assigned to treatment groups (108 control, 101 CDT). At completion of 24 months' follow-up, data for clinical status were available for 189 patients (90%; 99 control, 90 CDT). At 24 months, 37 (41·1%, 95% CI 31·5-51·4) patients allocated additional CDT presented with PTS compared with 55 (55·6%, 95% CI 45·7-65·0) in the control group (p=0·047). The difference in PTS corresponds to an absolute risk reduction of 14·4% (95% CI 0·2-27·9), and the number needed to treat was 7 (95% CI 4-502). Iliofemoral patency after 6 months was reported in 58 patients (65·9%, 95% CI 55·5-75·0) on CDT versus 45 (47·4%, 37·6-57·3) on control (p=0·012). 20 bleeding complications related to CDT included three major and five clinically relevant bleeds. Additional CDT should be considered in patients with a high proximal DVT and low risk of bleeding. South-Eastern Norway Regional Health Authority; Research Council of Norway; University of Oslo; Oslo University Hospital.


Abandoning sex: multiple origins of asexuality in the ciliate Tetrahymena.
By segregating somatic and germinal functions into large, compound macronuclei and small diploid micronuclei, respectively, ciliates can explore sexuality in ways other eukaryotes cannot. Sex, for instance, is not for reproduction but for nuclear replacement in the two cells temporarily joined in conjugation. With equal contributions from both conjugants, there is no cost of sex which theory predicts should favor asexuality. Yet ciliate asexuality is rare. The exceptional Tetrahymena has abandoned sex through loss of the micronucleus; its amicronucleates are abundant in nature where they reproduce by binary fission but never form conjugating pairs. A possible reason for their abundance is that the Tetrahymena macronucleus does not accumulate mutations as proposed by Muller's ratchet. As such, Tetrahymena amicronucleates have the potential to be very old. This study used cytochrome oxidase-1 barcodes to determine the phylogenetic origin and relative age of amicronucleates isolated from nature. Amicronucleates constituted 25% of Tetrahymena-like wild isolates. Of the 244 amicronucleates examined for cox1 barcodes, 237 belonged to Tetrahymena, seven to other genera. Sixty percent originated from 12 named species or barcoded strains, including the model Tetrahymena thermophila, while the remaining 40% represent 19 putative new species, eight of which have micronucleate counterparts and 11 of which are known only as amicronucleates. In some instances, cox1 haplotypes were shared among micronucleate and amicronucleates collected from the same source. Phylogenetic analysis showed that most amicronucleates belong to the "borealis" clade in which mating type is determined by gene rearrangement. Some amicronucleate species were clustered on the SSU phylogenetic tree and had longer branch lengths, indicating more ancient origin. Naturally occurring Tetrahymena amicronucleates have multiple origins, arising from numerous species. Likely many more new species remain to be discovered. Shared haplotypes indicate that some are of contemporary origin, while phylogeny indicates that others may be millions of years old. The apparent success of amicronucleate Tetrahymena may be because macronuclear assortment and recombination allow them to avoid Muller's ratchet, incorporate beneficial mutations, and evolve independently of sex. The inability of amicronucleates to mate may be the result of error(s) in mating type gene rearrangement.


[The Italian Group for the Study of Streptokinase in Myocardial Infarct: Changes in surface cardiac potentials. Study with chest electromaps].
The electrocardiographic changes during and after the thrombolytic treatment with streptokinase (SK) were assessed by means of body surface potential mapping. The aim of the study was to identify potential patterns suggesting reperfusion and revealing possible short-term effects on the infarct size of the recanalization. We studied 23 patients enrolled in the G.I.S.S.I. trial; 11 had an anterior and 12 had an inferior myocardial infarction; 14 were treated with SK and 9 were controls. Body surface maps were recorded from 105 lead points located on the anterior thoracic surface using an automated instrument. The maps were obtained immediately before the SK infusion (or at the time of randomization in the control patients), 30, 60, 120 minutes thereafter and then 24 hours and 7 days after the onset of the infarct symptoms. In each patient the surface potential distribution at 100 msec after the end of QRS was considered and the sum of all the positive potential values was calculated (sigma ST). In addition, the potential time integrals relating to two intervals of the cardiac cycle (first 100 msec of ST and first 40 msec of QRS) were calculated at each lead point and transferred to diagrams representing the chest surface explored (isointegral map). With respect to Q-40 maps, deviation index maps were calculated as follows: the mean Q-40 map (obtained from 30 normal subjects) was subtracted from the map of each patient; the value obtained at each lead point was then divided by the standard deviation of the normal values for that point. An area where the integral values were at least 2 SD lower than normal was considered a reliable index of infarct. By considering as index of reperfusion an early peak of CPK (less than 12 hours from the onset of infarct symptoms), we divided the patients into 2 subsets: reperfused (R) and not reperfused (NR). The mean values of sigma ST at 100 msec progressively decreased in all patients from the baseline to the subsequent recordings in both control and SK groups, without significant differences; nevertheless, the highest percent reductions of sigma ST were observed only in some R patients. The maximum on the ST-100 isointegral maps also showed a similar behaviour.(ABSTRACT TRUNCATED AT 250 WORDS)


Life histories of the seed bugs, Kleidocerys punctatus and Kleidocerys virescens.
The life cycles of the seed bugs, Kleidocerys punctatus Distant and Kleidocerys virescens F. (Hemiptera: Lygaeidae: Ischnorhynchinae), are reported for the first time. Description of all immature stages and adults are included. Adults and nymphs of K. punctatus are associated with several species of Alnus (Betulaceae), while those of K. virescens are associated with Nicotiana glauca Graham, Nicotiana tabacum L. (Solanaceae), and Buddleia crotonoides A. Gray and Buddleia sp. (Loganiaceae). Adults and nymphs feed mainly on the seeds, inside the dry fruit, but they also take plant juices from other reproductive and vegetative structures. Illustrations of the eggs, all nymphal instars, and the adults, as well as notes on their biology and their distribution in Mexico, are included.


Stress and pregnancy among African-American women.
Research on stress tends to support an adverse effect on pregnancy outcomes, and suggests that the impact of these stressors is modified by social class and/or race. This study explicitly examined social factors such as experiences of discrimination, either racial or sexual, and neighbourhood crime as predictors of stress. We also examined cortisol and stress as predictors of blood pressure. A subsample of 94 African-American pregnant women, aged 18-39 years, who were enrolled in a longitudinal study of pregnancy and exposure to lead in the environment were used in this analysis. The women were patients at an obstetrics clinic at Magee Women's Hospital in Pittsburgh, PA, USA. Younger age, higher income, lower education and experiences of discrimination, both racial and sexual, were related to greater perceived stress; however, life events were not related to perceived stress. Higher income and urinary cortisol adjusted for creatinine were related to systolic blood pressure after the 36th week. As a body of evidence suggests that stress can have deleterious effects in both pregnant and non-pregnant women, future research should examine these forms of discrimination, especially racial discrimination, as a possible reason for the disparity in adverse pregnancy outcomes between African-American and white women.


Basis of the gabamimetic profile of ethanol.
This article summarizes the proceedings of a symposium held at the 2005 Research Society on Alcoholism meeting. The initial presentation by Dr. Wallner provided evidence that selected GABA(A) receptors containing the delta subunit display sensitivity to low intoxicating ethanol concentrations and this sensitivity is further increased by a mutation in the cerebellar alpha6 subunit, found in alcohol-hypersensitive rats. Dr. Mameli reported that ethanol affects gamma-aminobutyric acid (GABA) function by affecting neural circuits that influence GABA release. Dr. Parsons presented data from electrophysiological and microdialysis investigations that ethanol is capable of releasing GABA from presynaptic terminals. Dr. Morrow demonstrated that systemic ethanol increases neuroactive steroids in brain, the absence of which alters various functional responses to ethanol. Dr. Criswell presented evidence that the ability of ethanol to increase GABA was apparent in some, but not all, brain regions indicative of regional specificity. Further, Dr. Criswell demonstrated that neurosteroids alone and when synthesized locally by ethanol act postsynaptically to enhance the effect of GABA released by ethanol in a region specific manner. Collectively, this series of reports support the GABAmimetic profile of acutely administered ethanol being dependent on several specific mechanisms distinct from a direct effect on the major synaptic isoforms of GABA(A) receptors.


Part-time physicians. Physician workload and patient-based assessments of primary care performance.
To examine the relationship between the number of hours physicians work and patients' assessment of the physician. Cross-sectional study with physician and patient surveys. Primary care practices in Massachusetts. A random sample of 6810 Massachusetts state employees in 15 different health plans. Eleven summary scales measuring 7 essential elements of primary care. Information was derived from the Primary Care Assessment Survey, a validated patient-completed questionnaire. Physicians were classified into 3 groups according to their reported hours of work: "overtime" (> 65 h/wk), "full time" (40-65 h/wk), and "part time" (< 40 h/wk). There was no statistically significant difference between the 3 groups of physicians in 10 of the 11 measures of primary care performance. Physicians who worked more than 65 hours per week were found to score significantly higher in the visit-based continuity of care category than physicians working fewer hours. Physicians working more than 65 hours per week were also found to be significantly less satisfied with the amount of time they had for family and personal life than the other 2 groups. Part-time physicians perform as well as full-time physicians in most aspects of primary care, including all interpersonal aspects of care, as reported by patients. Patients of physicians working more than 65 hours per week experienced higher levels of visit-based continuity of care than patients of physicians working fewer hours, but this appears to carry a cost to those physicians in the area of personal and professional satisfaction. Subsequent research should examine the relationship between physician workload and technical aspects of care.


AIDS meeting. Companies, donors pledge to close gap in AIDS treatment.
Although basic science and clinical research typically dominate at this biennial event, by far the most riveting sessions at the XIII International AIDS Conference here revolved around the thicket of issues that prevent poor people in sub-Saharan Africa--where a staggering 24.5 million people are estimated to be infected with HIV--from receiving the 14 anti-HIV treatments now used in wealthy countries. Pharmaceutical companies and donors last week loosed a flood of promises about removing barriers. Conference attendees welcomed the offers of help, but many stressed that much more will be needed.


An allelic series reveals essential roles for FY in plant development in addition to flowering-time control.
The autonomous pathway functions to promote flowering in Arabidopsis by limiting the accumulation of the floral repressor FLOWERING LOCUS C (FLC). Within this pathway FCA is a plant-specific, nuclear RNA-binding protein, which interacts with FY, a highly conserved eukaryotic polyadenylation factor. FCA and FY function to control polyadenylation site choice during processing of the FCA transcript. Null mutations in the yeast FY homologue Pfs2p are lethal. This raises the question as to whether these essential RNA processing functions are conserved in plants. Characterisation of an allelic series of fy mutations reveals that null alleles are embryo lethal. Furthermore, silencing of FY, but not FCA, is deleterious to growth in Nicotiana. The late-flowering fy alleles are hypomorphic and indicate a requirement for both intact FY WD repeats and the C-terminal domain in repression of FLC. The FY C-terminal domain binds FCA and in vitro assays demonstrate a requirement for both C-terminal FY-PPLPP repeats during this interaction. The expression domain of FY supports its roles in essential and flowering-time functions. Hence, FY may mediate both regulated and constitutive RNA 3'-end processing.


Examining the contemporaneous occurrence of bullying and teen dating violence victimization.
Teen dating violence (TDV) is a preventable public health issue that has been linked to other forms of aggression and violence victimization. It is also a growing concern for school psychologists who may be working to prevent TDV and related behavioral problems, like bullying. The current study examined various forms of bullying victimization (verbal, physical, and relational) and their association with physical and emotional TDV. Self-report data from 17,780 adolescents (33% African American, 54% White) in Grades 9-12 across 58 high schools were analyzed using 3-level models with dichotomous outcomes. Multilevel logistic regressions indicated that adolescents who had experienced bullying (physical, relational, and verbal) were more likely to have also experienced physical and emotional dating violence. Perceived norms about students' and adults' bullying interventions were associated with reduced odds of physical (OR(adults) = .82, p < .001) and emotional TDV (OR(adults) = .82, p < .001). Findings underscore the need to better understand the relationship between TDV and bullying victimization to design and enhance prevention efforts that address both forms of violence.


How to deal with multiple binding poses in alchemical relative protein-ligand binding free energy calculations.
Recent advances in improved force fields and sampling methods have made it possible for the accurate calculation of protein–ligand binding free energies. Alchemical free energy perturbation (FEP) using an explicit solvent model is one of the most rigorous methods to calculate relative binding free energies. However, for cases where there are high energy barriers separating the relevant conformations that are important for ligand binding, the calculated free energy may depend on the initial conformation used in the simulation due to the lack of complete sampling of all the important regions in phase space. This is particularly true for ligands with multiple possible binding modes separated by high energy barriers, making it difficult to sample all relevant binding modes even with modern enhanced sampling methods. In this paper, we apply a previously developed method that provides a corrected binding free energy for ligands with multiple binding modes by combining the free energy results from multiple alchemical FEP calculations starting from all enumerated poses, and the results are compared with Glide docking and MM-GBSA calculations. From these calculations, the dominant ligand binding mode can also be predicted. We apply this method to a series of ligands that bind to c-Jun N-terminal kinase-1 (JNK1) and obtain improved free energy results. The dominant ligand binding modes predicted by this method agree with the available crystallography, while both Glide docking and MM-GBSA calculations incorrectly predict the binding modes for some ligands. The method also helps separate the force field error from the ligand sampling error, such that deviations in the predicted binding free energy from the experimental values likely indicate possible inaccuracies in the force field. An error in the force field for a subset of the ligands studied was identified using this method, and improved free energy results were obtained by correcting the partial charges assigned to the ligands. This improved the root-mean-square error (RMSE) for the predicted binding free energy from 1.9 kcal/mol with the original partial charges to 1.3 kcal/mol with the corrected partial charges.


The comorbidity of hypertension and psychological distress: a study of nine countries in the former Soviet Union.
Mental health problems in those with physical ailments are often overlooked, especially in the former Soviet Union (fSU) where this comorbidity has received little attention. Our study examines the comorbidity of psychological distress and hypertension in the fSU. Nationally representative household survey data from Armenia, Azerbaijan, Belarus, Georgia, Kazakhstan, Kyrgyzstan, Moldova, Russia and Ukraine in 2001 and 2010 were analysed to compare the levels of psychological distress in people with and without self-reported hypertension. Multivariate regression analysed determinants of psychological distress in hypertensive respondents, and prevalence rate ratios were calculated to compare the change in distress between the two groups. There were significantly higher levels of psychological distress among hypertensive respondents (9.9%) than in the general population (4.9%), and a significant association between the two conditions [odds ratio (OR) = 2.27 (1.91; 2.70)]. Characteristics associated with distress among hypertensive respondents included residing in Armenia or Kyrgyzstan, being female, over age 50, with a poor economic situation, lower education, poor emotional support and limited access to medical drugs. Levels of distress declined between 2001 and 2010, but at a lesser rate in hypertensive respondents [rate ratio (RR) = 0.85 (0.75; 0.95)] than non-hypertensive respondents [RR = 0.65 (0.56; 0.75)]. There is a significant association between psychological distress and hypertension in the region.


Application of the DSM-5 Level of Personality Functioning Scale by Lay Raters.
One concern that has been expressed with the Alternative Model for Personality Disorders (AMPD) presented in DSM-5 is that the description of characteristic impairments in personality function uses concepts requiring considerable experience and clinical inference to apply. To examine this question, the individual indicators included in the AMPD's Level of Personality Functioning Scale (LPFS) that describes these core impairments were abstracted as individual items, and then rated on a target acquaintance by 194 undergraduate college students with minimal training in personality disorder and no training in the AMPD. Results indicated that the LPFS indicators were highly internally consistent as rated in this sample, and that the degree of discrimination between groups corresponded very well with the putative level of severity represented for each indicator in the LPFS. These findings support the contention that using the LPFS might not require any particular clinical experience or training.


The role of antiphospholipid autoantibodies in the cognitive deficits of patients with systemic lupus erythematosus.
To analyze the role of the antiphospholipid autoantibodies (aPL) on the neuropsychological deficits in systemic lupus erythematosus (SLE) patients, comparing groups of patients with antiphospholipid syndrome (APS; n = 15), SLE with aPL (n = 12), and SLE without aPL (n = 27), and a healthy control group (n = 31). Patients fulfilled the American College of Rheumatology SLE classification criteria or the Sydney criteria for APS. All participants were woman, and groups were matched on age and education. A standardized cognitive examination classified patients as cognitively declined or impaired according to the American College of Rheumatology. Differences between the groups were found in all of the studied variables, comprising attention and executive functions (sustained and selective attention, fluency, and inhibition), and memory (verbal and visual). Post-hoc analyses showed cognitive performance was equivalent between APS and SLE with aPL. Differences between SLE without aPL and control groups were found only in four of the 10 studied variables, while differences in all but two memory variables were found between SLE without aPL and control groups. Furthermore, cognitive deficit was three times more frequent in APS and SLE with aPL patients than for the control group (80%, 75%, and 16%, respectively), and two times more frequent compared to SLE patients without aPL (48%). Our results support the relationship between aPL and cognitive symptoms in SLE. Also, almost half of the patients with SLE and no aPL showed cognitive problems, pointing to the multifactorial causes of cognitive problems in SLE. Future research with larger sample size is guaranteed to replicate our results.


Spectral analysis of skin laser Doppler blood perfusion signal during cutaneous hyperemia in response to acetylcholine iontophoresis and ischemia in normal subjects.
Spectral analysis of skin laser Doppler (LD) signal was performed using Fourier transformation in twelve healthy subjects in order to compare post-ischemic hyperemia and acetylcholine (ACh) iontophoresis tests for the exploration of cutaneous microcirculatory endothelial function. Within the frequency range studied of 0.009-1.6 Hz, five different frequency intervals were considered, arising from local (endothelial and myogenic) and central (neural, respiratory and cardiac) regulatory mechanisms. The power density (PD) for each of the five frequency intervals considered was measured in perfusion units (PU)/Hz, in each subject, before and in response to ischemia and to ACh iontophoresis, using a dedicated software. The relative contribution of each frequency interval was calculated as per cent rate between each PD and the sum of the PD of the five frequency interval considered. PD of LD signals in response to ischemia and Ach iontophoresis significantly increased for all the five frequency intervals considered. A significant increase (from 25.0+/-6.3 to 48.3+/-17.8; p<0.01) in the relative contribution of 0.009-0.02 frequency interval, referred to endothelium activity, was observed only in response to ACh iontophoresis. No significant increase in the relative power contribution was observed in response to ischemia for any of the five frequency intervals considered. These data suggest that ACh iontophoresis test is more adequate than post-ischemic hyperemia test in exploring the skin microcirculatory endothelial function in normal subjects.


Distinct actions of cis and trans ATP within the double ring of the chaperonin GroEL.
The chaperonin GroEL is a double-ring structure with a central cavity in each ring that provides an environment for the efficient folding of proteins when capped by the co-chaperone GroES in the presence of adenine nucleotides. Productive folding of the substrate rhodanese has been observed in cis ternary complexes, where GroES and polypeptide are bound to the same ring, formed with either ATP, ADP or non-hydrolysable ATP analogues, suggesting that the specific requirement for ATP is confined to an action in the trans ring that evicts GroES and polypeptide from the cis side. We show here, however, that for the folding of malate dehydrogenase and Rubisco there is also an absolute requirement for ATP in the cis ring, as ADP and AMP-PNP are unable to promote folding. We investigated the specific roles of binding and hydrolysis of ATP in the cis and trans rings using mutant forms of GroEL that bind ATP but are defective in its hydrolysis. Binding of ATP and GroES in cis initiated productive folding inside a highly stable GroEL-ATP-GroES complex. To discharge GroES and polypeptide, ATP hydrolysis in the cis ring was required to form a GroEL-ADP-GroES complex with decreased stability, priming the cis complex for release by ATP binding (without hydrolysis) in the trans ring. These observations offer an explanation of why GroEL functions as a double-ring complex.


[The meadow tick Dermacentor reticulatus in natural foci of tick-borne encephalitis in Udmurtia].
In southern Udmurtia, where occur both Dermacentor reticulatus and Ixodes persulcatus, in the years of high abundance of small mammals feeding periods of larvae and nymphs of these ticks coincide completely. The duration of simultaneous parasitism of preimago of the above ixodid species on animals is of epidemiological significance as far as within this period the infection of young stages of D. reticulatus with the virus of tick-borne encephalitis occurs. This is confirmed by the isolation of I strain of the virus from D. reticulatus collected at the state of engorged nymphs on hosts common with I. persulcatus and moulted in the laboratory.


[Classification of children with specific language impairments: What can we think about them in 2016?]
Specific language impairments are characterized by a strong semiological heterogeneity. Numerous classifications have been used to bring out this heterogeneity and to create subgroups after outlining specific symptoms and deficits. However, semiological fixed profiles would not be adequate for these disorders related to neurodevelopmental trajectories. This heterogeneity is not only semiological: it is also found within underlying mechanisms, as well as in the etiopathogenesis, which is still not well understood today. The aim of this article is to introduce the main semiological classifications to highlight the fact that the symptomatic level alone is not sufficient to characterize specific language impairments.


Affinity of different local anesthetic drugs and catecholamines for the contraluminal transport system for organic cations in proximal tubules of rat kidneys.
To study the cellular uptake of [3H]tetraethylammonium (TEA) and its inhibition by different local anesthetic drugs and catecholamines, the capillary stopped flow microperfusion method was used. [3H]TEA concentration in the peritubular capillaries decreased in a time and concentration-dependent manner. This cellular uptake of [3H]TEA could be described by a facilitated diffusion model (Jmax = 0.57 +/- 0.08 pmol sec-1 cm-1, Km = 0.28 +/- 0.01 mmol/l). Between the pKa values of the local anesthetic drugs (range, 2.8-8.9) and their apparent Ki values, a significant correlation was found (r = -0.916, n = 12). In the case of catecholamines, hydrophobic substitution in the form of an alkyl chain in the alpha-position or at the nitrogen increased the inhibitory potency, whereas hydroxyl substitution in the beta-position decreased the affinity. Except for the catecholamines with complex substitutions at the benzene ring (--CH2OH,--NHCONH2,--OOCN[CH3)2, --Cl, --NH2) the apparent Ki values of the catecholamines were directly correlated with the hydrophobicity. It is concluded that hydrophobicity and pKa are two important physicochemical parameters which influence the affinity of a molecule for the transport system for organic cations at the contraluminal side of renal proximal tubules.


Microdissection of the Ah01 chromosome in upland cotton and microcloning of resistance gene anologs from the single chromosome.
Chromosome microdissection is one of the most important techniques in molecular cytogenetic research. Cotton (Gossypium Linnaeus, 1753) is the main natural fiber crop in the world. The resistance gene analog (RGA) cloning after its single chromosome microdissection can greatly promote cotton genome research and breeding. Using the linker adaptor PCR (LA-PCR) with the primers of rice disease-resistance homologues, three nucleotide sequences PS016 (KU051681), PS054 (KU051682), and PS157 (KU051680) were obtained from the chromosome Ah01 of upland cotton (cv. TM-1). The Blast results showed that the three sequences are the nucleotide binding site-leucine rich repeat (NBS-LRR) type RGAs. Clustering results indicated that they are homologous to these published RGAs. Thus, the three RGAs can definitely be confirmed as NBS-LRR class of RGAs in upland cotton. Using single chromosome microdissection technique, DNA libraries containing cotton RGAs were obtained. This technique can promote cotton gene cloning, marker development and even the improvement of cotton genome research and breeding.


Pseudoxanthoma elasticum in a patient with sickle cell disease: case report.
An 18 year female sickler (HbSS) presented with repeated history of epistaxis and bleeding gums. Features consistent with pseudoxanthoma elasticum were observed, such as hyper-extensile redundant skin folds in the neck, axilla, inguinal areas and abdomen. The skin biopsy showed swollen, clumped and fragmented elastic fibres and calcium deposits in the deep and mid reticular dermis, consistent with pseudoxanthoma elasticum. This is a well recognised complication of sickle cell disease which has not been described in Kenya.


Time to first consultation, diagnosis and treatment of TB among patients attending a referral hospital in Northwest, Ethiopia.
Early detection and treatment of TB is essential for the success of TB control program performance. The aim of this study was to determine the length and analyze predictors of patients', health systems' and total delays among patients attending a referral hospital in Bahir Dar, Ethiopia. A cross-sectional study was conducted among newly diagnosed TB cases ≥ 15 years of age. Delay was analyzed at three levels: the periods between 1) onset of TB symptoms and first visit to medical provider, i.e. patients' delay, 2) the first visit to a medical provider and the initiation of treatment i.e. health systems' delay and 3) onset of TB symptoms and initiation of treatment i.e. total delay. Uni- and multi-variate logistic regression analyses were performed to investigate predictors of patients', health systems' and total delays. The median time of patients' delay was 21 days [(interquartile range (IQR) (7 days, 60 days)]. The median health systems' delay was 27 days (IQR 8 days, 60 days) and the median total delay was 60 days (IQR 30 days, 121 days). Patients residing in rural areas had a three-fold increase in patients' delay compared to those from urban areas [Adjusted Odds Ratio (AOR) 3.4; 95% (CI 1.3, 8.9)]. Extra-pulmonary TB (EPTB) cases were more likely to experience delay in seeking treatment compared to pulmonary (PTB) cases [(AOR 2.6; 95% (CI 1.3, 5.4)]. Study subjects who first visited health centres [(AOR) 5.1; 95% (CI 2.1, 12.5)], private facilities [(AOR) 3.5; 95% (CI 1.3, 9.7] and health posts [(AOR) 109; 95% (CI 12, 958], were more likely to experience an increase in health systems' delay compared to those who visited hospitals. The majority of TB patients reported to medical providers within an acceptable time after the onset of symptoms. Rural residence was associated with patients' and total delays. Providing the population with information about TB symptoms and the importance of early health seeking may be an efficient way to decrease TB transmission, morbidity and mortality. Establishing efficient TB diagnostic and treatment facilities at the periphery level is imperative to reduce diagnostic delay and expedite TB treatment.


[A systematic review of the functional effectiveness of kinesiotaping in individuals with ankle instability].
Ankle instability often seen in people who have suffered repeated ankle sprains. Kinesio Tape is a technique widely applied in recent years, despite not having enough evidence to support its application. To determine the functional effectiveness of Kinesio Tape in individuals with ankle instability. Systematic review. A systematic search of articles was performed where Kinesio Tape was applied in subjects with ankle instability in the following databases: PubMed Central; PubMed/Medline; Cochrane; Embase; Sciencedirect; Biomed Central; CINAHL; Lippincott Williams and Wilkins; SAGE and Wiley-Blackwell. Only clinical trials were selected, in which patients with ankle instability were intervened by applying Kinesio Tape and which included at least one outcome variable associated with ankle function. 364 articles were found. Ten met the inclusion criteria; five presented high and five moderate methodological quality. In most cases, one article measured more than one variable. Two articles showed improvements in muscle strength. Of four articles that evaluated balance, two observed improvements; one article observe improvements in proprioception; two articles measured postural control, without significant improvements; one article did not observe significant improvements in neuromuscular control. Stability (one article), muscular activity (one article) and functionality (one article) did not report significant improvements with the use of Kinesio Tape. On the other hand, Kinesio tape does not report significant improvements with respect to postural control, neuromuscular control, muscle activity, joint stability and functionality in subjects suffering from ankle instability. Kinesio Tape is effective for improvement of muscle strength, balance and proprioception in subjects with ankle instability. However, Kinesio Tape does not report significant improvement of postural control, neuromuscular control, muscle activity, joint stability and functionality in people with ankle instability.


High Performance Nanofiltration Membrane for Effective Removal of Perfluoroalkyl Substances at High Water Recovery.
We demonstrate the fabrication of a loose, negatively charged nanofiltration (NF) membrane with tailored selectivity for the removal of perfluoroalkyl substances with reduced scaling potential. A selective polyamide layer was fabricated on top of a poly(ether sulfone) support via interfacial polymerization of trimesoyl chloride and a mixture of piperazine and bipiperidine. Incorporating high molecular weight bipiperidine during the interfacial polymerization enables the formation of a loose, nanoporous selective layer structure. The fabricated NF membrane possessed a negative surface charge and had a pore diameter of ∼1.2 nm, much larger than a widely used commercial NF membrane (i.e., NF270 with pore diameter of ∼0.8 nm). We evaluated the performance of the fabricated NF membrane for the rejection of different salts (i.e., NaCl, CaCl2, and Na2SO4) and perfluorooctanoic acid (PFOA). The fabricated NF membrane exhibited a high retention of PFOA (∼90%) while allowing high passage of scale-forming cations (i.e., calcium). We further performed gypsum scaling experiments to demonstrate lower scaling potential of the fabricated loose porous NF membrane compared to NF membranes having a dense selective layer under solution conditions simulating high water recovery. Our results demonstrate that properly designed NF membranes are a critical component of a high recovery NF system, which provide an efficient and sustainable solution for remediation of groundwater contaminated with perfluoroalkyl substances.


The role of magnetic resonance-guided laser ablation in neurooncology.
The use of magnetic resonance-guided laser-induced thermal therapy (MR-LITT) as a minimally invasive method of treating intra-cranial pathology is a rapidly growing field. The use of MR-LITT in neurooncology has shown promising results; however, there has been no review to date of the current literature. A review of the published literature regarding MR-LITT in neurooncology was performed. Studies on PubMed were included if at least one patient with a cerebral tumour or radiation necrosis was treated using quantitative MR thermography-guided LITT, as well as if either safety or outcomes were discussed. In treating recurrent Grade-III and -IV gliomas, we found improved median overall survival of 20.9 months from diagnosis of recurrence, which is comparable with that of 18.9 months for high-dose-rate brachytherapy and 24.4 months for repeated open surgery. Median progression-free survival (PFS) of recurrent glioma is noted to be 4.5 months. For metastatic lesions, we found a median overall survival (OS) to vary between 9.0 and 19.8 months with a PFS between 3.8 and 8.5 months. Current literature reports median OS in similar patients to lie between 7.0 and 28.6 months. Severe complication rates (with permanent deficits) are found to be between 12 and 16.7%, comparable with 11% found in literature for open surgery. The current literature shows that MR-LITT is safe and shows promising local tumour control rates. Larger randomised studies are warranted to further investigate this adjuvant therapy in the treatment of recurrent high-grade gliomas and metastases.


Role of glutathione in cell survival after hyperthermic treatment of Chinese hamster ovary cells.
Chinese hamster ovary cells cultured in vitro were exposed to hyperthermic treatment at 43 degrees C or 45 degrees C. Depletion of 99.5% of the intracellular glutathione (GSH) by addition of 0.1 mM diethylmaleate (DEM) increased thermal sensitivity. This effect, however, was dependent upon oxygen tension. Decreasing the oxygen tension reduced the effect. Thermal cytotoxicity was further augmented by continued depletion of GSH after hyperthermic treatment. Once cells became thermally tolerant, depletion of GSH did not affect thermal sensitivity. The development of thermotolerance was triggered by exposing Chinese hamster ovary cells to a thermal dose consisting of either 30 min at 43 degrees or 10 min at 45 degrees. After a 4-h incubation at 37 degrees, the cells were reheated at 43 degrees or 45 degrees, respectively. A triggering dose of 30 min at 43 degrees is a nonlethal treatment, while 10 min at 45 degrees kills 50% of the cells. Cells were exposed to 0.1 mM DEM during the triggering and development phases of tolerance. Depletion of GSH by addition of DEM did not inhibit the development of thermal tolerance when triggered by the 30-min-43 degrees thermal dose. However, exposure to DEM did inhibit the development of thermal tolerance if the triggering dose consisted of 10 min at 45 degrees. These results were interpreted to mean that GSH depletion will inhibit the development of thermotolerance only after a sufficiently toxic thermal dose.


Solar-microbial hybrid device based on oxygen-deficient niobium pentoxide anodes for sustainable hydrogen production.
Hydrogen gas is emerging as an attractive fuel with high energy density for the direction of energy resources in the future. Designing integrated devices based on a photoelectrochemical (PEC) cell and a microbial fuel cell (MFC) represents a promising strategy to produce hydrogen fuel at a low price. In this work, we demonstrate a new solar-microbial (PEC-MFC) hybrid device based on the oxygen-deficient Nb2O5 nanoporous (Nb2O5-x NPs) anodes for sustainable hydrogen generation without external bias for the first time. Owing to the improved conductivity and porous structure, the as-prepared Nb2O5-x NPs film yields a remarkable photocurrent density of 0.9 mA cm-2 at 0.6 V (vs. SCE) in 1 M KOH aqueous solution under light irradiation, and can achieve a maximum power density of 1196 mW m-2 when used as an anode in a MFC device. More importantly, a solar-microbial hybrid system by combining a PEC cell with a MFC is designed, in which the Nb2O5-x NPs electrodes function as both anodes. The as-fabricated PEC-MFC hybrid device can simultaneously realize electricity and hydrogen using organic matter and solar light at zero external bias. This novel design and attempt might provide guidance for other materials to convert and store energy.


Compendiums of cancer transcriptomes for machine learning applications.
There are massive transcriptome profiles in the form of microarray. The challenge is that they are processed using diverse platforms and preprocessing tools, requiring considerable time and informatics expertise for cross-dataset analyses. If there exists a single, integrated data source, data-reuse can be facilitated for discovery, analysis, and validation of biomarker-based clinical strategy. Here, we present merged microarray-acquired datasets (MMDs) across 11 major cancer types, curating 8,386 patient-derived tumor and tumor-free samples from 95 GEO datasets. Using machine learning algorithms, we show that diagnostic models trained from MMDs can be directly applied to RNA-seq-acquired TCGA data with high classification accuracy. Machine learning optimized MMD further aids to reveal immune landscape across various carcinomas critically needed in disease management and clinical interventions. This unified data source may serve as an excellent training or test set to apply, develop, and refine machine learning algorithms that can be tapped to better define genomic landscape of human cancers.


3K3A-activated protein C blocks amyloidogenic BACE1 pathway and improves functional outcome in mice.
3K3A-activated protein C (APC), a cell-signaling analogue of endogenous blood serine protease APC, exerts vasculoprotective, neuroprotective, and anti-inflammatory activities in rodent models of stroke, brain injury, and neurodegenerative disorders. 3K3A-APC is currently in development as a neuroprotectant in patients with ischemic stroke. Here, we report that 3K3A-APC inhibits BACE1 amyloidogenic pathway in a mouse model of Alzheimer's disease (AD). We show that a 4-mo daily treatment of 3-mo-old 5XFAD mice with murine recombinant 3K3A-APC (100 µg/kg/d i.p.) prevents development of parenchymal and cerebrovascular amyloid-β (Aβ) deposits by 40-50%, which is mediated through NFκB-dependent transcriptional inhibition of BACE1, resulting in blockade of Aβ generation in neurons overexpressing human Aβ-precursor protein. Consistent with reduced Aβ deposition, 3K3A-APC normalized hippocampus-dependent behavioral deficits and cerebral blood flow responses, improved cerebrovascular integrity, and diminished neuroinflammatory responses. Our data suggest that 3K3A-APC holds potential as an effective anti-Aβ prevention therapy for early-stage AD.


Organic dairy production systems in Pennsylvania: a case study evaluation.
The current market demand and price for organic milk is encouraging dairy producers, particularly those on smaller farms, to consider organic production as a means for improving the economic viability of their operations. Organic production systems vary widely in scale, in practices, and across agroclimatic settings. Within this context, case studies of 4 actual organic dairy farms were used to characterize existing systems in Pennsylvania. Based on data from these farms, a whole-farm simulation model (Integrated Farm System Model) was used to compare 4 production systems representing organic grass, organic crop, conventional crop with grazing, and conventional confinement production. The performance of each of these systems was simulated over each year of 25 yr of central Pennsylvania weather data. Simulation results indicated that farm level accumulation of soil P and K may be a concern on organic farms that use poultry manure as a primary crop nutrient source, and that erosion and runoff loss of P may be of concern on organic farms producing annual crops because more tillage is required for weed control. Whole-farm budgets with prices that reflect recent conditions showed an economic advantage for organic over conventional production. A sensitivity analysis showed that this economic advantage depended on a higher milk price for producers of organic milk and was influenced by the difference in milk production maintained by herds using organic and conventional systems. Factors found to have little effect on the relative profitability of organic over conventional production included the differences between organic and conventional prices for seed, chemicals, forage, and animals and the overall costs or prices assumed for organic certification, machinery, pasture fencing, fuel, and labor. Thus, at the current organic milk price, relative to other prices, the case study organic production systems seem to provide an option for improving the economic viability of dairy operations of the scale considered in Pennsylvania. To motivate transition to organic systems, the economic advantage found requires the persistence of a substantial difference between conventional and organic raw milk prices.


Photosynthetic Acclimation to Fluctuating Irradiance in Plants.
Unlike the short-term responses of photosynthesis to fluctuating irradiance, the long-term response (i.e., acclimation) at the chloroplast, leaf, and plant level has received less attention so far. The ability of plants to acclimate to irradiance fluctuations and the speed at which this acclimation occurs are potential limitations to plant growth under field conditions, and therefore this process deserves closer study. In the first section of this review, we look at the sources of natural irradiance fluctuations, their effects on short-term photosynthesis, and the interaction of these effects with circadian rhythms. This is followed by an overview of the mechanisms that are involved in acclimation to fluctuating (or changes of) irradiance. We highlight the chain of events leading to acclimation: retrograde signaling, systemic acquired acclimation (SAA), gene transcription, and changes in protein abundance. We also review how fluctuating irradiance is applied in experiments and highlight the fact that they are significantly slower than natural fluctuations in the field, although the technology to achieve realistic fluctuations exists. Finally, we review published data on the effects of growing plants under fluctuating irradiance on different plant traits, across studies, spatial scales, and species. We show that, when plants are grown under fluctuating irradiance, the chlorophyll a/b ratio and plant biomass decrease, specific leaf area increases, and photosynthetic capacity as well as root/shoot ratio are, on average, unaffected.


ETHY. A theory of fruit climacteric ethylene emission.
A theory of fruit climacteric ethylene emission was developed and used as the basis of a simulation model called ETHY. According to the theory, the biosynthetic pathway of ethylene is supplied by ATP and is regulated by 1-aminocyclopropane-1-carboxylic acid (ACC) synthase and ACC oxidase. The conjugation of ACC with malonate to form MACC was taken into account as a way to decrease the availability of ACC. Because of the seasonal increase of fruit volume, the dilution of biochemical compounds used in ETHY was taken into account. Finally, the ethylene diffusion across the skin was considered. The theory took into account the effect of temperature and O(2) and CO(2) internal concentrations on ethylene. The model was applied to peach (Prunus persica) fruit over 3 years, several leaf:fruit ratios, and irrigation conditions. An adequate ethylene increase was predicted without considering any increase in respiration during the ripening period, which suggests that the respiratory climacteric may not be required for ripening. Another important result of this study is the high sensitivity of ETHY to the parameters involved in the calculation of ACC oxidase and ACC synthase activities, ATP production, and skin surface and permeability. ETHY was also highly sensitive to changes in fruit growth and temperature.


Global issues in volatile substance misuse.
This special issue of Substance Use & Misuse addresses the public health issue of volatile substance misuse (VSM), the inhalation of gases or vapors for psychoactive effects, assessing the similarities and differences in the products misused, patterns, prevalence, etiologies, and impacts of VSM by examining it through sociocultural epidemiology, neuroscience, and interventions research. The Canadian, US, and Australian guest editors contend that, when compared with other drugs used at a similar prevalence, VSM has attracted relatively little research effort. The authors and editors call for further research to develop evidence-based policies and comprehensive interventions that respect culture and context-specific knowledge.


Is the issue of Chemsex changing?
In this letter, which is meant as a response to the letter titled "Sex enhancers: challenges, threats and the need for targeted measures", the Authors discuss the evolution of Chemsex phenomenon towards at least two directions: firstly, the use of psychoactive and non-psychoactive substances usually implicated in Chemsex, such as GHB/GBL, ketamine, mephedrone and other synthetic cathinones and erectile dysfunction medications, is currently accompanied by the use of illicit opioids, which have recently been indicated as a new serious health threat for consumers. In addition, as reported by the last European Drug report, the simultaneous use of illicit benzodiazepines with non-medical opioids misuse has also been observed. Secondly, strictly linked to the rising use of non-medical opioids is the risk of transition towards heroine followed by the adoption of risky injection practices frequently accompanied by high-risk sexual behaviors. In this sense, the current definition of the phenomenon as "the voluntary intake of certain psychoactive and non- psychoactive drugs in the context of sex parties and sexual intercourses with the intention of facilitating and/or enhancing the sexual encounter mostly among men who have sex with other men (MSM)" has been expanded to "heterosexual chemsex".


Biosynthesis in vitro of disialosylneolactotetraosylceramide by a solubilized sialyltransferase from embryonic chicken brain.
A sialyltransferase involved in the biosynthesis in vitro of LD1c (NeuAc alpha 2-8NeuGc alpha 2-3Gal beta 1-4Glc-NAc beta 1-3Gal beta 1-4Glc-Cer) has been characterized from 9 to 11-day-old embryonic chicken brains. The CMP-[14C]NeuAc:LM1(alpha 2-8)sialyltransferase (SAT-2) sedimented (75%) at the junction of 0.75 and 1.2 M on a discontinuous sucrose density gradient when still membrane bound. In addition to the biosynthesis of LD1c, the detergent-solubilized (0.4% Nonidet P-40) preparation also catalyzes the transfer of sialic acid to O-8 of sialic acid in GM3 to form GD3 (NeuAc alpha 2-8NeuAc alpha 2 - 3Gal beta 1 - 4Glc - Cer). Substrate inhibition studies indicated that these two reactions are probably catalyzed by the same enzyme, SAT-2. The kinetic parameters of SAT-2 activity were determined. The Km values were 70 and 63 microM with CMP-[14C]NeuAc and LM1, respectively, when the detergent-solubilized supernatant fraction was used as enzyme source. The (alpha 2-8)-linkage between the terminal and penultimate sialic acids was determined using nonradioactive CMP-NeuAc and [Ac-14C]LM1 as substrates (Higashi, H., and Basu, S. (1982) Anal. Biochem. 120, 159-164) for the enzyme, followed by identification of the permethylated [14C]sialic acid of the product by radioautography. At 0.5 mM N-ethylmaleimide, the SAT-2 activity was inhibited 50% whereas SAT-1 and SAT-3 activities (Basu, M., Basu, S., Stoffyn, A., and Stoffyn, P. (1982) J. Biol. Chem. 257, 12765-12769) remained uninhibited.


Selective Entrainment of Theta Oscillations in the Dorsal Stream Causally Enhances Auditory Working Memory Performance.
The implication of the dorsal stream in manipulating auditory information in working memory has been recently established. However, the oscillatory dynamics within this network and its causal relationship with behavior remain undefined. Using simultaneous MEG/EEG, we show that theta oscillations in the dorsal stream predict participants' manipulation abilities during memory retention in a task requiring the comparison of two patterns differing in temporal order. We investigated the causal relationship between brain oscillations and behavior by applying theta-rhythmic TMS combined with EEG over the MEG-identified target (left intraparietal sulcus) during the silent interval between the two stimuli. Rhythmic TMS entrained theta oscillation and boosted participants' accuracy. TMS-induced oscillatory entrainment scaled with behavioral enhancement, and both gains varied with participants' baseline abilities. These effects were not seen for a melody-comparison control task and were not observed for arrhythmic TMS. These data establish theta activity in the dorsal stream as causally related to memory manipulation. VIDEO ABSTRACT.


A systematic review and appraisal of the quality of practice guidelines for the management of Neisseria gonorrhoeae infections.
Clinical guidelines help ensure consistent care informed by current evidence. As shifts in antimicrobial resistance continue to influence first-line treatment, up-to-date guidelines are important for preventing treatment failure. A guideline's development process will influence its recommendations and users' trust. To assess the quality of current gonorrhoea guidelines' development processes. Multiple databases. Original and current English-language guidelines targeting health professionals and containing treatment recommendations for uncomplicated gonorrhoea in the general adult population. Two appraisers assessed the guidelines independently using the Appraisal of Guidelines for Research and Evaluation II (AGREE II) tool. Scores were combined as per the AGREE II users' manual. We identified 10 guidelines meeting the inclusion criteria. The quality of the gonorrhoea treatment guidelines varied. Most scored poorly on Rigour of Development; information on the evidence review process and methods for formulating recommendations was often missing. The WHO Guidelines for the Treatment of Neisseria gonorrhoeae and UK National Guideline for the Management of Gonorrhoea in Adults scored the highest on Rigour of Development. Methods to address conflicts of interest were often not described in the materials reviewed. Implementation of recommendations was often not addressed. By limiting our study to English-language guidelines, a small number of guidelines we identified were excluded. Our analysis was limited to either published or online materials that were readily available to users. We could not differentiate between items addressed in the development process but not documented from items that were not addressed. Gonorrhoea treatment guidelines may slow antimicrobial resistance. Many current guidelines are not in line with the current guideline development best practices; this might undermine the perceived trustworthiness of guidelines. By identifying current limitations, this study can help improve the quality of future guidelines.


[Protein synthesis in maize haploids and diploids during cultivation under normal conditions and in the presence of phenol].
14C-leucine incorporation in cytoplasmic protein was compared for three haploid forms, three derivative autodiploid lines and four hybrid forms of maize. It has been found that the precursor incorporation in the haploid forms was by 15-20% higher that in appropriate autodiploid lines. However, the level of protein synthesis wes practically the same for all investigated diploid forms. Thus, the increase of the ploidy level correlates with the decrease of the cytoplasmic protein synthesis. It was also shown that the protein content was practically equal in haploid and diploid forms. Proceeding from the authors' data one can conclude that the protein distraction may also have different intensity. The next step of the investigation was the study of the effects of the chemical factor of cytoplasmic protein synthesis in maize seedlings with different ploidy levels in vivo. It has been revealed, that the rate of 14C-leucine incorporation in the cytoplasmic proteins differ for the haploid forms which grew on the Knop media with the addition of phenol (0.05%) and without pollutant. Nevertheless, approximately equal level of 14C-leucine incorporation was observed for autodiploids. Recently it was demonstrated that the synthetic intensity of different RNA fractions is higher in the haploid maize forms than in the diploid ones and that the haploid and diploid forms of maize display the different reaction on phenol pollution. When comparing our data with the published results it is possible to conclude that the hypothesis about the low intensity of metabolism in haploid plants, existing in literature, is not true. The authors suppose that the high activity of synthesis of nuclear, transfer and ribosomal RNA and cytoplasmic proteins in the haploid plants may be connected with high mitotic activity. The authors' data are in agreement with the hypothesis that the ratio of nuclear volume and surface determines the metabolism rate.


The ghost of hosts past: impacts of host extinction on parasite specificity.
A growing body of research is focused on the extinction of parasite species in response to host endangerment and declines. Beyond the loss of parasite species richness, host extinction can impact apparent parasite host specificity, as measured by host richness or the phylogenetic distances among hosts. Such impacts on the distribution of parasites across the host phylogeny can have knock-on effects that may reshape the adaptation of both hosts and parasites, ultimately shifting the evolutionary landscape underlying the potential for emergence and the evolution of virulence across hosts. Here, we examine how the reshaping of host phylogenies through extinction may impact the host specificity of parasites, and offer examples from historical extinctions, present-day endangerment, and future projections of biodiversity loss. We suggest that an improved understanding of the impact of host extinction on contemporary host-parasite interactions may shed light on core aspects of disease ecology, including comparative studies of host specificity, virulence evolution in multi-host parasite systems, and future trajectories for host and parasite biodiversity. This article is part of the theme issue 'Infectious disease macroecology: parasite diversity and dynamics across the globe'.


Assessment of quality of life in children with peanut allergy.
Children with a peanut allergy (PA) are faced with food and social restrictions due to the potentially life-threatening nature of their disease, for which there is no cure or treatment. This inevitably impacts upon their quality of life (QoL). QoL of 20 children with PA and 20 children with insulin-dependent diabetes mellitus (IDDM) was measured using two disease-specific QoL questionnaires (higher scores correspond to a poorer QoL). One questionnaire was designed by us and the other was adapted from the Vespid Allergy QoL questionnaire. We gave subjects cameras to record how their QoL is affected over a 24-h period. Response rates for both questionnaires were 100%. Mean ages were 9.0 and 10.4 years for PA and IDDM subjects, respectively. Children with a PA reported a poorer quality of life than children with IDDM: mean scores were 54.85 for PA subjects and 46.40 for diabetics (p = 0.004) in questionnaire 1 and 54.30 and 34.50 (p</=0.001) in questionnaire 2. PA children reported more fear of an adverse event and more anxiety about eating, especially when eating away from home. Photographs fell into seven common categories: food, management, environment, away from home, physical activities, restaurant and people. Most photographs related to food and management issues and revealed difficulties for both groups regarding food restrictions. PA subjects felt more threatened by potential hazards within their environment, felt more restricted by their PA regarding physical activities, and worried more about being away from home. However, they felt safe when carrying epinephrine kits and were positive about eating at familiar restaurants. The QoL in children with PA is more impaired than in children with IDDM. Their anxiety may be considered useful in some situations, promoting better adherence to allergen avoidance advice and rescue plans.


Optimization of weaning strategy in the climbing perch (Anabas testudineus, Bloch 1792) larvae on growth, survival, digestive, metabolic and stress responses.
A 30-day experiment was carried out to know responses of different weaning approaches to the growth and survival of Anabas testudineus larvae. A total of 10800 larvae (Avg. weight 0.016 ± 0.03 mg; 3DPH) were randomly distributed in nine treatments (triplicates), including two controls. The strategies are as follows: C1 (Control I): feeding with live food (LF) for 30 days and C2 (Control II): feeding with microparticulate diet (MPD) for 30 days; T1: LF for 5 days and MPD for next 25 days; T2: LF for 10 days and MPD for next 20 days; T3: LF for 15 days and MPD for next 15 day; T4: LF for 20 days and MPD for next 10 days; T5: LF for 25 days and MPD for next 5 days; T6: LF for 5 days, then 25% LF replacement by MPD for next 5 days, 50% LF replacement by MPD for next 5 days, 75% LF replacement by MPD for next 5 days, and 100% LF replacement by MPD for last 10 days; and T7: LF for 10 days, then 25% LF replacement by MPD for next 5 days, 50% LF replacement by MPD for next 5 days, 75% LF replacement by MPD for next 5 days, and 100% LF replacement by MPD for last 5 days. Significantly (p < 0.05) higher WG and SGR were recorded in T2 (213.17 ± 0.32, 23.98 ± 0.02) followed by T6, whereas the lowest was found in C2. Significantly higher (p < 0.05) percentage survival was manifested in the T7 (31.83 ± 0.22), followed by T2 (24.75 ± 0.13), and the lowest survival was observed in the C2. The digestive enzyme activities were found to be non-significant (p > 0.05) between different treatment groups. The alanine aminotransferase (ALT), aspartate aminotransferase (AST), lactate dehydrogenase (LDH), and malate dehydrogenase (MDH) were reported to be significantly higher (p < 0.05) in C2 (68.52 ± 0.08, 19.55 ± 0.10, 21.79 ± 0.04, and 0.044 ± 0.01) followed by T1; however, their reduced level was observed in C1. The activity of superoxide dismutase (SOD), catalase (CAT), glucose, and cortisol levels was observed significantly (p < 0.05) higher in C2 and lower in C1 and T2. As per the finding, it can be recommended that the appropriate weaning time for A. testudineus larvae is from 13 DPH onwards, in which larvae can be fed an initial ten days LF afterward MPD and the best weaning strategy can be adopted as in the T7 group for higher survival percentage.


Performance of three severely mentally retarded adults on vocational response chains during extinction.
The intrachain performance patterns of three severely retarded adults were analyzed under continuous reinforcement and extinction schedules. The three subjects performed a nine-step operant chain that required the placement of eight electrical components in a circuit board blank. The time to perform the entire chain and the time taken at each step of the chain were electronically monitored. A combined multiple-baseline, ABA reversal design was used. Reinforcers were delivered following completion of the chain during A phases and withheld during the B phase. Data indicated that overall chain performance was slower during extinction, and this effect was due primarily to reduced response speed on the early steps in the chain. Implications of these data for vocational habilitation of retarded adults were discussed.


The landscape of Chlamydomonas histone H3 lysine 4 methylation reveals both constant features and dynamic changes during the diurnal cycle.
Chromatin modifications are epigenetic regulatory features with major roles in various cellular events, yet they remain understudied in algae. We interrogated the genome-wide distribution pattern of mono- and trimethylated histone H3 lysine 4 (H3K4) using chromatin-immunoprecipitation followed by deep-sequencing (ChIP-seq) during key phases of the Chlamydomonas cell cycle: early G1 phase, Zeitgeber Time 1 (ZT1), when cells initiate biomass accumulation, S/M phase (ZT13) when cells are replicating DNA and undergoing mitosis, and late G0 phase (ZT23) when they are quiescent. Tri-methylated H3K4 was predominantly enriched at transcription start sites of the majority of protein coding genes (85%). The likelihood of a gene being marked by H3K4me3 correlated with it being transcribed at some point during the life cycle but not necessarily by continuous active transcription, as exemplified by early zygotic genes, which may remain transcriptionally dormant for thousands of generations between sexual cycles. The exceptions to this rule were around 120 loci, some of which encode non-poly-adenylated transcripts, such as small nuclear RNAs and replication-dependent histones that had H3K4me3 peaks only when they were being transcribed. Mono-methylated H3K4 was the default state for the vast majority of histones that were bound outside of transcription start sites and terminator regions of genes. A small fraction of the genome that was depleted of any H3 lysine 4 methylation was enriched for DNA cytosine methylation and the genes within these DNA methylation islands were poorly expressed. Besides marking protein coding genes, H3K4me3 ChIP-seq data served also as a annotation tool for validation of hundreds of long non-coding RNA genes.


Design of Electrostatic Aberration Correctors for Scanning Transmission Electron Microscopy.
In a scanning transmission electron microscope (STEM), producing a high-resolution image generally requires an electron beam focused to the smallest point possible. However, the magnetic lenses used to focus the beam are unavoidably imperfect, introducing aberrations that limit resolution. Modern STEMs overcome this by using hardware aberration correctors comprised of many multipole elements, but these devices are complex, expensive, and can be difficult to tune. We demonstrate a design for an electrostatic phase plate that can act as an aberration corrector. The corrector is comprised of annular segments, each of which is an independent two-terminal device that can apply a constant or ramped phase shift to a portion of the electron beam. We show the improvement in image resolution using an electrostatic corrector. Engineering criteria impose that much of the beam within the probe-forming aperture be blocked by support bars, leading to large probe tails for the corrected probe that sample the specimen beyond the central lobe. We also show how this device can be used to create other STEM beam profiles such as vortex beams and probes with a high degree of phase diversity, which improve information transfer in ptychographic reconstructions.


Neuroprotective effect of menaquinone-4 (MK-4) on transient global cerebral ischemia/reperfusion injury in rat.
Cerebral ischemia/reperfusion (I/R) injury causes cognitive deficits, excitotoxicity, neuroinflammation, oxidative stress and brain edema. Vitamin K2 (Menaquinone 4, MK-4) as a potent antioxidant can be a good candidate to ameliorate I/R consequences. This study focused on the neuroprotective effects of MK-4 for cerebral I/R insult in rat's hippocampus. The rat model of cerebral I/R was generated by transient bilateral common carotid artery occlusion for 20 min. Rats were divided into control, I/R, I/R+DMSO (solvent (1% v/v)) and I/R+MK-4 treated (400 mg/kg, i.p.) groups. Twenty-four hours after I/R injury induction, total brain water content, superoxide dismutase (SOD) activity, nitrate/nitrite concentration and neuronal density were evaluated. In addition to quantify the apoptosis processes, TUNEL staining, as well as expression level of Bax and Bcl2, were assessed. To evaluate astrogliosis and induced neurotoxicity by I/R GFAP and GLT-1 mRNA expression level were quantified. Furthermore, pro-inflammatory cytokines including IL-1β, IL-6 and TNF-α were measured. Seven days post I/R, behavioral analysis to quantify cognitive function, as well as Nissl staining for surviving neuronal evaluation, were conducted. The findings indicated that administration of MK-4 following I/R injury improved anxiety-like behavior, short term and spatial learning and memory impairment induced by I/R. Also, MK-4 was able to diminish the increased total brain water content, apoptotic cell density, Bax/ Bcl2 ratio and GFAP mRNA expression following I/R. In addition, the high level of nitrate/nitrite, IL-6, IL-1β and TNF-α induced by I/R was reduced after MK-4 administration. However, MK-4 promotes the level of SOD activity and GLT-1 mRNA expression in I/R rat model. The findings demonstrated that MK-4 can rescue transient global cerebral I/R consequences via its anti-inflammatory and anti-oxidative stress features. MK-4 administration ameliorates neuroinflammation, neurotoxicity and neuronal cell death processes and leads to neuroprotection.


The use of otolith chemistry to characterize diadromous migrations.
Chemical constituents in otoliths have become a valuable tool for fish ecologists seeking to reconstruct migratory patterns and life-history diversity in a wide range of species worldwide. This approach has proved particularly effective with fishes that move across substantial salinity gradients over the course of their life, including many diadromous species. Freshwater endmembers of several elemental and isotope ratios (e.g. Sr:Ca, Ba:Ca and (87)Sr:(86)Sr) are typically identifiably distinct from marine values, and often differ among freshwater tributaries at fine spatial scales. Because these chemical tags are generally incorporated in proportion to their ambient dissolved concentrations, they can be effective proxies for quantifying the presence, duration and frequency of movements between freshwater and marine habitats. The development of high precision probe-based analytical techniques, such as laser ablation inductively coupled plasma mass spectrometry (ICP-MS) and microbeam methods, has allowed researchers to glean increasingly detailed life-history profiles of these proxies across otoliths. Researchers are also combining multiple chemical proxies in an attempt to refine interpretations of habitat residence patterns. A thorough understanding of the spatial and temporal variation in water chemistry as well as environmental and physiological controls on incorporation of specific elements into otoliths is required for confident estimation of lifetime salinity experience. First some assumptions, methodological considerations and data processing options that are particularly relevant to diadromous otolith chemistry studies are discussed. Insights into diadromous migrations obtained from decades of otolith chemistry research, highlighting the increasingly recognized importance of contingent behaviour and partial migration are then discussed. Finally, areas for future research and the need to integrate otolith chemistry studies into comprehensive assessments of the effects of global environmental change are identified.


Genetic Heterogeneity of Beta Globin Mutations among Asian-Indians and Importance in Genetic Counselling and Diagnosis.
There are an estimated 45 million carriers of β-thalassemia trait and about 12,000-15,000 infants with β-thalassemia major are born every year in India. Thalassemia major constitutes a significant burden on the health care system. The burden of thalassemia major can be decreased by premarital screening and prenatal diagnosis. The success of prenatal diagnosis requires proper knowledge of spectrum of β-thalassemia mutations. In present study, β-thalassemia mutations were characterized in 300 thalassemia cases from 2007 to 2010 using ARMS-PCR and DNA sequencing. The five most common mutations accounted 78.9% of the studied chromosomes that includes IVS1-5(G>C), Cod 41-42(-TCTT), Cod8-9(+G), Cod16(-C) and 619bp del. Though IVS1-5(G>C) is most common mutation in all the communities, the percentage prevalence were calculated on sub caste basis and found that IVS1-5(G>C) percentage prevalence varied from 25 to 60 in Aroras & Khatris and Thakur respectively. Interestingly Cod41-42(-TCTT) mutation which is the second commonest among the mutations reported was totally absent in Kayasthas and Muslim community. These findings have implications for providing molecular diagnosis, genetic counseling and prenatal diagnosis to high risk couples of β-thalassemia.


Specific high-affinity binding and biologic action of retinoic acid in human neuroblastoma cell lines.
Neuroblastoma cells are a good model for neuronal development because of their ability to extend neurites in response to various stimuli, including retinoic acid. In the present experiments, we have examined five human neuroblastoma cell lines (LA-N-1, IMR-32, LA-N-5, SK-N-MC, and CHP-100) for the presence of cellular retinoic acid binding protein (CRABP), a receptor-like protein implicated in the molecular functioning of vitamin A. CRABP is identified and quantitated by sucrose gradient centrifugation, selective inhibition by the mercurial reagent p-chloromercuribenzene sulfonic acid (PCMBS), and saturation analysis. All five lines contain significant levels of cytosolic CRABP (2.5-7.5 pmol/mg of protein), which display typical properties of specific high affinity retinoic acid binding, a sedimentation coefficient of 2 S, and inhibition by PCMBS. Three of the lines (LA-N-1, IMR-32, and LA-N-5) are strongly growth inhibited by 1 microM retinoic acid in monolayer culture, whereas two (LA-N-1 and LA-N-5) undergo marked differentiation to a stellate, fusiform morphology with characteristic neurite outgrowths. The SK-N-MC and CHP-100 lines are relatively resistant to the antiproliferative effects of retinoic acid under these conditions. Nevertheless, all five lines are effectively inhibited by retinoic acid in their ability to form anchorage-independent colonies in soft agar. Thus, although CRABP is not necessarily correlated with growth inhibition in monolayer culture, it is associated with retinoic acid's ability to inhibit neuroblastoma colony formation in soft agar. More experiments will be required to determine if this effect on growth in soft agar reflects the putative ability of retinoic acid to convert tumorigenic neuroblastoma cell lines into the normal differentiated phenotype.


Phase-Field Simulation of a Dynamic Protective Layer for the Inhibition of Dendrite Growth in Zinc Metal Batteries.
Metallic zinc (Zn) has been considered one of the most promising anode materials for next-generation aqueous Zn batteries due to its low redox potential and high storage capacity. However, excessive dendrite formation in Zn metal, corrosion, the evolution of hydrogen gas during the cycling process, and the poor Zn-ion (Zn2+) transport from the electrolyte to the electrode limit its practical application. One of the most effective strategies to suppress Zn dendrite growth and promote Zn2+ transport is to introduce suitable protective layers between the Zn metal electrode and the electrolyte. Herein, we mathematically simulated the dynamic interactions between the Zn deposition on the anode and the resulting displacement of a protective layer that covers the anode, the latter of which can simultaneously inhibit Zn dendrite growth and enhance the Zn2+ transport through the interface between the Zn anode and the protective layer. Our simulation results indicate that a protective layer of high Zn2+ diffusivity not only improves the deposition rate of the Zn metal but also prevents dendrite growth by homogenizing the Zn2+ concentration at the anode surface. In addition, it is revealed that the anisotropic Zn2+ diffusivity in the protective layer influences the 2D diffusion of Zn2+. Higher Zn2+ diffusivity perpendicular to the Zn metal surface inhibits dendrite growth, while higher diffusivity parallel to the Zn metal surface promotes dendrite growth. Our work thus provides a fundamental understanding and a design principle for controlling anisotropic Zn2+ diffusion in the protective layer for better suppression of dendrite growth in Zn metal batteries.


Comparative study of the ultrastructure and secretory dynamic of hypopharyngeal glands in queens, workers and males of Scaptotrigona postica Latreille (Hymenoptera, Apinae, Meliponini).
The secretory cycle of hypopharyngeal glands (HPGs) in Scaptotrigona postica resembles that of Apis mellifera: in newly emerged workers the HPGs are in prefunctional state, their maximum development happens in the nurse workers and in forager workers they show signs of reabsorption. In S. postica these glands are also present in queens and males where they are more developed in newly emerged individuals. The ultrastructural features of the HPG secretory cycle in workers of S. postica and A. mellifera are alike: granular endoplasmic reticulum well developed, large secretion masses around the intracellular canaliculus in nurse workers and extensive degenerative structures in forager workers. Then it is suggested that the HPG secrete similar substances in both species. A second secretory cycle seems to occur in early foragers, may be with production of enzymes. The role of the HPGs in queens and males remains unknown but one possibility is enzyme production.


Dietary fermented soybean suppresses UVB-induced skin inflammation in hairless mice via regulation of the MAPK signaling pathway.
Soybean may be a promising ingredient for regulating UVB-induced inflammatory damage to the skin. We investigated the anti-inflammatory effects of diets supplemented with fermented soybean on UVB-induced skin photodamage and the effectiveness of soybean (S) and fermented soybean (FS) dietary supplementation. To investigate the effects of two major isoflavones-daidzein and genistein-from FS, we used cocultures with keratinocytes and fibroblasts. Genistein treatment strongly inhibited the production of IL-6 and MAPK signaling. Forty hairless male mice divided into four groups were fed with a control diet (group N: normal, group C; +UVB) or diets with 2.5% S+UVB or 2.5% FS+UVB (group S, group FS) for 8 weeks. Macrophage infiltration to the dermis was reduced more in groups S and FS than in group C. The expression levels of iNOS and COX-2 were significantly decreased in group FS (by 7.7% ± 0.4% and 21.2% ± 0.3%, respectively [p < 0.05]).


Efficacy of topical permethrin as repellent against Aedes aegypti's bites.
Mosquitoes are the most important vectors of infectious diseases and their bites are related to several adverse skin reactions. Permethrin impregnated clothes are an efficient strategy against arthropods' bites; however, its topical efficacy as a repellent has not been well established. We studied the response to permethrin lotion 5 percent and N,N-Diethyl-meta-toluamide (DEET) spray 50 percent applied to the unprotected forearms of 10 volunteers. Each arm was exposed to 20 female mosquitoes of Aedes aegypti. We performed 71 bilateral comparative measurements evaluating the timing for the first bites. The average times for the arm without the product, with permethrin 5 percent, and with DEET 50 percent were: 7.9 seconds, 336.2 seconds and 7512.1 seconds. The results showed a significant difference between repellency times between either product and unprotected controls. In addition, there was a significant difference in time to first bite between permethrin and DEET treated arms (p<0.01). Permethrin affords some repellent activity against Aedes aegypti bites in this experimental setting. However, permethrin's profile of repellency was significantly inferior to that of DEET.


IgG3 production in MRL/lpr mice is responsible for development of lupus nephritis.
MRL/Mp-lpr/lpr (MRL/lpr) mice spontaneously develop lethal glomerulonephritis (GN) similar to human lupus nephritis, associated with the expression of lymphoproliferation gene lpr. To examine whether a particular IgG subclass is responsible for development of GN in these mice, first quantitative analysis of IgG subclasses in serum and in kidney eluates was performed. Although IgG2a was the dominant subclass in serum throughout the lifespan of mice, the IgG3 level in kidney eluates was three times higher than that of IgG2a at the 16 wk of age, which is the time of onset of development of severe GN. In sera of the 12-wk-old mice, half of the IgG3 was in immune complex form, whereas IgG2a in this form was only 17% of the total amount. Second, cyclosporin A, which ameliorates GN in MRL/lpr mice despite autoantibody production, was found to reduce serum IgG3 and mRNA levels, associated with the revision of cationic shift of the serum IgG3 spectrotype seen in isoelectric focusing. Third, among the hybrid mice with non-autoimmune-prone C3H/HeJ-lpr/lpr (C3H/lpr) mice, MRL/lpr x (MRL/lpr x C3H/lpr) F1, in which the genetic background for GN is likely segregated, the mRNA level for IgG3 correlated well with the degree of glomerular lesion. These findings indicate that production of IgG3 in MRL/lpr mice is one of the major factors responsible for development of GN in these mice, and that this is due to the genetic background of the MRL strain.