Questioning supports effective transmission of knowledge and increased exploratory learning in pre-kindergarten children.
How can education optimize transmission of knowledge while also fostering further learning? Focusing on children at the cusp of formal schooling (N = 180, age = 4.0-6.0 y), we investigate learning after direct instruction by a knowledgeable teacher, after questioning by a knowledgeable teacher, and after questioning by a naïve informant. Consistent with previous findings, instruction by a knowledgeable teacher allows effective information transmission but at the cost of exploration and further learning. Critically, we find a dual benefit for questioning by a knowledgeable teacher: Such pedagogical questioning both effectively transmits knowledge and fosters exploration and further learning, regardless of whether the question was directed to the child or directed to a third party and overheard by the child. These effects are not observed when the same question is asked by a naïve informant. We conclude that a teacher's choice of pedagogical method may differentially influence learning through their choices of how, and how not, to present evidence, with implications for transmission of knowledge and self-directed discovery. A video abstract of this article can be viewed at: https://www.youtube.com/watch?v=FJXH2b65wL8.


Type III secretion system and virulence markers highlight similarities and differences between human- and plant-associated pseudomonads related to Pseudomonas fluorescens and P. putida.
Pseudomonas fluorescens is commonly considered a saprophytic rhizobacterium devoid of pathogenic potential. Nevertheless, the recurrent isolation of strains from clinical human cases could indicate the emergence of novel strains originating from the rhizosphere reservoir, which could be particularly resistant to the immune system and clinical treatment. The importance of type three secretion systems (T3SSs) in the related Pseudomonas aeruginosa nosocomial species and the occurrence of this secretion system in plant-associated P. fluorescens raise the question of whether clinical isolates may also harbor T3SSs. In this study, isolates associated with clinical infections and identified in hospitals as belonging to P. fluorescens were compared with fluorescent pseudomonads harboring T3SSs isolated from plants. Bacterial isolates were tested for (i) their genetic relationships based on their 16S rRNA phylogeny, (ii) the presence of T3SS genes by PCR, and (iii) their infectious potential on animals and plants under environmental or physiological temperature conditions. Two groups of bacteria were delineated among the clinical isolates. The first group encompassed thermotolerant (41°C) isolates from patients suffering from blood infections; these isolates were finally found to not belong to P. fluorescens but were closely related and harbored highly conserved T3SS genes belonging to the Ysc-T3SS family, like the T3SSs from P. aeruginosa. The second group encompassed isolates from patients suffering from cystic fibrosis; these isolates belonged to P. fluorescens and harbored T3SS genes belonging to the Hrp1-T3SS family found commonly in plant-associated P. fluorescens.


Deficiency of 5'-deoxy-5'-methylthioadenosine phosphorylase activity in malignancy. Absence of the protein in human enzyme-deficient cell lines.
The absence of 5'-deoxy-5'-methylthioadenosine phosphorylase (MTAase) activity in malignant cells, and the putative localization of its gene, suggest that this enzyme deficiency might be due to a genomic alteration also involving a tumour-suppressor gene. We studied the possible occurrence of inactive forms of the protein in two MTAase-negative cell lines, namely K562 and Jurkat, by immunochemical methods. Two highly specific antisera, directed against different epitopes of the phosphorylase [Della Ragione, Oliva, Gragnaniello, Russo, Palumbo & Zappia (1990) J. Biol. Chem. 265, 6241-6246], were used to carry out immunotitration and immunoblotting analyses, as well as to investigate the biosynthesis of the enzyme. No MTAase protein was detected by Western-blotting technique performed under conditions where all the phosphorylase-positive samples gave a clear band at the MTAase subunit molecular mass. No cross-reacting material was observed by a sensitive immunotitration method which permitted the detection of as low as 0.5 ng of protein. Moreover, the results obtained by [35S]methionine-labelling experiments ruled out phosphorylase biosynthesis in the negative cell lines. Altogether, these data suggest that an alteration at the gene level hampering the specific mRNA biosynthesis or resulting in an untranslatable mRNA is the cause of the enzyme deficiency in the MTAase-negative cell lines studied.


3D modelling of gene expression patterns.
The current genome-sequencing projects provide "word indices" of the book of life. A central post-genomic question will be how these words are three-dimensionally deployed in the generation of organism form. Gene expression studies of developing organisms contribute an increasing wealth of snapshot data on the activation of individual genes at selected locations and single moments in the developmental process. However, a comprehensive understanding of the dynamic activation of multiple genes and their functional role in controlling the 3D processes of collective cell behaviour, pattern formation and morphogenesis, requires special tools for a systematic description of spatio-temporal patterns of gene activation and the ensuing phenotypic effects. This article concentrates on new, computer-based tools for the 3D analysis of gene expression patterns in embryonic development and their use for the systematic establishment of comprehensive gene expression maps.


Correlation of single-mode fiber radiation response and fabrication parameters.
Statistically significant correlations have been established between certain fabrication parameters of matched clad, single-mode optical fiber waveguides and their response to an ionizing radiation dose of 2000 rad. The reCOVE:ry data measured at -35 degrees C following exposure have been fit to nth-order kinetic behavior where the adjustable parameters are the initial and permanent incremental losses (A(o) and A(f), respectively), the half-life of attenuation tau, and the order of kinetics n. The set of fibers chosen for analysis had Ge-doped silica cores. In fibers with Ge-F-doped silica clads, A(o) correlates with the concentration of Ge-doped into the fiber core; A(f) correlates with the ratio of oxygen to reagents used during core deposition; and tau and n correlate with a two-way interaction of core oxygen and fiber draw speed. In P-F-doped clad fibers, the P concentration has been found to correlate with the order of the kinetics of recovery.


Incidence of bleeding events in patients on concomitant tyrosine kinase inhibitors and selective serotonin reuptake inhibitors.
In this study, we aim to determine the risk of bleeding or thrombosis with concurrent use of tyrosine kinase inhibitors (TKIs) used to treat CML, and serotonin reuptake inhibitors (SSRIs). We conducted a retrospective cohort study of patients with CP-CML cared for at Massachusetts General Hospital (MGH) between April 2016 to February 2021. Participants were included if diagnosed with CP-CML and began TKI treatment (imatinib, dasatinib, nilotinib, bosutinib, or ponatinib) after April 2016. One hundred patients were evaluated, eighty of whom were taking TKIs only (median age 55, 40% female), and twenty were taking TKI and SSRI concomitantly (median age 53.5, 55% female). Baseline demographics between these groups were similar across all variables. Patients in the TKI only group had 9 bleeding events and 3 thrombotic events. Patients in the combination group had 6 bleeding events and 1 thrombotic event. There was no difference between overall rates of major bleeding (4% v. 10%, p = 0.26) or thrombotic events (4% v. 5%, p = 1). However, patients in the combination group were more likely to have major intracranial bleeding events (0% v. 10%, p = 0.04), and there was a trend to significance for minor bleeding events (7.5% v. 20%, p = 0.11). Concomitant use of TKIs and SSRIs does not appear to increase the total risk of bleeding or thrombotic events compared to patients on TKIs only. However, concomitant use of TKIs and SSRIs may increase risk of intracranial bleeding. Further work is needed to fully assess this risk.


[Isolation and characteristics of endonuclease tightly bound to alpha-polymerase from the rat liver].
Three major polypeptides are found in purified DNA polymerase alpha from rat liver: 160, 77 and 58 kDa. The electrophoretic analysis has identified polypeptide 160 kDa as the catalytically active subunit of DNA polymerase alpha. The other two polypeptides showed no DNA polymerase activity. Individual polypeptide p77 kDa purified by sodium dodecyl sulfate polyacrylamide gel electrophoresis was used to produce antibodies in rabbits. Immunoblot analysis indicated that the complex DNA polymerase alpha-3'-5'-exonuclease contained polypeptide p77 kDa. To elucidate the function of the p77 kDa protein we have prepared an immunoabsorbent column with antibodies against the p77 kDa polypeptide. The antibody column purified p77 kDa protein was homogeneous according to sodium dodecyl sulfate gel electrophoresis. The activity of alpha-polymerase was increased approximately 10-fold as a result of purification of DNA polymerase alpha from the p77 kDa protein. The in vitro experiments showed the identity of the p77 kDa polypeptide to endonuclease. It cleaved both single-stranded and double-stranded DNA. The function of endonuclease p77 kDA in complex with DNA polymerase alpha remains obscure.


Flow cytometric evaluation of proliferative activity and ploidy in myelodysplastic syndromes and acute leukemias.
Propidium iodide (PI) DNA distribution of bone marrow (BM) cells was studied by flow cytometry (FCM) in 36 patients without hematologic or malignant disease (normal BM) and in 172 patients with anemias (36 pts), myelodysplastic syndromes (MDS) (33 pts) and acute leukemia (AL) at diagnosis (60 pts), remission (24 pts) and relapse (19 pts). White blood cells from normal male subjects were used as an external diploid reference standard (median CV = 3.8). Patients with normal BM, anemias, MDS and acute leukemia at diagnosis had tritiated thymidine labeling index (LI) and most with MDS and AL had also evaluable cytogenetics performed on the same BM sample used for FCM. In normal BM, median aliquot of cells with PI-DNA content intermediate between the diploid and the tetraploid value (2n-4n cells %) was 15.7. The ratio between the fluorescence intensity of the G0/1 peak of normal BM cells and the fluorescence intensity of the G0/1 peak of the reference standard (FI ratio) ranged from 93 to 1.05 (mean +/- 2SD). The 2n-4n cell % was higher than normal in anemias (p less than .001), lower in leukemias (p less than .001) and widely scattered in MDS. A linear correlation was found between 2n-4n cell % and LI, with 2n-4n cell % value higher than LI. The FI ratio was lower than normal in anemias (p less than .05), higher in AL with normal cytogenetics (p less than .02) and broadly scattered in MDS with normal cytogenetics. From our experience, PI-DNA-FCM is a simple and adequate method to evaluate proliferative activity in hematologic diseases. Nevertheless, caution must be taken in attributing small changes in FI ratio to aneuploidy, since they are found in anemias and in MDS and AL with normal cytogenetics, possibly due to differences in PI uptake by different cell types.


Spiroplasmas: cultivation in chemically defined medium.
A chemically defined medium, CC-494, supports the cultivation in vitro of several spiroplasmas belonging to three distinct serogroups. Medium CC-494 supports the growth of flower spiroplasmas FS 23-6 and FS SR-3 and of honey bee spiroplasma HBS AS-576. The maximal populations of the two flower spiroplasmas and honey bee spiroplasma are comparable to those cultured in the undefined medium C-3G. The growth rate for all three spiroplasmas is slower in the defined medium.


Aortic morphometry and microcephaly in hypoplastic left heart syndrome.
Microcephaly is a marker of abnormal fetal cerebral development, and a known risk factor for cognitive dysfunction. Patients with hypoplastic left heart syndrome have been found to have an increased incidence of abnormal neurodevelopmental outcomes. We hypothesized that reduced cerebral blood flow from the diminutive ascending aorta and transverse aortic arch in the setting of hypoplastic left heart syndrome may influence fetal growth of the brain. The purpose of our study, therefore, was to define the prevalence of microcephaly in full-term infants with hypoplastic left heart syndrome, and to investigate potential cardiac risk factors for microcephaly. We carried out a retrospective review of full-term neonates with hypoplastic left heart syndrome. Eligible patients had documented indexes of birth weight, and measurements of length, and head circumference, as well as adequate echocardiographic images for measurement of the diameters of the ascending aorta and transverse aortic arch. We used logistic regression for analysis of the data. A total of 129 neonates met the criteria for inclusion, with 15 (12%) proving to have microcephaly. The sizes of their heads were disproportionately smaller than their weights (p less than 0.001) and lengths (p less than 0.001) at birth. Microcephaly was associated with lower birth weight (p less than 0.001), lower birth length (p equal to 0.007), and a smaller diameter of the ascending aorta (p equal to 0.034), but not a smaller transverse aortic arch (p equal to 0.619), or aortic atresia (p equal to 0.969). We conclude that microcephaly was common in this cohort of neonates with hypoplastic left heart syndrome, with the size of the head being disproportionately smaller than weight and length at birth. Microcephaly was associated with a small ascending aorta, but not a small transverse aortic arch. Impairment of somatic growth may be an additional factor in the development of microcephaly in these neonates.


Evaluating the Performance of Different Large Language Models on Health Consultation and Patient Education in Urolithiasis.
To evaluate the effectiveness of four large language models (LLMs) (Claude, Bard, ChatGPT4, and New Bing) that have large user bases and significant social attention, in the context of medical consultation and patient education in urolithiasis. In this study, we developed a questionnaire consisting of 21 questions and 2 clinical scenarios related to urolithiasis. Subsequently, clinical consultations were simulated for each of the four models to assess their responses to the questions. Urolithiasis experts then evaluated the model responses in terms of accuracy, comprehensiveness, ease of understanding, human care, and clinical case analysis ability based on a predesigned 5-point Likert scale. Visualization and statistical analyses were then employed to compare the four models and evaluate their performance. All models yielded satisfying performance, except for Bard, who failed to provide a valid response to Question 13. Claude consistently scored the highest in all dimensions compared with the other three models. ChatGPT4 ranked second in accuracy, with a relatively stable output across multiple tests, but shortcomings were observed in empathy and human caring. Bard exhibited the lowest accuracy and overall performance. Claude and ChatGPT4 both had a high capacity to analyze clinical cases of urolithiasis. Overall, Claude emerged as the best performer in urolithiasis consultations and education. Claude demonstrated superior performance compared with the other three in urolithiasis consultation and education. This study highlights the remarkable potential of LLMs in medical health consultations and patient education, although professional review, further evaluation, and modifications are still required.


Towards an understanding of the enzymatic degradation of complex plant mannan structures.
Plant cell walls are composed of a heterogeneous mixture of polysaccharides that require several different enzymes to degrade. These enzymes are important for a variety of biotechnological processes, from biofuel production to food processing. Several classical mannanolytic enzyme functions of glycoside hydrolases (GH), such as β-mannanase, β-mannosidase and α-galactosidase activities, are helpful for efficient mannan hydrolysis. In this light, we bring three enzymes into the model of mannan degradation that have received little or no attention. By linking their three-dimensional structures and substrate specificities, we have predicted the interactions and cooperativity of these novel enzymes with classical mannanolytic enzymes for efficient mannan hydrolysis. The novel exo-β-1,4-mannobiohydrolases are indispensable for the production of mannobiose from the terminal ends of mannans, this product being the preferred product for short-chain mannooligosaccharides (MOS)-specific β-mannosidases. Second, the side-chain cleaving enzymes, acetyl mannan esterases (AcME), remove acetyl decorations on mannan that would have hindered backbone cleaving enzymes, while the backbone cleaving enzymes liberate MOS, which are preferred substrates of the debranching and sidechain cleaving enzymes. The nonhydrolytic expansins and swollenins disrupt the crystalline regions of the biomass, improving their accessibility for AcME and GH activities. Finally, lytic polysaccharide monooxygenases have also been implicated in promoting the degradation of lignocellulosic biomass or mannan degradation by classical mannanolytic enzymes, possibly by disrupting adsorbed mannan residues. Modelling effective enzymatic mannan degradation has implications for improving the saccharification of biomass for the synthesis of value-added and upcycling of lignocellulosic wastes.


Lack of bone stiffness/strength contribution to osteoarthritis--evidence for primary role of cartilage damage.
This study was performed to assess osseous contributions to osteoarthritis, obviating the analysis challenges presented by confounding factors in humans and rarity of osteoarthritis in free-ranging mammals. Frequency of osteoarthritis in 21 bird species was examined and contrasted with measures of afflicted element bone stiffness and strength and compression/tension-resistant characteristics. Osteoarthritis was present in the ankle of 0-16% of bird species analysed, independent of bone laminarity, cortical thickness, circularity, polarization, cross-sectional diameter, length and pneumatization. No correlation of frequency of osteoarthritis with parameters of bone strength and biomechanical parameters was found, suggesting that bone is only secondarily affected in osteoarthritis and that cartilage is the initial target of the disease.


Antibiotic treatment of biofilm infections.
Bacterial biofilms are associated with a wide range of infections, from those related to exogenous devices, such as catheters or prosthetic joints, to chronic tissue infections such as those occurring in the lungs of cystic fibrosis patients. Biofilms are recalcitrant to antibiotic treatment due to multiple tolerance mechanisms (phenotypic resistance). This causes persistence of biofilm infections in spite of antibiotic exposure which predisposes to antibiotic resistance development (genetic resistance). Understanding the interplay between phenotypic and genetic resistance mechanisms acting on biofilms, as well as appreciating the diversity of environmental conditions of biofilm infections which influence the effect of antibiotics are required in order to optimize the antibiotic treatment of biofilm infections. Here, we review the current knowledge on phenotypic and genetic resistance in biofilms and describe the potential strategies for the antibiotic treatment of biofilm infections. Of note is the optimization of PK/PD parameters in biofilms, high-dose topical treatments, combined and sequential/alternate therapies or the use antibiotic adjuvants.


Marijuana is not associated with progression of hepatic fibrosis in liver disease: a systematic review and meta-analysis.
An estimated 22 million adults use marijuana in the USA. The role of marijuana in the progression of hepatic fibrosis remains unclear. We carried out a systematic review and meta-analysis to evaluate the impact of marijuana on prevalence and progression of hepatic fibrosis in chronic liver disease. We searched several databases from inception through 10 November 2017 to identify studies evaluating the role of marijuana in chronic liver disease. Our main outcome of interest was prevalence/progression of hepatic fibrosis. Adjusted odds ratios (ORs) and hazards ratios (HRs) were pooled and analyzed using random-effects model. Nine studies with 5 976 026 patients were included in this meta-analysis. Prevalence of hepatic fibrosis was evaluated in nonalcoholic fatty liver disease (NAFLD), hepatitis C virus (HCV), and hepatitis C and HIV coinfection by two, four, and one studies. Progression of hepatic fibrosis was evaluated by two studies. Pooled OR for prevalence of fibrosis was 0.91 (0.72-1.15), I=75%. On subgroup analysis, pooled OR among NAFLD patients was 0.80 (0.75-0.86), I=0% and pooled OR among HCV patients was 1.96 (0.78-4.92), I=77%. Among studies evaluating HR, pooled HR for progression of fibrosis in HCV-HIV co-infected patients was 1.03 (0.96-1.11), I=0%. Marijuana use did not increase the prevalence or progression of hepatic fibrosis in HCV and HCV-HIV-coinfected patients. On the contrary, we noted a reduction in the prevalence of NAFLD in marijuana users. Future studies are needed to further understand the therapeutic impact of cannabidiol-based formulations in the management of NAFLD.


Long-term outcomes of endoscopic vs surgical drainage of the pancreatic duct in patients with chronic pancreatitis.
A randomized trial that compared endoscopic and surgical drainage of the pancreatic duct in patients with advanced chronic pancreatitis reported a significant benefit of surgery after a 2-year follow-up period. We evaluated the long-term outcome of these patients after 5 years. Between 2000 and 2004, 39 symptomatic patients were randomly assigned to groups that underwent endoscopic drainage or operative pancreaticojejunostomy. In 2009, information was collected regarding pain, quality of life, morbidity, mortality, length of hospital stay, number of procedures undergone, changes in pancreatic function, and costs. Analysis was performed according to an intention-to-treat principle. During the 79-month follow-up period, one patient was lost and 7 died from unrelated causes. Of the patients treated by endoscopy, 68% required additional drainage compared with 5% in the surgery group (P = .001). Hospital stay and costs were comparable, but overall, patients assigned to endoscopy underwent more procedures (median, 12 vs 4; P = .001). Moreover, 47% of the patients in the endoscopy group eventually underwent surgery. Although the mean difference in Izbicki pain scores was no longer significant (39 vs 22; P = .12), surgery was still superior in terms of pain relief (80% vs 38%; P = .042). Levels of quality of life and pancreatic function were comparable. In the long term, symptomatic patients with advanced chronic pancreatitis who underwent surgery as the initial treatment for pancreatic duct obstruction had more relief from pain, with fewer procedures, than patients who were treated endoscopically. Importantly, almost half of the patients who were treated with endoscopy eventually underwent surgery.


Collagen synthesis by cultured rabbit aortic smooth-muscle cells. Alteration with phenotype.
Enzymically isolated rabbit aortic smooth-muscle cells (SMC) in the first few days of primary culture express a 'contractile phenotype', but with time these cells modulate to a 'synthetic phenotype'. Synthetic-state SMC are able to proliferate, and, provided that they undergo fewer than 5 cumulative population doublings, return to the contractile phenotype after reaching confluency [Campbell, Kocher, Skalli, Gabbiani & Campbell (1989) Arteriosclerosis 9, 633-643]. The present study has determined the synthesis of collagen, at the protein and mRNA levels, by cultured SMC as they undergo a change in phenotypic state. The results show that, upon modulating to the synthetic phenotype, SMC synthesized 25-30 times more collagen than did contractile cells. At the same time, non-collagen-protein synthesis increased only 5-6-fold, indicating a specific stimulation of collagen synthesis. Steady-state mRNA levels are also elevated, with alpha 2(I) and alpha 1(III) mRNA levels 30 times and 20 times higher respectively, probably reflecting increased transcriptional activity. Phenotypic modulation was also associated with an alteration in the relative proportions of type I and III collagens synthesized, contractile SMC synthesizing 78.1 +/- 3.6% (mean +/- S.D.) type I collagen and 17.5 +/- 4.7% type III collagen, and synthetic cells synthesizing 90.3 +/- 2.0% type I collagen and 5.8% +/- 1.8% type III collagen. Enrichment of type I collagen was similarly noted at the mRNA level. On return to the contractile state, at confluency, collagen production and the percentage of type I collagen decreased. This further illustrates the close association between the phenotypic state of SMC and their collagen-biosynthetic phenotype.


Absence of residues from crops irrigated with water treated with trifenmorph for schistosomiasis control.
The fate of the molluscicide trifenmorph (N-tritylmorpholine) following its use in irrigation systems to control the intermediate hosts of schistosomes was studied by laboratory and field experiments. Laboratory studies with (14)C-trifenmorph indicated that bean plants did not take up detectable residues when the soil in which they were grown was irrigated with water containing trifenmorph in concentrations likely to occur in the field. These laboratory studies have been complemented by field irrigation studies of growing crops in southern Africa and the Sudan. Residues of trifenmorph and its breakdown product triphenylmethanol could not be detected in the crops and the limit of detectability was 0.01-0.02 ppm. These results indicate that residues are not likely to be present in irrigated crops following the treatment of the irrigation water systems with trifenmorph.


From mutagenic to non-mutagenic nitroarenes: effect of bulky alkyl substituents on the mutagenic activity of nitroaromatics in Salmonella typhimurium. Part II. Substituents far away from the nitro group.
Derivatives of 4-nitrobiphenyl, 4-nitrosobiphenyl, 2-phenyl-5-nitropyridine (2-aza-4-nitrobiphenyl) and 2-nitrofluorene, bearing various alkyl substituents far away from the nitro group (4'-position in nitrobiphenyls, 7-position in 2-nitrofluorenes) were synthesised and tested for mutagenic potency in strains TA98 and TA100 of Salmonella typhimurium. In the absence of S9 in both strains, mutagenicity of all4'-Ad (Ad=adamantyl). Changes of the molecular shape from 'planar' to non-planar caused by the bulk of the introduced substituents (without influencing the twisting of the nitro substituent or the phenyl rings in the biphenyl compounds) may be responsible for this effect by interfering with an efficient intercalation into DNA.A comparison between experimental and theoretical values as calculated from recently developed equations (QSAR) confirmed our previous results (see the preceding paper) that mutagenicity of alkyl-substituted nitroaromatics cannot be predicted by hydrophobicity and LUMO-energies alone without including steric parameters.


Is cancer risk associated with anger control and negative affect? Findings from a prospective cohort study.
To examine the associations between anger control and negative affect and the risk of five common cancers and total cancers. Possible associations between emotional states and the risk of cancer have long been postulated. Prospective cohort study with average follow-up of 9 years. A total of 19,730 adults (99% aged between 40 and 69 years) answered questions on negative affect and anger control at baseline. A total of 1952 cancers were diagnosed, including 352 breast cancers, 318 prostate cancers, 88 lung cancers, 280 colorectal cancers, and 261 melanomas. After adjustment for potential confounders, there was no significant association between anger control or negative affect and risk of breast cancer, melanoma, or total cancers. Weak associations were identified between anger control and prostate cancer, hazards ratio (HR) 1.17 (1.04-1.30) for a 1-unit increase in the standardized scale, negative affect, and lung cancer, HR 1.24 (1.01-1.52) and colorectal cancer, HR 1.14 (1.01-1.28). There was no evidence of an interaction effect between anger control and negative affect. Results suggest that anger control and negative affect are not associated with breast cancer, melanoma, or total cancer risk, although they may have a small role in risk of prostate, colorectal, and lung cancer. Although more research is needed to confirm these latter associations, the results suggest that if affective states are associated with cancer development, the association may differ for different cancers and argue against the use of total cancer as an outcome measure for studies in this area.


Metabolism of thalidomide in liver microsomes of mice, rabbits, and humans.
Thalidomide is increasingly important in clinical treatment, not only of various inflammatory conditions but also in multiple myeloma and other malignancies. Moreover, the metabolism of thalidomide varies considerably among different species, indicating a need to understand its mechanistic basis. Our previous in vivo studies showed the plasma half-life of thalidomide to be much shorter in mice than in humans, with rabbits showing intermediate values. We were unable to detect hydroxylated thalidomide metabolites in humans and suggested that interspecies differences in thalidomide hydroxylation might account for the differences in plasma half-life. We sought here to establish whether these species differences in the formation of hydroxylated thalidomide metabolites could be discerned from in vitro studies. Liver microsomes of mice, rabbit, and human donors were incubated with thalidomide and analyzed using liquid chromatography-mass spectrometry. Hydrolysis products were detected for all three species, and the rates of formation were similar to those for spontaneous hydrolysis, except in rabbits where phthaloylisoglutamine formation increased linearly with microsomal enzyme concentration. Multiple hydroxylation products were detected, including three dihydroxylated metabolites not observed in vivo. Thalidomide-5-O-glucuronide, detected in vivo, was absent in vitro. The amount of 5-hydroxythalidomide formed was high in mice, lower in rabbits, and barely detectable in humans. We conclude that major interspecies differences in hepatic metabolism of thalidomide relate closely to the rate of in vivo metabolite formation. The very low rate of in vitro and in vivo hydroxylation in humans strongly suggests that thalidomide hydroxylation is not a requirement for clinical anticancer activity.


Genomics of human health and aging.
Despite notable progress of the candidate-gene and genome-wide association studies (GWAS), understanding the role of genes contributing to human health and lifespan is still very limited. We use the Framingham Heart Study to elucidate if recognizing the role of evolution and systemic processes in an aging organism could advance such studies. We combine throughput methods of GWAS with more detail methods typical for candidate-gene analyses and show that both lifespan and ages at onset of CVD and cancer can be controlled by the same allelic variants. The risk allele carriers are at highly significant risk of premature death (e.g., RR=2.9, p=5.0 × 10(-66)), onset of CVD (e.g., RR=1.6, p=4.6 × 10(-17)), and onset of cancer (e.g., RR=1.6, p=1.5 × 10(-6)). The mechanism mediating the revealed genetic associations is likely associated with biological aging. These aging-related phenotypes are associated with a complex network which includes, in this study, 62 correlated SNPs even so these SNPs can be on non-homologous chromosomes. A striking result is three-fold, highly significant (p=3.6 × 10(-10)) enrichment of non-synonymous SNPs (N=27) in this network compared to the entire qualified set of the studied SNPs. Functional significance of this network is strengthened by involvement of genes for these SNPs in fundamental biological processes related to aging (e.g., response to stimuli, protein degradation, apoptosis) and by connections of these genes with neurological (20 genes) and cardio-vascular (nine genes) processes and tumorigenesis (10 genes). These results document challenging role of gene networks in regulating human health and aging and call for broadening focus on genomics of such phenotypes.


'One receptor' rules in sensory neurons.
With the recent explosion in the characterization of different sensory systems, a general rule is emerging: only one type of sensory receptor molecule is expressed per receptor neuron. The visual system is no exception and, in most cases, photoreceptors express only one visual pigment per cell. However, the mechanisms underlying the exclusion of sensory receptors are poorly understood. As expression of a given receptor in a given cell is often stochastic, a decision must first be made to express one of the many receptors of the same family (i.e. one particular rhodopsin) and this expression must correlate with the silencing of the other receptors. Furthermore, the projection center for the receptors in the brain must be informed of the decision in order to process this information. Although cells can choose from up to hundreds of sensory receptors (e.g. in the olfactory system), they make almost no mistakes. Evidence has recently emerged that the exclusion mechanism involves the sensory receptor molecules themselves. Here, we describe the findings from various systems in mammals and Drosophila, and review evidence that in the simple visual system of the fly, rhodopsin molecules play an important role in sensory receptor exclusion.


Two-year bond strengths of "all-in-one" adhesives to dentine.
To test the long-term in vitro bonding performance of "all-in-one" self-etch adhesives to dentine. Adper Prompt (3M ESPE), Bond Force (Tokuyama), Brush & Bond (Parkell), iBond Self Etch (Heraeus Kulzer), OptiBond All-In-One (Kerr), and Xeno IV (Dentsply Caulk) were tested. The two-step self-etch adhesive Clearfil SE Bond (Kuraray) served as the control. Extracted human teeth were assigned to groups after creation of smear layer on occlusal dentine. Adhesives were applied according to manufacturers' instructions, and resin composite (Filtek Z250, 3M ESPE) build-ups were placed. The bonded specimens were sectioned into beams after 24h of storage in water. Microtensile bond strengths (μTBS) was determined at baseline, and following one and two years of water storage using an EZ-Test device (Shimadzu). Representative specimens were examined under field emission scanning electron microscope. Fracture mode was recorded. The data were analyzed using linear mixed effects and least square means analyses. The significance level was set at 0.01 for the pairwise comparisons and at 0.05 for the overall model effects. The mean μTBS decreased significantly over time for Adper Prompt, iBond Self Etch, and OptiBond All-In-One. After two years of water ageing, Bond Force, Brush & Bond, OptiBond All-In-One, and Xeno IV had mean μTBS values not statistically different from Clearfil SE Bond. After two years of water ageing, Clearfil SE Bond outperformed two of six all-in-one adhesives. That indicates that the composition of the adhesives rather than their application approach is accountable for their performance. Despite the data available suggesting that most all-in-one adhesives are severely affected by hydrolytic degradation, results of the present in vitro study suggest that some may perform as well as the clinically proven Clearfil SE Bond in the longer term.


Bandwidth analysis of waveguide grating coupler.
The bandwidth of planar waveguide grating couplers is theoretically investigated based on the rigorous grating theory. We observe that the bandwidth behavior is not only determined by the grating coupler intrinsic properties, but also affected by the fiber parameters such as position, beam waist and Numerical Aperture. The rigorous bandwidth formula is derived. By analyzing the formula, several practical guidelines are proposed for grating coupler design and fiber operation in order to achieve wideband performance.


Increasing the clinical utility of the BESTest, mini-BESTest, and brief-BESTest: normative values in Canadian adults who are healthy and aged 50 years or older.
Balance is a composite ability requiring the integration of multiple systems. The Balance Evaluation Systems Test (BESTest) and 2 abbreviated versions (the Mini-BESTest and the Brief-BESTest) are balance assessment tools that target these systems. To date, no normative data exist for any version of the BESTest. The purpose of this study was to determine the age-related normative scores on the BESTest, Mini-BESTest, and Brief-BESTest for Canadians who are healthy and 50 to 89 years of age. A cross-sectional study design was used. Seventy-nine adults who were healthy and aged 50 to 89 years (mean age=68.9 years; 50.6% women) participated. Normative scores were reported by age decade. Mean BESTest scores were 95.7 (95% confidence interval [CI]=94.4-97.1) for adults who were aged 50 to 59 years, 91.4 (95% CI=89.8-93.0) for those who were aged 60 to 69 years, 85.4 (95% CI=82.5-88.2) for those who were aged 70 to 79 years, and 79.4 (95% CI=74.3-84.5) for those who were aged 80 to 89 years. Similar results were obtained for the Mini-BESTest and the Brief-BESTest, and all 3 tests showed statistically significant differences in scores among the age cohorts. Because only adults who were 50 to 89 years of age were tested, there are still no normative data for people outside this age range. Also, the scores presented may not be generalizable to all countries. These normative data enhance the clinical utility of the BESTest, Mini-BESTest, and Brief-BESTest by providing clinicians with reference points to guide treatment.


Young Children's Motor Interference Is Influenced by Novel Group Membership.
From early childhood onward, individuals use behavior copying to communicate liking and belonging. This non-verbal signal of affiliation is especially relevant in the context of social groups and indeed both children and adults copy in-group more than out-group members. Given the societal importance of inter-group interactions, it is imperative to understand the mechanistic level at which group modulations of copying occur early in development. The current study was designed to investigate the effect of novel group membership on young children's motor behavior during a simultaneous movement-observation and -execution task. Four- to six-year-olds (n = 65) first gained membership to one of two novel groups based on their color preference and put on a vest in their chosen color. Subsequently, they were instructed to draw a straight line back-and-forth on a tablet computer that was concurrently displaying a stimulus video in which a model moved her arm congruently or incongruently to the child's instructed direction. In half of the stimulus videos the model belonged to the in-group, while in the other half the model belonged to the out-group, as identified by the color of her dress. The deviations into the uninstructed direction of the children's drawings were quantified as a measure of how much observing the models' behaviors interfered with executing their own behaviors. The motor interference effect, namely higher deviations in the incongruent trials than in the congruent trials, was found only for the out-group condition. An additional manipulation of whether the models' arms followed a biological or non-biological velocity profile had little effect on children's motor interference. The results are interpreted in the context of the explicit coordinative nature of the task as an effect of heightened attention toward interacting with an out-group member. This study demonstrates that already during early childhood, novel group membership dynamically influences behavior processing as a function of interaction context.


Gastrointestinal absorption of metals.
Estimating gastrointestinal absorption remains a significant challenge in the risk assessment of metals. This presentation reviews our current understanding of the gastrointestinal absorption of lead (Pb) to illustrate physiological mechanisms involved in metal absorption, new approaches that are being applied to the problem of estimating metal absorption in humans, and issues related to integrating this information into risk assessment. Absorption of metals can be highly variable in human populations because it is influenced by a variety of factors that include the chemical form of the metal, environmental matrix in which the ingested metal is contained, gastrointestinal tract contents, diet, nutritional status, age, and, in some cases, genotype. Thus, in risk assessment models, gastrointestinal absorption is best described as a variable whose distribution is determined in part by the above multiple influences. Although we cannot expect to evaluate empirically each of the above factors in human populations, we can expect to achieve a sufficiently detailed understanding of absorption mechanisms to develop conceptual and, eventually, quantitative models of absorption that account for some aspects of individual variability. A conceptual model is presented of the physiological processes involved in the transfer of ingested metals from the lumen of the gastrointestinal tract to the blood circulation. Components of the model include delivery of the metal to the site(s) of absorption; distribution of metal among intracellular and extracellular ligands and transcellular and paracellular pathways of transfer across the gastrointestinal tract epithelium. The gastrointestinal absorption of Pb is discussed in the context of this model.


Regulation of ecdysone biosynthesis in the tobacco hornworm, Manduca sexta: titre of the haemolymph stimulatory factor during the last larval instar.
A recently isolated haemolymph protein appears to be an important regulator of ecdysone biosynthesis by prothoracic glands in Manduca sexta. Using a dose-response titration protocol, the haemolymph titre of this stimulatory factor was determined during the last larval instar. The titre was high (greater than 2.0 U ml-1) on days 0 and 1, then dropped significantly to 0.55 U ml-1 on day 2, and remained depressed until day 4. The titre of the stimulatory factor then increased to a peak of 1.62 U ml-1 on day 7, and remained elevated (approx. 1.1 U ml-1) until the end of the instar. A set of physical and biochemical criteria was used to confirm that the stimulatory activity present in haemolymph on different days of the instar represented the presence of the factor. The data are consistent with the hypothesis that fluctuations in the titre of the haemolymph stimulatory factor play a critical role in regulating ecdysone biosynthesis during larval-pupal development.


A rapid analytical method for cholecalciferol (vitamin D3) in fortified infant formula, milk and milk powder using Diels-Alder derivatisation and liquid chromatography-tandem mass spectrometric detection.
A method for analysing vitamin D(3) (VD3, cholecalciferol) has been established and validated. This method is rapid and cost effective and is intended for use in quality control in the manufacture of fortified infant formulae and milk powders. Milk or reconstituted milk powder was solubilised in methanol and extracted in one step into isooctane, which was separated by centrifugation. A portion of the isooctane layer was then transferred, and an aliquot of 4-phenyl-1,2,4-triazoline-3,5-dione was added to derivatise VD3. The analyte was then re-extracted into a small volume of acetonitrile and analysed by reverse-phase chromatography. Detection was by triple quadrupole mass spectrometer using a selective transition, m/z 560 → 298. An internal standard, deuterium-labelled VD3, was used to correct for losses in extraction and any variation in derivatisation and ionisation efficiencies. The method has been subjected to a single-laboratory validation and has been found to be linear, highly selective and accurate with respect to National Institute of Standards and Technology Standard Reference Material 1849, analyte spiking experiments and comparison with an LC-UV-based method. The repeatability standard deviation was 4.23 %. Significantly for routine laboratories, the method returns results within 2 h, generates minimal waste and minimises health and safety concerns to the analyst.


Home automation in the workplace.
Environmental control units and home automation devices contribute to the independence and potential of individuals with disabilities, both at work and at home. Devices currently exist that can assist people with physical, cognitive, and sensory disabilities to control lighting, appliances, temperature, security, and telephone communications. This article highlights several possible applications for these technologies and discusses emerging technologies that will increase the benefits these devices offer people with disabilities.


Cereal pollen sensitisation in pollen allergic patients: to treat or not to treat?
Pollen allergens of the Poaceae family comprise one of the main causes of pollinosis worldwide. Although most of cereals are included in this family, certain pollination characteristics and aerobiological features differentiate them from common wild grass pollen. Cereal pollen grains cannot be easily characterised as potential sources of aero allergens, because of their pollination mode (most of them are autogamous plants), the consecutive low numbers of pollen they produce and their pollen's large volume and consequent high weight which further prevent dispersion and pollen transport. However, various epidemiologicalstudies concluded in comparable sensitisation patterns between common grass and cereal pollen. This fact can be attributed to the common epitopes shared between grass and cereal pollen allergens, which are responsible for the high cross reactivity observed not only in vitro but also in vivo. Therefore, the sensitisation patterns do not usually reflect a genuine, cereal-specific IgE recognition. On the contrary, genuine cereal sensitisation and allergy are referred to only in rare cases of occupational exposure to high amounts of these pollen grains (farmers and field workers) or in patients staying in the vicinity of cereal fields. Thus, cautious considerations should be taken into account when diagnostically approaching patients in which cereal pollen allergy could be considered in the differential diagnosis.


Radiological and functional outcomes of modified Metaizeau technique in displaced radial neck fractures.
The management of displaced radial neck fractures in children is still a controversial topic. The objective of this study is to examine the outcomes of modified Metaizeau technique in the children with displaced radius neck fractures. The retrospective study included 15 children with displaced radial neck fracture with an angulation of more than 30° who were managed with the use of leverage technique by mosquito clamps and internal fixation with elastic stable intramedullary nailing (ESIN). Radiological and functional assessments were performed during follow-up. Additionally, the patients were evaluated using Mayo Elbow Per- formance Score (MEPS). All the children could be managed with clamp-assisted closed reduction. The average duration of follow up was 25.5 ± 6.1 months (15-36 months). An excellent elbow function was achieved in all but one patient. Based on Metaizeau classification, excellent, good, fair, and poor outcomes were achieved in 11, 1, 2, and 1 patients, respectively. The average postoperative MEPS score was 98.7 ± 5.1 (80-100). Clamp-assisted closed reduction and fixation with ESIN is a good choice in the children with displaced radial neck fractures. This technique is associated with good functional and radiologic outcomes in the medium-term. Further studies are warranted with larger sample sizes.


[Possibilities of radionuclide study of lower-limb muscles perfusion in diagnosis of chronic arterial insufficiency].
The authors have hereby proposed a method to diagnose lower-limb muscular ischaemia by means of scintigraphy, used in 35 patients suffering from atherosclerosis obliterans with stage IIB and III A chronic arterial insufficiency of limbs. To do so, administration of the radioactive agent "Myoview" (⁹⁹m)Tc-tetrophosmine) manufactured by the "Nycomed" Company into a patient's peripheral vein was followed by assessing its accumulation in lower-limb muscles at rest and after standard physical load. Alterations in the parameters studied relative to the norm made it possible to judge on the presence of lower-limb circulatory impairments. The method proved highly efficient, thus making it possible to use it for both therapeutic decision-making and probably in future for dynamic control after the treatment performed. Minimally invasive nature and simplicity in use of the methodology open wide possibilities for its use.


Capsaicin cough sensitivity is decreased in smokers.
Although capsaicin provocation has been used to evaluate treatment against cough, which is one of the most common respiratory symptoms, there are still methodological considerations that are not fully known. Capsaicin stimulates the unmyelinated slow C-fibres of the sensory nervous system, which leads to coughing. Smoking often leads to respiratory symptoms with cough and phlegm. The aim of this study was to examine the effect of smoking on capsaicin provocation. Subjectively healthy smokers and non-smokers were challenged with capsaicin in increasing doses. The coughs were counted and irritation of the lower airways was graded on a symptom score. Smokers reacted to provocation with significantly fewer coughs, but there was no difference regarding other symptoms. These results are in agreement with the hypothesis that nicotine inhibits or blocks C-fibres of the sensory nervous system of the lower respiratory tract. Clinically, this may serve to explain increasing airway symptoms that are often seen after cessation of smoking.


Circulating maternal corticotropin-releasing hormone and gonadotropin-releasing hormone in normal and abnormal pregnancies.
Corticotropin-releasing hormone and gonadotropin-releasing hormone are produced by the human placenta and have been measured in the maternal circulation during pregnancy. Our objective was to determine concentrations of these substances in maternal plasma throughout normal pregnancies and in early pregnancy loss. Fifty-one pregnant women were followed up prospectively and plasma samples were drawn at 8, 10, 12, 14, 16, 28, and 36 weeks' gestation and during labor. Specific and sensitive radioimmunoassays were used to determine corticotropin-releasing hormone and gonadotropin-releasing hormone concentrations in these samples. Blood samples were drawn at all time points and outcome data were available from 33 women who completed their pregnancies at term without complications. In this normal group circulating corticotropin-releasing hormone concentrations increased from low or undetectable concentrations at 8 weeks (< or = 23.2 +/- 1.3 pg/ml, mean +/- SEM) to measurable values at 16 weeks (34.3 +/- 2.2 pg/ml). Thereafter there was a significant increase to 1294 +/- 113 pg/ml in labor. Gonadotropin-releasing hormone demonstrated a trimodal distribution, increasing significantly from 8 to 14 weeks, decreasing at 16 weeks, and increasing again by term. The ratio of corticotropin-releasing hormone to gonadotropin-releasing hormone in the normal group demonstrated a 30-fold increase from 8 weeks to term. In eight cases of early pregnancy loss corticotropin-releasing hormone and gonadotropin-releasing hormone concentrations were not significantly different from those of the normal group in early pregnancy. In two cases of premature delivery gonadotropin-releasing hormone concentrations and ratios were within the normal range; corticotropin-releasing hormone levels were normal in both cases of premature delivery. In this study we determined maternal concentrations of corticotropin-releasing hormone and gonadotropin-releasing hormone in normal pregnancies and in labor at term. Neither maternal concentrations of corticotropin-releasing hormone nor gonadotropin-releasing hormone were useful in identifying pregnant women at risk for early pregnancy loss.


Spectral phase induced by the reflection on a mirror-based waveguide grating in the neighborhood of modal resonance.
A linearly polarized plane wave impinging on a mirror-based waveguide grating is shown to experience a 2pi phase shift of controllable slope in the reflection spectrum. Such a zeroth-order resonant grating effect is adequately confirmed by means of an ellipsometer. The close agreement between the analytical representation of grating coupled waveguide resonances with the experimental results confirms the relevance of the underlying phenomenological understanding of resonant gratings.


Emerging trends of HIV drug resistance in Chinese HIV-infected patients receiving first-line highly active antiretroviral therapy: a systematic review and meta-analysis.
Highly active antiretroviral therapy (HAART) has led to a dramatic decrease in AIDS-related morbidity and mortality through sustained suppression of human immunodeficiency virus (HIV) replication and reconstitution of the immune response. Settings like China that experienced rapid HAART rollout and relatively limited drug selection face considerable challenges in controlling HIV drug resistance (DR). We conducted a systematic review and meta-analysis to describe trends in emergent HIV DR to first-line HAART among Chinese HIV-infected patients, as reflected in the point prevalence of HIV DR at key points and fixed intervals after treatment initiation, using data from cohort studies and cross-sectional studies respectively. Pooled prevalence of HIV DR from longitudinal cohorts studies was 10.79% (95% confidence interval [CI], 5.85%-19.07%) after 12 months of HAART and 80.58% (95% CI, 76.6%-84.02%) after 72 months of HAART. The HIV DR prevalence from cross-sectional studies was measured in treatment intervals; during the 0-12-month HAART treatment interval, the pooled prevalence of HIV DR was 11.1% (95% CI, 7.49%-16.14%), which increased to 22.92% at 61-72 months (95% CI, 9.45%-45.86%). Stratified analyses showed that patients receiving a didanosine-based regimen had higher HIV DR prevalence than those not taking didanosine (15.82% vs 4.97%). Patients infected through former plasma donation and those receiving AIDS treatment at village clinics had higher HIV DR prevalence than those infected through sexual transmission or treated at a county-level hospital. Our findings indicate higher prevalence of HIV DR for patients with longer cumulative HAART exposure, highlighting important subgroups for future HIV DR surveillance and control.


Chemotactic signaling via carbohydrate phosphotransferase systems in Escherichia coli.
Chemotaxis allows bacteria to follow gradients of nutrients, environmental stimuli, and signaling molecules, optimizing bacterial growth and survival. Escherichia coli has long served as a model of bacterial chemotaxis, and the signal processing by the core of its chemotaxis pathway is well understood. However, most of the research so far has focused on one branch of chemotactic signaling, in which ligands bind to periplasmic sensory domains of transmembrane chemoreceptors and induce a conformational change that is transduced across the membrane to regulate activity of the receptor-associated kinase CheA. Here we quantitatively characterize another, receptor-independent branch of chemotactic signaling that is linked to the sugar uptake through a large family of phosphotransferase systems (PTSs). Using in vivo characterization of intracellular signaling and protein interactions, we demonstrate that signals from cytoplasmic PTS components are transmitted directly to the sensory complexes formed by chemoreceptors, CheA and an adapter protein CheW. We further conclude that despite different modes of sensing, the PTS- and receptor-mediated signals have similar regulatory effects on the conformation of the sensory complexes. As a consequence, both types of signals become integrated and undergo common downstream processing including methylation-dependent adaptation. We propose that such mode of signaling is essential for efficient chemotaxis to PTS substrates and may be common to most bacteria.


The fractionation of the fatty acid synthetase activities of avocado mesocarp plastids.
1. The range of fatty acids formed by preparations of ultrasonically ruptured avocado mesocarp plastids was dependent on the substrate. Whereas [1-14C]palmitate and [14C]oleate were the major products obtained from [-14C]acetate and [1-14C]acetyl-CoA, the principal product from [2-14C]malonyl-CoA was [14-C]stearate. 2. Ultracentrifugation of the ruptured plastids at 105000g gave a supernatant that formed mainly stearate from [2-14C]malonyl-CoA and to a lesser extent from [1-14C]acetate. The incorporation of [1-14C]acetate into stearate by this fraction was inhibited by avidin. 3. The 105000g precipitate of the disrupted plastids incorporated [1-14C]acetate into a mixture of fatty acids that contained largely [14C]plamitate and [14C]oleate. The formation of [14C]palmitate and [14C]oleate by disrupted plastids was unaffected by avidin. 4. The soluble fatty acid synthetase was precipitated from the 105000g supernatant in the 35-65%-saturated-(NH4)2SO4 fraction and showed an absolute requirement for acyl-carrier protein. 5. Both fractions synthesized fatty acids de novo.


First report of Creutzfeldt-Jakob disease occurring in 2 siblings unexplained by PRNP mutation.
Sibling concurrence of pathologically confirmed prion disease has only been reported in association with pathogenic mutation of the prion protein gene (PRNP). Here, we report 2 siblings with classic neuropathologic features of sporadic Creutzfeldt-Jakob disease unexplained by PRNP mutation or known risk factors for iatrogenic transmission of prion infection. Possible explanations include coincidental occurrence, common exposure to an unidentified environmental source of prions, horizontal transmission of disease, or the presence of unknown shared genetic predisposition.


Recommendations for advancing health literacy measurement.
Health literacy has become a national priority in the United States. Although less is known about the rate, outcomes, and costs associated with health literacy globally relative to the United States, the subject has received increasing attention internationally as well. Definitions, conceptual models, and health literacy measures have proliferated in recent years, and consensus does not exist regarding which of these to use. This article offers the following 5 recommendations for setting a research agenda to advance the science of health literacy measurement: (a) develop a comprehensive unified conceptual framework, (b) leverage the measurement knowledge the field has gained thus far, (c) empirically test frameworks and measures using robust research methods, (d) use a tiered approach to measuring health literacy, and (e) advocate for ongoing research and dissemination. These recommendations seek to ensure clarity, rigor, and transparency as part of a systematic approach to health literacy measurement. Once these steps are taken, the field of health literacy can move forward more effectively.


Novel Thiosemicarbazone Derivatives: In Vitro and In Silico Evaluation as Potential MAO-B Inhibitors.
MAO-B inhibitors are frequently used in the treatment of neurodegenerative diseases such as Parkinson's and Alzheimer's. Due to the limited number of compounds available in this field, there is a need to develop new compounds. In the recent works, it was shown that various thiosemicarbazone derivatives show hMAO inhibitory activity in the range of micromolar concentration. It is thought that benzofuran and benzothiophene structures may mimic structures such as indane and indanone, which are frequently found in the structures of such inhibitors. Based on this view, new benzofuran/benzothiophene and thiosemicarbazone hybrid compounds were synthesized, characterized and screened for their hMAO-A and hMAO-B inhibitory activity by an in vitro fluorometric method. The compounds including methoxyethyl substituent (2b and 2h) were found to be the most effective agents in the series against MAO-B enzyme with the IC50 value of 0.042 ± 0.002 µM and 0.056 ± 0.002 µM, respectively. The mechanism of hMAO-B inhibition of compounds 2b and 2h was investigated by Lineweaver-Burk graphics. Compounds 2b and 2h were reversible and non-competitive inhibitors with similar inhibition features as the substrates. The Ki values of compounds 2b and 2h were calculated as 0.035 µM and 0.046 µM, respectively, with the help of secondary plots. The docking study of compound 2b and 2h revealed that there is a strong interaction between the active sites of hMAO-B and analyzed compound.


Intraoperative touch imprint cytology for the diagnosis of sentinel lymph node metastases in breast cancer.
Intraoperative detection of sentinel lymph node (SLN) metastases enables the surgeon to take an immediate decision to proceed to completion axillary lymph node dissection (ALND). The aim of this study was to determine the accuracy of touch imprint cytology (TIC) for the diagnosis of SLN metastases in sentinel nodes from women with breast cancer. Touch imprints from 235 sentinel nodes in 133 women with breast cancer were diagnosed by cytopathology and compared with definitive histopathology results. After a feasibility study, a real-time study was performed with the surgeon proceeding to ALND based on the TIC diagnosis. The clinical opinion of the operating surgeon as to whether the SLN appeared to contain metastases was recorded, as was the time taken for the result to be available. TIC detected metastases with a sensitivity of 81.1 per cent and a specificity of 100 per cent. False-negative TIC diagnoses were associated with micrometastases and lobular carcinoma. The majority of false-negative diagnoses were due to sampling rather than interpretation errors. Clinical assessment of sentinel nodes had a sensitivity of 64.3 per cent and a specificity of 87.6 per cent. TIC is feasible and enables the rapid diagnosis of SLN metastases with an acceptable accuracy for clinical use in ductal carcinoma of the breast.


The impact of osteoarthritis: implications for research.
Osteoarthritis, the clinical syndrome of joint pain and dysfunction caused by joint degeneration, affects more people than any other joint disease. There are no consistently effective methods for preventing osteoarthritis or slowing its progression, and symptomatic treatments provide limited benefit for many patients. Osteoarthritis disables about 10% of people who are older than 60 years, compromises the quality of life of more than 20 million Americans, and costs the United States economy more than $60 billion per year. The incidence of osteoarthritis rises precipitously with age; as a result, the prevalence and burden of this disorder is increasing rapidly. Study of the patterns osteoarthritis incidence and prevalence shows that it occurs frequently in the hand, foot, knee, spine and hip, but rarely in the ankle, wrist, elbow, and shoulder, and the most important universal risk factors are age, excessive joint loading, and joint injury. Analysis of the impact of osteoarthritis raises questions that include: Why does the incidence increase progressively with age? Why are some joints rarely affected? How do mechanical forces cause joint degeneration? What biologic and mechanical factors slow or accelerate the rate of joint degeneration? Answering these questions could lead to effective methods of preventing osteoarthritis and slowing its progression.


Clinical and Economic Impact of a Two-layer Compression System for the Treatment of Venous Leg Ulcers: A Systematic Review.
This study evaluates the clinical and cost effectiveness of a 2-layer compression system (2LBA; 3M Coban Two-Layer Compression System; 3M, St Paul, MN) compared with other 2-layer (2LB) and 4-layer (4LB) compression systems in patients with noninfected venous leg ulcers (VLUs). The MEDLINE, EMBASE, CINAHL, Cochrane Library, National Health Service (NHS) Economic Evaluation, and EconLit databases were searched from inception up to January 2017. The MEDLINE search was updated on March 31, 2017. Study selection, quality assessment, and data synthesis were undertaken in accordance with recommended standards. Findings were presented narratively. In total, 5 studies (N = 1509 patients) of mixed methodological quality were included. At 6 months, 2LBA achieved better ulcer healing in comparison with 2LBB (odds ratio [OR], 1.57; 95% confidence interval [CI], 1.10-2.24; P = .03) and 4LBA (OR, 1.93, 95% CI, 1.26-2.97; P = .05) in patients with newly diagnosed ulcers only. For a combined population with newly diagnosed and existing VLUs, healing outcomes were OR, 2.87; 95% CI, 1.06-7.77; P = .04, and OR, 16.51; 95% CI, 2.08-131.37; P = .008, for 2LBs and 4LBs, respectively. Results on slippage were inconclusive. Adverse events were infrequent and did not differ significantly between interventions. Lower 6-month NHS costs for the combined population (£2413 vs. £2707 or £2648) and for newly diagnosed patients (£3045 vs. £3842 or £4480) were observed comparing 2LBA with 2LBB or 4LBA. Also, 2LBA was associated with better health-related quality of life (HRQoL) at 6 months. Based on these findings, 2LBA may result in lower treatment costs and better ulcer healing and HRQoL compared with other multicomponent therapies, especially in patients with newly diagnosed VLUs. However, further high-quality research is needed, especially for outcomes such as slippage and bandage wear time.


Atypical development in plant and soil nematodes.
Observations of atypical developmental and anatomical characteristics have been recorded for many taxa of soil nematodes. They include the unusual occurrence of extra feeding structures, aberrant configuration of features of both male and female reproductive systems, and the occurrence of intersexes assumed to be functionally female, functionally male, or non-functional. In many cases, hypotheses have been advanced regarding the genetic or developmental mechanisms and environmental stimuli that control, regulate, or facilitate abnormalities, but many are quite speculative and lack experimental verification. Further, the fitness costs or advantages, and the heritability of aberrant characters are largely unknown, except where they clearly preclude reproduction, either apomictic or amphimictic. Underlying mechanisms and ecological consequences may be difficult to study in organisms that are not readily cultured under axenic or sterile laboratory conditions, however information on developmental processes in Caenorhabditis elegans represents an important resource in which to seek homologies.


Respiratory muscle pacing with chronically implanted intramuscular Permaloc electrodes: A feasibility study.
We tested the feasibility of stimulating upper-intercostal and abdominal muscles plus the diaphragm by using chronically implanted intramuscular electrodes.During two survival surgeries with six dogs, intramuscular electrodes were implanted bilaterally in the three respiratory muscles. Standard stimulation of the diaphragm was conducted. The dorsolateral and ventrolateral abdominal wall areas were stimulated with a 25 mA current. The second to fourth intercostal spaces were stimulated to elicit the largest tidal volume associated with the least coactivation of the serratus and latissimus muscles. Lone diaphragm and upper-intercostal muscle pacing produced inhaled tidal volumes (mean +/- standard error of the mean) of 293 +/- 36 mL and 59 +/- 17 mL, respectively. Lone abdominal muscle pacing produced an exhaled volume of 55 +/- 17 mL. Combined pacing of diaphragm and intercostal muscles increased the inhaled volume to 389 +/- 39 mL. The addition of abdominal pacing following the combined stimulation of diaphragm and intercostals increased the exhaled volume to 472 +/- 54 mL. During autopsy, dislodgement of the electrodes overlying the ribs was a concern and probably resulted from loose animal jackets. Chronic intramuscular Permaloc electrodes can be implanted in several respiratory muscles and increase tidal volumes more than diaphragm stimulation alone.


Sympathetic activity controls fat-induced oleoylethanolamide signaling in small intestine.
Ingestion of dietary fat stimulates production of the small-intestinal satiety factors oleoylethanolamide (OEA) and N-palmitoyl-phosphatidylethanolamine (NPPE), which reduce food intake through a combination of local (OEA) and systemic (NPPE) actions. Previous studies have shown that sympathetic innervation of the gut is necessary for duodenal infusions of fat to induce satiety, suggesting that sympathetic activity may engage small-intestinal satiety signals such as OEA and NPPE. In the present study, we show that surgical resection of the sympathetic celiac-superior mesenteric ganglion complex, which sends projections to the upper gut, abolishes feeding-induced OEA production in rat small-intestinal cells. These effects are accounted for by suppression of OEA biosynthesis, and are mimicked by administration of the selective β2-adrenergic receptor antagonist ICI-118,551. We further show that sympathetic ganglionectomy or pharmacological blockade of β2-adrenergic receptors prevents NPPE release into the circulation. In addition, sympathetic ganglionectomy increases meal frequency and lowers satiety ratio, and these effects are corrected by pharmacological administration of OEA. The results suggest that sympathetic activity controls fat-induced satiety by enabling the coordinated production of local (OEA) and systemic (NPPE) satiety signals in the small intestine.


Hyperadrenergic postural tachycardia syndrome in mast cell activation disorders.
Postural tachycardia syndrome (POTS) is a disabling condition that commonly affects otherwise normal young females. Because these patients can present with a flushing disorder, we hypothesized that mast cell activation (MCA) can contribute to its pathogenesis. Here we describe POTS patients with MCA (MCA+POTS), diagnosed by episodes of flushing and abnormal increases in urine methylhistamine, and compared them to POTS patients with episodic flushing but normal urine methylhistamine and to normal healthy age-matched female controls. MCA+POTS patients were characterized by episodes of flushing, shortness of breath, headache, lightheadedness, excessive diuresis, and gastrointestinal symptoms such as diarrhea, nausea, and vomiting. Triggering events include long-term standing, exercise, premenstrual cycle, meals, and sexual intercourse. In addition, patients were disabled by orthostatic intolerance and a characteristic hyperadrenergic response to posture, with orthostatic tachycardia (from 79+/-4 to 114+/-6 bpm), increased systolic blood pressure on standing (from 117+/-5 to 126+/-7 mm Hg versus no change in POTS controls), increased systolic blood pressure at the end of phase II of the Valsalva maneuver (157+/-12 versus 117+/-9 in normal controls and 119+/-7 mm Hg in POTS; P=0.048), and an exaggerated phase IV blood pressure overshoot (50+/-10 versus 17+/-3 mm Hg in normal controls; P<0.05). In conclusion, MCA should be considered in patients with POTS presenting with flushing. These patients often present with a typical hyperadrenergic response, but beta-blockers should be used with great caution, if at all, and treatment directed against mast cell mediators may be required.


Is high-dose rate RapidArc-based radiosurgery dosimetrically advantageous for the treatment of intracranial tumors?
In linac-based stereotactic radiosurgery (SRS) and radiotherapy (SRT), circular cone(s) or conformal arc(s) are conventionally used to treat intracranial lesions. However, when the target is in close proximity to critical structures, it is frequently quite challenging to generate a quality plan using these techniques. In this study, we investigated the dosimetric characteristics of using high-dose rate RapidArc (RA) technique for radiosurgical treatment of intracranial lesions. A total of 10 intracranial SRS/SRT cases previously planned using dynamic conformal arc (DCA) or cone-based techniques have been included in this study. For each case, 3 treatment plans were generated: (1) a DCA plan with multiple noncoplanar arcs, (2) a high-dose rate RA plan with arcs oriented the same as DCA (multiple-arc RA), and 3) a high-dose rate RA plan with a single coplanar arc (single-arc RA). All treatment plans were generated under the same prescription and similar critical structure dose limits. Plan quality for different plans was evaluated by comparing various dosimetric parameters such as target coverage, conformity index (CI), homogeneity index (HI), critical structures, and normal brain tissue doses as well as beam delivery time. With similar critical structure sparing, high-dose rate RA plans can achieve much better target coverage, dose conformity, and dose homogeneity than the DCA plans can. Plan quality indices CI and HI, for the DCA, multiple-arc RA, and single-arc RA techniques, were measured as 1.67 ± 0.39, 1.32 ± 0.28, and 1.38 ± 0.30 and 1.24 ± 0.11, 1.10 ± 0.04, and 1.12 ± 0.07, respectively. Normal brain tissue dose (V12Gy) was found to be similar for DCA and multiple-arc RA plans but much larger for the single-arc RA plans. Beam delivery was similar for DCA and multiple-arc RA plans but shorter with single-arc RA plans. Multiple-arc RA SRS/SRT can provide better treatment plans than conventional DCA plans, especially for complex cases.


Conformational selection of the intrinsically disordered plant stress protein COR15A in response to solution osmolarity - an X-ray and light scattering study.
The plant stress protein COR15A stabilizes chloroplast membranes during freezing. COR15A is an intrinsically disordered protein (IDP) in aqueous solution, but acquires an α-helical structure during dehydration or the increase of solution osmolarity. We have used small- and wide-angle X-ray scattering (SAXS/WAXS) combined with static and dynamic light scattering (SLS/DLS) to investigate the structural and hydrodynamic properties of COR15A in response to increasing solution osmolarity. Coarse-grained ensemble modelling allowed a structure-based interpretation of the SAXS data. Our results demonstrate that COR15A behaves as a biomacromolecule with polymer-like properties which strongly depend on solution osmolarity. Biomacromolecular self-assembly occurring at high solvent osmolarity is initiated by the occurrence of two specific structural subpopulations of the COR15A monomer. The osmolarity dependent structural selection mechanism is an elegant way for conformational regulation and assembly of COR15A. It highlights the importance of the polymer-like properties of IDPs for their associated biological function.


Prospective study of blunt aortic injury: helical CT is diagnostic and antihypertensive therapy reduces rupture.
There were two aims of this study. The first was to evaluate the application of helical computed tomography of the thorax (HCTT) for the diagnosis of blunt aortic injury (BAI). The second was to evaluate the efficacy of beta-blockers with or without nitroprusside in preventing aortic rupture. Aortography has been the standard for diagnosing BAI for the past 4 decades. Conventional chest CT has not proven to be of significant value. Helical CT scanning is faster and has higher resolution than conventional CT. Retrospective studies have suggested the efficacy of antihypertensives in preventing aortic rupture. A prospective study comparing HCTT to aortography in the diagnosis of BAI was performed. A protocol of beta-blockers with or without nitroprusside was also examined for efficacy in preventing rupture before aortic repair and in allowing delayed repair in patients with significant associated injuries. Over a period of 4 years, 494 patients were studied. BAI was diagnosed in 71 patients. Sensitivity was 100% for HCTT versus 92% for aortography. Specificity was 83% for HCTT versus 99% for aortography. Accuracy was 86% for HCTT versus 97% for aortography. Positive predictive value was 50% for HCTT versus 97% for aortography. Negative predictive value was 100% for HCTT versus 97% for aortography. No patient had spontaneous rupture in this study. HCTT is sensitive for diagnosing intimal injuries and pseudoaneurysms. Patients without direct HCTT evidence of BAI require no further evaluation. Aortography can be reserved for indeterminate HCTT scans. Early diagnosis with HCTT and presumptive treatment with the antihypertensive regimen eliminated in-hospital aortic rupture.


MicroRNA-708-5p regulates mycobacterial vitality and the secretion of inflammatory factors in Mycobacterium tuberculosis-infected macrophages by targeting TLR4.
Tuberculosis (TB), a major public health problem worldwide, is induced by Mycobacterium tuberculosis (M.tb) infection. Macrophages serve as the cellular home in immunoreaction against M.tb infection, which is tightly adjusted by host microRNAs (miRNAs) expression. The purpose of this research was to investigate the function mechanism of miR-708-5p in mycobacterial vitality and immunoreaction in human macrophages (HTP-1 and U937 cells) after M.tb infection. Colony-forming unit (CFU) assay was used to measure mycobacterial survival. The interferon-γ (IFN-γ), interleukin-6 (IL-6), IL-1β, and tumor necrosis factor-α (TNF-α) expression in cell supernatants were detected by enzyme-linked immunosorbent assay (ELISA). The relationship between miR-708-5p and toll-like receptor 4 (TLR4) was predicted and revealed by TargetScan and Dual-Luciferase Reporter Assay. Our results suggested that the miR-708-5p level was increased in a concentration-dependent and time-dependent manner in M.tb-infected human macrophages. Compared with the control group, miR-708-5p mimic enhanced the intracellular mycobacterial survival during M.tb infection, while miR-708-5p downregulation suppressed the mycobacteria survival. Moreover, the secretion of the pro-inflammatory factors, including IFN-γ, IL-6, IL-1β, and TNF-α significantly enhanced in M.tb-induced macrophages, while miR-708-5p mimic reduced these inflammatory cytokines. Conversely, miR-708-5p inhibitor dramatically promoted the accumulation of the inflammatory factors in macrophages after M.tb treatment. In addition, evidence indicated that TLR4 was a direct and functional target of miR-708-5p. MiR-708-5p negatively regulated the TLR4 level in macrophages. The findings indicated that miR-708-5p level was upregulated in macrophages after M.tb infection. And miR-708-5p could regulate mycobacterial vitality and inflammatory response to M.tb infection in human macrophages by targeting TLR4.


Phenotypical characterization of lymphocytes infiltrating regressing papillomas.
Papillomavirus-induced lesions often regress spontaneously in both humans and animals. Papilloma regression is deemed to be due to a cell-mediated immune response, the nature of which is still ill defined, and is accompanied by immune cell infiltrates. To gain further information on the nature and role of the immune cells present in regressing papillomas, we have analyzed biopsies of papillomas induced in the soft palate of cattle by bovine papillomavirus type 4 (BPV-4) and have phenotypically characterized and quantified the lymphocytes present in these lesions. Eleven papilloma biopsies and seven biopsies of noninfected palate were analyzed for the presence of activated CD4+, CD8+, and gamma delta(WC1+) lymphocytes. We found large numbers of lymphocytes in the subepithelial derma of papillomas but not in normal palate tissue; these cellular masses consisted predominantly of CD4+ lymphocytes, with only a few CD8+ and gamma delta(WC1+) lymphocytes, generally positioned at the periphery of these masses. All three subtypes of lymphocytes were found interdigitated with the cells of the basal layer both in papillomas and in normal palate tissue, but while basal layer CD8+ and gamma delta(WC1+) T cells were detected with similar frequencies in papillomas and uninfected palate, basal layer CD4+ T cells were much more frequent in papillomas. CD4+, CD8+, and gamma delta(WC1+) lymphocytes were found in the suprabasal layers of papillomas, but the CD8+ and gamma delta(WC1+) T cells were more numerous and had migrated further into the differentiating keratinocytes of the papilloma fronds than the CD4+ T cells. We conclude that T-cell infiltration is characteristic of regressing BPV-4 papillomas, that CD4+ lymphocytes are specifically and massively recruited into the regressing papillomas, and that although all three lymphocyte subsets can penetrate the papilloma, only the CD8+ and gamma delta(WC1+) lymphocytes are able to migrate into the fronds. These results suggest that all three lymphocyte subsets have an important role to fulfill during natural regression of papillomas.


Hexacene generated on passivated silicon.
On-surface synthesis represents a successful strategy to obtain designed molecular structures on an ultra-clean metal substrate. While metal surfaces are known to favor adsorption, diffusion, and chemical bonding between molecular groups, on-surface synthesis on non-metallic substrates would allow the electrical decoupling of the resulting molecule from the surface, favoring application to electronics and spintronics. Here, we demonstrate the on-surface generation of hexacene by surface-assisted reduction on a H-passivated Si(001) surface. The reaction, observed by scanning tunneling microscopy and spectroscopy, is probably driven by the formation of Si-O complexes at dangling bond defects. Supported by density functional theory calculations, we investigate the interaction of hexacene with the passivated silicon surface, and with single silicon dangling bonds.


The effects of low-copper diets with or without supplemental molybdenum on specific immune responses of stressed cattle.
Angus bull calves (n = 42; 7 mo of age; 254 kg initial BW) were used to investigate the effects of dietary Cu and Mo on immune function of stressed cattle. Randomly selected calves (n = 22) were injected with 90 mg of Cu as Cu glycinate 28 d before weaning and castrated at weaning. These calves received 7.5 and 5 mg of supplemental Cu/kg of DM during a 41-d receiving phase and a 196-d growing phase, respectively. The remainder of the steers received no supplemental Cu during the experiment. Copper-supplemented steers had adequate Cu status at weaning, whereas unsupplemented calves were marginally Cu-deficient. Cell-mediated response to intradermal injection of phytohemagglutinin was not affected by dietary treatment during the receiving phase. During the growing phase, half of the steers in each Cu treatment were given 5 mg of supplemental Mo/kg of DM. Copper supplementation increased (P<.05) humoral response to ovalbumin injected on d 133 of the growing phase. On d 168 of the growing phase, calves receiving only supplemental Mo were severely Cu-deficient based on plasma and liver Cu concentrations. The other treatment groups had adequate Cu status. Before feeding on d 168 of the growing phase, half of the steers were loaded onto trailers and transported 2.5 h, and they remained on the trailers an additional 9.5 h. Humoral response to porcine erythrocytes (PRBC) and delayed-type hypersensitivity (DTH) to dinitrochlorobenzene was tested at the end of the stress period. There was a Cu x stress interaction for humoral response to PRBC, with Cu decreasing antibody titers in unstressed calves and increasing titers in stressed steers. Stressed steers had lower (P = .03) ADG during the 28 d following stress. The results of this study indicate that Cu deficiency and 5 mg of supplemental Mo/kg of DM do not dramatically alter the specific immunity of stressed cattle.


Experimental Characterization of Hb Flurlingen (HBA2: c.177 C > G, p.His > Gln) and Hb Boghé (HBA2: c.177 C > A, p.His > Gln) Reveals Contradictory HBA2 Expression and Translation Patterns Despite Identical Amino Acid Substitutions.
In this study, we describe the clinical features and provide experimental analyses of Hb Flurlingen (HBA2: c.177 C > G, p.His > Gln) that contrasted with Hb Boghé (HBA2: c.177 C > A, p.His > Gln). Despite the identical amino acid substitution in both variants, Hb Flurlingen shows the phenotype of α-thalassemia (α-thal), whereas Hb Boghé has no impact on α2-globin (HBA2) production. For in vitro transcription analysis, HBA2 expression constructs carrying the HBA2-WT (wild type), Hb Flurlingen and Hb Boghé sequences were generated and expressed in human bladder carcinoma 5637 cells for downstream analyses by quantitative real time-polymerase chain reaction (qReTi-PCR) and immunofluorochemistry (IFC). In silico analysis of secondary folding structures of the HBA2-WT, Hb Flurlingen and Hb Boghé mRNA sequences was performed using Mfold software. The gene transcription and translation analyses revealed that cells transfected with the Hb Flurlingen construct had significantly lower HBA2 transcription (-55.4%, p ≤ 0.01) and reduced protein synthesis when compared to the wild type group. In contrast, cells transfected with the Hb Boghé construct showed no significant changes in HBA2 transcription or translation activities when compared to the wild type group. The in silico prediction of possible effects of these mutations on the folding structures of the HBA2 transcripts showed a change of secondary folding pattern in the Hb Flurlingen transcript when compared to those of HBA2-WT and Hb Boghé. Our experimental findings support the clinical presentation of an α-thalassemic phenotype for Hb Flurlingen in contrast with Hb Boghé, despite identical amino acid substitutions. The results confirm the importance of experimental analysis in establishing the impact of novel base substitutions.


Cultured hepatocytes adopt progenitor characteristics and display bipotent capacity to repopulate the liver.
Clinical studies have proved the therapeutic potential of hepatocyte transplantation as a promising alternative to whole organ liver transplantation in the treatment of hereditary or end-stage liver disease. However, donor shortage seriously restricts cell availability, and the lack of appropriate cell culture protocols for the storage and maintenance of donor cells constitutes a significant obstacle. The aim of this study was to stimulate mature hepatocytes in culture to multiply in vitro and track their fate on transplantation. Rat hepatocytes isolated nonenzymatically were cultured serum free for up to 10 days. They were stimulated into proliferation in the presence of growth factors and conditioned media from nonparenchymal and hepatocyte culture supernatants, as well as 10 mM lithium chloride (LiCl). Cell proliferation was assessed by determining DNA content. Additionally, the extent of cell differentiation was estimated using immunofluorescence staining of hepatic, biliary, progenitor, and mesenchymal markers and gene expression analyses. Transplantation studies were performed on the Fischer CD26-mutant rat following pretreatment with retrorsine and partial hepatectomy. Proliferating hepatocytes increasingly adopted precursor characteristics, expressing progenitor (OV6, CD133), hepatic lineage (CK18), biliary (CD49f, CK7, CK19), and mesenchymal (vimentin) markers. The supplement of LiCl further enhanced the proliferative capacity by 30%. Transplantation studies revealed extensive repopulation by large donor hepatocyte clusters. Furthermore, bile duct-like structures deriving from donor cells proved to be immunoreactive to ductular markers and formed in close proximity to endogenous bile ducts. Mature hepatocytes reveal their potential to "switch" between phenotypes, adopting progenitor characteristics during proliferation in vitro. Following transplantation, these "retrodifferentiated" cells further expanded in vivo, thereby generating bipotentially differentiated progenies (hepatocytes and bile duct-like structures). This apparent plasticity of mature hepatocytes may open new approaches for cell-based strategies to treat liver disease.


Occupational asthma in a hairdressing salon.
Occupational asthma among hairdressers has been recognised for some years and cases of work related asthma due to hair bleaches containing persulphates and hair dyes have been reported. The extent of the disease among hairdressers remains unknown. An investigation was carried out on an entire hairdressing salon, which specialised in hair bleaching and colouring and which employed 23 staff. On the basis of history and specific and non-specific bronchial provocation testing, four out of 23 staff were found to have occupational asthma due to the persulphate salts contained in hair bleaches. Only one of these had a positive skinprick test response to persulphate salts. Tests for non-specific bronchial reactivity to histamine in this work force were more sensitive for the diagnosis of asthma than simple lung function tests or recordings of peak flow rates performed four times daily for three weeks. The response to these agents was studied in greater detail by specific bronchial provocation tests in 14 members of the salon as well as one hairdresser from elsewhere with occupational asthma, three individuals with non-occupational asthma, and four normal subjects. Only those with a history of work related asthma and bronchial hyperreactivity responded positively, confirming that the response to bleach powders was specific. Studies of pulmonary mechanics after challenge showed that the response arose from changes in airway calibre not lung volumes. Measurement of neutrophil chemotactic activity after challenge showed significant rises in those affected, suggesting that mast cells may play a part in the pathogenesis of occupational asthma due to persulphates.


Pneumococcal carriage and antibiotic resistance in young children before 13-valent conjugate vaccine.
We sought to measure trends in Streptococcus pneumoniae carriage and antibiotic resistance in young children in Massachusetts communities after widespread adoption of heptavalent 7-valent pneumococcal conjugate vaccine (PCV7) and before the introduction of the 13-valent PCV (PCV13). We conducted a cross-sectional study including collection of questionnaire data and nasopharyngeal specimens among children aged <7 years in primary care practices from 8 Massachusetts communities during the winter season of 2008-2009 and compared with similar studies performed in 2001, 2003-2004, and 2006-2007. Antimicrobial susceptibility testing and serotyping were performed on pneumococcal isolates, and risk factors for colonization in recent seasons (2006-2007 and 2008-2009) were evaluated. We collected nasopharyngeal specimens from 1011 children, 290 (29%) of whom were colonized with pneumococcus. Non-PCV7 serotypes accounted for 98% of pneumococcal isolates, most commonly 19A (14%), 6C (11%), and 15B/C (11%). In 2008-2009, newly targeted PCV13 serotypes accounted for 20% of carriage isolates and 41% of penicillin-nonsusceptible S. pneumoniae. In multivariate models, younger age, child care, young siblings, and upper respiratory illness remained predictors of pneumococcal carriage, despite near-complete serotype replacement. Only young age and child care were significantly associated with penicillin-nonsusceptible S. pneumoniae carriage. Serotype replacement post-PCV7 is essentially complete and has been sustained in young children, with the relatively virulent 19A being the most common serotype. Predictors of carriage remained similar despite serotype replacement. PCV13 may reduce 19A and decrease antibiotic-resistant strains, but monitoring for new serotype replacement is warranted.


Nutritional Interventions in the Management of Fibromyalgia Syndrome.
Fibromyalgia (FM) is a multifactorial syndrome of unknown etiology, characterized by widespread chronic pain and various somatic and psychological manifestations. The management of FM requires a multidisciplinary approach combining both pharmacological and nonpharmacological strategies. Among nonpharmacological strategies, growing evidence suggests a potential beneficial role for nutrition. This review summarizes the possible relationship between FM and nutrition, exploring the available evidence on the effect of dietary supplements and dietary interventions in these patients. Analysis of the literature has shown that the role of dietary supplements remains controversial, although clinical trials with vitamin D, magnesium, iron and probiotics' supplementation show promising results. With regard to dietary interventions, the administration of olive oil, the replacement diet with ancient grains, low-calorie diets, the low FODMAPs diet, the gluten-free diet, the monosodium glutamate and aspartame-free diet, vegetarian diets as well as the Mediterranean diet all appear to be effective in reducing the FM symptoms. These results may suggest that weight loss, together with the psychosomatic component of the disease, should be taken into account. Therefore, although dietary aspects appear to be a promising complementary approach to the treatment of FM, further research is needed to provide the most effective strategies for the management of FM.


Transcription factor-dependent chromatin remodeling of Il18r1 during Th1 and Th2 differentiation.
The IL-18Ralpha-chain is expressed on Th1 but not Th2 cells. We have recently shown that Stat4 is an important component of programming the Il18r1 locus (encoding IL-18Ralpha) for maximal expression in Th1 cells. Il18r1 is reciprocally repressed during Th2 development. In this report, we demonstrate the establishment of DH patterns that are distinct among undifferentiated CD4 T, Th1, and Th2 cells. Stat6 is required for the repression of Il18r1 expression and in Stat6-deficient Th2 cultures, mRNA levels, histone acetylation, and H3K4 methylation levels are intermediate between levels observed in Th1 and Th2 cells. Despite the repressive effects of IL-4 during Th2 differentiation, we observed only modest binding of Stat6 to the Il18r1 locus. In contrast, we observed robust GATA-3 binding to a central region of the locus where DNase hypersensitivity sites overlapped with conserved non-coding sequences in Il18r1 introns. Ectopic expression of GATA-3 in differentiated Th1 cells repressed Il18r1 mRNA and surface expression of IL-18Ralpha. These data provide further mechanistic insight into transcription factor-dependent establishment of Th subset-specific patterns of gene expression.


Lower extremity peripheral vascular disease. Part 1: Scanning electron microscopy.
The surgical loss of the foot or leg is vigorously resisted by podiatrists who are committed to the conservation of limbs at risk because of peripheral vascular disease. Pathologic changes in blood vessels may, however, progress to a point where no other option is available to the patient. This study assesses amputation of the extremities and investigates tissue alterations that can be identified in diseased blood vessels implicated in these circumstances. Gross pathology and scanning electron microscopy are examined in this, the first of a two-part study.


[Donatori di Musica: when oncology meets music].
Donatori di Musica is a network of musicians - both physicians and volunteers - that was initially founded in 2009 with the aim to set up and coordinate classical music concerts in hospitals. This activity was initially started and led by the Oncology Departments at Carrara and Bolzano Hospitals, where high profile professional musicians make themselves available for concerts in support of Oncological in/out-patients of that specific Hospital. A live classical music performance is a deeply touching experience - particularly for those who live a critical condition like cancer. Main characteristics of Donatori di Musica concerts are: continuity (concerts are part of a regular and non-stopping music season); quality (concerts are held by well-established professional musicians); philanthropic attitude (musicians do not wear a suit and usually chat with patients; they also select an easy-to-listen program; a convivial event is usually organized after the performance with the aim of overcoming distinctions and barriers between physician and patient); no profit: musicians perform for free - travel expenses and/or overnight staying only can be claimed; concerts have free access for patients, their families and hospital staff.Patients and musicians therefore do get in close contact and music is able to merge each other experiences - with patients being treated by the beauty of music and musicians being treated theirselves by patients daily-life feedback. The Donatori di Musica experience is therefore able to help Medicine to retrieve its very first significance - the medical act regain that human and cultural dimension that seems to be abandoned in the last decades in favour of a mere technicism. This is the spirit and the deep significance of Donatori di Musica - «[…] the hope that Music can become a key support to medical treatments in every Oncology department» (by Gian Andrea Lodovici).