Dementia Prevalence in Greece: The Hellenic Longitudinal Investigation of Aging and Diet (HELIAD).
Study of the epidemiology of dementia to gain insight into putative predisposing and prophylactic factors is the first step toward establishing effective preventive and therapeutic strategies for this ever-growing public health problem. Relevant data in Greece are scattered and outdated. We investigated dementia prevalence as part of a population-representative epidemiological study [Hellenic Longitudinal Investigation of Aging and Diet (HELIAD)] in 2 Greek regions. Our sample comprised 1792 adults 65 years of age or older, who received a full neurological and neuropsychological evaluation that led to a consensus diagnosis. The overall prevalence of dementia was 5.0%, with 75.3% of the cases attributed to Alzheimer disease. Dementia odds were 15.8% higher for every year of advancing age and 9.4% lower for every additional year of education. Carrying at least 1 APOE-ε4 allele doubled the risk of dementia, whereas sex did not exert a statistically significant effect. Our results are consistent with previous research in Southern European countries; dementia prevalence in Greece is in the lower range of what has been reported globally.


An in vitro model of hippocampal sharp waves: regional initiation and intracellular correlates.
During slow wave sleep and consummatory behaviors, electroencephalographic recordings from the rodent hippocampus reveal large amplitude potentials called sharp waves. The sharp waves originate from the CA3 circuitry and their generation is correlated with coherent discharges of CA3 pyramidal neurons and dependent on activities mediated by AMPA glutamate receptors. To model sharp waves in a relatively large hippocampal circuitry in vitro, we developed thick (1 mm) mouse hippocampal slices by separating the dentate gyrus from the CA2/CA1 areas while keeping the functional dentate gyrus-CA3-CA1 connections. We found that large amplitude (0.3-3 mV) sharp wave-like field potentials occurred spontaneously in the thick slices without extra ionic or pharmacological manipulation and they resemble closely electroencephalographic sharp waves with respect to waveform, regional initiation, pharmacological manipulations, and intracellular correlates. Through measuring tissue O2, K+, and synaptic and single cell activities, we verified that the sharp wave-like potentials are not a consequence of anoxia, nonspecific elevation of extracellular K+ and dissection-related tissue damage. Our data suggest that a subtle but crucial increase in the CA3 glutamatergic activity effectively recruits a population of neurons thus responsible for the generation of the sharp wave-like spontaneous field potentials in isolated hippocampal circuitry.


Purification and properties of 3-deoxy-d-arabinoheptulosonic acid-7-phosphate synthetase (phe) from a lambda aroG+ transductant of Escherichia coli.
By incorporating aroG, the structural gene for 3-deoxy-d-arabinoheptulosonic acid-7-phosphate (DAHP) synthetase (phe), into the genome of a heat-inducible susR60 mutant of phage lambda, it has been possible to increase the intracellular levels of DAHP synthetase (phe) in a lysogenized strain of Escherichia coli some 15-fold over levels found in the wild-type strain. By using this strain, the enzyme has been purified approximately 2,000-fold compared with wild type, and various kinetic parameters of the purified enzyme have been studied. In contrast to previous reports, the inhibition by phenylalanine was found to exhibit sigmoidal kinetics, suggestive of cooperative interactions between phenylalanine binding sites. Stimulation of enzyme activity by Co(2+) was minimal (14%).


A simplified liquid nitrogen vapour method for the cryopreservation of human semen.
A simple manually operated liquid nitrogen vapour freezing apparatus has been developed for the cryopreservation of human semen. A test method using this apparatus was evaluated by comparison with a conventional freezing method using an ultra cold (-70 degrees C) freezer. Twenty different unselected fresh semen samples were analysed, diluted with cryoprotective medium and then divided into equal numbers of straws prior to freezing by the two methods. A comparison of the average sperm motility before freezing and after thawing using these two methods demonstrated a statistically significant superiority of the simplified test method.


The diagnostic yield of exome sequencing in liver diseases from a curated gene panel.
Exome sequencing (ES) has been used in a variety of clinical settings but there are limited data on its utility for diagnosis and/or prediction of monogenic liver diseases. We developed a curated list of 502 genes for monogenic disorders associated with liver phenotypes and analyzed ES data for these genes in 758 patients with chronic liver diseases (CLD). For comparison, we examined ES data in 7856 self-declared healthy controls (HC), and 2187 patients with chronic kidney disease (CKD). Candidate pathogenic (P) or likely pathogenic (LP) variants were initially identified in 19.9% of participants, most of which were attributable to previously reported pathogenic variants with implausibly high allele frequencies. After variant annotation and filtering based on population minor allele frequency (MAF ≤ 10-4 for dominant disorders and MAF ≤ 10-3 for recessive disorders), we detected a significant enrichment of P/LP variants in the CLD cohort compared to the HC cohort (X2 test OR 5.00, 95% CI 3.06-8.18, p value = 4.5e-12). A second-level manual annotation was necessary to capture true pathogenic variants that were removed by stringent allele frequency and quality filters. After these sequential steps, the diagnostic rate of monogenic disorders was 5.7% in the CLD cohort, attributable to P/LP variants in 25 genes. We also identified concordant liver disease phenotypes for 15/22 kidney disease patients with P/LP variants in liver genes, mostly associated with cystic liver disease phenotypes. Sequencing results had many implications for clinical management, including familial testing for early diagnosis and management, preventative screening for associated comorbidities, and in some cases for therapy. Exome sequencing provided a 5.7% diagnostic rate in CLD patients and required multiple rounds of review to reduce both false positive and false negative findings. The identification of concordant phenotypes in many patients with P/LP variants and no known liver disease also indicates a potential for predictive testing for selected monogenic liver disorders.


Simmons' tamponade shell for leaking filtration blebs.
We used the Simmons' tamponade shell to successfully treat four of five patients who had leaking filtration blebs after glaucoma surgery. Bleb leaks in both the immediate postoperative period as well as many years following surgery were treated. In each case, previous pressure patching had been unsuccessful. Permanent closure of the leaks with the shell tamponade occurred within three days. Use of this technique is an effective alternative to surgical repair.


Bridge populations in the spread of HIV/AIDS in Thailand.
To determine the extent to which men provide a bridge population between commercial sex workers (CSW) and the general female population in Thailand. Sexual network and serological data were collected from a systematic quota sample of low income men and truckers during 1992 in three Thailand provinces. Completed sample size was 1075 men aged 17-45 years and 330 truckers. Sexual network information was used to identify those men who have sex with both female CSW and non-CSW partners (the 'bridge population'). A new method was used for calculating the partner acquisition rate and to establish the potential number of women exposed to HIV via inconsistent condom use among the bridge population. Approximately 17% of men and 25% of truckers can be included in the bridge population. These men are more likely to be HIV-positive and to have had at least one other sexually transmitted infection in the past year (odds ratio, 2.2 and 3.4, respectively). Consistent condom use with CSW is less than 30%, and is less than 1% with non-CSW partners. As a result, 30 women in the general population were potentially exposed to HIV per 100 sexually active men in the last year:nine women each additional year. Younger men and truckers expose almost twice as many women to HIV; more female peers than wives are exposed. Bridge populations may be as important as "core groups' for the spread of HIV into the general Thai population. Young men and women are strategic intervention targets because they have more partners, are more likely to be in bridging networks, and are more receptive to condom use.


T cell specificity in twice-irradiated F1----parent bone marrow chimeras: failure to detect a role for immigrant marrow-derived cells in imprinting intrathymic H-2 restriction.
In an attempt to resolve the issue of whether H-2-restricted T cell specificity is controlled by thymic epithelial cells or by cells of the macrophage/dendritic cell (M phi/DC) lineages, long-term F1----parent chimeras were subjected to secondary irradiation and reconstitution with F1 marrow cells. The rationale was that if F1 M phi/DC enter the thymus only quite slowly after irradiation, as claimed by other investigators, leaving F1----parent chimeras for a period of several months before re-irradiation would ensure that the new wave of T cells generated in the thymus of the chimeras would have no difficulty in making contact with donor-derived F1 M phi/DC. According to the view that M phi/DC rather than epithelial cells control H-2 restriction, the T cells differentiating in these chimeras would be expected to show H-2 restriction to both parental strains. In practice, T cells from twice-irradiated (1000 + 800 rad) chimeras showed strong restriction to host (thymic) H-2 determinants, the degree of restriction to host determinants being as marked as with T cells from once-irradiated chimeras. This finding applied both to T proliferative responses to KLH assayed in vitro and to T helper function for sheep erythrocytes measured in vivo. Preliminary experiments established that the initial dose of irradiation used for preparing the chimeras (1000 rad) resulted in almost total replacement of intrathymic M phi/DC by donor-derived cells within 4 wk of irradiation; M phi/DC were typed by determining their capacity to stimulate mixed-lymphocyte reactions. Collectively, the data imply that, at least under the conditions used, H-2-restricted T cell specificity is controlled by epithelial cells rather than by M phi/DC.


Creating Designer Engineered Extracellular Vesicles for Diverse Ligand Display, Target Recognition, and Controlled Protein Loading and Delivery.
Efficient and targeted delivery of therapeutic agents remains a bottleneck in modern medicine. Here, biochemical engineering approaches to advance the repurposing of extracellular vesicles (EVs) as drug delivery vehicles are explored. Targeting ligands such as the sugar GalNAc are displayed on the surface of EVs using a HaloTag-fused to a protein anchor that is enriched on engineered EVs. These EVs are successfully targeted to human primary hepatocytes. In addition, the authors are able to decorate EVs with an antibody that recognizes a GLP1 cell surface receptor by using an Fc and Fab region binding moiety fused to an anchor protein, and they show that this improves EV targeting to cells that overexpress the receptor. The authors also use two different protein-engineering approaches to improve the loading of Cre recombinase into the EV lumen and demonstrate that functional Cre protein is delivered into cells in the presence of chloroquine, an endosomal escape enhancer. Lastly, engineered EVs are well tolerated upon intravenous injection into mice without detectable signs of liver toxicity. Collectively, the data show that EVs can be engineered to improve cargo loading and specific cell targeting, which will aid their transformation into tailored drug delivery vehicles.


Parent-child interaction during the first year of life in infants at elevated likelihood of autism spectrum disorder.
Autism spectrum disorder (ASD) likely emerges from a complex interaction between pre-existing neurodevelopmental vulnerabilities and the environment. The interaction with parents forms a key aspect of an infant's social environment, but few prospective studies of infants at elevated likelihood (EL) for ASD (who have an older sibling with ASD) have examined parent-child interactions in the first year of life. As part of a European multisite network, parent-child dyads of free play were observed at 5 months (62 EL infants, 47 infants at typical likelihood (TL)) and 10 months (101 EL siblings, 77 TL siblings). The newly-developed Parent-Infant/Toddler Coding of Interaction (PInTCI) scheme was used, focusing on global characteristics of infant and parent behaviors. Coders were blind to participant information. Linear mixed model analyses showed no significant group differences in infant or parent behaviors at 5 or 10 months of age (all ps≥0.09, d≤0.36), controlling for infant's sex and age, and parental educational level. However, without adjustments, EL infants showed fewer and less clear initiations at 10 months than TL infants (p = 0.02, d = 0.44), but statistical significance was lost after controlling for parental education (p = 0.09, d = 0.36), which tended to be lower in the EL group. Consistent with previous literature focusing on parent-infant dyads, our findings suggest that differences between EL and TL dyads may only be subtle during the first year of life. We discuss possible explanations and implications for future developmental studies.


A comparison of DNA damage in testicular and proximal epididymal spermatozoa in obstructive azoospermia.
Testicular and epididymal spermatozoa are used routinely for intracytoplasmic sperm injection (ICSI) to treat men with obstructive azoospermia. Little is known of the effects of obstruction and stasis on the DNA of these spermatozoa, particularly in the epididymis where spermatozoa have been retained for long periods. Surgical epididymal aspiration for ICSI could provide spermatozoa that are senescent or dying. Using the Comet assay, the percentage of undamaged DNA of testicular spermatozoa from 20 men with obstructive azoospermia was significantly better (83.0 +/- 1. 2%) than from proximal epididymal spermatozoa (75.4 +/- 2.3%; P < 0. 05). There was no difference between the percentage of undamaged DNA of testicular spermatozoa from 39 men with obstructive azoospermia (84.0 +/- 0.9) or from 10 fertile men at vasectomy (86.8 +/- 1.8) or from ejaculated spermatozoa from five of the controls (78.9 +/- 3.9; P > 0.05). In nine subjects, a second biopsy was carried out 6 months later. There was no significant difference in undamaged DNA on these two occasions (83.5 +/- 5.6 and 84.1 +/- 4.2; P > 0.05). This confirms the reproducibility of the Comet assay for non-ejaculated spermatozoa. Our data suggest that testicular sperm DNA appears to be significantly less damaged than epididymal sperm DNA, and so testicular spermatozoa should be used in preference for ICSI to treat men with obstructive azoospermia.


Development and characterization of an innovative heparin coating to stabilize and protect liposomes against adverse immune reactions.
Liposomes have been recognized as excellent drug delivery systems, but when they come in direct contact with different blood components they may trigger an immediate activation of the innate immune system. The aim of the present study was to produce long-circulating, blood-compatible liposomes by developing a construct of liposomes covered by a novel unique heparin complex (CHC; 70 heparin molecules per complex) to avoid recognition by the innate immune system. Unilamellar, cationic liposomes were produced by hand extrusion through a 100-nm polycarbonate membrane. Coating of liposomes with the macromolecular CHC was accomplished by electrostatic interactions. Dynamic light scattering as well as QCM-D measurements were used to verify the electrostatic deposition of the negatively charged CHC to cationic liposomes. The CHC-coated liposomes did not aggregate when in contact with lepirudin anti-coagulated plasma. Unlike previous attempts to coat liposomes with heparin, this technique produced freely moveable heparin strands sticking out from the liposome surface, which exposed AT binding sites reflecting the anticoagulant potentials of the liposomes. In experiments using lepirudin-anticoagulated plasma, CHC-coated liposomes, in contrast to non-coated control liposomes, did not activate the complement system, as evidenced by low C3a and sC5b-9 generation and reduced leakage from the liposomes. In conclusion, we show that liposomes can be successfully coated with the biopolymer CHC, resulting in biocompatible and stable liposomes that have significant application potential.


Health service utilization and reported satisfaction among adolescents in Dejen District, Ethiopia: a cross-sectional study.
There is no adequate health service or counseling specifically suitable for adolescents in Ethiopia. Adolescents' satisfaction on the health service provided is important to increase utilization and quality of care. The objective of this study was to assess health service utilization, reported satisfaction and predictors of satisfaction among adolescents of 15-19 years in Dejen District. A community based cross-sectional study was done from February 05 to 17, 2012. Interview method was used to collect data from 690 adolescents. Following stratification into urban and rural, six kebeles were selected by lottery method. Study participants allocated proportionally to households' size of kebele. Households were selected randomly, and one from each household was used. Descriptive measures and binary logistic regression were used to identify independent predictors for health service satisfaction. Among 690 adolescents, 313(45%) used health service. Of these, 190 (60.7%) were satisfied. Physical proximity (AOR=3.6, 95% CI: 1.8, 7.3), drug availability (AOR=2.7, 95% CI: 1.3, 5.8), health services availability (AOR=2.5, 95% CI: 1.1, 6.0), treatment in separate room (AOR=2.9, 95% CI: 1.4, 5.6), checked all adolescents problem (AOR=4.0, 95% CI: 2.0, 8.5), treated with respect (AOR=3.0, 95% CI: 1.4, 5.7) and opportunity to explain feeling (AOR=3.3, 95% CI: 1.7, 6.6) were predictors of satisfaction. Adolescents' health service utilization and satisfaction were low. Adolescents' perception of accessibility, acceptability and interaction with health workers' had significant influence on health services satisfaction. Therefore, health professionals and administrators should work on availing services with close proximity and acceptable behavior in order to increase adolescents' satisfaction.


Enhanced cellular uptake of remnant high-density lipoprotein particles: a mechanism for high-density lipoprotein lowering in insulin resistance and hypertriglyceridemia.
A low level of high-density lipoprotein (HDL) cholesterol is characteristic of insulin resistance and hypertriglyceridemia and likely contributes to the increased risk of cardiovascular disease associated with these conditions. One pathway involves enhanced clearance of lipolytically modified HDL particles, but the underlying mechanisms remain poorly understood. Here, we examine the effect of triglyceride enrichment and hepatic lipase hydrolysis on HDL binding, internalization, and degradation in cultured liver and kidney cells. Maximal binding of remnant HDL (HDL enriched with triglycerides followed by hepatic lipase hydrolysis), but not binding affinity, was markedly higher than native and triglyceride-rich HDL in both HepG2 cells and HEK293 cells. Compared with native and triglyceride-rich HDL, remnant HDL was internalized to a greater extent in both cell types and was more readily degraded in HEK293 cells. The increased binding of remnant HDL was not mediated by the low-density lipoprotein receptor or scavenger receptor class B type I (SR-BI), because enhanced remnant HDL binding was observed in low-density lipoprotein receptor-deficient cells with or without SR-BI overexpression. Disruption of cell surface heparan sulfate proteoglycans or blockage of apolipoprotein E-mediated lipoprotein binding also did not abolish the enhanced remnant HDL binding. Our observations indicate that remodeling of triglyceride-enriched HDL by hepatic lipase may result in enhanced binding, internalization, and degradation in tissues involved in HDL catabolism, contributing to rapid clearance and overall lowering of plasma HDL cholesterol in insulin resistance and hypertriglyceridemia.


Dominant positive and negative selection using a hygromycin phosphotransferase-thymidine kinase fusion gene.
The hygromycin phosphotransferase gene was fused in-frame with the herpes simplex virus type 1 thymidine kinase gene. The resulting fusion gene (termed HyTK) confers hygromycin B resistance for dominant positive selection and ganciclovir sensitivity for negative selection and provides a means by which these selectable phenotypes may be expressed and regulated as a single genetic entity.


Biotransformation in carcinogen-induced diploid and polyploid hepatocytes separated by centrifugal elutriation.
Biotransformation in carcinogen-induced diploid and polyploid hepatocytes was studied using isozyme-selective substrates for several enzyme pathways. Diploid hepatocytes were induced by partial hepatectomy, a single injection of diethylnitrosamine, and 4 weeks of 2-acetylaminofluorene (2-AAF) feeding. Then, after an additional 3-5 weeks on the control diet, diploid and polyploid hepatocytes were separated from freshly isolated hepatocytes by centrifugal elutriation. Benzo(a)pyrene hydroxylase, ethoxyresorufin O-deethylase, and methoxycoumarin O-demethylase activities were approximately 15-40% lower in the diploid hepatocyte fraction than in the polyploid cell fraction. Activities of 1-chloro-2,4-dinitrobenzene, glutathione S-transferase, 3-hydroxy-benzo(a)pyrene or 4-hydroxybiphenyl UDP-glucuronosyltransferase, and DT-diaphorase were not different in the two cell fractions. Determination of activity during the 2-AAF treatment indicated that 2-AAF increased 7-ethoxyresorufin O-deethylase and 3-hydroxybenzo(a)pyrene glucuronosyltransferase activities by 300 and 200%, respectively, in both the diploid and polyploid hepatocyte fractions. Administration of phenobarbital for 4 days at the end of the control diet period increased ethoxyresorufin and methoxycoumarin dealkylations by 2- and 4-fold, and 3-hydroxybenzo(a)pyrene glucuronidation and 1-chloro-2,4-dinitrobenzene conjugation with glutathione by 1.5- to 2-fold in both hepatocyte fractions. Slight increases in benzo(a)pyrene hydroxylation and 4-hydroxybiphenyl glucuronidation were also evident in diploid cells. Although there is a slight decrease in cytochrome P-450-dependent monooxygenase activities, these data indicate that carcinogen-induced diploid hepatocytes do not show the typical toxicant-resistant phenotype observed in preneoplastic hepatocytes of altered liver foci, which are characterized by large decreases in monooxygenase biotransformations as well as increased activities of several phase II enzymes. This finding is compatible with the hypothesis that 2-AAF-induced nonploidizing growth of diploid hepatocytes is caused by nontoxic mechanisms in the present experimental paradigm. In addition, carcinogen-induced diploid cells respond to phenobarbital in a manner similar to that of polyploid hepatocytes.


Genome-wide analysis of transposable elements and tandem repeats in the compact placozoan genome.
The placozoan Trichoplax adhaerens has a compact genome with many primitive eumetazoan characteristics. In order to gain a better understanding of its genome architecture, we conducted a detailed analysis of repeat content in this genome. The transposable element (TE) content is lower than that of other metazoans, and the few TEs present in the genome appear to be inactive. A new phylogenetic clade of the gypsy-like LTR retrotransposons was identified, which includes the majority of gypsy-like elements in Trichoplax. A particular microsatellite motif (ACAGT) exhibits unexpectedly high abundance, and also has strong association with its nearby genes.


Open resource metagenomics: a model for sharing metagenomic libraries.
Both sequence-based and activity-based exploitation of environmental DNA have provided unprecedented access to the genomic content of cultivated and uncultivated microorganisms. Although researchers deposit microbial strains in culture collections and DNA sequences in databases, activity-based metagenomic studies typically only publish sequences from the hits retrieved from specific screens. Physical metagenomic libraries, conceptually similar to entire sequence datasets, are usually not straightforward to obtain by interested parties subsequent to publication. In order to facilitate unrestricted distribution of metagenomic libraries, we propose the adoption of open resource metagenomics, in line with the trend towards open access publishing, and similar to culture- and mutant-strain collections that have been the backbone of traditional microbiology and microbial genetics. The concept of open resource metagenomics includes preparation of physical DNA libraries, preferably in versatile vectors that facilitate screening in a diversity of host organisms, and pooling of clones so that single aliquots containing complete libraries can be easily distributed upon request. Database deposition of associated metadata and sequence data for each library provides researchers with information to select the most appropriate libraries for further research projects. As a starting point, we have established the Canadian MetaMicroBiome Library (CM(2)BL [1]). The CM(2)BL is a publicly accessible collection of cosmid libraries containing environmental DNA from soils collected from across Canada, spanning multiple biomes. The libraries were constructed such that the cloned DNA can be easily transferred to Gateway® compliant vectors, facilitating functional screening in virtually any surrogate microbial host for which there are available plasmid vectors. The libraries, which we are placing in the public domain, will be distributed upon request without restriction to members of both the academic research community and industry. This article invites the scientific community to adopt this philosophy of open resource metagenomics to extend the utility of functional metagenomics beyond initial publication, circumventing the need to start from scratch with each new research project.


Steric and electrostatic interactions govern nanofiltration of amino acids.
Crossflow nanofiltration experiments were performed to investigate the factors influencing the removal of amino acids by a commercially available polymeric thin-film composite membrane. The removals of five monoprotic (Ala, Val, Leu, Gly, and Thr), one diprotic (Asp), and one dibasic (Arg) amino acids in a range of permeate fluxes, feed pH values, and ionic strengths were analyzed using a phenomenological model of membrane transport. At any given pH and ionic strength, reflection coefficients (rejection at asymptotically infinite flux) of monoprotic amino acids increased with molar radius demonstrating the role of steric interactions on their removal. Additionally, consistent with Donnan exclusion, higher reflection coefficients were obtained when the membrane and the amino acids both carried the same nature of charge (positive or negative). In other words, both co-ion repulsion and molecular size determined amino acids removal. Importantly, the removal of effectively neutral amino acids were significantly higher than neutral sugars and alcohols of similar size demonstrating that even near their isoelectric point, zwitterionic characteristics preclude them from being considered as strictly neutral.


The long-term effect of a plaque control program on tooth mortality, caries and periodontal disease in adults. Results after 30 years of maintenance.
The biofilm that forms and remains on tooth surfaces is the main etiological factor in caries and periodontal disease. Prevention of caries and periodontal disease must be based on means that counteract this bacterial plaque. To monitor the incidence of tooth loss, caries and attachment loss during a 30-year period in a group of adults who maintained a carefully managed plaque control program. In addition, a comparison was made regarding the oral health status of individuals who, in 1972 and 2002, were 51-65 years old. In 1971 and 1972, more than 550 subjects were recruited. Three hundred and seventy-five subjects formed a test group and 180 a control group. After 6 years of monitoring, the control group was discontinued but the participants in the test group was maintained in the preventive program and was finally re-examined after 30 years. The following variables were studied at Baseline and after 3, 6, 15 and 30 years: plaque, caries, probing pocket depth, probing attachment level and CPITN. Each patient was given a detailed case presentation and education in self-diagnosis. Once every 2 months during the first 2 years, once every 3-12 months during years 3-30, the participants received, on an individual need basis, additional education in self-diagnosis and self-care focused on proper plaque control measures, including the use of toothbrushes and interdental cleaning devices (brush, dental tape, toothpick). The prophylactic sessions that were handled by a dental hygienist also included (i) plaque disclosure and (ii) professional mechanical tooth cleaning including the use of a fluoride-containing dentifrice/paste. Few teeth were lost during the 30 years of maintenance; 0.4-1.8 in different age cohorts. The main reason for tooth loss was root fracture; only 21 teeth were lost because of progressive periodontitis or caries. The mean number of new caries lesions was 1.2, 1.7 and 2.1 in the three groups. About 80% of the lesions were classified as recurrent caries. Most sites, buccal sites being the exception, exhibited no sign of attachment loss. Further, on approximal surfaces there was some gain of attachment between 1972 and 2002 in all age groups. The present study reported on the 30-year outcome of preventive dental treatment in a group of carefully monitored subjects who on a regular basis were encouraged, but also enjoyed and recognized the benefit of, maintaining a high standard of oral hygiene. The incidence of caries and periodontal disease as well as tooth mortality in this subject sample was very small. Since all preventive and treatment efforts during the 30 years were delivered in one private dental office, caution must be exercised when comparisons are made with longitudinal studies that present oral disease data from randomly selected subject samples.


Linezolid resistance in Mycobacterium tuberculosis isolates at a tertiary care centre in Mumbai, India.
Linezolid (LZD) is increasingly being used in tuberculosis (TB) treatment. However, LZD resistance has already been reported, which is highly alarming, given its critical therapeutic role. This study was aimed to phenotypically and genotypically assess LZD resistance in Mycobacterium tuberculosis (MTB) isolates at a laboratory in a tertiary care centre in Mumbai, India. A sample of 32 consecutive LZD-resistant MTB isolates identified by liquid culture susceptibility testing was subjected to whole-genome sequencing (WGS) on the Illumina NextSeq platform. Sequences were analyzed using BioNumerics software to predict resistance for 12 antibiotics within 15 min. Sixty eight of the 2179 isolates tested for LZD resistance by MGIT-based susceptibility testing (June 2015 to June 2016) were LZD-resistant. Thirty two consecutive LZD-resistant isolates were analyzed by WGS to screen for known mutations conferring LZD resistance. WGS of 32 phenotypically LZD-resistant isolates showed that C154R in the rplC gene and G2814T in the rrl gene were the major resistance determinants. LZD resistance poses an important risk to the success of treatment regimens, especially those designed for resistant isolates; such regimens are extensively used in India. As LZD-containing regimens increase in prominence, it is important to support clinical decision-making with an improved understanding of the common mutations conferring LZD resistance and their frequency in different settings.


Pressure-Induced Collapse Transition in BaTi2Pn2O (Pn = As, Sb) with an Unusual Pn-Pn Bond Elongation.
Making and breaking bonds in a solid-state compound greatly influences physical properties. A well-known playground for such bonding manipulation is the ThCr2Si2-type structure AT2X2, allowing a collapse transition where a X-X dimer forms by a chemical substitution or external stimuli. Here, we report a pressure-induced collapse transition in the structurally related BaTi2Pn2O (Pn = As, Sb) at a transition pressure Pc of ∼15 GPa. The Pn-Pn bond formation is related with Pn-p band filling, which is controlled by charge transfer from the Ti-3d band. At Pc, the Sb-Sb distance in BaTi2Sb2O shrinks due to bond formation, but interestingly, the Sb-Sb expands with increasing pressure above Pc. This expansion, which was not reported in ThCr2Si2-type compounds, may arise from heteroleptic coordination geometry around titanium, where a compression of the Ti-O bond plays a role. Electrical resistivity measurements of BaTi2Sb2O up to 55 GPa revealed an increasing trend of the superconducting transition temperature with pressure. This study presents structure motifs that allow flexible bonding manipulation and property control with heteroleptic coordination geometry.


Long-term monitoring (1960-2008) of the river-sediment transport in the Red River Watershed (Vietnam): temporal variability and dam-reservoir impact.
The Red River (China/Vietnam, A=155,000 km(2)) is a typical humid tropics river originating from the mountainous area of Yunnan Province in China. Based on information on daily discharge (Q) and suspended particulate matter (SPM) concentration between 1960-2008 for the SonTay gauging station (outlet of the River and entry to the Delta) provided by the National Institute IMHE-MONRE, the mean annual SPM flux was estimated at 90 Mt/yr, corresponding to a sediment yield of 600 t/km(2)/yr. The temporal variability of annual SPM fluxes (ranging from 24 to 200 Mt/yr) is strongly related to the interannual hydrological conditions. However, some years of high water flow were not associated with high sediment fluxes, especially after 1989 when the HoaBinh dam came into operation. Therefore, the median discharge pre- (3389 m(3)/s) and post 1989 (3495 m(3)/s) are similar indicating there was little or no change between both periods. Sediment rating curves (power law-type; SPM=aQ(b)) were fitted for both periods (1960-1989; 1990-2008). The analysis of the pre- and post-1989 sediment rating parameters (a, b) suggests a downshift of b-parameter values after 1989, attributed to a decrease of the sediment supply due to the commissioning of the HoaBinh dam. A single sediment rating curve derived from 1960-1989 data was used to simulate the annual variability of former sediment delivery, generating excellent cumulative flux estimates (error ~1%). In contrast, applying the same rating curve to the 1990-2008 data resulted in systematic and substantial (up to 109%) overestimation. This suggests that the HoaBinh dam reduces annual SPM delivery to the delta by half, implying important metal/metalloid storage behind the HoaBinh dam.


Estimation of water quality variables based on machine learning model and cluster analysis-based empirical model using multi-source remote sensing data in inland reservoirs, South China.
Reservoirs play important roles in the drinking water supply for urban residents, agricultural water provision, and the maintenance of ecosystem health. Satellite optical remote sensing of water quality variables in medium and micro-sized inland waters under oligotrophic and mesotrophic status is challenging in terms of the spatio-temporal resolution, weather conditions and frequent nutrient status changes in reservoirs, etc., especially when quantifying non-optically active components (non-OACs). This study was based on the surface reflectance products of unmanned aerial vehicle (UAV) multispectral images, Sentinel-2B Multispectral instrument (MSI) images and Landsat 7 Enhanced Thematic Mapper Plus (ETM+) by utilizing fuzzy C-means (FCM) clustering algorithm was combined with band combination (BC) model to construct the FCM-BC empirical model, and used mixed density network (MDN), extreme gradient boosting (XGBoost), deep neural network (DNN) and support vector regression (SVR) machine learning (ML) models to invert 12 kinds of optically active components (OACs) and non-OACs. Compared with the unclustered BC (UC) model, the mean coefficient of determination (MR) of the FCM-BC models was improved by at least 46.9%. MDN model showed best accuracy (R2 in the range of 0.60-0.98) and stability (R2 decreased by up to 13.2%). The accuracy of UAV was relatively higher in both empirical methods and machine learning methods. Additionally, the spatio-temporal distribution maps of four water quality variables were mapped based on the MDN model and UAV images, all platforms showed good consistency. An inversion strategy of water quality variables in various monitoring frequencies and weather conditions were proposed finally. The purpose of introducing the UAV platform was to cooperate with the satellite to improve the monitoring response ability of OACs and non-OACs in small and micro-sized oligotrophic and mesotrophic water bodies.


[Immunomodulators and cytokines in the treatment of internal diseases, associated with immunologic status disturbance in elderly patients].
The last years witnessed significant change in therapeutic and infectious pathology of man, its immunoreactivity changed, especially in elderly patients. Thus, older people show the increase of chronic infectious-inflammatory diseases, characterized by flaccid, constantly recurring course, torpid to adequate etiotopic therapy. They are caused by conventionally-pathogenic or opportunistic microbes, often with atypical biological properties, having multiple resistance to antibiotics. About 30 new pathogenic microorganisms were revealed for the last 20-30 years, including HIV-infection agent. The number of patients with allergic and autoimmune diseases increased, thus resulting in high urgency of this problem in gerontology. Decrease of immunologic reactivity, observed practically in all parts of the world, but mostly manifested in Russia, is one of the causes of infectious pathology, including pathology of inner organs, manifestation of allergic and autoimmune diseases. General practitioners, gerontologists understand that to cope with the increased rate of infectious, allergic or autoimmune diseases only by antibiotics or anti-inflammatory medicines is next to impossible. It's clear now that the use of only chemotherapeutic drugs in treatment of elderly and therapeutic patients is a mistake. Immunocorrective medicines, which stimulate reparative processes, allow to accelerate the processes of involution and shorten the general period of treatment. Antibiotic inhibits the agent multiplication, but its complete elimination from the body is a result of immune factors activity. In elderly patients with inner organ pathology and chronic pathology on the background of the suppressed immunoreactivity the effect of antibiotics, as well as antifungal, antiviral and other chemotherapeutic medicines is ineffective or of low effect. Optimal clinical effect in elderly patients can obviously be achieved only if there is synergism in the activity of the body protective forces and antimicrobial drugs. Thus, immunoreactivity dysfunction has a significant part in the pathogenesis of inner organ diseases in elderly patients with the disturbance of immunological status. So in complex of medical means, indicated for this pathology, adequate immunotherapy plays a special role, and the main problem is the choice of particular drugs.


[The role of atherosclerosis risk factors in development of ischemic heart disease in young men].
To determine distribution of different atherosclerosis risk factors and their correlation with severity of coronary arteries (CA) affection in young men. A total of 235 males aged 28-45 years were examined with coronarography which detected more than 50% narrowing in one or several major CA in 200 patients (the study group with coronary artery disease), 35 males without coronary stenosis served control. Atherosclerosis risk factors, serum lipids, lipoprotein(a), fibrinogen, C-reactive protein (CRP) were assessed. Smoking and hyperlipidemia (HLE) were registered in the study group more frequently. The level of lipoprotein(a) above 30 mg/dl was recorded in 98 (49%) and 7 (20%) patients of the study and control group, respectively (p = 0.001). As shown by a correlation analysis, the number of affected CA was associated with smoking (r = 0.2; p < 0.001), HLE (r = 0.23; p < 0.001), concentration of total cholesterol (r = 0.23; p < 0.0.001), lipoprotein (a) (r = 0.26; p < 0.001) and CRP (r = 0.26; p < 0.05). The level of lipoprotein(a) in patients with myocardial infarction and occluded artery was significantly higher than in patients without myocardial infarction and non-occlusive CA affection. The presence and severity of coronary atherosclerosis in young men with coronary artery disease are associated with smoking, HLE and high concentration of lipoprotein (a).


Targeting interferon-alpha to dendritic cells enhances a CD8+ T cell response to a human CD40-targeted cancer vaccine.
Targeting antigens to antigen presenting cells (APC) enhances the potency of recombinant protein CD8+ T cell vaccines. Recent comparisons of recombinant protein-based dendritic cell (DC) targeting vaccines revealed differences in cross-presentation and identified CD40 as a potent human DC receptor target for antigen cross-presentation. Contrary to in vitro-derived monocyte (mo)DC, we found that interferon-alpha (IFNα) stimulation of human blood-derived DC was necessary for an antigen-specific IFNγ CD8+ T cell response to a CD40 targeted cancer vaccine. Importantly, targeting an adjuvant in the form of IFNα to DC increased their potency to elicit antigen-specific production of IFNγ by CD8+ T cells. Thus, we introduce the concept of DC adjuvant targeting to enhance the potency of vaccination.


Collision energy dependence of state-to-state differential cross sections for rotationally inelastic scattering of H2O by He.
The inelastic scattering of H2O by He as a function of collision energy in the range 381 cm-1 to 763 cm-1 at an energy interval of approximately 100 cm-1 has been investigated in a crossed beam experiment using velocity map imaging. Change in collision energy was achieved by varying the collision angle between the H2O and He beam. We measured the state-to-state differential cross section (DCS) of scattered H2O products for the final rotational states JKaKc = 110, 111, 221 and 414. Rotational excitation of H2O is probed by (2 + 1) resonance enhanced multiphoton ionization (REMPI) spectroscopy. DCS measurements over a wide range of collision energies allowed us to probe the H2O-He potential energy surface (PES) with greater detail than in previous work. We found that a classical approximation of rotational rainbows can predict the collision energy dependence of the DCS. Close-coupling quantum mechanical calculations were used to produce DCS and partial cross sections. The forward-backward ratio (FBR), is introduced here to compare the experimental and theoretical DCS. Both theory and experiments suggest that an increase in the collision energy is accompanied with more forward scattering.


Hesperidin Is a Potential Inhibitor against SARS-CoV-2 Infection.
Hesperidin (HD) is a common flavanone glycoside isolated from citrus fruits and possesses great potential for cardiovascular protection. Hesperetin (HT) is an aglycone metabolite of HD with high bioavailability. Through the docking simulation, HD and HT have shown their potential to bind to two cellular proteins: transmembrane serine protease 2 (TMPRSS2) and angiotensin-converting enzyme 2 (ACE2), which are required for the cellular entry of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Our results further found that HT and HD suppressed the infection of VeroE6 cells using lentiviral-based pseudo-particles with wild types and variants of SARS-CoV-2 with spike (S) proteins, by blocking the interaction between the S protein and cellular receptor ACE2 and reducing ACE2 and TMPRSS2 expression. In summary, hesperidin is a potential TMPRSS2 inhibitor for the reduction of the SARS-CoV-2 infection.


Overcoming the Limits of Hypoxia in Photodynamic Therapy: A Carbonic Anhydrase IX-Targeted Approach.
A major challenge in photodynamic cancer therapy (PDT) is avoiding PDT-induced hypoxia, which can lead to cancer recurrence and progression through activation of various angiogenic factors and significantly reduce treatment outcomes. Reported here is an acetazolamide (AZ)-conjugated BODIPY photosensitizer (AZ-BPS) designed to mitigate the effects of PDT-based hypoxia by combining the benefits of anti-angiogenesis therapy with PDT. AZ-BPS showed specific affinity to aggressive cancer cells (MDA-MB-231 cells) that overexpress carbonic anhydrase IX (CAIX). It displayed enhanced photocytotoxicity compared to a reference compound, BPS, which is an analogous PDT agent that lacks an acetazolamide unit. AZ-BPS also displayed an enhanced in vivo efficacy in a xenograft mouse tumor regrowth model relative to BPS, an effect attributed to inhibition of tumor angiogenesis by both PDT-induced ROS generation and CAIX knockdown. AZ-BPS was evaluated successfully in clinical samples collected from breast cancer patients. We thus believe that the combined approach described here represents an attractive therapeutic approach to targeting CAIX-overexpressing tumors.


Controlled release matrix tablets of zidovudine: effect of formulation variables on the in vitro drug release kinetics.
The purpose of this research was to design oral controlled release (CR) matrix tablets of zidovudine (AZT) using hydroxypropyl methylcellulose (HPMC), ethyl cellulose (EC) and carbopol-971P (CP) and to study the effect of various formulation factors on in vitro drug release. Release studies were carried out using USP type 1 apparatus in 900 ml of dissolution media. Release kinetics were analyzed using zero-order, Higuchi's square root and Ritger-Peppas' empirical equations. Release rate decreased with increase in polymer proportion and compression force. The release rate was lesser in formulations prepared using CP (20%) as compared to HPMC (20%) as compared to EC (20%). No significant difference was observed in the effect of pH of dissolution media on drug release from formulations prepared using HPMC or EC, but significant difference was observed in CP based formulations. Decrease in agitation speed from 100 to 50 rpm decreased release rate from HPMC and CP formulations but no significant difference was observed in EC formulations. Mechanism of release was found to be dependent predominantly on diffusion of drug through the matrix than polymer relaxation incase of HPMC and EC formulations, while polymer relaxation had a dominating influence on drug release than diffusion incase of CP formulations. Designed CR tablets with pH independent drug release characteristics and an initial release of 17-25% in first hour and extending the release up to 16-20 h, can overcome the disadvantages associated with conventional tablets of AZT.


Rapid diffusion of CH4/H2 mixtures in single-walled carbon nanotubes.
Equilibrium molecular dynamics (EMD) are used to examine the self-diffusion and macroscopic diffusion of CH4/H2 mixtures adsorbed inside a (10,10) single-walled carbon nanotube. EMD can be used to determine the macroscopic diffusion coefficients of adsorbed mixtures by evaluating the matrix of Onsager transport coefficients. Earlier studies have indicated the diffusion of light gases adsorbed as single components in carbon nanotubes is extremely rapid compared to that in other known nanoporous materials. The results presented here indicate that extremely rapid diffusion can also occur for mixtures of adsorbed molecules. The rapid diffusion of adsorbed molecules and the strong coupling between the fluxes of the adsorbed species in a mixture have interesting implications for uses of carbon nanotubes in membrane-based applications.


A superfast muscle in the complex sonic apparatus of Ophidion rochei (Ophidiiformes): histological and physiological approaches.
In teleosts, superfast muscles are generally associated with the swimbladder wall, whose vibrations result in sound production. In Ophidion rochei, three pairs of muscles were named 'sonic' because their contractions affect swimbladder position: the dorsal sonic muscle (DSM), the intermediate sonic muscle (ISM), and the ventral sonic muscle (VSM). These muscles were investigated thanks to electron microscopy and electromyography in order to determine their function in sound production. Fibers of the VSM and DSM were much thinner than the fibers of the ISM and epaxial musculature. However, only VSM fibers had the typical ultrastructure of superfast muscles: low proportion of myofibrils, and high proportions of sarcoplasmic reticulum and mitochondria. In females, each sound onset was preceded by the onset of electrical activity in the VSM and the DSM (ISM was not tested). The electromyograms of the VSM were very similar to the waveforms of the sounds: means for the pulse period were 3.6±0.5 and 3.6±0.7 ms, respectively. This shows that the fast VSM (ca. 280 Hz) is responsible for the pulse period and fundamental frequency of female sounds. DSM electromyograms were generally characterized by one or two main peaks followed by periods of lower electrical activity, which suggests a sustained contraction over the course of the sound. The fiber morphology of the ISM and its antagonistic position relative to the DSM are not indicative of a muscle capable of superfast contractions. Overall, this study experimentally shows the complexity of the sound production mechanism in the nocturnal fish O. rochei.


[Establishment of platelet-mediated transmitochondrial cell model].
To establish a transmitochondrial cell model for further researches on molecular genetics of mitochondrial related disease. The fusion process was conducted between mitochondrial DNA-lacking rho degrees cell (a gift from NIH) and platelet using polyethylene glycol as fusion promoting reagent. The fusion cells were confirmed by PCR and electronic microscopic cytochemistry. The mitochondrial morphology and function of 3 families of Rett syndrome were investigated. The platelet-mediated transmitochondrial cell model was constructed successfully. The frequency of transformation ranged from 0.5 to 1.6 clones in 10(4) recipient cells. The mitochondrial vacuolation was occasionally observed in 2 cases of Rett syndrome. Transmitochondrial cell model can be applied to assessment of the mitochondrial morphology and function of fusion cells and is found to be of great use in evaluating the gene expression of mitochondrial genome at different levels.


Dominant suppressors of a muscle mutant define an essential gene of Caenorhabditis elegans.
The sup-11 1 locus of C. elegans was defined by rare dominant suppressors of unc-93(e1500) III, a mutation that affects muscle structure. All ten of these dominant suppressors have a recessive "scrawny" phenotype. Two additional classes of sup-11 alleles were identified. One class, null alleles, was obtained by reversion of the dominant suppressor activity. These null alleles are recessive embryonic lethals, indicating that sup-11 is an essential gene. Members of the second class, rare semidominant revertants of the "scrawny" phenotype, are partial suppressors of unc-93(e1500). The genetic properties of the dominant suppressor mutations suggest that they are rare missense mutations that confer a novel activity to the sup-11 protein. We consider some of the ways that sup-11 alleles might suppress unc-93(e1500), including the possibilities that the altered sup-11 proteins restore function to a protein complex or are modified products of a gene that is a member of an unc-93 gene family.


Enhancement of class II-restricted T cell responses by costimulatory NK receptors for class I MHC proteins.
An important feature of the human immune system is the ability of T cells to respond to small quantities of antigen. Class II major histocompatibility complex (MHC)-restricted T cells that expressed a costimulatory natural killer (NK) cell receptor for class I MHC proteins were cloned. In the presence of low doses of superantigen, the proliferative response of these T cell clones was three- to ninefold greater when the T cells were costimulated by way of the NK receptor. Thus, the action of costimulatory NK receptors on T cells may play a significant role in initiating and sustaining immune responses.


EXPERIMENTAL RUNT DISEASE IN MICE CAUSED BY SALMONELLA TYPHIMURIUM, VAR. COPENHAGEN.
The strain of Salmonella typhimurium isolated from the subcutaneous abscess of a runted mouse and used in this study was somewhat unusual, but not unique, in that it had a high virulence for young mice, yet low infectivity. This strain could mimic many of the features, signs, and symptoms of immunological runting when injected into neonates, either in pure culture, or when mixed with spleen cells, or when present in infected isologous or F(1) hybrid spleen cells. Thus, the incidence of Salmonella runting was dose-dependent and related to the age of the neonate. Runts failed to gain weight, were sickly, and usually died within 30 days. They had a marked splenomegaly and hepatomegaly associated with areas of necrosis. However, in marked contrast to immunological runts they did not have lymphoid atrophy. The incidence of runting was diminished when frozen-thawed spleen cell suspensions were used, but not with sonicated or heated suspensions or spleen cells from lethally irradiated mice. Runting could be prevented by immunizing breeders with S. typhimurium, and serum from mice immunized against S. typhimurium protected neonates injected with this organism. Isologous adult spleen cells did not protect against Salmonella runting. It is suggested that in studies on runting only the intravenous route be used and that heated cells serve as a control. More rigid criteria should be applied to runting than those frequently accepted and mice should be autopsied whenever possible.


Association of multiple cellular stress pathways with accelerated atherosclerosis in hyperhomocysteinemic apolipoprotein E-deficient mice.
A causal relation between hyperhomocysteinemia (HHcy) and accelerated atherosclerosis has been established in apolipoprotein E-deficient (apoE-/-) mice. Although several cellular stress mechanisms have been proposed to explain the atherogenic effects of HHcy, including oxidative stress, endoplasmic reticulum (ER) stress, and inflammation, their association with atherogenesis has not been completely elucidated. ApoE-/- mice were fed a control or a high-methionine (HM) diet for 4 (early lesion group) or 18 (advanced lesion group) weeks to induce HHcy. Total plasma homocysteine levels and atherosclerotic lesion size were significantly increased in early and advanced lesion groups fed the HM diet compared with control groups. Markers of ER stress (GRP78/94, phospho-PERK), oxidative stress (HSP70), and inflammation (phospho-IkappaB-alpha) were assessed by immunohistochemical staining of these atherosclerotic lesions. GRP78/94, HSP70, and phospho-IkappaB-alpha immunostaining were significantly increased in the advanced lesion group fed the HM diet compared with the control group. HSP47, an ER-resident molecular chaperone involved in collagen folding and secretion, was also increased in advanced lesions of mice fed the HM diet. GRP78/94 and HSP47 were predominantly localized to the smooth muscle cell-rich fibrous cap, whereas HSP70 and phospho-IkappaB-alpha were observed in the lipid-rich necrotic core. Increased HSP70 and phospho-IkappaB-alpha immunostaining in advanced lesions of mice fed the HM diet are consistent with enhanced carotid artery dihydroethidium staining. Interestingly, GRP78/94 and phospho-PERK were markedly increased in macrophage foam cells from early lesions of mice fed the control or the HM diet. Multiple cellular stress pathways, including ER stress, are associated with atherosclerotic lesion development in apoE-/- mice.


Genotoxic activity of nickel subsulphide alpha-Ni3S2.
Four mutagenicity tests of alpha-Ni3S2 were performed: the Ames test on five Salmonella typhimurium strains, HPRT test on V79 cells, in vitro chromosomal aberrations on human lymphocytes and the in vivo micronucleus test in mice. The in vitro tests were carried out without metabolic activation. (i) The Ames test (5-1500 micrograms/plate) demonstrated no mutagenic activity, thus confirming previous observations by other authors. (ii) The HPRT test was carried out under standard conditions (3 h exposure) with alpha-Ni3S2 concentrations from 30 to 1000 micrograms/ml. No significant difference was observed with the control cells. Ultrastructural examination revealed alpha-Ni3S2 binding to the cell membrane. Very few particles were found in the cytoplasm but not in the nucleus. (iii) In vitro metaphase analysis in human lymphocytes were performed after exposure to total alpha-Ni3S2 suspension and to the soluble fraction in culture medium with or without fetal calf serum (FCS) (3-100 micrograms/ml, 24 h exposure). Nickel concentrations in the soluble fraction were determined by electrothermal atomic absorption spectrometry. A clastogenic effect of alpha-Ni3S2 became evident under all experimental conditions with a significant additional increase of chromosomal aberrations in 20% FCS complemented medium. No difference was observed between total suspension and soluble fraction. (iv) The micronucleus test confirmed the clastogenic effect of alpha-Ni3S2 in vivo after administration of 250 mg/kg (i.p.). This test revealed a clear increase of micronuclei frequency in polychromatic erythrocytes 24, 48 and 72 h after the treatment. In addition, we observed a statistically significant decrease in the number (%) of polychromatic erythrocytes after 24 and 48 h exposure. The soluble, i.e. cell-entering, fraction seems to play a great part in the clastogenic effect, as has also been shown with other test methods by other authors.


Inhibiting cancer cell hallmark features through nuclear export inhibition.
Treating cancer through inhibition of nuclear export is one of the best examples of basic research translation into clinical application. Nuclear export factor chromosomal region maintenance 1 (CRM1; Xpo1 and exportin-1) controls cellular localization and function of numerous proteins that are critical for the development of many cancer hallmarks. The diverse actions of CRM1 are likely to explain the broad ranging anti-cancer potency of CRM1 inhibitors observed in pre-clinical studies and/or clinical trials (phase I-III) on both advanced-stage solid and hematological tumors. In this review, we compare and contrast the mechanisms of action of different CRM1 inhibitors, and discuss the potential benefit of unexplored non-covalent CRM1 inhibitors. This emerging field has uncovered that nuclear export inhibition is well poised as an attractive target towards low-toxicity broad-spectrum potent anti-cancer therapy.


Sonic Hedgehog signaling pathway as a potential target to inhibit the progression of hepatocellular carcinoma.
Hepatocellular carcinoma (HCC) is one of the leading causes of cancer-associated mortality worldwide. Hepatocarcinogenesis involves numerous interlinked factors and processes, including the Sonic hedgehog (Shh) signaling pathway, which participates in the carcinogenesis, progression, invasiveness, recurrence and cancer stem cell maintenance of HCC. The Shh signaling pathway is activated by ligands that bind to their receptor protein, Protein patched homolog (Ptch). The process of Shh ligand binding to Ptch weakens the inhibition of smoothened homolog (SMO) and activates signal transduction via glioma-associated oncogene homolog (Gli) transcription factors. The overexpression of Shh pathway molecules, including Shh, Ptch-1, Gli and SMO has been indicated in patients with HCC. It has also been suggested that the Shh signaling pathway exhibits cross-talk between numerous other signaling pathways. The inactivation of the Shh signaling pathway reduces HCC growth, increases radio-sensitivity and increases the beneficial effect of chemotherapy in HCC treatment. Therefore, inhibition of the Shh pathway may be an effective target therapy that can be used in the treatment of HCC.


Phase I study of alpelisib (BYL-719) and trastuzumab emtansine (T-DM1) in HER2-positive metastatic breast cancer (MBC) after trastuzumab and taxane therapy.
Activation of the phosphoinositide 3-kinase (PI3K) pathway is an important resistance mechanism to anti-HER2 therapies. This study aimed to assess the safety and activity of alpelisib (a PI3Kα isoform-specific inhibitor) with T-DM1 in trastuzumab- and taxane-resistant HER2-positive MBC. Patients with HER2-positive MBC that had progressed on trastuzumab-based therapy were treated with alpelisib daily and T-DM1 3.6 mg/kg every 3 weeks. The dose-limiting toxicity (DLT), maximum tolerated dose (MTD), adverse events, overall response rate (ORR), and clinical benefit rate (CBR = CR + PR + SD > 6 months) were assessed with descriptive statistics. Progression-free survival (PFS) was calculated by the Kaplan-Meier method. Seventeen patients were enrolled with a median of 3 prior therapies for metastatic disease. The DLT was a maculopapular rash and MTD was 250 mg alpelisib daily. The most frequently occurring toxicities included fatigue, rash, gastrointestinal side effects, thrombocytopenia, anemia, elevated liver enzymes, and hyperglycemia. Fourteen patients were evaluable for response with an ORR of 43%. In patients with prior treatment and progression on T-DM1 (n = 10), the ORR was 30%. The CBR was 71% in evaluable patients and 60% in those with prior T-DM1. The median PFS was 8.1 months. The combination of alpelisib and T-DM1 is tolerable and demonstrates activity in trastuzumab-resistant HER2-positive MBC. Furthermore, activity was observed in T-DM1-resistant disease. These data suggest that PIK3CA inhibition targets an important resistance pathway to anti-HER2 therapy, providing rationale for further study of PI3K inhibition in refractory HER2-positive MBC to validate these results.


Habitat Specialization by Wildlife Reduces Pathogen Spread in Urbanizing Landscapes.
AbstractUrban areas are expanding globally with far-reaching ecological consequences, including for wildlife-pathogen interactions. Wildlife show tremendous variation in their responses to urbanization; even within a single population, some individuals can specialize on urban or natural habitat types. This specialization could alter pathogen impacts on host populations via changes to wildlife movement and aggregation. Here, we build a mechanistic model to explore how habitat specialization in urban landscapes affects interactions between a mobile host population and a density-dependent specialist pathogen that confers no immunity. We model movement on a network of resource-stable urban sites and resource-fluctuating natural sites, where hosts are urban specialists, natural specialists, or generalists that use both patch types. We find that for generalists, natural and partially urban landscapes produce the highest infection prevalence and mortality, driven by high movement rates at natural sites and high densities at urban sites. However, habitat specialization protects hosts from these negative effects of partially urban landscapes by limiting movement between patch types. These findings suggest that habitat specialization can benefit populations by reducing infectious disease transmission, but by reducing movement between habitat types it could also carry the cost of reducing other movement-related ecosystem functions, such as seed dispersal and pollination.


Dental agenesis: genetic and clinical perspectives.
Dental agenesis is the most common developmental anomaly in humans and is frequently associated with several other oral abnormalities. Whereas the incidence of missing teeth may vary considerably depending on dentition, gender, and demographic or geographic profiles, distinct patterns of agenesis have been detected in the permanent dentition. These frequently involve the last teeth of a class to develop (I2, P2, M3) suggesting a possible link with evolutionary trends. Hypodontia can either occur as an isolated condition (non-syndromic hypodontia) involving one (80% of cases), a few (less than 10%) or many teeth (less than 1%), or can be associated with a systemic condition or syndrome (syndromic hypodontia), essentially reflecting the genetically and phenotypically heterogeneity of the condition. Based on our present knowledge of genes and transcription factors that are involved in tooth development, it is assumed that different phenotypic forms are caused by different genes involving different interacting molecular pathways, providing an explanation not only for the wide variety in agenesis patterns but also for associations of dental agenesis with other oral anomalies. At present, the list of genes involved in human non-syndromic hypodontia includes not only those encoding a signaling molecule (TGFA) and transcription factors (MSX1 and PAX9) that play critical roles during early craniofacial development, but also genes coding for a protein involved in canonical Wnt signaling (AXIN2), and a transmembrane receptor of fibroblast growth factors (FGFR1). Our objective was to review the current literature on the molecular mechanisms that are responsible for selective dental agenesis in humans and to present a detailed overview of syndromes with hypodontia and their causative genes. These new perspectives and future challenges in the field of identification of possible candidate genes involved in dental agenesis are discussed.


Eyetracking reveals multiple-category use in induction.
Category information is used to predict properties of new category members. When categorization is uncertain, people often rely on only one, most likely category to make predictions. Yet studies of perception and action often conclude that people combine multiple sources of information near-optimally. We present a perception-action analog of category-based induction using eye movements as a measure of prediction. The categories were objects of different shapes that moved in various directions. Experiment 1 found that people integrated information across categories in predicting object motion. The results of Experiment 2 suggest that the integration of information found in Experiment 1 were not a result of explicit strategies. Experiment 3 tested the role of explicit categorization, finding that making a categorization judgment, even an uncertain one, stopped people from using multiple categories in our eye-movement task. Experiment 4 found that induction was indeed based on category-level predictions rather than associations between object properties and directions. (PsycINFO Database Record


Revisiting the Helical Cooperativity of Synthetic Polypeptides in Solution.
Using synthetic polypeptides as a model system, the theories of helix-coil transition were developed into one of the most beautiful and fruitful subjects in macromolecular science. The classic models proposed by Schellman and Zimm-Bragg more than 50 years ago, differ in the assumption on whether the configuration of multiple helical sequences separated by random coil sections is allowed in a longer polypeptide chain. Zimm also calculated the critical chain lengths that facilitate such interrupted helices in different solvent conditions. The experimental validation of Zimm's prediction, however, was not carefully examined at that time. Herein, we synthesize a series of homopolypeptide samples with different lengths, to systematically examine their helix-coil transition and folding cooperativity in solution. We find that for longer chains, polypeptides do exist as interrupted helices with scattered coil sections even in helicogenic solvent conditions, as predicted in the Zimm-Bragg model. The critical chain lengths that facilitate such interrupted helices, however, are substantially smaller than Zimm's original estimation. The inaccuracy is in part due to an approximation that Zimm made in simplifying the calculation. But more importantly, we find there exist intramolecular interactions between different structural segments in the longer polypeptides, which are not considered in the classic helix-coil theories. As such, even the Zimm-Bragg model in its exact form cannot fully describe the transition behavior and folding cooperativity of longer polypeptides. The results suggest that long "all-helix" chains may be much less prevalent in solution than previously imagined, and a revised theory is required to accurately account for the helix-coil transition of the longer chains with potential "non-local" intramolecular interactions.


A Fine-Scale and Minimally Invasive Marking Method for Use with Conventional Tungsten Microelectrodes.
In neurophysiology, achieving precise correlation between physiological responses and anatomic structures is a significant challenge. Therefore, the accuracy of the electrode marking method is crucial. In this study, we describe a tungsten-deposition method, in which tungsten oxide is generated by applying biphasic current pulses to conventional tungsten electrodes. The electrical current used was 40-50 μA, which is similar to that used in electrical microstimulation experiments. The size of the markings ranged from 10 to 100 μm, corresponding to the size of the electrode tip, which is smaller than that of existing marking methods. Despite the small size of the markings, detection is easy as the marking appears in bright red under dark-field observation after Nissl staining. This marking technique resulted in low tissue damage and was maintained in vivo for at least two years. The feasibility of this method was tested in mouse and macaque brains.


Complete brachial plexus lesions: a ten-year follow-up of twenty cases.
Twenty patients with complete brachial plexus lesions were reviewed approximately nine and a half years after injury. Thirteen were amputees and seven had received no surgical treatment. Amputation did not alleviate pain and a prosthesis was frequently of no greater use of the patient than the useless limb it replaced: only two of the thirteen amputees were true prosthetic users and they both had dominant limb involvement, the rest adapting easily to being one-handed. Initial treatment should therefore be conservative, with intensive rehabilitation and retraining. It is recommended that amputation should not be considered until a year after injury and only if the flail limb causes repulsion, prevents sporting activities or if the patient has difficulty in converting to the non-dominant limb. In no instance should smputation be done for relief of pain.


Localization of intestinal intraepithelial T lymphocytes involves regulation of alphaEbeta7 expression by transforming growth factor-beta.
Induction of alphaEbeta7 expression on T cells by transforming growth factor (TGF)-beta is thought to be important for intestinal intraepithelial T lymphocyte (IEL) entry into the epithelial compartment. However, there has been no in vivo evidence that up-regulation of alphaEbeta7 expression on T cells by TGF-beta is critical for the selective localization of intestinal IEL in the epithelial area. We have recently established transgenic mice expressing Smad7 under the control of a distal lck promoter where TGF-beta/Smad signaling is specifically blocked in mature T cells. Here we showed that TGF-beta-mediated up-regulation of alphaEbeta7 was impaired on T cells isolated from the Smad7 transgenic mice associated with reduced numbers of intestinal IEL when compared with that in wild-type littermates. These results indicated that failure to induce alphaEbeta7 on T cells by TGF-beta resulted in reduced numbers of intestinal IEL, suggesting the importance of alphaEbeta7 expression by TGF-beta in selective localization of intestinal IEL.


DNA strand breaks and poly(ADP-ribose) synthetase activation in pancreatic islets--a new aspect to development of insulin-dependent diabetes and pancreatic B-cell tumors.
Alloxan and streptozotocin, which produce diabetes mellitus in experimental animals, have been known to inhibit various functions of pancreatic islets including proinsulin synthesis. However, little is known about the mechanisms underlying the action of these agents in pancreatic islets. Our recent in vivo and in vitro study using rats and isolated islets showed that one of the primary targets of the diabetogenic agents is the DNA of pancreatic islets. The first step is the generation of hydroxyl radical by alloxan which attacks DNA to produce strand breaks. In the case of streptozotocin, the alkylating activity of this compound may be causally related to its ability to induce DNA strand breaks. Subsequently, the fragmented DNA activates poly(ADP-ribose) synthetase which depletes cellular NAD. Since NAD is the most abundant of cellular coenzymes and participates in many biological reactions in mammalian cells, the reduction in intracellular NAD to such a nonphysiological level may severely affect islet cell functions including proinsulin synthesis. These results, in turn, raise the possibility that insulin-dependent diabetes may be preventable by inhibiting the occurrence of DNA strand breaks or the poly(ADP-ribose) synthetase. In fact, by poly(ADP-ribose) synthetase inhibitors such as nicotinamide and picolinamide, alloxan- and streptozotocin-induced NAD depletion was completely prevented, and B-cell functions including proinsulin synthesis proceeded normally. However, poly(ADP-ribose) synthetase inhibitors did not prevent the DNA strand breaks at all. Therefore, B-cells may survive with the residual DNA damage within their genome. About one year after the combined administration to rats of alloxan or streptozotocin with poly(ADP-ribose) synthetase inhibitors, diabetes did not develop but islet B-cell tumors were found frequently. This suggests that insulin-dependent diabetes and B-cell tumors are closely related with respect to their developmental processes. In other words, DNA breaks initiate two kinds of pathological state in B-cells, one is degeneratively and the other is oncogenically expressed.


The heterogeneous dynamics of economic complexity.
What will be the growth of the Gross Domestic Product (GDP) or the competitiveness of China, United States, and Vietnam in the next 3, 5 or 10 years? Despite this kind of questions has a large societal impact and an extreme value for economic policy making, providing a scientific basis for economic predictability is still a very challenging problem. Recent results of a new branch--Economic Complexity--have set the basis for a framework to approach such a challenge and to provide new perspectives to cast economic prediction into the conceptual scheme of forecasting the evolution of a dynamical system as in the case of weather dynamics. We argue that a recently introduced non-monetary metrics for country competitiveness (fitness) allows for quantifying the hidden growth potential of countries by the means of the comparison of this measure for intangible assets with monetary figures, such as GDP per capita. This comparison defines the fitness-income plane where we observe that country dynamics presents strongly heterogeneous patterns of evolution. The flow in some zones is found to be laminar while in others a chaotic behavior is instead observed. These two regimes correspond to very different predictability features for the evolution of countries: in the former regime, we find strong predictable pattern while the latter scenario exhibits a very low predictability. In such a framework, regressions, the usual tool used in economics, are no more the appropriate strategy to deal with such a heterogeneous scenario and new concepts, borrowed from dynamical systems theory, are mandatory. We therefore propose a data-driven method--the selective predictability scheme--in which we adopt a strategy similar to the methods of analogues, firstly introduced by Lorenz, to assess future evolution of countries.


Hydrogen sulfide attenuates mitochondrial dysfunction-induced cellular senescence and apoptosis in alveolar epithelial cells by upregulating sirtuin 1.
Hydrogen sulfide (H2S), an endogenous gaseous signal molecule, regulates many pathologies related to aging. Sirtuin 1 (SIRT1) has been shown to protect against mitochondrial dysfunction and other pathological processes, including premature senescence. This study was aimed to investigate whether and how H2S attenuates senescence and apoptosis of alveolar epithelial cells via a SIRT1-dependent mechanism. Our results showed that treatment with sodium hydrosulfide (NaHS), a donor of H2S, attenuated cigarette smoke extract (CSE)-induced oxidative stress, mitochondrial dysfunction, cellular senescence and apoptosis in A549 cells. This was associated with SIRT1 upregulation. SIRT1 activation by a pharmacological activator, SRT1720, attenuated CSE-induced oxidative stress and mitochondrial dysfunction in A549 cells. While SIRT1 inhibition by EX 527 or silencing by siRNA transfection significantly attenuated or abolished the ability of NaHS to reverse the CSE-induced oxidative stress, mitochondrial dysfunction and the imbalance of mitochondrial fusion and fission. Also, SIRT1 inhibition or silencing abolished the protection of NaHS against CSE-induced cellular senescence and apoptosis. In conclusion, H2S attenuates CSE-induced cellular senescence and apoptosis by improving mitochondrial function and reducing oxidative stress in alveolar epithelial cells in a SIRT1-dependent manner. These findings provide novel mechanisms underlying the protection of H2S against cigarette smoke-induced COPD.


Lung cancer: from single-gene methylation to methylome profiling.
DNA methylation as part of the epigenetic gene-silencing complex is a universal occurring change in lung cancer. Numerous studies investigated methylation of specific genes in primary tumors, in serum or plasma samples, and in specimens from the aerodigestive tract epithelium of lung cancer patients. In most studies, single genes or small numbers of genes were analyzed. Moreover, it has been observed that methylation of certain genes can already be detected in samples from the upper aerodigestive tract epithelium of cancer-free heavy smokers. These findings indicated that methylation of certain genes may be a useful biomarker for prognosis, disease recurrence, early detection, and lung cancer risk assessment. So far, several genes were identified which seem to be of worse prognostic relevance when they were found to be methylated. In addition, it has been shown that a panel of markers may be relevant to predict disease recurrence after surgery. In comparison to analysis of single or small numbers of genes, methods for genome-wide detection of methylation were developed recently. These approaches are focused on either pharmacological re-activation of methylated genes followed by expression microarray analysis or on microarray analysis of sodium bisulfite-treated or affinity-enriched methylated DNA sequences. With currently available methods for the simultaneous detection of methylation, up to 28,000 CpG islands can be analyzed. Overall, we are just at the beginning of translating these findings into the clinic and there is hope that future patients will benefit from these results.


Increased mortality among patients sustaining hip fractures requiring surgery in a district general hospital.
Fractures of the femoral neck are a common condition the incidence of which continues to rise. Associated mortality remains very high and knowledge of factors contributing to this high mortality is crucial in understanding outcome for both patient and surgeon. A retrospective review of 1254 patients operated on for fracture neck of femur was conducted to identify mortality at 30 days, 90 days and one year. Age, type of fracture, sex and waiting time for surgery were evaluated along with mortality for 47 patients who sustained their fracture while already inpatients with other concurrent illnesses. Those patients who sustained an inpatient fracture were found to have significantly higher mortality at 30 days, 90 days and one year. We concluded that these patients represent a very high-risk group that needs special consideration in management of their fracture and concurrent medical condition to reduce their high mortality.


Metabolic evaluation of cooled equine spermatozoa.
Microscopy has been used in the routine evaluation of sperm metabolism. Nevertheless, it has limited capacity to preview male fertility. As calorimetry may be used to evaluate directly the metabolic activity of a biological system, the aim of this study was to use microcalorimetry as an additive method for sperm metabolism evaluation of cooled equine semen. Two ejaculates of four stallions were collected and motility, viability (eosin 3%) and membrane functional integrity (hyposmotic swelling test) of spermatozoa were evaluated. Sperm samples were processed following different protocols and the metabolism of these samples was accessed by calorimetry. Centrifugation is part of some of these processing protocols and although this procedure has been deleterious for sperm viability and plasma membrane integrity, no decrease in sperm motility was observed. Microcalorimetry was capable of detecting the positive effect of re-suspending the sperm pellet with Kenney extender. Thus, the use of microcalorimetry offered additional information for equine sperm metabolism evaluation and was efficient in detecting important information from sperm cell metabolism.


Shock in acute myocardial infarction: the Cape Horn for trials?
Despite therapeutic improvements, cardiogenic shock (CS) remains the most common cause of death in patients with acute myocardial infarction (AMI). In addition to percutaneous coronary intervention, inotropes, fluids, adjunctive medication, intra-aortic balloon counterpulsation, and also assist devices are widely used for treatment. However, currently, there is only limited evidence for any of the above treatments. This review will therefore outline the underlying causes, pathophysiology, and treatment of CS complicating AMI with major focus on interventional techniques and advancement of new therapeutical arsenals, both pharmacological and mechanical.


A comparison of sLASER and MEGA-sLASER using simultaneous interleaved acquisition for measuring GABA in the human brain at 7T.
γ-Aminobutyric acid (GABA), the major inhibitory neurotransmitter, is challenging to measure using proton spectroscopy due to its relatively low concentration, J-coupling and overlapping signals from other metabolites. Currently, the prevalent methods for detecting GABA at ultrahigh field strengths (≥ 7 T) are GABA-editing and model fitting of non-editing single voxel spectra. These two acquisition approaches have their own advantages: the GABA editing approach directly measures the GABA resonance at 3 ppm, whereas the fitting approach on the non-editing spectrum allows the detection of multiple metabolites, and has an SNR advantage over longer echo time (TE) acquisitions. This study aims to compare these approaches for estimating GABA at 7 T. We use an interleaved sequence of semi-LASER (sLASER: TE = 38 ms) and MEGA-sLASER (TE = 80 ms). This simultaneous interleaved acquisition minimizes the differential effect of extraneous factors, and enables an accurate comparison of the two acquisition methods. Spectra were acquired with an 8 ml isotropic voxel at six different brain regions: anterior-cingulate cortex, dorsolateral-prefrontal cortex, motor cortex, occipital cortex, posterior cingulate cortex, and precuneus. Spectral fitting with LCModel quantified the GABA to total Cr (tCr: Creatine + Phosphocreatine) concentration ratio. After correcting the T2 relaxation time variation, GABA/tCr ratios were similar between the two acquisition approaches. GABA editing showed smaller spectral fitting error according to Cramér-Rao lower bound than the sLASER approach for all regions examined. We conclude that both acquisition methods show similar accuracy but the precision of the MEGA-editing approach is higher for GABA measurement. In addition, the 2.28 ppm GABA resonance was found to be important for estimating GABA concentration without macromolecule contamination in the GABA-edited acquisition, when utilizing spectral fitting with LCModel.


[Multiple gliomas. Illustrative cases of 4 different presentations].
Multiple gliomas are uncommon and may be classified according to: a) the time of presentation in early (at diagnosis) or late (during treatment); b) the characteristics of computed tomography or magnetic resonance imaging (CT/MRI) in multifocal (with evidence of spread) and multicentric (without evidence of spread). From 212 patients with histopathologic diagnosis of glioma evaluated from March/90 to September/99, 15 (7%) had multiple lesions. We describe 4 patients: early multicentric, late multicentric, early multifocal and late multifocal, with emphasis on characteristics of CT/MRI and possible differential diagnosis. The differential diagnosis of multiple lesions in the central nervous system includes mainly infectious/inflammatory diseases and metastasis, however multiple gliomas should always be considered, even in patients with known systemic cancer, as described by others. Considering that CT/MRI features are not definite, the diagnosis should always be confirmed by histopathologic examination.


Application of single-stage stereotactic radiosurgery for cerebral arteriovenous malformations >10 cm3.
Stereotactic radiosurgery (SRS) is a safe and effective treatment for small arteriovenous malformations (AVMs), the use of this modality for the treatment of large AVMs is still controversial, although it has been used in difficult cases. The aim of this study was to evaluate the treatment outcomes of patients who underwent single-stage SRS for large AVMs and to discuss the role of SRS in the treatment of these challenging lesions. Between 1998 and 2010, 65 patients with AVMs >10 cm(3) underwent single-stage SRS using the Leksell Gamma Knife. Patients who had prospective volume-staged SRS were excluded from this series. Outcomes including the rates of obliteration, hemorrhage after treatment, and adverse events were retrospectively evaluated. The mean nidus volume was 14.9 cm(3) (±3.8 cm(3)), and a mean margin dose of 20 Gy (±1.5 Gy) was applied. The mean observation period was 60 months (range, 7-178 months). The nidus obliteration rates after SRS were 44%, 76%, and 81% at 3, 5, and 6 years, respectively. The annual hemorrhage rate after SRS was 1.94% and permanent adverse events were observed in 2 patients (3%). For large AVMs <20 cm(3), single-stage radiosurgery by applying >16 Gy marginal dose presented favorable obliteration rates with relatively low rate of morbidity. Further accumulation of cases is awaited to fully evaluate the results of single-stage radiosurgery for large AVMs.


Environmental health and nursing practice: a survey of registered nurses.
The purpose of this study was to develop and implement a survey for Registered Nurses (RNs) that explored beliefs regarding environmental health (EH) and nursing practice, perceived preparation related to EH, barriers to addressing EH in practice, and factors that may facilitate integrating EH into nursing practice. Five hundred surveys were mailed, with a 39% return rate. Results indicate that RNs believe the environment and human health are related, but feel poorly prepared to address EH in practice. Barriers and facilitators to integrating EH into practice were found. The need for programs and resources to ensure basic EH education for all nurses was substantiated.


Genomic insights into acute alcohol tolerance.
Alcohol "sensitivity" has been proposed as a predictive factor for development of alcohol dependence (Schuckit et al., 2005). Most measures of alcohol sensitivity in humans and animals include a component that can be ascribed to acute functional tolerance (AFT). AFT is a form of tolerance that develops within a single period of alcohol exposure and has a genetic component. We used microarray technology as well as quantitative trait locus analysis of phenotypic and gene expression data across 30 BXD recombinant inbred strains of mice, 20 inbred strains of mice, and two replicate lines of mice selectively bred for differences in AFT, to identify differentially expressed candidate genes that contribute to predisposition to AFT. Eight candidate genes were identified by our statistical and filtering methods. The location of brain expression of these genes was mapped using the Allen Brain Atlas (http://www.brain-map.org), and the transcript location and molecular pathway analysis indicated that brain structures and biochemical pathways implicated in long-term potentiation and memory might also participate in the generation of acute functional alcohol tolerance.


A process evaluation of a stroke-specific follow-up care model for stroke patients and caregivers; a longitudinal study.
There is a need for follow-up care after stroke, but there is no consensus about the way to organise it. An intervention providing follow-up care for stroke patients and caregivers showed favourable effects on the level of social activities, but no other effects were found. The intervention consists of a maximum of five home visits to patients and caregivers during a period of 18 months post-discharge. The home visits are conducted by a stroke care coordinator (SCC) using a structured assessment tool. The objective of this study was to examine process-related factors that could have influenced the effectiveness of the intervention. 77 stroke patients, 59 caregivers and 4 SCCs participated in the study. Data on the organisational characteristics of and the satisfaction with the intervention were collected by means of structured assessments, interviews and self-administered questionnaires at 1, 6, 12 and 18 months of follow-up. The intervention was provided between April 2008 and June 2011. Patients received an average of 3.8 home visits (SD 1.4) and 55% of them had a follow-up period of a maximum of 18 months. There were 1074 problems identified and the SCCs initiated 363 follow-up care and referral options. Stroke patients and caregivers were very satisfied with the intervention. The SCCs were satisfied with the assessment tool, but would like to see a structured referral system. The intervention was only partially performed in accordance with the protocol and was positively evaluated by patients, caregivers and SCCs. It is recommended to add a structured referral system to the intervention.


Profiling Tryptophan Catabolites of Human Gut Microbiota and Acute-Phase Protein Levels in Neonatal Dried Blood Specimens.
National screening programs use dried blood specimens to detect metabolic disorders or aberrant protein functions that are not clinically evident in the neonatal period. Similarly, gut microbiota metabolites and immunological acute-phase proteins may reveal latent immune aberrations. Microbial metabolites interact with xenobiotic receptors (i.e., aryl hydrocarbon and pregnane-X) to maintain gastrointestinal tissue health, supported by acute-phase proteins, functioning as sensors of microbial immunomodulation and homeostasis. The delivery (vaginal or cesarean section) shapes the microbial colonization, which substantially modulates both the immune system's response and mucosal homeostasis. This study profiled microbial metabolites of the kynurenine and tryptophan pathway and acute-phase proteins in 134 neonatal dried blood specimens. We newly established neonatal blood levels of microbial xenobiotic receptors ligands (i.e., indole-3-aldehyde, indole-3-butyric acid, and indole-3-acetamide) on the second day of life. Furthermore, we observed diverse microbial metabolic profiles in neonates born vaginally and via cesarean section, potentially due to microbial immunomodulatory influence. In summary, these findings suggest the supportive role of human gut microbiota in developing and maintaining immune system homeostasis.


Medical costs associated with use of systemic therapy in adults with colorectal cancer.
New cytotoxic agents and regimens, as well as immunotherapeutics, have recently been introduced for treatment of colorectal cancer (CRC). To identify the patient-related and clinical and treatment-related factors associated with higher total health care expenditures in newly diagnosed patients with CRC who are receiving systemic therapy (biologic or chemotherapy) from a commercially insured population. A longitudinal, retrospective analysis was employed to estimate costs and determinants of CRC treatment in a U.S. claims database for health care services used by commercial patients aged 18 to 64 years, who were diagnosed with CRC between January 1, 2005, and June 30, 2009. Generalized linear regression modeling was used to estimate the influence of demographic, clinical, and treatment factors on medical expenditures. Among the 5,160 patients newly diagnosed with CRC, 99.6% of patients had chemotherapy; 32.6% had biologics; and 85.6% had other pharmaceuticals (excluding the chemotherapy and biologics of interest). The average annualized per patient cost of CRC treatment was $97,400 and consisted of chemotherapy ($17,500), biologics ($30,400), other pharmaceuticals ($2,300), inpatient treatment ($26,300), and outpatient treatment ($42,900). From first line only, first and second lines only, and third+ lines, the cost per patient was $70,500, $100,100, and $152,900, respectively. After adjusting for health care inflation, the average treatment cost of CRC patients increased by 73% from 2005 to 2009. Adjusted analyses showed that the higher medical cost for CRC patients was associated with use of new regimens, metastasis, comorbidities, surgery, radiation, insurance plan, age, sex, and region. The health care cost of CRC treatment is increasing significantly over time, which is most likely caused by the use of new regimens, higher chances of surgery and radiation, and occurrence of various comorbidities and metastatic diseases due to increasing survival time.


Definition of a commonly deleted region in ovarian cancers to a 300-kb segment of chromosome 6q27.
Allelic deletions of chromosome 6q that occur frequently in ovarian cancers imply the presence of a putative tumor suppressor gene in this chromosomal vicinity. We analyzed DNA from 32 patients with ovarian carcinomas for loss of heterozygosity at loci on the distal portion of chromosome 6q and constructed a detailed deletion map. The map indicated a commonly deleted region between loci D6S149 (defined by CI6-24) and A2, which are estimated to be 300 kb apart on the basis of our cosmid contig map. By means of exon trapping, we found that the human AF-6 gene, which is disrupted in acute myeloid leukemia cells that carry a (6;11)(q27;q23) translocation, is located within the commonly deleted region. Subsequent screening of the AF-6 gene in ovarian carcinomas revealed no mutations. However, our mapping results, which narrowed the region containing the putative tumor suppressor gene to a 300-kb segment of 6q27, will facilitate further efforts to identify a gene associated with ovarian cancer.


A comparison of methods used for measuring the accumulation of quinolones by Enterobacteriaceae, Pseudomonas aeruginosa and Staphylococcus aureus.
Accumulation of norfloxacin by Escherichia coli was studied with a range of published procedures that used either radioactively-labelled norfloxacin (14C and 3H) or the natural fluorescence of the quinolone for detection. All methods except bioassay generated comparable data. A method involving the detection of fluorescence was found to be the method of choice. This method was used to study the accumulation kinetics of ciprofloxacin, lomefloxacin, fleroxacin, norfloxacin, and enoxacin by several species of Gram-negative bacteria, and a Staphylococcus aureus strain. Saturation and efflux kinetics were also studied. There was no saturation at a concentration of norfloxacin less than 50 mg/L. Norfloxacin efflux was minimal during the uptake assay as the samples were withdrawn into ice-cold buffer; however, when the cells were sampled into buffer at 37 degrees C, up to 50% of cell-associated quinolone effluxed within 5 min.


Geo-Located Tweets. Enhancing Mobility Maps and Capturing Cross-Border Movement.
Capturing human movement patterns across political borders is difficult and this difficulty highlights the need to investigate alternative data streams. With the advent of smart phones and the ability to attach accurate coordinates to Twitter messages, users leave a geographic digital footprint of their movement when posting tweets. In this study we analyzed 10 months of geo-located tweets for Kenya and were able to capture movement of people at different temporal (daily to periodic) and spatial (local, national to international) scales. We were also able to capture both long and short distances travelled, highlighting regional connections and cross-border movement between Kenya and the surrounding countries. The findings from this study has broad implications for studying movement patterns and mapping inter/intra-region movement dynamics.