Distinct trajectories of multimorbidity in primary care were identified using latent class growth analysis.
To investigate the use of latent class growth analysis (LCGA) in understanding onset and changes in multimorbidity over time in older adults. This study used primary care consultations for 42 consensus-defined chronic morbidities over 3 years (2003-2005) by 24,615 people aged >50 years at 10 UK general practices, which contribute to the Consultations in Primary Care Archive database. Distinct groups of people who had similar progression of multimorbidity over time were identified using LCGA. These derived trajectories were tested in another primary care consultation data set with linked self-reported health status. Five clusters of people representing different trajectories were identified: those who had no recorded chronic problems (40%), those who developed a first chronic morbidity over 3 years (10%), a developing multimorbidity group (37%), a group with increasing number of chronic morbidities (12%), and a multi-chronic group with many chronic morbidities (1%). These trajectories were also identified using another consultation database and associated with self-reported physical and mental health. There are distinct trajectories in the development of multimorbidity in primary care populations, which are associated with poor health. Future research needs to incorporate such trajectories when assessing progression of disease and deterioration of health.


Environmental constraints underpin the distribution and phylogenetic diversity of nifH in the Yellowstone geothermal complex.
Biological nitrogen fixation is a keystone process in many ecosystems, providing bioavailable forms of fixed nitrogen for members of the community. In the present study, degenerate primers targeting the nitrogenase iron protein-encoding gene (nifH) were designed and employed to investigate the physical and chemical parameters that underpin the distribution and diversity of nifH as a proxy for nitrogen-fixing organisms in the geothermal springs of Yellowstone National Park (YNP), Wyoming. nifH was detected in 57 of the 64 YNP springs examined, which varied in pH from 1.90 to 9.78 and temperature from 16°C to 89°C. This suggested that the distribution of nifH in YNP is widespread and is not constrained by pH and temperature alone. Phylogenetic and statistical analysis of nifH recovered from 13 different geothermal spring environments indicated that the phylogeny exhibits evidence for both geographical and ecological structure. Model selection indicated that the phylogenetic relatedness of nifH assemblages could be best explained by the geographic distance between sampling sites. This suggests that nifH assemblages are dispersal limited with respect to the fragmented nature of the YNP geothermal spring environment. The second highest ranking explanatory variable for predicting the phylogenetic relatedness of nifH assemblages was spring water conductivity (a proxy for salinity), suggesting that salinity may constrain the distribution of nifH lineages in geographically isolated YNP spring ecosystems. In summary, these results indicate a widespread distribution of nifH in YNP springs, and suggest a role for geographical and ecological factors in constraining the distribution of nifH lineages in the YNP geothermal complex.


Dynamic optimal control of homeostasis: an integrative system approach for modeling of the central nitrogen metabolism in Saccharomyces cerevisiae.
The theory of dynamic optimal metabolic control (DOMC), as developed by Giuseppin and Van Riel (Metab. Eng., 2000), is applied to model the central nitrogen metabolism (CNM) in Saccharomyces cerevisiae. The CNM represents a typical system encountered in advanced metabolic engineering. The CNM is the source of the cellular amino acids and proteins, including flavors and potentially valuable biomolecules; therefore, it is also of industrial interest. In the DOMC approach the cell is regarded as an optimally controlled system. Given the metabolic genotype, the cell faces a control problem to maintain an optimal flux distribution in a changing environment. The regulation is based on strategies and balances feedback control of homeostasis and feedforward regulation for adaptation. The DOMC approach is an integrative, holistic approach, not based on mechanistic descriptions and (therefore) not biased by the variation present in biochemical and molecular biological data. It is an effective tool to structure the rapidly increasing amount of data on the function of genes and pathways. The DOMC model is used successfully to predict the responses of pulses of ammonia and glutamine to nitrogen-limited continuous cultures of a wild-type strain and a glutamine synthetase-negative mutant. The simulation results are validated with experimental data.


Mixed-method approach incorporating Geographic information system (GIS) tools for optimizing collection costs and convenience of the biowaste separate collection.
Nowadays, dealing with organic waste (or biowaste) remains a global phenomenon. Especially developing countries worldwide generate more than 50 percent organicwaste. In the European Union (EU) with a share of 34%, biowaste is a dominantfraction of the municipal waste (EEA, 2020). Therefore, separate collection at source and environmentally sound treatment of biowaste are of key importance. An intensive optimisation of biowaste separate collection is needed to balance demands of municipal representatives and households' needs. Based on the mixed-method approach we developed a MCDA model complemented by expert-based weighting assessment and combined with the GIS localisation tools aimed at the optimisation of biowaste container locations that reflects various spatial conditions, preconditions for the localisation of containers and its cost intensity. We concluded that changing the density of containers, distance between the address point and container, and selecting container locations that respect the habits of households and demands of the collection technology significantly affect the total and collection costs. We confirmed that the decreases in the total costs were not significant for maximum walking distances of over 95 m, and would approach zero for distances of over 230 m. When the maximum walking distance exceeds 268 m, 40% of all inhabitants would not participate in the system as it would be inconvenient for them. A recycling campaign is needed to increase their willingness to participate in the system. We provided arguments for decision-makers how to balance convenience of the biowaste separation system and collection costs by proper localisation of biowaste containers.


Time course of the hemoglobin mass response to natural altitude training in elite endurance cyclists.
To determine the time course of hemoglobin mass (Hb(mass)) to natural altitude training, Hb(mass), erythropoietin [EPO], reticulocytes, ferritin and soluble transferrin receptor (sTfR) were measured in 13 elite cyclists during, and 10 days after, 3 weeks of sea level (n=5) or altitude (n=8, 2760 m) training. Mean Hb(mass), with a typical error of ∼2%, increased during the first 11 days at altitude (mean ± standard deviation 2.9 ± 2.0%) and was 3.5 ± 2.5% higher than baseline after 19 days. [EPO] increased 64.2 ± 18.8% after 2 nights at altitude but was not different from baseline after 12 nights. Hb(mass) and [EPO] did not increase in sea level. Reticulocytes (%) were slightly elevated in altitude at Days 5 and 12 (18.9 ± 17.7% and 20.4 ± 25.3%), sTfR was elevated at Day 12 (18.9 ± 15.0%), but both returned to baseline by Day 20. Hb(mass) and [EPO] decreased on descent to sea level while ferritin increased. The mean increase in Hb(mass) observed after 11 days (∼300 h) of altitude training was beyond the measurement error and consitent with the mean increase after 300 h of simulated live high:train low altitude. Our results suggest that in elite cyclists, Hb(mass) increases progressively with 3 weeks of natural altitude exposure, with greater increases expected as exposure persists.


Taxonomic and regional uncertainty in species-area relationships and the identification of richness hotspots.
Species-area relationships (SARs) are fundamental to the study of key and high-profile issues in conservation biology and are particularly widely used in establishing the broad patterns of biodiversity that underpin approaches to determining priority areas for biological conservation. Classically, the SAR has been argued in general to conform to a power-law relationship, and this form has been widely assumed in most applications in the field of conservation biology. Here, using nonlinear regressions within an information theoretical model selection framework, we included uncertainty regarding both model selection and parameter estimation in SAR modeling and conducted a global-scale analysis of the form of SARs for vascular plants and major vertebrate groups across 792 terrestrial ecoregions representing almost 97% of Earth's inhabited land. The results revealed a high level of uncertainty in model selection across biomes and taxa, and that the power-law model is clearly the most appropriate in only a minority of cases. Incorporating this uncertainty into a hotspots analysis using multimodel SARs led to the identification of a dramatically different set of global richness hotspots than when the power-law SAR was assumed. Our findings suggest that the results of analyses that assume a power-law model may be at severe odds with real ecological patterns, raising significant concerns for conservation priority-setting schemes and biogeographical studies.


Intracellular distribution of serum albumin and its possible precursors in rat liver.
1. The fractionation of intracellular albumin labelled with radioactive l-leucine was studied in rat liver by means of isoelectric focusing. 2. Isoelectric fractionation was compared with ion-exchange chromatography for purification of radioactive intracellular albumin obtained by antibody precipitation. Similar results were obtained with both methods of separation. Purified albumin contains only a minor amount of the radioactivity. The remainder is associated with albumin-like protein(s). 3. The albumin-like protein has the properties of a precursor of plasma albumin. 4. The distribution and turnover of radioactive albumin in rough and smooth microsomal fractions and in a Golgi-rich fraction were studied. 5. It is concluded that newly synthesized albumin, as such, appears only momentarily if at all in any intracellular structure before its appearance in the plasma. 6. It is also concluded that the rate-limiting step in the secretion of plasma albumin is the conversion of precursor(s) into albumin. We can find no evidence to suggest that there is any significant transport of albumin, as such, during the course of secretion.


Incidental isolation of Setaria equina microfilariae in preparations of equine peripheral blood mononuclear cells.
In the course of a vaccine experiment on horses, microfilariae were observed in cultures of peripheral blood mononuclear cells (PBMCs) isolated from eleven of fifteen study horses. The microfilariae were clearly viable as evidenced by their vigorous movements in the cultures, thus indicating that they had survived the Ficoll gradient purification and the cryopreservation method used for retaining the PBMCs. The microfilariae were identified as Setaria equina, which is a vector-borne filarial nematode that causes a relatively benign infection of equids in which the adult worms reside in the peritoneal cavity. Although it is not possible to definitely state where the infections were acquired, the horses originated from Saskatchewan, Canada and spent a relatively short period of time in the United States prior to blood sampling. Therefore, it is likely that the infections occurred in Canada. Interestingly, assays conducted to determine levels of cytokine mRNA transcripts in the isolated PBMCs seemed to be largely unaltered by the presence of the microfilariae in the cell cultures. These findings demonstrate that a standard method used to purify and cryopreserve PBMCs from blood can result in the unintended co-isolation of worms from microfilaremic animals. Furthermore, the presence of the microfilariae did not appear to alter significantly the results of our immunologic assays, suggesting either that the nematode antigens were not recognized or that immunological tolerance may have developed in these horses. Although notable effects on the assays were not observed in this study, it seems possible that microfilarial contamination could represent a confounding variable for experiments examining cellular immunity.


Biomarkers in Progressive Chronic Kidney Disease. Still a Long Way to Go.
The traditional chronic kidney disease (CKD) biomarkers (eGFR based on serum creatinine, sex and age and albuminuria) cannot predict a patient's individual risk for developing progressive CKD. For this reason, it is necessary to identify novel CKD biomarkers that will be able to predict which patients are prone to develop progressive disease and discriminate between disease processes in different parts of the nephron (glomeruli or tubules). A good biomarker should change before or simultaneously with lesion development and its changes should correlate strongly with lesion development. Also, there should be a close relationship between severity of injury and amount of detectable biomarker and its levels should decrease with diminishing injury. Among the large number of molecules under investigation, we have reviewed the most promising ones: NGAL and KIM-1, MCP-1, MMP-9, clusterin, MMP-9, TIMP-1, Procollagen I alpha 1 and suPAR. All these, have been studied as biomarkers for prediction of CKD progression in cohorts of patients with chronic kidney disease of different stages and various aetiologies (proteinuric and non-proteinuric, glomerulonephritides, diabetic, hypertensive and polycystic kidney disease). There is evidence that these molecules could be useful as biomarkers for progressive chronic kidney disease, however, the available data are not enough to draw final conclusions. Further studies with large cohorts and long follow-up are required to identify appropriate biomarkers, that will be able to accurately and reliably define the risk for progressive chronic kidney disease.


Binding to gangliosides containing N-acetylneuraminic acid is sufficient to mediate the immunomodulatory properties of the nontoxic mucosal adjuvant LT-IIb(T13I).
By use of a mouse mucosal immunization model, LT-IIb(T13I), a nontoxic mutant type II heat-labile enterotoxin, was shown to have potent mucosal and systemic adjuvant properties. In contrast to LT-IIb, which binds strongly to ganglioside receptors decorated with either N-acetylneuraminic acid (NeuAc) or N-glycolylneuraminic acid (NeuGc), LT-IIb(T13I) binds NeuAc gangliosides much less well. Rather, LT-IIb(T13I) binds preferentially to NeuGc gangliosides. To determine if the adjuvant properties of LT-IIb(T13I) are altered in the absence of NeuGc ganglioside receptors, experiments were conducted using a Cmah-null mouse line which is deficient in the synthesis of NeuGc gangliosides. Several immunomodulatory properties of LT-IIb(T13I) were shown to be dependent on NeuGc gangliosides. LT-IIb(T13I) had reduced binding activity for NeuGc-deficient B cells and macrophages; binding to NeuGc-deficient T cells and dendritic cells (DC) was essentially undetectable. Treatment of Cmah-null macrophages with LT-IIb(T13I), however, upregulated the transcription of interleukin-4 (IL-4), IL-6, IL-17, and gamma interferon (IFN-gamma), four cytokines important for promoting immune responses. The production of mucosal IgA and serum IgG against an immunizing antigen was augmented in NeuGc-deficient mice administered LT-IIb(T13I) as a mucosal adjuvant. Notably, NeuGc gangliosides are not expressed in humans. Still, treatment of human monocytes with LT-IIb(T13I) induced the secretion of IL-6, an inflammatory cytokine that mediates differential control of leukocyte activation. These results suggested that NeuAc gangliosides are sufficient to mediate the immunomodulatory properties of LT-IIb(T13I) in mice and in human cells. The nontoxic mutant enterotoxin LT-IIb(T13I), therefore, is potentially a new and safe human mucosal adjuvant.


Navigation of brain networks.
Understanding the mechanisms of neural communication in large-scale brain networks remains a major goal in neuroscience. We investigated whether navigation is a parsimonious routing model for connectomics. Navigating a network involves progressing to the next node that is closest in distance to a desired destination. We developed a measure to quantify navigation efficiency and found that connectomes in a range of mammalian species (human, mouse, and macaque) can be successfully navigated with near-optimal efficiency (>80% of optimal efficiency for typical connection densities). Rewiring network topology or repositioning network nodes resulted in 45-60% reductions in navigation performance. We found that the human connectome cannot be progressively randomized or clusterized to result in topologies with substantially improved navigation performance (>5%), suggesting a topological balance between regularity and randomness that is conducive to efficient navigation. Navigation was also found to (i) promote a resource-efficient distribution of the information traffic load, potentially relieving communication bottlenecks, and (ii) explain significant variation in functional connectivity. Unlike commonly studied communication strategies in connectomics, navigation does not mandate assumptions about global knowledge of network topology. We conclude that the topology and geometry of brain networks are conducive to efficient decentralized communication.


Association between Cryptosporidium infection and human leukocyte antigen class I and class II alleles.
Cryptosporidium species are a common cause of diarrhea, which can be severe and protracted in young children and immunocompromised individuals. A cohort of 226 Bangladeshi children aged 2-5 years was prospectively followed for >3 years to study the role of host genetics in susceptibility to infection, as well as the community impact of cryptosporidiosis on this population. Ninety-six children (42.5%) received a diagnosis of Cryptosporidium infection. A total of 51 (22.6%) had asymptomatic infection. Fifty-eight (25.7%) had cryptosporidiosis, of whom 17 (29.3%) had recurrent disease. Children with cryptosporidiosis presented early, and most had abdominal pain and a short course of diarrhea. Infected children were more likely to carry the human leukocyte antigen (HLA) class II DQB1*0301 allele, particularly those with both asymptomatic and symptomatic infection (P = .009); a strong association was found between carriage of the DQB1*0301/DRB1*1101 haplotype and development of both asymptomatic and symptomatic infection (P = .009). Infected children were also more likely to carry the B*15 HLA class I allele. This is the first study to describe a possible genetic component of the immune response to Cryptosporidium infection, which includes HLA class I and II alleles. Cryptosporidiosis in Bangladeshi children aged 2-5 year is common and often recurrent, but the duration is shorter and the abdominal pain greater than that described in children aged <2 years.


Risk factors for bladder cancer recurrence after nephroureterectomy for upper tract urothelial tumors: results from the Canadian Upper Tract Collaboration.
To evaluate risk factors for bladder cancer recurrence in a cohort of patients treated with radical nephroureterectomy (RNU). At 10 Canadian University Centers, we retrospectively evaluated data, between 1990 and 2010, from 743 patients who were free from bladder cancer and were previously treated with RNU for upper tract urothelial cancer. Of 743 patients, 167 (22.5%) developed bladder tumors after a median time of 17.2 months after RNU. Multivariable analysis detected age (hazard ratio [HR] = 1.028; 95% CI: 1.010-1.046; P = 0.0018), tumor location in both the renal pelvis and the ureter (HR = 2.205; 95% CI: 1.355-3.589; P = 0.0015), the use of adjuvant systemic chemotherapy (HR = 2.309; 95% CI: 1.439-3.705; P = 0.0005), and laparoscopic surgery (HR = 1.876; 95% CI: 1.226-2.87; P = 0.0037) as risk factors for bladder cancer recurrence. Open excision of a bladder cuff (HR = 0.661; 95% CI: 0.453-0.965; P = 0.0319) and transurethral resection of the intramural ureter (HR = 0.548; 95% CI: 0.306-0.981; P = 0.0429) on comparison with extravesical resection decreased the risk of bladder cancer recurrence significantly. Major limitations were the retrospective design and partially missing data, although the significance of variables did not change in the imputation analysis. Older patients, those with tumor location in both the renal pelvis and the ureter, and those treated with adjuvant systemic chemotherapy were found at higher risk for intravesical recurrence, as were those having undergone extravesical ureterectomy or laparoscopic RNU.


Full-length archaeal Rad51 structure and mutants: mechanisms for RAD51 assembly and control by BRCA2.
To clarify RAD51 interactions controlling homologous recombination, we report here the crystal structure of the full-length RAD51 homolog from Pyrococcus furiosus. The structure reveals how RAD51 proteins assemble into inactive heptameric rings and active DNA-bound filaments matching three-dimensional electron microscopy reconstructions. A polymerization motif (RAD51-PM) tethers individual subunits together to form assemblies. Subunit interactions support an allosteric 'switch' promoting ATPase activity and DNA binding roles for the N-terminal domain helix-hairpin-helix (HhH) motif. Structural and mutational results characterize RAD51 interactions with the breast cancer susceptibility protein BRCA2 in higher eukaryotes. A designed P.furiosus RAD51 mutant binds BRC repeats and forms BRCA2-dependent nuclear foci in human cells in response to gamma-irradiation-induced DNA damage, similar to human RAD51. These results show that BRCA2 repeats mimic the RAD51-PM and imply analogous RAD51 interactions with RAD52 and RAD54. Both BRCA2 and RAD54 may act as antagonists and chaperones for RAD51 filament assembly by coupling RAD51 interface exchanges with DNA binding. Together, these structural and mutational results support an interface exchange hypothesis for coordinated protein interactions in homologous recombination.


Risk Assessment and Deployment for Safety Showers and Eyewash Stations in the Process Plant Industry.
Safety showers and eyewash stations are equipment used for primary washing if their operator is exposed to hazardous chemicals. Therefore, safety showers and eyewash stations should be installed to ensure operator safety in process plants with excessive hazardous chemicals. International guidelines related to safety showers and eyewash stations are introduced in ANSI Z358.1, BS EN 15154, and German DIN 12899-3:2009, but only mechanical specifications regarding safety showers and eyewash stations are suggested. As such, there are currently no engineering guidelines, books, or technical journal papers requiring safety showers or eyewash stations and their efficient deployment. Thus, this study conducted risk assessment from an industrial hygiene perspective, suggesting which process equipment requires a safety shower and eyewash, including their economical and efficient deployment for operator safety. In industry, safety showers and eyewash stations are considered part of the process safety field; this study attempted to contribute to the safety improvement of operators by applying risk assessment of the industrial hygiene field. More studies are needed that contribute to operators' safety by incorporating industrial hygiene fields for other process safety fields, including safety showers and eyewash stations.


Changes in plasma lipids during renin-angiotensin system blockade by combination therapy (enalapril plus valsartan) in patients with diabetes and hypertension.
There is experimental evidence of an interaction between the angiotensin system and lipid metabolism. The aim of this study was to evaluate whether a block of the angiotensin system achieved both by ACE inhibition and angiotensin II-AT1 receptor blockade could affect the plasma lipid profile and, if so, what relationship exists between these possible changes and glucose metabolism and blood pressure. In 50 patients with type 2 diabetes and hypertension, treated with diabetes drugs and enalapril, we evaluated the glycemic and lipid profile together with the HOMA insulin-resistance index, blood pressure and microalbuminuria at baseline and 3 months after the addition of valsartan. At the second evaluation, blood pressure was reduced as expected, whereas the glycemic profile, the HOMA index, and the body mass index were unchanged. Total cholesterol, LDL-c, and apoprotein B were reduced during combination therapy (P = 0.003, P = 0.001, and P = 0.004, respectively), plasma HDL-c was slightly though significantly increased (P = 0.024), whereas apoprotein A and triglyceride levels did not change. After adjustment for the insulin resistance index and for blood pressure, the reduction of LDL-c and apoprotein B and the increase in HDL-c remained significant. The variation in lipid profile was not related to the changes in blood pressure. Moreover, the addition of valsartan to enalapril was associated with a reduction in microalbuminuria, which remained significant after adjustment for LDL-c or blood pressure changes. Thus, the greater degree of renin-angiotensin system blockade or specific pharmacodynamic effects of valsartan could account for the changes in plasma lipid profile observed in this study.


Mutations in FARS2 and non-fatal mitochondrial dysfunction in two siblings.
Recently, mutations in FARS2, which encodes for mitochondrial phenylalanyl-tRNA synthetase, have been implicated in autosomal recessive combined oxidative phosphorylation deficiency 14. Associated clinical features in three previously reported patients with confirmed FARS2 mutations include infantile onset epilepsy, and a fatal Alpers-like encephalopathy. Herein, we report on two siblings with global developmental delay, dysarthria and tremor and compound heterozygous FARS2 abnormalities. They have a heterozygous missense mutation, c.1255C>T which predicts p.Arg419Cys in exon 7 of FARS2, inherited from their father and uncovered on exome sequencing, and an interstitial deletion of chromosome 6p25.1 inherited from their mother and uncovered on SNP array. This interstitial deletion includes all of exon 6 and parts of introns 5 and 6 of FARS2. Biochemical studies were also consistent with a mitochondrial disorder. While these siblings had considerable developmental difficulties, they are making consistent developmental progress and appear to be considerably less severely affected than the other patients reported in the literature with FARS2 associated mitochondrial disease. Thus, this study expands the phenotypic spectrum of FARS2 related disease and emphasizes intragenic deletion in the list of causative mutations.


Vibrio fischeri siderophore production drives competitive exclusion during dual-species growth.
When two or more bacterial species inhabit a shared niche, often, they must compete for limited nutrients. Iron is an essential nutrient that is especially scarce in the marine environment. Bacteria can use the production, release, and re-uptake of siderophores, small molecule iron chelators, to scavenge iron. Siderophores provide fitness advantages to species that employ them by enhancing iron acquisition, and moreover, by denying iron to competitors incapable of using the siderophore-iron complex. Here, we show that cell-free culture fluids from the marine bacterium Vibrio fischeri ES114 prevent the growth of other vibrio species. Mutagenesis reveals the aerobactin siderophore as the inhibitor. Our analysis reveals a gene, that we name aerE, encodes the aerobactin exporter, and LuxT is a transcriptional activator of aerobactin production. In co-culture, under iron-limiting conditions, aerobactin production allows V. fischeri ES114 to competitively exclude Vibrio harveyi, which does not possess aerobactin production and uptake genes. In contrast, V. fischeri ES114 mutants incapable of aerobactin production lose in competition with V. harveyi. Introduction of iutA, encoding the aerobactin receptor, together with fhuCDB, encoding the aerobactin importer are sufficient to convert V. harveyi into an "aerobactin cheater."


Site-specific glycosylation of secretory immunoglobulin A from human colostrum.
Secretory immunoglobulin A (sIgA) is a major glycoprotein in milk and plays a key role in mediating immune protection of the gut mucosa. Although it is a highly glycosylated protein, its site-specific glycosylation and associated glycan micro-heterogeneity have still not been fully elucidated. In this study, the site-specific glycosylation of sIgA isolated from human colostrum (n = 3) was analyzed using a combination of LC-MS and LC-MS/MS and in-house software (Glycopeptide Finder). The majority of the glycans found are biantennary structures with one or more acidic Neu5Ac residues; however, a large fraction belonged to truncated complex structures with terminal GlcNAc. Multiple glycosites were identified with nearly 30 glycan compositions located at seven sites on the secretory component, six compositions at a single site on the J chain, and 16 compositions at five sites on the IgA heavy (H) chain. Site-specific heterogeneity and relative quantitation of each composition and the extent of occupation at each site were determined using nonspecific proteases. Additionally, 54 O-linked glycan compositions located at the IgA1 hinge region (HR) were identified by comparison against a theoretical O-glycopeptide library. This represents the most comprehensive report to date detailing the complexity of glycan micro-heterogeneity with relative quantitation of glycoforms for each glycosylation site on milk sIgA. This strategy further provides a general method for determining site-specific glycosylation in large protein complexes.


Soluble form of canine transferrin receptor inhibits canine parvovirus infection in vitro and in vivo.
Canine parvovirus (CPV) disease is an acute, highly infectious disease threatening the dog-raising industry. So far there are no effective therapeutic strategies to control this disease. Although the canine transferrin receptor (TfR) was identified as a receptor for CPV infection, whether extracellular domain of TfR (called soluble TfR (sTfR)) possesses anti-CPV activities remains elusive. Here, we used the recombinant sTfR prepared from HEK293T cells with codon-optimized gene structure to investigate its anti-CPV activity both in vitro and in vivo. Our results indicated that codon optimization could significantly improve sTfR expression in HEK293T cells. The prepared recombinant sTfR possessed a binding activity to both CPV and CPV VP2 capsid proteins and significantly inhibited CPV infection of cultured feline F81 cells and decreased the mortality of CPV-infected dogs, which indicates that the sTfR has the anti-CPV activity both in vitro and in vivo.


Impact of school-based health promotion interventions aimed at different behavioral domains: a systematic review.
Given that lifestyleshave similar determinants and that school-based interventions are usually targeted at all the risks that affect adolescents, the objective of this systematic review was to summarize the characteristics and effects of school-based interventions acting on different behavioral domains of adolescent health promotion. The review process was conducted by two independent reviewers who searched PubMed, Scopus, PsycINFO, and ERIC databases for experimental or observational studies with at least two measures of results published from 2007 to 2011, given that the research information available doubles every 5 years. Methodological quality was assessed with a standardized tool. Information was extracted from 35 studies aiming to prevent risk behaviors and promote healthy nutrition, physical activity, and mental and holistic health. Activities were based on theoretical models and were classified into interactive lessons, peer mediation, environmental changes, parents' and community activities, and tailored messages by computer-assisted training or other resources, usually including multiple components. In some cases, we identified some moderate to large, short- and long-term effects on behavioral and intermediate variable. This exhaustive review found that well-implemented interventions can promote adolescent health. These findings are consistent with recent reviews. Implications for practice, public health, and research are discussed.


A pharmacokinetic study of a topical anesthetic (EMLA® ) in mouse soft tissue laceration.
The use of topical anesthesia instead of injection of local anesthetics for managing soft tissue lacerations in the emergency situations may be a relief for both patients and surgeons. Topical anesthesia in the form of a cream eutectic mixture of local anesthetics (EMLA®) containing 2.5% lidocaine and 2.5% prilocaine has been reported as an efficient anesthetic on skin before venipuncture anesthesia and as an alternative to injection anesthesia in some minor surgery situations. The aim of this study was to compare the pharmacokinetics of EMLA® when applied in a laceration with topical skin application in the mouse. A total of 120 Albino Laboratory-bred strain mouse (BALB-c) male mice were divided into three groups with regard to application mode of EMLA®. Group A: with laceration, 48 mice; Group B: on intact shaved skin, 48 mice; Group C: control group (24 mice) with same procedures but without application of EMLA®. Blood levels were collected at 0, 10, 20, 30, 45, 60, 75, and 90 min post-EMLA® application. Plasma sample analysis was carried out by employing liquid chromatography coupled with tandem mass spectrometric (LC-MS/MS) method, and the pharmacokinetic analysis of the mouse plasma samples was estimated by standard non-compartmental methods. The pharmacokinetic parameters of lidocaine and prilocaine were significantly altered following EMLA® application to lacerated mouse skin in contrast to intact skin. The absorption of lidocaine and prilocaine was rapid following application of EMLA® to lacerated and intact mouse skin. Maximum drug plasma concentration (C(max) ) and area under the drug plasma concentration-time curve (AUC) values of lidocaine were significantly increased by 448.6% and 161.5%, respectively, following application of EMLA to lacerated mouse skin in comparison with intact mouse skin. Similarly, prilocaine's C(max) and AUC values were also increased by 384% and 265.7%, respectively, following EMLA application to lacerated mouse skin, in contrast to intact skin. Further pharmacokinetic studies on different carriers of lidocaine/prilocaine are warranted before any firm conclusions for the clinic can be drawn.


CT of the normal brain in preterm infants.
Twenty-two normal preterm and 10 full-term infants were studied by cranial computed tomography. The study included correlation of the gestational age and birth weight to the ventricular size, brain size, and the area of the sylvian fissure. The most relevant parameters were the bifrontal diameter, bifrontal index, and the transverse diameter of the brain. The incidence of visualization of the sylvian fissure diminished with gestational age. Density measurements revealed the high cortical sections had a higher average density than the whole brain sections. The parietooccipital region showed an increasing average attenuation during the preterm period until term.


Effects of exogenous progesterone following ovariectomy on the metrial glands of pregnant mice.
The numbers of granulated metrial gland cells, the percentage of them incorporating tritiated thymidine and the numbers of granulated metrial gland cell precursors in a defined area of the metrial gland, and the cross sectional area of the metrial gland, have been determined in mice ovariectomised on Day 8 of pregnancy. The effects, on these parameters and on fetal survival, of progesterone treatment, given at two levels (0.5 mg or 1 mg twice daily) and started immediately after ovariectomy or after 6, 10 or 24 hours delay, have been studied. Granulated metrial gland cell number, DNA synthetic activity and metrial gland size decreased rapidly after ovariectomy but there was a significant increase in the number of granulated metrial gland cell precursors in the first 24 hours. Progesterone treatment, at the higher dose level started immediately after ovariectomy, resulted in significantly more granulated metrial gland cells in the area analysed two days after ovariectomy although a normal proportion of them were in DNA synthesis. Immediate progesterone treatment, at the higher dose level, was able to prolong the DNA synthetic activity of granulated metrial gland cells until Day 19 and this probably accounted for the significantly greater numbers of granulated metrial gland cells in these mice than in control mice, at this time. Delaying progesterone treatment at the higher level by 24 hours significantly reduced the size of the metrial glands at Day 10 but there was no effect on the cellular composition of the area analysed. Delaying the lower dose of progesterone treatment by 24 hours also resulted in small metrial glands and was associated with significantly fewer granulated metrial gland cells in the area analysed but significantly more of them were synthesising DNA. The observations are discussed and an attempt is made to relate the effects on individual fetal and placental survival to the effects on the corresponding metrial glands.


Frailty Prevalence and Association with Health-Related Quality of Life Impairment among Rural Community-Dwelling Older Adults in Vietnam.
Measuring health-related quality of life (HRQOL) is critical to evaluate the burden of frailty in the older population.This study explored the prevalence of frailty among Vietnamese older people in rural communities, determined the factors associated with frailty, and examined the differences in HRQOL between non-frail, pre-frail, and frail people. A cross-sectional study was conducted on older adults (≥60 years old) residing in Soc Son district, northern Vietnam. Non-frailty, pre-frailty, and frailty conditions were evaluated using Fried's frailty criteria. The EuroQol-5 Dimensions-5 Levels(EQ-5D-5L) instrument was employed to measure HRQOL. Socioeconomic, behavioral, health status, and healthcare utilization characteristics were collected as covariates. Among 523 older adults, 65.6% were pre-frail, and 21.7% were frail. The mean EQ-5D-5L indexes of the non-frailty, pre-frailty, and frailty groups were 0.70 (SD = 0.18), 0.70 (SD = 0.19), and 0.58 (SD = 0.20), respectively. The differences were found between non-frailty and frailty groups (p < 0.01), as well as the pre-frailty and frailty groups (p<0.01). After adjusting for covariates, the estimated mean difference in the HRQOL between the non-frailty and frailty groups was -0.10 (95%CI= -0.17; -0.02) (R2 = 45.2%), showing a 10% reduction of the maximum EQ-5D-5L index.This study emphasized the high prevalence of frailty among older adults in the rural communities of Vietnam. Frailty was found to be associated with a small reduction of HRQOL in this population.


Conditional deletion of cytochrome p450 reductase in osteoprogenitor cells affects long bone and skull development in mice recapitulating antley-bixler syndrome: role of a redox enzyme in development.
NADPH-cytochrome P450 oxidoreductase (POR) is the primary electron donor for cytochromes P450, dehydrocholesterol reductase, heme oxygenase, and squalene monooxygenase. Human patients with specific mutations in POR exhibit severe developmental malformations including disordered steroidogenesis, sexual ambiguities and various bone defects, similar to those seen in patients with Antley-Bixler syndrome (ABS). To probe the role of POR during bone development, we generated a conditional knockout mouse (CKO) by cross breeding Por (lox/lox) and Dermo1 Cre mice. CKO mice were smaller than their littermate controls and exhibited significant craniofacial and long bone abnormalities. Differential staining of the CKO mice skull bases shows premature fusion of the sphenooccipital and basioccipital-exoccipital synchondroses. Class III malocclusion was noted in adult knockout mice with an unusual overgrowth of the lower incisors. Shorter long bones were observed along with a reduction in the bone volume fraction, measured by microCT, in the Por-deleted mice compared to age- and sex-matched littermate controls. Concerted up- or down-regulation of proteins in the FGF signaling pathway observed by immunohistochemistry in the tibia samples of CKO mice compared to wild type controls shows a decrease in the FGF signaling pathway. To our knowledge, this is the first report of a mouse model that recapitulates both skull and long bone defects upon Por deletion, offering an approach to study the sequelae of POR mutations. This unique model demonstrates that P450 metabolism in bone itself is potentially important for proper bone development, and that an apparent link exists between the POR and FGF signaling pathways, begging the question of how an oxidation-reduction flavoprotein affects developmental and cellular signaling processes.


Acute otitis media: clinical course among children who received a short course of high dose antibiotic.
A prospective study was carried out in 274 children aged 3 to 10 years with acute otitis media. They were randomly allocated to one of two treatment regimens: (a) a seven day course of amoxycillin 125 mg three times a day, and (b) a two day course of amoxycillin 750 mg twice a day. They were followed up by symptom diaries and clinical examination. The findings in the 243 children who completed the trial showed that the short course of treatment was as effective as the seven day course in the speed of resolution of symptoms and signs, irrespective of previous history of otitis media or of episodes in which bulging of the eardrums was observed at presentation. A subgroup of 185 children was followed up for one year after entry to the trial. During this period no appreciable differences emerged between the two antibiotic regimens, either in recurrence rate of otitis media or in the frequency of hearing loss at one month and six months after entry to the study. Side effects of treatment were few, and those that could be attributed to antibiotic use occurred with equal frequency in the two treatment groups.


Adipokines and ghrelin in gastric cancer cachexia.
To investigate the roles of the adipocytokines, ghrelin and leptin in gastric cancer cachexia. Resistin, ghrelin, leptin, adiponectin, insulin and insulin-like growth factor (IGF-I), were measured in 30 healthy subjects, and 60 gastric cancer patients of which 30 suffered from cancer-induced cachexia and 30 served as a control group. The relationships between hormones, body mass index (BMI) loss ratio, age, gender, and Glasgow Prognostic Score (GPS) were investigated. Cachexia patients had higher tumor stage and GPS when compared with non-cachexia patients (P < 0.05). Ghrelin, resistin, leptin, adiponectin and IGF-I, showed a significant correlation with BMI loss ratio and GPS (P < 0.05). A strong correlation was seen between GPS and BMI loss (R = -0.570, P < 0.0001). Multivariate analysis indicated that BMI loss was significantly independent as a predictor of ghrelin, resistin, leptin and IGF-I (P < 0.05). Existence of an important significant relationship between resistin and insulin resistance was also noted. These results showed that serum ghrelin, leptin, adiponectin, and IGF-I play important roles in cachexia-related gastric cancers. No relationship was found between resistin and cancer cachexia. Also, because of the correlation between these parameters and GPS, these parameters might be used as a predictor factor.


Uterine contractions preceding labor.
To evaluate whether 12 contractions in 1 hour is a meaningful signal that spontaneous labor has begun or is imminent. This prospective observational cohort study includes all women reporting contractions who presented to a labor and delivery triage unit between August 1 and October 31, 2006, who met the following criteria: 1) 36 0/7 to 41 6/7 weeks of gestation, 2) cervical dilation less than 4 cm, 3) intact membranes, and 4) no other medical or obstetric complications that might influence admission. Each woman received external fetal monitoring for a minimum of 1 hour. Women were discharged home if cervical dilation did not advance and the fetal heart rate pattern was reassuring. Women who progressed to a cervical dilation of 4 cm were admitted with the diagnosis of labor. Among 768 women studied, labor was diagnosed within 24 hours in 268 (76%) with 12 or more contractions per hour compared with 216 (52%) of 416 women with fewer than 12 contractions per hour, P<.001. Cervical condition and fetal station were more advanced on presentation in women with 12 or more contractions per hour. Twelve contractions or more per hour at term is a meaningful signal that true labor has either begun or is imminent. II.


Low-temperature CeCoMnOx spinel-type catalysts prepared by oxalate co-precipitation for selective catalytic reduction of NO using NH3: A structure-activity relationship study.
CeCoMnOx spinel-type catalysts for the selective catalytic reduction of NO using NH3 (NH3-SCR) are usually prepared by alkaline co-precipitation. In this paper, a series of CeCoMnOx spinel-type catalysts with different calcination temperatures were prepared by acidic oxalate co-precipitation. The physicochemical structures and NH3-SCR activities of the CeCoMnOx spinel-type catalysts prepared by oxalate co-precipitation and conventional ammonia co-precipitation were systematically compared. The results show that the CeCoMnOx spinel-type catalysts prepared by the oxalate precipitation method (CeCoMnOx-C) have larger specific surface area, more mesopores and surface active sites, stronger redox properties and adsorption activation properties than those prepared by the traditional ammonia co-precipitation method at 400 °C (CeCoMnOx-N-400), and thus CeCoMnOx-C have better low-temperature NH3-SCR performance. At the same calcination temperature of 400 °C, the NO conversion of CeCoMnOx-C-400 exceeds 89 % and approaches 100 % within the reaction temperature of 100-125 °C, which is 14.8 %-2.5 % higher than that of CeCoMnOx-N-400 at 100-125 °C. In addition, the enhanced redox and acid cycle matching mechanisms on the CeCoMnOx-C surface, as well as the enhanced monoadsorption Eley-Rideal (E-R) and double adsorption Langmuir-Hinshelwood (L-H) reaction mechanisms, are also derived from XPS and in situ DRIFTS characterization.


Hypophysis surgery with or without endoscopy.
Hardy's operation with microscope has long been the standard method for pituitary adenoma. But a new approach via the nasal cavity using an endoscope has been adopted recently. In this study, the postoperative outcome as well as the preoperative evaluation of endoscopic hypophysectomy and non-endoscopic one were compared at our faculty. We performed the non-endoscopic transnasal hypophysectomy on 18 patients and the endoscopic transnasal hypophysectomy on thirteen patients who had a pituitary lesions from February 1996 to October 1999. As to these patients the situations from preoperation through postoperation such as chief complaints, serum hormone level, final diagnosis, tumor size, as well as operating time or blood loss during the operation were discussed precisely. Then the merits and demerits of endoscopic hypophysectomy were discussed. Five PRL-producing adenoma, three GH-producing adenoma, nine non-functioning adenoma, and two ACTH or TSH-producing adenoma were included in this discussion as endoscopic group. The age of non-endoscopic group are from 23 to 73 (49.4 in average), and they include ten males and eight females. On the other hand three PRL-producing adenoma, two GH-producing adenoma, two non-functioning adenoma, and one Rathke's cyst were included in this discussion as endoscopic group. The age of endoscopic group are from 19 to 73 (49.1 in average), and they include seven males and six females. As to non-endopscopic group the blood loss during each operation is 568 ml and operating time is 256 min in average. For endoscopic group the blood loss is 296 ml and operating time is 234 min in average. By microsurgery in the pituitary operation with endoscopy, the minimal invasive surgery becomes possible by reducing blood loss and shortening operating time. During the operation cooperation between neurosurgeon and ENT surgeon is indispensable in order to perform hypophysectomy smoothly. The development of optical better aids and operation instruments for endonasal hypophysectomy is desired in the future. The navigation system was more useful than X-ray fluoroscopy to obtain the detailed information.


Genetic homogeneity of North-African goats.
North Africa represents a rich and early reservoir of goat genetic diversity, from which the main African breeds have been derived. In this study, the genetic diversity of four indigenous Algerian goat breeds (i.e., Arabia, Makatia, M'Zabite and Kabyle, with n = 12 for each breed) has been investigated for the first time by genome-wide SNP genotyping; moreover in a broader context, genetic structuration of Algerian and Moroccan goats was explored (via FST, MDS, STRUCTURE, FineSTRUCTURE, BAPS, sPCA and DAPC analyses). At national level, the study revealed high level of genetic diversity and a significant phenomenon of admixture affecting all the Algerian breeds. At broader scale, clear global genetic homogeneity appeared considering both Algerian and Moroccan stocks. Indeed, genetic structuration was almost nonexistent among Arabia (from Algeria), Draa, Black and Nord (from Morocco), while the ancestral Kabyle and M'Zabite breeds, reared by Berber peoples, showed genetic distinctness. The study highlighted the threat to the Maghrebin stock, probably induced by unsupervised cross-breeding practices which have intensified in recent centuries. Moreover, it underlined the necessity to deepen our understanding of the genetic resources represented by the resilient North-African goat stock.


Expression of claudins 1, 4, 5, and 7 and occludin, and relationship with prognosis in squamous cell carcinoma of the tongue.
Claudins and occludin are tight junctional proteins known to play a role in normal tissues and epithelial tumors. We analyzed the distribution patterns of claudins 1, 4, 5, and 7 and occludin in the superficial and invasive front of squamous cell carcinoma of the tongue of 97 patients and their relationship to cause-specific (squamous cell carcinoma of the tongue) patient survival (median follow-up period of 33.5 months; range, 1-234 months). Claudins 1 and 7 were strongly expressed, claudin 4 had moderate expression, whereas claudin 5 was least expressed. Occludin staining was mostly negative or very weak. Western blot analysis of tongue carcinoma (HSC-3) cells showed that all these proteins are also expressed in vitro. In cause-specific survival analysis, strong and low immunoreactivity of claudin 7 tended to be associated with decreased survival compared with medium immunoreactivity. We suggest that analyzing the immunohistologic staining levels of claudin 7 could be used for the prognostic purposes in patients with tongue squamous cell carcinoma.


Rapid, estrogen receptor-mediated signaling: why is the endothelium so special?
Classical, ligand-activated genomic effects of estrogen receptors (ERs) were once thought to mediate all estrogen responses. It is now accepted that rapid, nongenomic responses, mediated by ER-containing membrane complexes, occur in many tissues. The endothelium is a major target of such responses and is the critical regulatory tissue that, when normally functional, determines a state of "vascular health." When dysfunctional, the phenotypic and functional alterations result in vascular pathology, the most common form of which is atherosclerosis. Nitric oxide (NO) is a vascular protective substance generated by endothelial NO synthase (eNOS) in endothelial cells. The engagement of membrane ERs by 17beta-estradiol (E2) is a potent stimulus to eNOS activation and NO release. Here, we describe the multimolecular components of ER-containing membrane complex assembly and the mechanisms directing ER targeting to caveolae microdomains in the plasma membrane. We discuss the possibility that various ERalpha splice forms, expressed in endothelial cells, may be particularly efficient signal transducers and may use classical receptor domains for membrane targeting and insertion. Finally, we discuss the biomedical ramifications of ER-mediated endothelial activation, including the controversies surrounding hormone replacement therapy and cardiovascular disease.


Amplification of IL-21 signalling pathway through Bruton's tyrosine kinase in human B cell activation.
B cells play an important role in the pathogenesis of autoimmune diseases. The role of Bruton's tyrosine kinase (Btk) in cytokine-induced human B cell differentiation and class-switch recombination remains incompletely defined. This study analysed the effect of Btk on human activated B cells. Purified B cells from healthy subjects were stimulated with B cell receptor (BCR) and other stimuli with or without a Btk inhibitor and gene expression was measured. The B cell line BJAB was used to assess Btk-associated signalling cascades. Phosphorylated Btk (p-Btk) in peripheral blood B cells obtained from 10 healthy subjects and 41 patients with RA was measured by flow cytometry and compared with patient backgrounds. IL-21 signalling, in concert with BCR, CD40 and BAFF signals, led to robust expression of differentiation- and class-switch DNA recombination-related genes and IgG production in human B cells, all of which were significantly suppressed by the Btk inhibitor. Although phosphorylation of STAT1 and STAT3 was induced by co-stimulation with IL-21, BCR and CD40, STAT1 phosphorylation in the nucleus, but not in the cytoplasm, was exclusively impaired by Btk blockade. High levels of p-Btk were noted in B cells of RA patients compared with controls and they correlated significantly with titres of RF among RF-positive patients. The findings elucidate a model in which Btk not only plays a fundamental role in the regulation of BCR signalling, but may also mediate crosstalk with cytokine signalling pathways through regulation of IL-21-induced phosphorylation of STAT1 in the nuclei of human B cells. Btk appears to have pathological relevance in RA.


Evaluation of Hyperparameter Optimization in Machine and Deep Learning Methods for Decoding Imagined Speech EEG.
Classification of electroencephalography (EEG) signals corresponding to imagined speech production is important for the development of a direct-speech brain-computer interface (DS-BCI). Deep learning (DL) has been utilized with great success across several domains. However, it remains an open question whether DL methods provide significant advances over traditional machine learning (ML) approaches for classification of imagined speech. Furthermore, hyperparameter (HP) optimization has been neglected in DL-EEG studies, resulting in the significance of its effects remaining uncertain. In this study, we aim to improve classification of imagined speech EEG by employing DL methods while also statistically evaluating the impact of HP optimization on classifier performance. We trained three distinct convolutional neural networks (CNN) on imagined speech EEG using a nested cross-validation approach to HP optimization. Each of the CNNs evaluated was designed specifically for EEG decoding. An imagined speech EEG dataset consisting of both words and vowels facilitated training on both sets independently. CNN results were compared with three benchmark ML methods: Support Vector Machine, Random Forest and regularized Linear Discriminant Analysis. Intra- and inter-subject methods of HP optimization were tested and the effects of HPs statistically analyzed. Accuracies obtained by the CNNs were significantly greater than the benchmark methods when trained on both datasets (words: 24.97%, p < 1 × 10-7, chance: 16.67%; vowels: 30.00%, p < 1 × 10-7, chance: 20%). The effects of varying HP values, and interactions between HPs and the CNNs were both statistically significant. The results of HP optimization demonstrate how critical it is for training CNNs to decode imagined speech.


miRNA-193a-3p Regulates the AKT2 Pathway to Inhibit the Growth and Promote the Apoptosis of Glioma Cells by Targeting ALKBH5.
Emerging evidence indicates that microRNA (miR)-193a-3p is involved in the tumor progression of various cancers. However, the biological functions and precise molecular mechanisms of miR-193a-3p in gliomas have not been well documented. Accordingly, this study focused on the tumor suppressor role and molecular mechanisms of miR-193a-3p in glioma cells. miR-193a-3p expression was determined by qRT-PCR in glioma tissues and cell lines. U251 and U87 glioma cells were transfected with a miR-193a-3p mimic. The effects of miR-193a-3p on cell growth and apoptosis were investigated using MTT, colony-forming, and flow cytometry assays. Overexpression of miR-193a-3p in U87 cells also significantly suppressed tumorigenicity and induced apoptosis in the xenograft mouse model. Luciferase assays were conducted to determine if ALKBH5 is a direct target of miR-193a-3p in glioma cells. Immunoprecipitation was used to explore the interaction between ALKBH5 and RAC-serine/threonine-protein kinase 2 (AKT2) in glioma cells. miR-193a-3p was downregulated in glioma tissues and cell lines. miR-193a-3p treatment suppressed proliferation and promoted apoptosis in both U251 and U87 cells. Bioinformatics analysis and luciferase reporter assay identified a novel miR-193a-3p target, ALKBH5. Notably, the antitumor effect of miR-193a-3p transfection in glioma cells may be due to the miR-193a-3p-induced inhibition of AKT2 expression caused by the suppression of ALKBH5 expression. Furthermore, immunoprecipitation indicated that ALKBH5 physically interacted with AKT2 through an RNA-independent mechanism in glioma cells. miR-193a-3p directly targets ALKBH5 to inhibit the growth and promote the apoptosis of glioma cells by suppressing the AKT2 pathway both in vitro and in vivo, and the physical interaction between ALKBH5 and AKT2 is essential for suppressing cell apoptosis by upregulating miR-193a-3p in glioma cells. Our study revealed that the antitumor effects of miR-193a-3p on glioma cells is due to ALKBH5 mediation of the AKT2-induced intrinsic apoptosis signaling pathway.


[Effects of incubation temperature on the results of passive hemagglutination test].
Passive hemagglutination test was carried out at several incubation temperatures: 4, 21, 37, and 56 degrees C. Blood sera of donors, salmonellosis enteritidis patients, and specific nonadsorbed sera were used in the study. The least fluctuations of the results were seen in tests with the sera with the least manifest immune properties. The optimal incubation temperatures were found 4 and 21 degrees C, and in 1/3 of cases cold incubation was found preferable. The relationship between antibody titers and incubation temperature may be described by the formula: T4 > or = T21 > or = T37 > or = T56, with Tn representing the reverse value of antibody titer in certain incubation temperature.


The Kaposi's Sarcoma-Associated Herpesvirus (KSHV) gH/gL Complex Is the Predominant Neutralizing Antigenic Determinant in KSHV-Infected Individuals.
Kaposi's sarcoma-associated herpesvirus (KSHV) is the etiological agent of Kaposi's sarcoma (KS), one of the most prevalent cancers of people living with HIV/AIDS in sub-Saharan Africa. The seroprevalence for KSHV is high in the region, and no prophylactic vaccine against the virus is available. In this study, we characterized the antigenic targets of KSHV-specific neutralizing antibodies (nAbs) in asymptomatic KSHV-infected individuals and KS patients with high nAbs titers. We quantified the extent to which various KSHV envelope glycoproteins (gB, ORF28, ORF68, gH, gL, gM, gN and gpK8.1) adsorbed/removed KSHV-specific nAbs from the plasma of infected individuals. Our study revealed that plasma from a majority of KSHV neutralizers recognizes multiple viral glycoproteins. Moreover, the breadth of nAbs responses against these viral glycoproteins varies among endemic KS, epidemic KS and asymptomatic KSHV-infected individuals. Importantly, among the KSHV glycoproteins, the gH/gL complex, but neither gH nor gL alone, showed the highest adsorption of KSHV-specific nAbs. This activity was detected in 80% of the KSHV-infected individuals regardless of their KS status. The findings suggest that the gH/gL complex is the predominant antigenic determinant of KSHV-specific nAbs. Therefore, gH/gL is a potential target for development of KSHV prophylactic vaccines.


Trypanosoma cruzi: RNA structure and post-transcriptional control of tubulin gene expression.
Changes in tubulin expression are among the biochemical and morphological adaptations that occur during the life cycle of Trypanosomatids. To investigate the mechanism responsible for the differential accumulation of tubulin mRNAs in Trypanosoma cruzi, we determine the sequences of alpha- and beta-tubulin transcripts and analyzed their expression during the life cycle of the parasite. Two beta-tubulin mRNAs of 1.9 and 2.3 kb were found to differ mainly by an additional 369 nucleotides at the end of the 3' untranslated region (UTR). Although their transcription rates are similar in epimastigotes and amastigotes, alpha- and beta-tubulin transcripts are 3- to 6-fold more abundant in epimastigotes than in trypomastigotes and amastigotes. Accordingly, the half-lives of alpha- and beta-tubulin mRNAs are significantly higher in epimastigotes than in amastigotes. Transient transfection experiments indicated that positive regulatory elements occur in the 3' UTR plus downstream intergenic region of the alpha-tubulin gene and that both positive and negative elements occur in the equivalent regions of the beta-tubulin gene.


Plant phenolics: recent advances on their biosynthesis, genetics, and ecophysiology.
Land-adapted plants appeared between about 480 and 360 million years ago in the mid-Palaeozoic era, originating from charophycean green algae. The successful adaptation to land of these prototypes of amphibious plants - when they emerged from an aquatic environment onto the land - was achieved largely by massive formation of "phenolic UV light screens". In the course of evolution, plants have developed the ability to produce an enormous number of phenolic secondary metabolites, which are not required in the primary processes of growth and development but are of vital importance for their interaction with the environment, for their reproductive strategy and for their defense mechanisms. From a biosynthetic point of view, beside methylation catalyzed by O-methyltransferases, acylation and glycosylation of secondary metabolites, including phenylpropanoids and various derived phenolic compounds, are fundamental chemical modifications. Such modified metabolites have altered polarity, volatility, chemical stability in cells but also in solution, ability for interaction with other compounds (co-pigmentation) and biological activity. The control of the production of plant phenolics involves a matrix of potentially overlapping regulatory signals. These include developmental signals, such as during lignification of new growth or the production of anthocyanins during fruit and flower development, and environmental signals for protection against abiotic and biotic stresses. For some of the key compounds, such as the flavonoids, there is now an excellent understanding of the nature of those signals and how the signal transduction pathway connects through to the activation of the phenolic biosynthetic genes. Within the plant environment, different microorganisms can coexist that can establish various interactions with the host plant and that are often the basis for the synthesis of specific phenolic metabolites in response to these interactions. In the rhizosphere, increasing evidence suggests that root specific chemicals (exudates) might initiate and manipulate biological and physical interactions between roots and soil organisms. These interactions include signal traffic between roots of competing plants, roots and soil microbes, and one-way signals that relate the nature of chemical and physical soil properties to the roots. Plant phenolics can also modulate essential physiological processes such as transcriptional regulation and signal transduction. Some interesting effects of plant phenolics are also the ones associated with the growth hormone auxin. An additional role for flavonoids in functional pollen development has been observed. Finally, anthocyanins represent a class of flavonoids that provide the orange, red and blue/purple colors to many plant tissues. According to the coevolution theory, red is a signal of the status of the tree to insects that migrate to (or move among) the trees in autumn.


Public health preparedness for mass-casualty events: a 2002 state-by-state assessment.
The ongoing threat of a terrorist attack places public agencies under increasing pressure to ensure readiness in the event of a disaster. Yet, little published information exists regarding the current state of readiness, which would allow local and regional organizations to develop disaster preparedness plans that would function seamlessly across service areas. The objective of this study is to characterize state-level disaster readiness soon after September 2001 and correlate readiness with existing programs providing an organized response to medical emergencies. During the first quarter of 2002, a cross-sectional survey assessing five components of disaster readiness was administered in all 50 states. The five components of disaster readiness included: (1) statewide disaster planning; (2) coordination; (3) training; (4) resource capacity; and (5) preparedness for biological/chemical terrorism. Most states reported the presence of a statewide disaster plan (94%), but few are tested by activation (48%), and still fewer contain a bioterrorism component (38%). All states have designated disaster operations centers (100%), but few states have an operating communications system linking health and medical resources (36%). Approximately half of states offer disaster training to medical professionals; about 10% of states require the training. Between 22-48% of states have various contingency plans to treat victims when service capacity is exceeded. Biochemical protective equipment for health professionals is lacking in all but one state, and only 10% of states indicate that all hospitals have decontamination capabilities. States with a functioning statewide trauma system were significantly more likely to possess key attributes of a functioning disaster readiness plan. These findings suggest that disaster plans are prevalent among states. However, key programs and policies were noticeably absent. Communication systems remain fragmented and adequate training programs and protective equipment for health personnel are markedly lacking. Statewide trauma systems may provide a framework upon which to build future medical disaster readiness capacity.


Pre-hospital antibiotic therapy preceded by blood cultures in a physician-manned mobile emergency care unit.
Rapid recognition and antibiotic treatment, preferably preceded by blood cultures (BCs), is a mainstay in sepsis therapy. The objective of this investigation was to determine if pre-hospital BCs were feasible and drawn with an acceptably low level of contamination and to investigate whether pre-hospital antibiotics were administered on correct indications. We performed a register-based study in a pre-hospital physician-manned mobile emergency care unit (MECU) operating in a mixed urban/rural area in Denmark. All patients who received pre-hospital antibiotics by the MECU from November 2013 to October 2018 were reviewed. Outcome measures were characterisation of microbial findings and subsequent in-hospital confirmation of the pre-hospital indication for antibiotics. One-hundred-and-nineteen patients received antibiotics pre-hospitally. Six were excluded. One-hundred-and-thirteen patients were included in the study. BCs were drawn in 107 of the 113 patients (94.7% [88.8%-98.0%]). We found a true pathogen of sepsis in 29 (27.1% [19.0%-36.6%]) of these 107 patients. Nine (8.4% [3.9%-15.4%]) patients had contaminated pre-hospital BCs. Forty-nine of all patients (36.3% [27.4%-45.9%]) had causative pathogens in either their BCs or other samples confirming the pre-hospital tentative diagnosis. Eighty-two (72.6% [63.4%-80.5%]) patients received antibiotic therapy in-hospitally, while 27 (23.9% [16.4%-32.8%]) were assigned an in-hospital diagnosis not associated with infection. Four (3.5% [1.0%-8.8%]) patients died in hospital before a diagnosis was established. Pre-hospital administration of antibiotics preceded by BCs is feasible, although with somewhat high blood culture contamination rates. Antibiotics are administered on reasonable indications.


Food preferences of wild mountain gorillas.
Determining the nutritional and phenolic basis of food preference is important for understanding the nutritional requirements of animals. Preference is a measure of which foods would be consumed by an animal if there was no variation in availability among food items. From September 2004 to August 2005, we measured the food preferences of four wild mountain gorilla groups that consume foliage and fruit in Bwindi Impenetrable National Park, Uganda, to determine what nutrients and phenols are preferred and/or avoided. To do so, we asked the following questions: (1) Which plant species do the gorillas prefer? (2) Considering the different plant parts consumed of these preferred species, what nutrients and/or phenols characterize them? (3) Do the nutritional and phenolic characteristics of preferred foods differ among gorilla groups? We found that although some species were preferred and others were not, of those species found in common among the different group home ranges, the same ones were generally preferred by all groups. Second, all groups preferred leaves with relatively high protein content and relatively low fiber content. Third, three out of four groups preferred leaves with relatively high sugar amounts. Fourth, all groups preferred pith with relatively high sugar content. Finally, of the two groups tested, we found that the preferred fruits of one group had relatively high condensed tannin and fiber/sugar contents, whereas the other group's preferred fruits were not characterized by any particular nutrient/phenol. Overall, there were no differences among gorilla groups in nutritional and phenolic preferences. Our results indicate that protein and sugar are important in the diets of gorillas, and that the gorillas fulfil these nutritional requirements through a combination of different plant parts, shedding new light on how gorillas balance their diets in a variable environment.


Metabolomic profiling for the identification of novel diagnostic markers in prostate cancer.
Metabolomic profiling offers a powerful methodology for understanding the perturbations of biochemical systems occurring during a disease process. During neoplastic transformation, prostate cells undergo metabolic reprogramming to satisfy the demands of growth and proliferation. An early event in prostate cell transformation is the loss of capacity to accumulate zinc. This change is associated with a higher energy efficiency and increased lipid biosynthesis for cellular proliferation, membrane formation and cell signaling. Moreover, recent studies have shown that sarcosine, an N-methyl derivative of glycine, was significantly increased during disease progression from normal to localized to metastatic prostate cancer. Mapping the metabolomic profiles to their respective biochemical pathways showed an upregulation of androgen-induced protein synthesis, an increased amino acid metabolism and a perturbation of nitrogen breakdown pathways, along with high total choline-containing compounds and phosphocholine levels. In this review, the role of emerging biomarkers is summarized, based on the current understanding of the prostate cancer metabolome.


Mitochondrial DNA defects and selective extraocular muscle involvement in CPEO.
PURPOSE. Chronic progressive external ophthalmoplegia (CPEO) is a prominent, and often the only, presentation among patients with mitochondrial diseases. The mechanisms underlying the preferential involvement of extraocular muscles (EOMs) in CPEO were explored in a comprehensive histologic and molecular genetic study, to define the extent of mitochondrial dysfunction in EOMs compared with that in skeletal muscle from the same patient. METHODS. A well-characterized cohort of 13 CPEO patients harboring a variety of primary and secondary mitochondrial (mt)DNA defects was studied. Mitochondrial enzyme function was determined in EOM and quadriceps muscle sections with cytochrome c oxidase (COX)/succinate dehydrogenase (SDH) histochemistry, and the mutation load in single muscle fibers was quantified by real-time PCR and PCR-RFLP assays. RESULTS. CPEO patients with mtDNA deletions had more COX-deficient fibers in EOM (41.6%) than in skeletal muscle (13.7%, P > 0.0001), and single-fiber analysis revealed a lower mutational threshold for COX deficiency in EOM. Patients with mtDNA point mutations had a less severe ocular phenotype, and there was no significant difference in the absolute level of COX deficiency or mutational threshold between these two muscle groups. CONCLUSIONS. The more pronounced mitochondrial biochemical defect and lower mutational threshold in EOM compared with skeletal muscle fibers provide an explanation of the selective muscle involvement in CPEO. The data also suggest that tissue-specific mechanisms are involved in the clonal expansion and expression of secondary mtDNA deletions in CPEO patients with nuclear genetic defects.


Kinesin's backsteps under mechanical load.
Kinesins move processively toward the plus end of microtubules by hydrolyzing ATP for each step. From an enzymatic perspective, the mechanism of mechanical motion coupled to the nucleotide chemistry is often well explained using a single-loop cyclic reaction. However, several difficulties arise in interpreting kinesin's backstepping within this framework, especially when external forces oppose the motion of kinesin. We review evidence, such as an ATP-independent stall force and a slower cycle time for backsteps, that has emerged to challenge the idea that kinesin backstepping is due to ATP synthesis, i.e., the reverse cycle of kinesin's forward-stepping chemomechanics. Supplementing the conventional single-loop chemomechanics with routes for ATP-hydrolyzing backward steps and nucleotide-free steps, especially under load, gives a better physical interpretation of the experimental data on backsteps.


Controllable Coalescence Dynamics of Nanodroplets on Textured Surfaces Decorated with Well-Designed Wrinkled Nanostructures: A Molecular Dynamics Study.
The control of coalescence and motion of droplets play a significant role in advanced technology and our daily life, and one of the most important approaches is surface design. Here, the microtextured surfaces decorated with wrinkled substrates are designed and studied for coalescence behaviors. The simulation results show that the coalescence speed would be slower on such surfaces due to the downward movement of the droplets induced by the penetration of atoms into the grooves that delays their movement along the coalescence direction, which is considered as a restriction effect. With the increase of the angles and the interval distance of wrinkled structures, the coalescence time becomes longer and the coalescing process becomes unfavorable, resulting from the enhanced restriction effect. More importantly, a composite substrate possessing the original and sub-wrinkled structures has been designed to tune the coalescence dynamics. The results of this work may not only help shed light on understanding the coalescence behaviors on the wrinkled substrates but also propose a feasible method for controlling the liquid coalescence behavior, which is expected to provide some useful implications for practical applications.


IgA nephropathy with crescents in kidney transplant recipients.
Crescentic glomerulonephritis is an uncommon finding in renal allografts. Recurrence or de novo mesangial deposition of immunoglobulin A (IgA) in renal allografts most often is clinically benign, but some case reports have shown that IgA nephropathy in renal allografts can present as crescentic glomerulonephritis and may lead to rapid deterioration of graft function and/or graft loss. We reviewed diagnoses of all allograft biopsies at University of Washington Medical Center (Seattle, WA) from 1989 to 2003 and found 33 cases of glomerulonephritis with crescents. Eight of these cases were the result of recurrent or de novo IgA nephropathy. Clinical and pathological features of these patients were reviewed. Six of 8 cases with crescents were the result of recurrent IgA nephropathy, and 2 cases were presumptive de novo IgA nephropathy. Of the 8 patients with IgA nephropathy with crescents, 6 patients presented clinically with increasing serum creatinine levels; 4 patients, with proteinuria; and 4 patients, with hematuria. In 6 patients, there was 10% to 30% involvement of glomeruli, with crescents partially or completely filling urinary spaces. The other patients showed lesser (approximately 7% of sampled glomeruli) involvement. Four patients with IgA nephropathy with crescents developed renal failure and returned to hemodialysis therapy. Three patients had a benign clinical course, with stabilization of renal function. One patient was lost to follow-up. We identified a cohort of patients with glomerulonephritis with crescents in renal allografts with IgA nephropathy as the cause. In half the affected patients, this led to early progressive renal insufficiency and return to hemodialysis therapy.


The role of Melancholia in prostate cancer patients' depression.
Although it is well established that prostate cancer (PCa) patients are more likely to experience clinical depression than their age-matched non-prostate cancer peers, and that such depression can have negative effects upon survival, little is known about the underlying nature of the depressive symptomatology that these men experience. In particular, the incidence of melancholic symptoms of depression, which are signs of increased risk of suicide and resistance to treatment, has not previously been reported in PCa patients. The present study aimed to measure the incidence and nature of Melancholia in PCa depression. A sample of 507 PCa patients in Queensland, Australia, completed anonymous and confidential questionnaires about their background, treatment status, and depression. Data were analysed to select depressive symptoms that were part of the definition of Melancholia vs those which were not. Regression was used to determine the links between Melancholia and overall depressive status, and factor analysis revealed the underlying components of Melancholia, which were mapped over time since diagnosis for 3 years. Psychometric data were satisfactory. Melancholia significantly predicted depressive status for the most depressed subset of patients, but not for the total sample. Melancholia was factored into its components of Anhedonia and Agitation, and the first of these was more powerful in predicting Melancholia. Variability over the 3 years following diagnosis was noted for each of these two components of Melancholia. The strong presence of Melancholia in the depressive symptomatology of this sample of PCa patients suggests that some forms of treatment for depression may be more likely to succeed than others. The dominance of Anhedonia and Agitation over other symptoms of Melancholia also holds implications for treatment options when assisting these men to cope with their depression.


Analysis of psychometric properties of the brazilian portuguese version of the community attitudes towards the mentally ill (CAMI-BR).
To perform an analysis of the psychometric properties of the Brazilian Portuguese version of the COMMUNITY ATTITUDES TOWARDS THE MENTALLY ILL (CAMI-BR), a 40-item scale divided into four sub-scales. The study was conducted in a non-probability sample of 230 households located close to therapeutic residences in the west area of Rio de Janeiro. Reliability was assessed by test-retest and the Principal Component Analysis (PCA) was used to test the internal structure of the questionnaire. Most participants were women, married, with children aged 18 years or over. The overall score was 27.72 (SD = 3.31), showing attitudes ranging from neutral to positive stereotypes. The scale showed a high internal consistency (? = 0.842), consistent with other international studies. In the factor analysis, the sample was adequate (KMO = 0.800). The strength of the correlations among subscales and the factors of factor analysis were highly satisfactory. The version in Brazilian Portuguese suggests a better distinction among sub-scales through the lower correlation among them (between 0.336 and 0.441) as compared to higher values (between 0.630 and 0.770) found in the original scale. The community mental health ideology sub-scale showed a strong relation to factor 1 (? = 0.910). Benevolence had a strong relationship with factor 2 (? = 0.847); Authoritarianism and Social Restrictiveness had the highest correlation with factor 3 (? = 0.631 and 0.577 respectively). The scale psychometric properties were maintained after adjustment. Having registered a lower correlation between the scales the factor analysis further suggests that the Brazilian Portuguese version conveys more clearly the differences between the sub-scales.


Prolyl hydroxylase PHD3 enhances the hypoxic survival and G1 to S transition of carcinoma cells.
Hypoxia restricts cell proliferation and cell cycle progression at the G1/S interface but at least a subpopulation of carcinoma cells can escape the restriction. In carcinoma hypoxia may in fact select for cells with enhanced hypoxic survival and increased aggressiveness. The cellular oxygen sensors HIF proline hydroxylases (PHDs) adapt the cellular functions to lowered environmental oxygen tension. PHD3 isoform has shown the strongest hypoxic upregulation among the family members. We detected a strong PHD3 mRNA expression in tumors of head and neck squamous cell carcinoma (HNSCC). The PHD3 expression associated with expression of hypoxic marker gene. Using siRNA in cell lines derived from HNSCC we show that specific inhibition of PHD3 expression in carcinoma cells caused reduced cell survival in hypoxia. The loss of PHD3, but not that of PHD2, led to marked cell number reduction. Although caspase-3 was activated at early hypoxia no induction of apoptosis was detected. However, hypoxic PHD3 inhibition caused a block in cell cycle progression. Cell population in G1 phase was increased and the population in S phase reduced demonstrating a block in G1 to S transition under PHD3 inhibition. In line with this, the level of hyperphosphorylated retinoblastoma protein Rb was reduced by PHD3 knock-down in hypoxia. PHD3 loss led to increase in cyclin-dependent kinase inhibitor p27 expression but not that of p21 or p16. The data demonstrated that increased PHD3 expression under hypoxia enhances cell cycle progression and survival of carcinoma cells.


Identification of novel mutations including a double mutation in patients with inherited cardiomyopathy by a targeted sequencing approach using the Ion Torrent PGM system.
Inherited cardiomyopathy is the major cause of sudden cardiac death (SCD) and heart failure (HF). The disease is associated with extensive genetic heterogeneity; pathogenic mutations in cardiac sarcomere protein genes, cytoskeletal protein genes and nuclear envelope protein genes have been linked to its etiology. Early diagnosis is conducive to clinical monitoring and allows for presymptomatic interventions as needed. In the present study, the entire coding sequences and flanking regions of 12 major disease (cardiomyopathy)-related genes [namely myosin, heavy chain 7, cardiac muscle, β (MYH7); myosin binding protein C, cardiac (MYBPC3); lamin A/C (LMNA); troponin I type 3 (cardiac) (TNNI3); troponin T type 2 (cardiac) (TNNT2); actin, α, cardiac muscle 1 (ACTC1); tropomyosin 1 (α) (TPM1); sodium channel, voltage gated, type V alpha subunit (SCN5A); myosin, light chain 2, regulatory, cardiac, slow (MYL2); myosin, heavy chain 6, cardiac muscle, α (MYH6); myosin, light chain 3, alkali, ventricular, skeletal, slow (MYL3); and protein kinase, AMP-activated, gamma 2 non-catalytic subunit (PRKAG2)] in 8 patients with dilated cardiomyopathy (DCM) and in 8 patients with hypertrophic cardiomyopathy (HCM) were amplified and then sequenced using the Ion Torrent Personal Genome Machine (PGM) system. As a result, a novel heterozygous mutation (MYH7, p.Asn885Thr) and a variant of uncertain significance (TNNT2, p.Arg296His) were identified in 2 patients with HCM. These 2 missense mutations, which were absent in the samples obtained from the 200 healthy control subjects, altered the amino acid that was evolutionarily conserved among a number of vertebrate species; this illustrates that these 2 non-synonymous mutations play a role in the pathogenesis of HCM. Moreover, a double heterozygous mutation (PRKAG2, p.Gly100Ser plus MYH7, p.Arg719Trp) was identified in a patient with severe familial HCM, for the first time to the best of our knowledge. This patient provided us with more information regarding the genotype-phenotype correlation between mutations of MYH7 and PRKAG2. Taken together, these findings provide insight into the molecular mechanisms underlying inherited cardiomyopathy. The mutations identified in this study may be further investigated in the future in order to improve the diagnosis and treatment of patients with inherited cardiomyopathy. Furthermore, our findings indicated that sequencing using the Ion Torrent PGM system is a useful approach for the identification of pathogenic mutations associated with inherited cardiomyopathy, and it may be used for the risk evaluation of individuals with a possible susceptibility to inherited cardiomyopathy.


The Relationship Between HIV Infection and Cardiovascular Disease.
Over 30 million people are currently living with human immunodeficiency virus (HIV) infection, and over 2 million new infections occur per year. HIV has been found to directly affect vascular biology resulting in an increased risk of cardiovascular disease compared to uninfected persons. Although HIV infection can now be treated effectively with combination antiretroviral medications, significant toxicities such as hyperlipidemia, diabetes, and excess cardiovascular co-morbidity; as well as the potential for significant drug-drug interactions between HIV and cardiovascular medications, present new challenges for the management of persons infected with HIV. We first review basic principles of HIV pathogenesis and treatment and then discuss relevant clinical management strategies that will be useful for cardiologists who might be involved in the care of HIV infected patients.


Risk factors affecting outcome of metal-on-metal surface arthroplasty of the hip.
We evaluated radiologic and clinical features affecting the outcome of hybrid metal-on-metal surface arthroplasty of the hip in 119 hips in patients 40 years and younger. Only the hips that had either failed or had minimum 2-year followup were reviewed. Ninety-four hips in 83 patients with a mean age of 34.2 years (range, 15-40 years) were reviewed. Seventy-one percent of the patients were males and 29% of the patients were females; 14% had previous surgery. The Chandler index and surface arthroplasty risk index were calculated. The mean followup at 3 years (range, 2-5 years) showed that three hips were converted to a total hip replacement at a mean of 27 months (range, 2-50 months) after the original surgery, and 10 hips had significant radiologic changes. The mean surface arthroplasty risk index for these 13 problematic hips versus the remaining hips was significantly higher, 4.7 and 2.6, respectively. The mean angle between the prosthesis stem and femoral shaft in the problematic group was significantly smaller than in the remaining hips: 133 degrees and 139 degrees, respectively. With a surface arthroplasty risk index score greater than 3 the relative risk of early problems is 12 times greater than if surface arthroplasty risk index less than or equal to 3.


Polyps as biomarkers for colorectal neoplasia.
Current understanding of colorectal carcinogenesis suggests a series of genetic changes occurring pari passu with morphological changes ultimately resulting in a cancerous lesion. The adenomatous polyp was originally the prototype of the preneoplastic lesion but recently, other colonic polyps, primarily the hamartomas, have been clinically characterized as colorectal cancer biomarkers with genetic changes found mainly in the mesenchymal component as opposed to the ectodermal, or epithelial element. This, with the current interest in angiogenesis playing a role in the propagation of neoplastic lesions, has now encompassed every tissue element and opened the way for an understanding of the oncogenic process. This has suggested that considerable interaction occurs between all tissue elements, including what was previously described as epithelial-matrix interactions. While hyperplastic polyps are thought not to confer risk for cancer, they may offer clues as to the first steps of the overall process. Microadenomas have introduced new clinical as well as biological considerations, as unique risk factors. Investigation of these lesions has moved from purely morphological correlations to mechanistic dissections of important biological pathways using both genetic and protein chemistry tools. This review explores the microcosm of the colonic polyp and its relation to cancer as the quintessential premalignant biomarker.


[Lower cranial nerve involvement as the initial manifestation of Lyme borreliosis].
Lyme disease or Lyme borreliosis is an infectious disease transmitted by ticks and caused by Borrelia burgdorferi. Being clinically different from Relapsing Fever it may cause an array of symptoms, specially cutaneous and neurological but also musculoskeletal and cardiac ones. Within the neurologic manifestations of Lyme disease the affectation of low cranial nerves is, to our knowledge, extremely infrequent. We present the clinical case of a 35 years old male whose initial symptoms were low cranial nerve dysfunction with a cerebrospinal fluid compatible with meningitis. Serology against Borrelia burgdorferi both in serum and cerebrospinal fluid was positive.


[Risk factors for adverse course of gastric and duodenal peptic ulcer].
Introduction: High morbidity rate, frequent relapses, and significant economic losses give reasons for highlighting the peptic ulcer disease as the most topical medical-statistical problem. The aim of the study is to assess the influence of the main risk factors on the course of gastric and duodenal peptic ulcer. Materials and methods: We formed up the risk groups consisted of patients with 12 modified (4) and regular (8) factors, each characterized with its own signs and gradations. We performed the quantitative evaluation of the factors and scored the signs thereof, the results of which were used for determination of the most informative ones. Results: Among the regular factors, we placed emphasis on gender, age, burdened heredity, and 0(I), Rh+ blood type. The risk of peptic ulcer in hereditary tainted young men of the working age with parental lineage (+2.3) and in males with 0(I) Rh+ blood type (+1.4) was proved. Helicobacter pylori (Hp) infection is a key contributor (a predictor of) to severity of the disease course (+9.7) among the modified risk factors. Negative effect of a concomitant pathology (+5.0), including hepatobiliary lesions (+3.8), hypertension (+4.0), and diabetes mellitus (+1.3) is also significant. Diet violation (+3.7), tobacco smoking (+3.2) and stress (+3.0) were ranked third. Conclusions: The results of quantitative evaluation of the factors scoring suggest of the underlying H.pylori infection (the significance of which is growing along with the growth of the disease incidence) and irrational diet as the most informatively important ones. We have established the direct dependence between the most important peptic ulcer risk factors, severity of the disease, and duration and periodicity of treatment thereof.


Vitamin D: a potential role in reducing suicide risk?
Suicide attempts are known to peak in the spring, overlapping with the time of year when 25-hydroxyvitamin D [25(OH)D] levels are at their nadir in the northern hemisphere because of negligible skin production of vitamin D owing to low levels of ultraviolet B radiation. Low levels of 25(OH)D, the vitamin D metabolite used to diagnose vitamin D deficiency, have been associated with certain pro-suicidal factors such as exacerbation of depression, anxiety, psychosis, and certain medical conditions. Therefore, we hypothesize that vitamin D deficiency could also be associated with increased risk of completed suicides. Here, we briefly review the literature on vitamin D, its deficiency, and its reported association with certain risk factors for suicide.


Effect of Mass Fraction on Leaching Kinetics of Hydrophobic Ultraviolet Stabilizers in Low-Density Polyethylene.
The leaching kinetics of five hydrophobic ultraviolet (UV) stabilizers from low-density polyethylene (LDPE) (micro)fibers into water was evaluated in this study, with variation of the mass fraction (ω = 0.1-2.0 wt %) of the stabilizers. A one-dimensional convection-diffusion model for a cylindrical geometry, requiring partitioning between the LDPE fibers and water (KLDPEw) and the internal diffusion coefficients (DLDPE), was used to evaluate the leaching process and the leaching half-life of the target UV stabilizers at ω < 0.5 wt % (Case I). Diffusion through the aqueous boundary layer is the rate-determining step, and the leaching half-life is predicted to be very long (a few months to years) under unaffected conditions. When the UV stabilizers are supersaturated within LDPE fibers (i.e., ω > 0.5 wt %, Case II), the possible formation of a surficial crystal layer of the additives on the LDPE fiber extends the time scale for leaching compared to that in Case I due to the requirement of overcoming the crystallization energy. This study provides a fundamental understanding of the leaching profiles of plastic additives for assessing their potential chemical risks in aquatic environments; further studies under the relevant environmental conditions are required.


Rapid end-point quantitation of prion seeding activity with sensitivity comparable to bioassays.
A major problem for the effective diagnosis and management of prion diseases is the lack of rapid high-throughput assays to measure low levels of prions. Such measurements have typically required prolonged bioassays in animals. Highly sensitive, but generally non-quantitative, prion detection methods have been developed based on prions' ability to seed the conversion of normally soluble protease-sensitive forms of prion protein to protease-resistant and/or amyloid fibrillar forms. Here we describe an approach for estimating the relative amount of prions using a new prion seeding assay called real-time quaking induced conversion assay (RT-QuIC). The underlying reaction blends aspects of the previously described quaking-induced conversion (QuIC) and amyloid seeding assay (ASA) methods and involves prion-seeded conversion of the alpha helix-rich form of bacterially expressed recombinant PrP(C) to a beta sheet-rich amyloid fibrillar form. The RT-QuIC is as sensitive as the animal bioassay, but can be accomplished in 2 days or less. Analogous to end-point dilution animal bioassays, this approach involves testing of serial dilutions of samples and statistically estimating the seeding dose (SD) giving positive responses in 50% of replicate reactions (SD(50)). Brain tissue from 263K scrapie-affected hamsters gave SD(50) values of 10(11)-10(12)/g, making the RT-QuIC similar in sensitivity to end-point dilution bioassays. Analysis of bioassay-positive nasal lavages from hamsters affected with transmissible mink encephalopathy gave SD(50) values of 10(3.5)-10(5.7)/ml, showing that nasal cavities release substantial prion infectivity that can be rapidly detected. Cerebral spinal fluid from 263K scrapie-affected hamsters contained prion SD(50) values of 10(2.0)-10(2.9)/ml. RT-QuIC assay also discriminated deer chronic wasting disease and sheep scrapie brain samples from normal control samples. In principle, end-point dilution quantitation can be applied to many types of prion and amyloid seeding assays. End point dilution RT-QuIC provides a sensitive, rapid, quantitative, and high throughput assay of prion seeding activity.


Structure-activity relationship study of a series of N-substituted piperazinyl-fluoroquinolones as anti-Helicobacter pylori agents.
Helicobacter pylori is now recognized as the primary etiological factor associated with gastritis, peptic ulcer disease and gastric cancers. Fluoroquinolones have been shown to be active against H. pylori. For develop new anti-H. pylori agents, we have investigated the SAR of a series of N-(phenethyl)piperazinyl quinolones for their antimicrobial activity against H. pylori. The anti-H. pylori activity of synthesized compounds along with commercially available anti-H. pylori agents such as metronidazole, and parent quinolones was evaluated by the disc diffusion bioassay. The results indicated that the potency and anti-H. pylori activity profile of the quinolones is highly dependent on the type of substituent at N-1 and the structure of phenethyl unit on piperazine ring. Most compounds containing a cyclopropyl at N-1 exhibited good activity against H. pylori strains. Among them, ciprofloxacin derivative 13 containing 2-methoxyimino-2-(2-chlorophenyl)ethyl moiety was the most active compound.