Physical Activity Across the Cancer Journey: Experiences and Recommendations From People Living With and Beyond Cancer.
The majority of individuals living with and beyond cancer are not sufficiently active to achieve health benefits. The aim of this study was to explore individuals' experiences of physical activity (PA) behavior across the cancer journey and to ask individuals living with and beyond cancer to identify strategies to support habitual PA. An exploratory, descriptive, qualitative design was used. Purposive sampling methods were used to recruit individuals living with and beyond cancer who had been referred to, and/or participated in, a community-based exercise program or were attending a cancer support center. The focus group discussions were audio recorded, transcribed verbatim, and analyzed using a thematic analysis approach. Seven focus groups were conducted with 41 participants. Many individuals reported that regular PA provided a vehicle for recovery that created a sense of "self-power," defined as taking ownership and control of one's health to increase well-being. Barriers to PA participation included environmental-, patient-, and treatment-related challenges. Recommendations to support long-term adherence to PA included completion of fitness assessments at regular intervals and provision of a home exercise program. The benefits and barriers to PA participation for individuals diagnosed with cancers that were not represented may not have been identified. The strategies recommended to support habitual PA may be salient only to individuals whose cancer diagnoses were represented. Exercise is seen as a vehicle for recovery from cancer but long-term adherence for individuals is complex. The findings from this study can inform the development of exercise oncology rehabilitation programs and could support a greater likelihood of program success, thereby optimizing the health, well-being, and quality of life of survivors of cancer.


Inhibition of tumor growth by plant-derived mAb.
The tumor-associated antigen EpCAM (GA733-2) is a highly expressed target on adenocarcinoma cells, as defined by murine mAb CO17-1A. We recently developed a transgenic plant system for the safe and inexpensive production of large quantities of mAb CO17-1A as a future source of clinical-grade protein. Although the glycosylation pattern of plant-derived mAb (mAb(P)) CO17-1A differs considerably from that of the mammalian-derived mAb (mAb(M)), we show here that the biological activity of both mAbs is quite similar. mAb(P) heavy and light chains assembled to bind the recombinant antigen GA733-2E and specifically bound to human SW948 colorectal carcinoma cells expressing the antigen GA733-2 to the same extent as mAb(M). mAb(P) was as effective as mAb(M) CO17-1A in inhibiting tumor growth of xenotransplanted SW948 cells in nude mice. These results suggest the promise of transgenic plants as a useful alternative way to produce full-size mAb for cancer immunotherapy.


Survival after hospital discharge for ST-segment elevation and non-ST-segment elevation acute myocardial infarction: a population-based study.
Limited recent data are available describing differences in long-term survival, and factors affecting prognosis, after ST-segment elevation myocardial infarction (STEMI) and non-ST-segment elevation myocardial infarction (NSTEMI), especially from the more generalizable perspective of a population-based investigation. The objectives of this study were to examine differences in post-discharge prognosis after hospitalization for STEMI and NSTEMI, with a particular focus on factors associated with reduced long-term survival. We reviewed the medical records of residents of the Worcester, MA, USA metropolitan area hospitalized at eleven central Massachusetts medical centers for acute myocardial infarction (AMI) during 2001, 2003, 2005, and 2007. A total of 3762 persons were hospitalized with confirmed AMI; of these, 2539 patients (67.5%) were diagnosed with NSTEMI. The average age of study patients was 70.3 years and 42.9% were women. Patients with NSTEMI experienced higher post-discharge death rates with 3-month, 1-year, and 2-year death rates of 12.6%, 23.5%, and 33.2%, respectively, compared to 6.1%, 11.5%, and 16.4% for patients with STEMI. After multivariable adjustment, patients with NSTEMI were significantly more likely to have died after hospital discharge (adjusted hazards ratio 1.28; 95% confidence interval 1.14-1.44). Several demographic (eg, older age) and clinical (eg, history of stroke) factors were associated with reduced long-term survival in patients with NSTEMI and STEMI. The results of this study in residents of central Massachusetts suggest that patients with NSTEMI are at higher risk for dying after hospital discharge, and several subgroups are at particularly increased risk.


NanoLC-timsTOF-Assisted Analysis of Glycated Albumin in Diabetes-Affected Plasma and Tears.
Glycation is a spontaneous and nonenzymatic glycosylation. Glycated albumin (GA), which serves as an important biomarker in plasma in the diagnosis and characterization of diabetes, can be passively filtered from the plasma to tears. Tears are important targets for research in clinical diagnostics due to the ability to collect this biofluid noninvasively and repeatably. Therefore, the analysis of GA in tear film provides information for monitoring diabetes progression independent of blood pathologies. Due to the limited volume (1-5 μL) of natural tear film, we developed a small volume assay using a nano liquid chromatography-trapped ion mobility spectrometry-time-of-flight MS (nanoLC-timsTOF) platform for the analysis of glycated albumin in human plasma and tear films affected by diabetes. The peptides containing lysine 525, which is the main glycation site in GA, were relatively quantified and represented as the GA level. The results of the measurements showed that GA levels were significantly higher in diabetes-affected plasma and tears compared to controls with a p-value < 0.01. A strong correlation of glycated albumin levels was observed for the plasma and tear film in diabetes samples (Pearson coefficient 0.92 with a p-value 0.0012). Moreover, the number of GA glycation sites was significantly higher in diabetes-affected plasma and tear comparatively to controls. Among all the glycation sites in plasma albumin, the GA level quantified by lysine 136/137 had a strong correlation with more commonly used lysine 525, suggesting that lysine 136 /137 is an alternative diabetes biomarker in plasma. Overall, our findings demonstrate GA in tears as a biomarker for monitoring diabetes progression, highlighting new possibilities for quick and noninvasive diabetes detection and monitoring.


The impact of the Internet on quality measurement.
Consumers are eager for information about health. However, their use of such data has been limited to date. When consumers do consider data in making health care choices, they rely more on word-of-mouth reputation than on traditional quality measures, although this information has not necessarily been readily accessible. The Internet changes the exercise of quality measurement in several ways. First, quality information--including reputation--will be more readily available. Second, consumers will increasingly use it. Third, the Internet provides a low-cost, standard platform that will make it vastly easier for providers to collect quality information and pass it on to others. However, major barriers still stand in the way of public access to quality information on the Internet as well as of having that access actually improve patients' care.


Influence of the playing surface on changes of direction and plantar pressures during an agility test in youth futsal players.
The ability to effect a change of direction (COD) when performing high-speed actions is essential in team sports like futsal. Nevertheless, the interaction effect of sports surfaces on this ability remains unknown. This research aimed to analyse the plantar pressures and time performance of 15 youth futsal players when performing CODs on two playing surfaces with different mechanical properties. The shock absorption and vertical deformation of one synthetic flooring surface and one wooden flooring surface were assessed. CODs were evaluated using a modified version of the Agility T-Test, while total time of the test, time of CODs, and plantar pressure in CODs were all recorded. The wooden flooring surface displayed higher values of shock absorption (35.70% ± 2.87%) and vertical deformation (2.77 ± 0.38 mm) compared with the synthetic flooring (p < 0.01). Faster CODs were performed on the wooden flooring than on the synthetic flooring (-0.05 s, 95% confidence interval [CI]: -0.10 to -0.007, effect size [ES]: 0.07, p < 0.05), but no differences in total test time were found (p > 0.05). Finally, no differences in plantar pressures by playing surface were found. In sum, the differences in the mechanical properties of the two futsal surfaces affected the performance of futsal players in the modified agility test. However, these differences were not great enough to generate different plantar pressures on players, probably due to playerś own adaptations.


Congenital duplication of the foot and toes.
Congenital duplication of the foot and toes is a commonly diagnosed condition in newborn nurseries across the United States. The foot involvement varies from simple fleshy tags to complex attempts at whole extremity duplication. We reviewed 59 patients (82 feet) with a variety of duplications (excluding skin tags). During our review, we devised a new descriptive system of classification of congenital duplications of the foot based primarily on the type and complexity of the skeletal duplication. Variables related to sex, bilaterality, age at surgery, family history, and follow-up results were documented, and an exact classification of each foot was accomplished. Results and prognosis vary with the class of duplication being treated and must be individualized case by case.


Spine cancer: a summary and review.
Spine pain is a common presenting complaint of patients who visit physicians and although spine cancer accounts for a very small percentage of cases of back pain, metastasis is a relatively common cause of spine pain in the elderly. The presentation of patients with spine cancers is highly variable, and in many cases the clinical findings of benign and cancerous causes of spine pain can be similar, often confounding the clinical picture. This can create difficulties in interpreting the clinical data available to the physician, particularly with a disease with such a devastating prognosis. This manuscript discusses the more common causes of malignant lesions of the spine, including an overview of the incidence/prevalence data and clinical features of both primary and secondary malignancies. It also provides the reader with a clinical overview of patients with spine cancer. It is important to appreciate the myriad epidemiologic and clinical features of primary and secondary spine cancers. Patients with malignant skeletal lesions may be asymptomatic in the area of cancerous bone disease and, as a consequence, these lesions can be overlooked. This may result in dismal consequences for the patient, given the generally poor prognosis associated with spine cancers. Knowledge of the features discussed in this paper will assist the clinician in appropriately raising his/her index of suspicion for spine cancer in suitable clinical circumstances.


The use of the ALOPEX process in extracting normal and abnormal visual evoked potentials.
The ALOPEX method, a multiparameter stochastic optimization technique, is used for the optimal extraction of parameters in fitting randomly chosen action potentials to normal and abnormal visual evoked potential (VEP) waveforms. It is assumed that conformational changes in the abnormal VEPs can be attributed to an asynchrony in the arrival of action potentials at an integration point in the visual cortex. The results indicate that the assumption is valid in all five of the five records processed, of which two were normal and three abnormal. Further experimentation is needed to see if the method can be used as a diagnostic tool.


Reproductive hormones in the early menopausal transition: relationship to ethnicity, body size, and menopausal status.
We measured serum reproductive hormone concentrations in a community-based, multiethnic population of premenopausal and early perimenopausal women to determine whether there are ethnic differences in hormones that can be explained by host factors. We studied 2930 participants in the Study of Women's Health Across the Nation who were aged 42-52 yr and self-identified as African-American (27.6%), Caucasian (47.1%), Chinese (7.4%), Hispanic (8.8%), or Japanese (9.0%) at 7 clinical sites. Outcome measures from this baseline assessment of a longitudinal study were serum estradiol (E2), FSH, testosterone (T), dehydroepiandrosterone sulfate, and SHBG concentrations and calculated estimates of free steroid availability, free testosterone index, and free E2 index from serum collected primarily in the early follicular phase of a spontaneous menstrual cycle. The primary explanatory variables were race/ethnicity, menopausal status, age, body mass index, day of the cycle, smoking, alcohol use, and physical activity. Chinese women had lower unadjusted E2 and SHBG levels, and Hispanic women had lower unadjusted T levels than other ethnic groups. Unadjusted serum FSH levels did not differ by race/ethnicity. E2 levels adjusted for host characteristics, particularly body size, did not differ by race/ethnicity. Adjusted FSH levels were higher, and adjusted T levels were lower in African-American and Hispanic women. Serum E2 and FSH concentrations were highly variable. Serum FSH levels, but no other hormone concentrations, were positively correlated with menopausal status. Serum dehydroepiandrosterone sulfate levels were negatively correlated with age, but not menopausal status. All hormone concentrations were significantly correlated with body mass index. We conclude that serum sex steroid, FSH, and SHBG levels vary by ethnicity, but are highly confounded by ethnic disparities in body size.


The ATP-dependent chromatin remodeling enzyme Fun30 represses transcription by sliding promoter-proximal nucleosomes.
The evolutionarily conserved ATP-dependent chromatin remodeling enzyme Fun30 has recently been shown to play important roles in heterochromatin silencing and DNA repair. However, how Fun30 remodels nucleosomes is not clear. Here we report a nucleosome sliding activity of Fun30 and its role in transcriptional repression. We observed that Fun30 repressed the expression of genes involved in amino acid and carbohydrate metabolism, the stress response, and meiosis. In addition, Fun30 was localized at the 5' and 3' ends of genes and within the open reading frames of its targets. Consistent with its role in gene repression, we observed that Fun30 target genes lacked histone modifications often associated with gene activation and showed an increased level of ubiquitinated histone H2B. Furthermore, a genome-wide nucleosome mapping analysis revealed that the length of the nucleosome-free region at the 5' end of a subset of genes was changed in Fun30-depleted cells. In addition, the positions of the -1, +2, and +3 nucleosomes at the 5' end of target genes were shifted significantly, whereas the position of the +1 nucleosome remained largely unchanged in the fun30Δ mutant. Finally, we demonstrated that affinity-purified, single-component Fun30 exhibited a nucleosome sliding activity in an ATP-dependent manner. These results define a role for Fun30 in the regulation of transcription and indicate that Fun30 remodels chromatin at the 5' end of genes by sliding promoter-proximal nucleosomes.


Genome-wide profiling of stored mRNA in Arabidopsis thaliana seed germination: epigenetic and genetic regulation of transcription in seed.
To reveal the transcriptomes of Arabidopsis seed, comprehensive expression analysis was performed using ATH1 GeneChips (Affymetrix, Santa Clara, CA, USA). In the dry seed, more than 12 000 stored mRNA species were detected, including all ontological categories. Statistical analysis revealed that promoters of highly expressed genes in wild-type dry seeds overrepresented abscisic acid-responsive elements (ABREs) containing the core motif ACGT. Although the coupling element and seed-specific enhancer RY motif alone were not prominently overrepresented in genes with high expression, the presence of these elements in combination with ABRE was associated with particularly high gene expression. The transcriptome of the imbibed seeds differed from that of the dry seed even at 6 h after seed imbibition. After imbibition many upregulated and downregulated genes were co-regulated in clusters of three to five genes. Genes for which expression was affected by the abi5 mutation tended to be located in clusters, suggesting that transactivation by ABI5 is not restricted to a single gene, but affects other proximal genes. Furthermore, cytosine methylation was observed not only in large silent retrotransposon clusters in centromeric regions, but also in non-centromeric silent gene clusters in the seed. These results suggest that such regions might be transcriptionally silenced by methylation or heterochromatin structures. Our analyses reveal that transcriptomes of Arabidopsis seed are characterized by multiple regulatory mechanisms: epigenetic chromatin structures, chromosomal locations (e.g. co-regulated gene clusters) and cis-acting elements.


Mitochondrial DNA diversity of Anatolian indigenous domestic goats.
Anatolia has been an important region for civilizations and agricultural revolution as a major domestication centre for livestock species. Goats (Capra hircus) were among the earliest domesticated animals in this region. In this study, genetic diversity of Anatolian goat breeds was characterized by comparison of mitochondrial DNA hypervariable region 1. A total of 295 individuals, including 99 Anatolian Black goats, 96 Angora goats and 100 Kilis goats, were used. Haplogroup A was found to be the dominant haplogroup in all three breeds. The highest haplogroup diversity, including haplogroups A, B2, C and G, was observed in the Anatolian Black breed. Haplogroup D was only observed in Kilis and Angora goats. Haplogroup G was found in Angora and Anatolian Black breeds. The Anatolian goat breeds had high genetic diversity values and a weak phylogeographical structure. The nucleotide diversity values were found to be higher than those in previously studied goat breeds. The fact that Anatolia is a domestication centre and its geographical position as a junction of trade routes may have caused the higher genetic diversity of Anatolian goat breeds.


The theoretical foundations of enaction: Precariousness.
Enaction is an increasingly influential approach to cognition that grew out of Maturana and Varela's earlier work on autopoiesis and the biology of cognition. As with any relatively new scientific discipline, the enactive approach would benefit greatly from a careful analysis of its theoretical foundations. Here we initiate such an analysis for one of the core concepts of enaction, precariousness. Specifically, we consider three types of fragility: systemic, processual and thermodynamic. Using a glider in the Game of Life as a toy model, we illustrate each of these fragilities and examine the relationships between them. We also argue that each type of fragility is characterized by which aspects of a system are hardwired into its definition from the outset and which aspects are emergent and hence vulnerable to disintegration without ongoing maintenance.


A systems approach to delineate functions of paralogous transcription factors: role of the Yap family in the DNA damage response.
Duplication of genes encoding transcription factors plays an essential role in driving phenotypic variation. Because regulation can occur at multiple levels, it is often difficult to discern how each duplicated factor achieves its regulatory specificity. In these cases, a "systems approach" may distinguish the role of each factor by integrating complementary large-scale measurements of the regulatory network. To explore such an approach, we integrate growth phenotypes, promoter binding profiles, and gene expression patterns to model the DNA damage response network controlled by the Yeast-specific AP-1 (YAP) family of transcription factors. This analysis reveals that YAP regulatory specificity is achieved by at least three mechanisms: (i) divergence of DNA-binding sequences into two subfamilies; (ii) condition-specific combinatorial regulation by multiple Yap factors; and (iii) interactions of Yap 1, 4, and 6 with chromatin remodeling proteins. Additional microarray experiments establish that Yap 4 and 6 regulate gene expression through interactions with the histone deacetylase, Hda1. The data further highlight differences among Yap paralogs in terms of their regulatory mode of action (activation vs. repression). This study suggests how other large TF families might be disentangled in the future.


Role of the clinical nurse specialist: an integrative review of the literature.
This review of the literature related to the role of the clinical nurse specialist was commissioned by the AACN in order to provide a comprehensive resource for advanced practice nurses. This review provides a historical overview of clinical nurse specialists and some of the gaps or barriers to practice that have been experienced. A summary of these gaps shows that many have repeated themselves over the years. Also provided are potential solutions to facilitate the progression of this role in the future.


Pancreatic and extrapancreatic galanin release during sympathetic neural activation.
To address the hypothesis that the neutropeptide, galanin, functions as a sympathetic neurotransmitter in the endocrine pancreas, we sought to determine if galanin is released from pancreatic sympathetic nerves during their direct electrical stimulation in halothane-anesthetized dogs. During bilateral thoracic splanchnic nerve stimulation (BTSNS), both peripheral arterial and pancreatic venous levels of galanin-like immunoreactivity (GLIR) increased (delta at 10 min = +92 +/- 31 and +88 +/- 25 fmol/ml, respectively). Systemic infusions of synthetic galanin demonstrated that 1) the increment of arterial GLIR observed during BTSNS was sufficient to modestly restrain basal insulin secretion and 2) only 25% of any given increment of arterial GLIR appears in the pancreatic vein, suggesting that the pancreas extracts galanin, as it does other neurotransmitters. By use of 75% for pancreatic extraction of circulating galanin, it was calculated that pancreatic galanin spillover (output) increased by 410 +/- 110 fmol/min during BTSNS. To reinforce the conclusion that pancreatic sympathetic nerves release galanin, GLIR spillover was next measured during direct local stimulation of the pancreatic sympathetic input produced by electrical stimulation of the mixed autonomic pancreatic nerves (MPNS) in the presence of the ganglionic blocker, hexamethonium. During this local pancreatic sympathetic nerve stimulation, arterial GLIR remained unchanged, but pancreatic venous GLIR increased by 123 +/- 34 fmol/ml. Thus pancreatic GLIR spillover increased by 420 +/- 110 fmol/min during MPNS in the presence of hexamethonium. We conclude that galanin is released from both pancreatic and extrapancreatic sources during sympathetic neural activation in dogs.


DNA binding restricts the intrinsic conformational flexibility of methyl CpG binding protein 2 (MeCP2).
Mass spectrometry-based hydrogen/deuterium exchange (H/DX) has been used to define the polypeptide backbone dynamics of full-length methyl CpG binding protein 2 (MeCP2) when free in solution and when bound to unmethylated and methylated DNA. Essentially the entire MeCP2 polypeptide chain underwent H/DX at rates faster than could be measured (i.e. complete exchange in ≤10 s), with the exception of the methyl DNA binding domain (MBD). Even the H/DX of the MBD was rapid compared with that of a typical globular protein. Thus, there is no single tertiary structure of MeCP2. Rather, the full-length protein rapidly samples many different conformations when free in solution. When MeCP2 binds to unmethylated DNA, H/DX is slowed several orders of magnitude throughout the MBD. Binding of MeCP2 to methylated DNA led to additional minor H/DX protection, and only locally within the N-terminal portion of the MBD. H/DX also was used to examine the structural dynamics of the isolated MBD carrying three frequent mutations associated with Rett syndrome. The effects of the mutations ranged from very little (R106W) to a substantial increase in conformational sampling (F155S). Our H/DX results have yielded fine resolution mapping of the structure of full-length MeCP2 in the absence and presence of DNA, provided a biochemical basis for understanding MeCP2 function in normal cells, and predicted potential approaches for the treatment of a subset of RTT cases caused by point mutations that destabilize the MBD.


Role of zinc in plasma membrane function.
The concentration of plasma zinc is the generally accepted index of zinc status. Although low plasma zinc is an essential criterion of deficiency, alone it is inadequate. To supplement this index, we sought to determine the first limiting biochemical defect in animals fed zinc-deficient diets and concluded that the limiting function is associated with a posttranslational change in plasma membrane proteins. Among the signs of zinc deficiency in rats is a bleeding tendency associated with failure of platelet aggregation, a phenomenon that correlates with impaired uptake of Ca(2+) when stimulated. Zinc-deficient guinea pigs exhibit signs of peripheral neuropathy, and their brain synaptic vesicles exhibit impaired Ca(2+) uptake when they are stimulated with glutamate. Red cells from zinc-deficient rats show increased osmotic fragility associated with decreased plasma membrane sulfhydryl concentration. Both phenomena are readily reversed (2 d) by dietary zinc repletion. Volume recovery is dependent on Ca-dependent K channels and the sulfhydryl redox state. Both the impaired aggregation and calcium uptake of zinc-deficient platelets are corrected by in vitro incubation of blood with glutathione. Considering the fact that plasma membranes from several cell types show impaired function that is associated with a decreased rate of calcium uptake, it is postulated that a defect in calcium channels is the first limiting biochemical defect in zinc deficiency. The calcium uptake defect and consequent impaired second-messenger function likely results from an abnormal sulfhydryl redox state in the membrane channel protein.


Arabidopsis cop8 and fus4 mutations define the same gene that encodes subunit 4 of the COP9 signalosome.
The pleiotropic constitutive photomorphogenic/deetiolated/fusca (cop/det/fus) mutants of Arabidopsis exhibit features of light-grown seedlings when grown in the dark. Cloning and biochemical analysis of COP9 have revealed that it is a component of a multiprotein complex, the COP9 signalosome (previously known as the COP9 complex). Here, we compare the immunoaffinity and the biochemical purification of the COP9 signalosome from cauliflower and confirm its eight-subunit composition. Molecular cloning of subunit 4 of the complex revealed that it is a proteasome-COP9 complex-eIF3 domain protein encoded by a gene that maps to chromosome 5, near the chromosomal location of the cop8 and fus4 mutations. Genetic complementation tests showed that the cop8 and fus4 mutations define the same locus, now designated as COP8. Molecular analysis of the subunit 4-encoding gene in both cop8 and fus4 mutants identified specific molecular lesions, and overexpression of the subunit 4 cDNA in a cop8 mutant background resulted in complete rescue of the mutant phenotype. Thus, we conclude that COP8 encodes subunit 4 of the COP9 signalosome. Examination of possible molecular interactions by using the yeast two-hybrid assay indicated that COP8 is capable of strong self-association as well as interaction with COP9, FUS6/COP11, FUS5, and Arabidopsis JAB1 homolog 1, the latter four proteins being previously defined subunits of the Arabidopsis COP9 signalosome. A comparative sequence analysis indicated that COP8 is highly conserved among multicellular eukaryotes and is also similar to a subunit of the 19S regulatory particle of the 26S proteasome.


Heat-shock response transcriptional program enables high-yield and high-quality recombinant protein production in Escherichia coli.
The biosynthesis of soluble, properly folded recombinant proteins in large quantities from Escherichia coli is desirable for academic research and industrial protein production. The basal E. coli protein homeostasis (proteostasis) network capacity is often insufficient to efficiently fold overexpressed proteins. Herein we demonstrate that a transcriptionally reprogrammed E. coli proteostasis network is generally superior for producing soluble, folded, and functional recombinant proteins. Reprogramming is accomplished by overexpressing a negative feedback deficient heat-shock response transcription factor before and during overexpression of the protein-of-interest. The advantage of transcriptional reprogramming versus simply overexpressing select proteostasis network components (e.g., chaperones and co-chaperones, which has been explored previously) is that a large number of proteostasis network components are upregulated at their evolved stoichiometry, thus maintaining the system capabilities of the proteostasis network that are currently incompletely understood. Transcriptional proteostasis network reprogramming mediated by stress-responsive signaling in the absence of stress should also be useful for protein production in other cells.


Psychometric evaluation of a self-reported physical activity questionnarie used in the pilot phase of the AZAR Cohort Study.
The purpose of this study was to assess the psychometric properties of a self-reported physical activity (PA) questionnaire based on data from the pilot phase of the AZAR Cohort Study. In this cross-sectional study, all 35-70 years old people living in Khameneh, a city in East Azarbaijan, Iran were invited to take part in the pilot phase of the AZAR Cohort Study. A total of 952 people completed the self-reported PA questionnaire and the International Physical Activity Questionnaire (IPAQ). Construct validity was evaluated by exploratory and confirmatory factor analyses (EFA and CFA). Spearman's correlation coefficient between the scores of the two instruments was used to examine the concurrent validity. Reliability was measured using intraclass correlation coefficient (ICC) and Cronbach's alpha coefficient. In EFA applying principal component analysis with varimax rotation, four factors were identified including recreational leisure time (variance = 52.73%), sedentary leisure time (variance = 38.68%), household/gardening work (variance = 38.66%), and occupation work (variance = 12.67%). The extracted factors were also supported by the CFA (CFI = 0.98, GFI =0.936, RMSEA=0.057). The results indicated moderate concurrent validity (ρ = 0.62, P < 0.001). ICC and Cronbach's alpha were 0.59 and 0.7, respectively. These results showed acceptable and moderate psychometric properties for the self-reported PA questionnaire to assess PA in this population-based study.


FISHIS: fluorescence in situ hybridization in suspension and chromosome flow sorting made easy.
The large size and complex polyploid nature of many genomes has often hampered genomics development, as is the case for several plants of high agronomic value. Isolating single chromosomes or chromosome arms via flow sorting offers a clue to resolve such complexity by focusing sequencing to a discrete and self-consistent part of the whole genome. The occurrence of sufficient differences in the size and or base-pair composition of the individual chromosomes, which is uncommon in plants, is critical for the success of flow sorting. We overcome this limitation by developing a robust method for labeling isolated chromosomes, named Fluorescent In situ Hybridization In suspension (FISHIS). FISHIS employs fluorescently labeled synthetic repetitive DNA probes, which are hybridized, in a wash-less procedure, to chromosomes in suspension following DNA alkaline denaturation. All typical A, B and D genomes of wheat, as well as individual chromosomes from pasta (T. durum L.) and bread (T. aestivum L.) wheat, were flow-sorted, after FISHIS, at high purity. For the first time in eukaryotes, each individual chromosome of a diploid organism, Dasypyrum villosum (L.) Candargy, was flow-sorted regardless of its size or base-pair related content. FISHIS-based chromosome sorting is a powerful and innovative flow cytogenetic tool which can develop new genomic resources from each plant species, where microsatellite DNA probes are available and high quality chromosome suspensions could be produced. The joining of FISHIS labeling and flow sorting with the Next Generation Sequencing methodology will enforce genomics for more species, and by this mightier chromosome approach it will be possible to increase our knowledge about structure, evolution and function of plant genome to be used for crop improvement. It is also anticipated that this technique could contribute to analyze and sort animal chromosomes with peculiar cytogenetic abnormalities, such as copy number variations or cytogenetic aberrations.


Diversification and molecular evolution of ATOH8, a gene encoding a bHLH transcription factor.
ATOH8 is a bHLH domain transcription factor implicated in the development of the nervous system, kidney, pancreas, retina and muscle. In the present study, we collected sequence of ATOH8 orthologues from 18 vertebrate species and 24 invertebrate species. The reconstruction of ATOH8 phylogeny and sequence analysis showed that this gene underwent notable divergences during evolution. For those vertebrate species investigated, we analyzed the gene structure and regulatory elements of ATOH8. We found that the bHLH domain of vertebrate ATOH8 was highly conserved. Mammals retained some specific amino acids in contrast to the non-mammalian orthologues. Mammals also developed another potential isoform, verified by a human expressed sequence tag (EST). Comparative genomic analyses of the regulatory elements revealed a replacement of the ancestral TATA box by CpG-islands in the eutherian mammals and an evolutionary tendency for TATA box reduction in vertebrates in general. We furthermore identified the region of the effective promoter of human ATOH8 which could drive the expression of EGFP reporter in the chicken embryo. In the opossum, both the coding region and regulatory elements of ATOH8 have some special features, such as the unique extended C-terminus encoded by the third exon and absence of both CpG islands and TATA elements in the regulatory region. Our gene mapping data showed that in human, ATOH8 was hosted in one chromosome which is a fusion product of two orthologous chromosomes in non-human primates. This unique chromosomal environment of human ATOH8 probably subjects its expression to the regulation at chromosomal level. We deduce that the great interspecific differences found in both ATOH8 gene sequence and its regulatory elements might be significant for the fine regulation of its spatiotemporal expression and roles of ATOH8, thus orchestrating its function in different tissues and organisms.


The wide genetic landscape of clinical frontotemporal dementia: systematic combined sequencing of 121 consecutive subjects.
PurposeTo define the genetic spectrum and relative gene frequencies underlying clinical frontotemporal dementia (FTD).MethodsWe investigated the frequencies and mutations in neurodegenerative disease genes in 121 consecutive FTD subjects using an unbiased, combined sequencing approach, complemented by cerebrospinal fluid Aβ1-42 and serum progranulin measurements. Subjects were screened for C9orf72 repeat expansions, GRN and MAPT mutations, and, if negative, mutations in other neurodegenerative disease genes, by whole-exome sequencing (WES) (n = 108), including WES-based copy-number variant (CNV) analysis.ResultsPathogenic and likely pathogenic mutations were identified in 19% of the subjects, including mutations in C9orf72 (n = 8), GRN (n = 7, one 11-exon macro-deletion) and, more rarely, CHCHD10, TARDBP, SQSTM1 and UBQLN2 (each n = 1), but not in MAPT or TBK1. WES also unraveled pathogenic mutations in genes not commonly linked to FTD, including mutations in Alzheimer (PSEN1, PSEN2), lysosomal (CTSF, 7-exon macro-deletion) and cholesterol homeostasis pathways (CYP27A1).ConclusionOur unbiased approach reveals a wide genetic spectrum underlying clinical FTD, including 11% of seemingly sporadic FTD. It unravels several mutations and CNVs in genes and pathways hitherto not linked to FTD. This suggests that clinical FTD might be the converging downstream result of a delicate susceptibility of frontotemporal brain networks to insults in various pathways.


[The congruence of nursing diagnoses, interventions, and outcomes between care observations, patient perceptions, and nursing records: a qualitative multiple case study].
The congruence of nursing diagnoses, interventions, and outcomes between care observations, patient perceptions, and nursing records: a qualitative multiple case study Abstract. Background: Nurses with good decision-making competencies state more relevant and accurate nursing diagnoses, perform more effective nursing interventions, and achieve better nursing-sensitive patient outcomes. It was reported that the content of nursing records sometimes doesn't match with statements of patients and nurses. In what extent do the recorded nursing diagnoses, interventions, and outcomes match with observed care situations and patients' statements? A multiple case study with structured observations, guided interviews, and document analyses was performed. The congruence of nursing diagnoses, interventions, and outcomes between the three data sources was investigated by within- and cross-case-analysis in a sample of 24 patients. In total, 114 nursing diagnoses were identified of which 66 were recorded, 37 were found by observations, and 11 by patient interviews. A high congruence between the three perspectives was determined in 59 % of the recorded nursing diagnoses, in 41 % of nursing interventions, and in 33 % of nursing-sensitive patient outcomes. Almost all documented nursing diagnoses (89 %) were supported by most codes from all three sources. To increase the congruence of the three perspectives, nurses' clinical decision-making competencies should be fostered, so that nurses choose more effective nursing interventions that lead to better nursing-sensitive patient outcomes. Patient participation should be fostered.


Family-based treatment of eating disorders.
The current article provides a brief description of the theory and empirical support for family treatment of eating disorders. The main literature related to family treatment for anorexia nervosa (AN) and bulimia nervosa (BN) is reviewed and the findings highlighted. Family treatment, particularly as devised by researchers at the Maudsley Hospital, appears to be an effective treatment for adolescents with short-term AN. It also may be an appropriate treatment for BN in the same age group, although evidence for this is in much shorter supply. Data support the use of family treatments for adolescents with eating disorders. Controlled trials and other systematic research are needed to determine whether family treatment is the best approach.


The impact of HLA genotyping on survival following unrelated donor haematopoietic stem cell transplantation.
One of the major factors that have contributed to improving the outcomes of Stem Cell Transplantation is progress made in the field of human leucocyte antigen(s) (HLA). This is evident not only in developing techniques for rapid and accurate tissue typing, but also in the greatly improved understanding of the HLA system and the impact of HLA matching on transplant complications. It is now accepted that high-resolution HLA matching for transplant recipients and unrelated donors is associated with the best clinical outcomes. The most important HLA determinants are the six 'classical' polymorphic HLA loci: HLA-A, -B, -C, -DRB1,-DQB1, -DPB1. For several years, based on the outcome of numerous studies, a 10/10 matched donor (HLA-A, -B, -C, -DRB1, -DQB1) was considered the ideal. The impact of HLA-DPB1 has been less clear, in view of reduced likelihood of patient/donor matching for this locus. More recently, several large studies have questioned the importance of HLA-DQB1 matching on outcome. Based on the findings of recent studies, the current gold standard unrelated donor is believed to be one matched for 8/8 alleles at high resolution i.e. matched for HLA-A, -B, -C, -DRB1, however, in certain circumstances, mismatches may be tolerated and/or permissive.


Ferroptosis as a Novel Therapeutic Target for Friedreich's Ataxia.
Friedreich ataxia (FRDA) is a progressive neuro- and cardio-degenerative disorder characterized by ataxia, sensory loss, and hypertrophic cardiomyopathy. In most cases, the disorder is caused by GAA repeat expansions in the first introns of both alleles of the FXN gene, resulting in decreased expression of the encoded protein, frataxin. Frataxin localizes to the mitochondrial matrix and is required for iron-sulfur-cluster biosynthesis. Decreased expression of frataxin is associated with mitochondrial dysfunction, mitochondrial iron accumulation, and increased oxidative stress. Ferropotosis is a recently identified pathway of regulated, iron-dependent cell death, which is biochemically distinct from apoptosis. We evaluated whether there is evidence for ferroptotic pathway activation in cellular models of FRDA. We found that primary patient-derived fibroblasts, murine fibroblasts with FRDA-associated mutations, and murine fibroblasts in which a repeat expansion had been introduced (knockin/knockout) were more sensitive than normal control cells to erastin, a known ferroptosis inducer. We also found that the ferroptosis inhibitors ethyl 3-(benzylamino)-4-(cyclohexylamino)benzoate (SRS11-92) and ethyl 3-amino-4-(cyclohexylamino)benzoate, used at 500 nM, were efficacious in protecting human and mouse cellular models of FRDA treated with ferric ammonium citrate (FAC) and an inhibitor of glutathione synthesis [L-buthionine (S,R)-sulfoximine (BSO)], whereas caspase-3 inhibitors failed to show significant biologic activity. Cells treated with FAC and BSO consistently showed decreased glutathione-dependent peroxidase activity and increased lipid peroxidation, both hallmarks of ferroptosis. Finally, the ferroptosis inhibitor SRS11-92 decreased the cell death associated with frataxin knockdown in healthy human fibroblasts. Taken together, these data suggest that ferroptosis inhibitors may have therapeutic potential in FRDA.


Neuroprotective effects of metallothionein against rotenone-induced myenteric neurodegeneration in parkinsonian mice.
Parkinson's disease (PD) is a neurodegenerative disease with motor symptoms as well as non-motor symptoms that precede the onset of motor symptoms. Mitochondrial complex I inhibitor, rotenone, has been widely used to reproduce PD pathology in the central nervous system (CNS) and enteric nervous system (ENS). We reported previously that metallothioneins (MTs) released from astrocytes can protect dopaminergic neurons against oxidative stress. The present study examined the changes in MT expression by chronic systemic rotenone administration in the striatum and colonic myenteric plexus of C57BL mice. In addition, we investigated the effects of MT depletion on rotenone-induced neurodegeneration in CNS and ENS using MT-1 and MT-2 knockout (MT KO) mice, or using primary cultured neurons from MT KO mice. In normal C57BL mice, subcutaneous administration of rotenone for 6 weeks caused neurodegeneration, increased MT expression with astrocytes activation in the striatum and myenteric plexus. MT KO mice showed more severe myenteric neuronal damage by rotenone administration after 4 weeks than wild-type mice, accompanied by reduced astroglial activation. In primary cultured mesencephalic neurons from MT KO mice, rotenone exposure induced neurotoxicity in dopaminergic neurons, which was complemented by addition of recombinant protein. The present results suggest that MT seems to provide protection against neurodegeneration in ENS of rotenone-induced PD model mice.


Cytotoxicity of nickel-chromium alloys: bulk alloys compared to multiple ion salt solutions.
Nickel-based alloys have been in use since the 1930s; however, there are concerns regarding the biocompatibility of the metallic ions released from these alloys to surrounding tissues. The objective of this study was to better understand nickel-based alloy cytotoxicity as well as determine if multiple ion salt solutions can be used to model the cytotoxic effects of bulk implant alloys. This study evaluated cellular morphology, viability, membrane integrity, and alterations in metabolic activity, including DNA synthesis, RNA synthesis, protein synthesis, oxygen consumption, intracellular ATP levels, and glucose-6-phosphate dehydrogenase in response to bulk alloys and multiple ion salt solutions. Over a 24- or 72-h exposure time, the nickel-based alloys released a total ion concentration in the parts per billion range and caused alterations in DNA, RNA, and protein synthesis, intracellular ATP levels, and glucose-6-phosphate dehydrogenase activity. Interestingly, cellular responses to the salt solutions representing the ions released from the alloys were not consistently significantly similar to those elicited from the alloys. From these studies, it was shown that a number of cellular functions are altered in response to ions released from these implant alloys. However, cellular functions were not similarly altered in response to salt solutions representing the ions released from the alloys. These results demonstrated salt solutions cannot be easily used to represent alloy cytotoxicity, and ionic release from alloys is a complex process dependent on variables including ion chemistry, ion valence, and dose-time dependence. This study provides a better understanding of the metabolic response of fibroblasts to ions released from dental alloys; and is a good first step towards developing a more reliable cell culture model of cytotoxicity.


In vitro antimalarial activity of extracts of some plants from a biological reserve in Costa Rica.
Treatment with the usual antimalarial drugs, have induced parasite resistance, reinforcing the need to finding natural antimalarial components that would be found on plants from the forest. Therefore, we decided to look for these components in Costa Rican plants from a protected forest area. Fresh and dry extracts of roots, bark, leaves, flowers and fruits of 25 plants from a biological reserve in Costa Rica, Reserva Biol6gica Alberto Manuel Brenes (REBAMB), were studied in vitro for the presence of substances with antimalarial activity. By studying the inhibition of P berghei schizogony, we assessed the antimalarial activity of several plant extracts: Aphelandra aurantiaca, A. tridentata (Acanthaceae); Xanthosoma undipes (Araceae); Iriartea deltoidea (Arecaceae); Neurolaena lobata (Asteraceae); Senna papillosa, Pterocarpus hayessi, Lonchocarpus pentaphyllus (Fabaceae); Nectandra membranacea, Persea povedae, Cinamomum chavarrianum (Lauraceae); Hampea appendiculata (Malvaceae); Ruagea glabra, Guarea glabra (Meliaceae); Psidium guajava (Myrtaceae); Bocconia frutescens (Papaveraceae); Piper friedrichsthalii (Piperaceae); Clematis dioica (Ranunculaceae); Prunus annularis (Rosaceae); Siparuna thecaphora (Siparunaceae); Solanum arboreum, Witheringia solanacea (Solanaceae); Ticodendrum incognitum (Ticodendraceae); Heliocarpus appendiculatus (Tiliaceae) and Myriocarpa longipes (Urticaceae). We used different parts of the plants as well as fresh and dried extracts for testing IC50. The solid content of the extracts ranged from 1-71.9 microg/mL. The fresh extracts showed stronger activity than the dry ones. Since the plants showing the strongest antimalarial activity are very common in Central America, and some similar genera of these plants have shown positives results in South America, we considered important to present these findings for discussion. On the other hand, this is the first systematic study of this kind ever realized in a circumscribed and protected area of Costa Rica.


Variations in mating-type-like (MTL) loci direct PCR-based tracking of Zygosaccharomyces strains formed by mating.
Variations of chromosomal structures and nucleotide sequences around mating-type-like (MTL) loci among Zygosaccharomyces species have been reported. We have analyzed these differences in more detail and, on the basis of PCR- and next-generation sequencing data, we describe the MTL loci on chromosomes C and F for Z. rouxii type-strain NBRC1130, Z. rouxii NBRC0740 and Zygosaccharomyces sp. NBRC1876. We developed a mating strategy for Zygosaccharomyces sp. NBRC1876 and Z. rouxii NBRC0740, and found that the mated stains could be identified from parental strains on the basis of nucleotide sequence variations of the MTL loci. We further obtained evidence that Zygosaccharomyces sp. NBRC1876 is a natural interspecies hybrid between Z. rouxii and a related species.


Direct projection from the visual associative cortex to the caudate nucleus in the feline brain.
Recent morphological and physiological studies support the assumption that the extrageniculate ascending tectofugal pathways send visual projection to the caudate nucleus (CN) in amniotes. In the present study we investigate the anatomical connection between the visual associative cortex along the anterior ectosylvian sulcus (AES) and the CN in adult domestic cats. An anterograde tracer - fluoro-dextrane-amine - was injected into the AES cortex. The distribution of labeled axons was not uniform in the CN. The majority of labeled axons and terminal like puncta was found only in a limited area in the dorsal part of the CN between the coordinates anterior 12-15. Furthermore, a retrograde tracer - choleratoxin-B - was injected into the dorsal part of the CN between anterior 12 and 13. We detected a large number of labeled neurons in the fundus and the dorsal part of the AES between the coordinates anterior 12-14. Based upon our recent results we argue that there is a direct monosynaptic connection between the visual associative cortex along the AES and the CN. Beside the posterior thalamus, the AES cortex should also participate in the transmission of the tectal visual information to the CN. This pathway is likely to convey complex information containing both sensory and motor components toward the basal ganglia, which supports their integrative function in visuomotor actions such as motion and novelty detection and saccade generation.


Rapid ornithine decarboxylase test for the identification of enterobacteriaceae.
Conventional methods for detecting ornithine decarboxylase activity require an extended period of incubation. However, with a few simple modifications, accurate results were obtained within a few hours rather than several days. The broth medium was modified, primarily by omitting glucose and by decreasing the pH to 5.5. A 1-ml amount of this broth was inoculated with one colony and then overlaid with sterile mineral oil. Within 2 to 4 hr, the pH increased if ornithine was decarboxylated, thus changing the color of the internal pH indicator to a dark purple. If the amino acid was not decarboxylated, the pH decreased to pH 5.0 to 5.2, enough to give a definite yellow color. With 347 selected clinical isolates, the rapid test gave results identical to those obtained in 1 to 4 days with Moeller's decarboxylase medium. Less reliable results were obtained with Difco's decarboxylase medium with 0.3% agar which was stabinoculated and read after 18 to 24 hr without a mineral oil seal. The rapid ornithine decarboxylase test represents a simple, accurate technique which is well suited for the clinical microbiology laboratory.


Effect of computer-generated nurse/physician reminders on influenza immunization among seniors.
Despite long-standing recommendations by the Canadian and U.S. national immunization advisory committees that all persons aged 65 or older be vaccinated annually against influenza, vaccination rates among North American seniors remain approximately 20%. This paper reports a study of the effect of computer-generated nurse/physician reminders to offer influenza vaccine to elderly patients seen during the pre-influenza season in a community based group family practice. The study employed a pretest-post-test design strengthened by the use of a no-intervention comparison practice and repeated pretest measures of vaccine coverage. Outcome was measured by clinical records review. The influenza vaccination rate increased by 165% from 10.1% to 26.8% following introduction of the computer-generated reminders. There was no increase in influenza immunization in the comparison practice during the corresponding time period. Over 60% of eligible patients were seen in the practice during the pre-influenza season; 41.6% of those received influenza vaccinations.


Nek8 mutation causes overexpression of galectin-1, sorcin, and vimentin and accumulation of the major urinary protein in renal cysts of jck mice.
The jck murine model, which results from a double point mutation in the nek8 gene, has been used to study the mechanism of autosomal recessive polycystic kidney disease (ARPKD). The renal proteome of jck mice was characterized by two-dimensional gel electrophoresis combined with mass spectrometry (MALDI-TOF/TOF). Four newly identified proteins were found to accumulate in the kidneys of jck mice with polycystic kidney disease (PKD) compared with their wild-type littermates. The proteins galectin-1, sorcin, and vimentin were found to be induced 9-, 9-, and 25-fold, respectively, in the PKD proteome relative to the wild type. The identity of these proteins was established by peptide mass fingerprinting and de novo MS/MS sequencing of selected peptides. Up-regulation of these three proteins may be due to the nek8 mutation, and their function may be related to the signaling and structural processes in the primary cilium. Additionally a series of protein isoforms observed only in the ARPKD kidney was identified as the major urinary protein (MUP). Peptide sequencing demonstrated that the isoforms MUP1, MUP2, and MUP6 are contained in this series. The MUP series showed a number of male-specific isoforms and a phosphorylation of the entire series with an increasing degree of phosphorylation of the acidic isoforms. In addition, the MUP series was localized to the cyst fluid of PKD mice, and a cellular mislocalization of galectin-1, sorcin, and vimentin in PKD tubular epithelial cells was shown. The abnormal and extremely high accumulation of the MUPs in the ARPKD kidney may be linked to a defect in protein transport and secretion. The discovery of these proteins will provide new information on the molecular and cellular processes associated with the mechanism of ARPKD.


Multiple transisthmian divergences, extensive cryptic diversity, occasional long-distance dispersal, and biogeographic patterns in a marine coastal isopod with an amphi-American distribution.
Excirolana braziliensis is a coastal intertidal isopod with a broad distribution spanning the Atlantic and Pacific tropical and temperate coasts of the American continent. Two separate regional studies (one in Panama and one in Chile) revealed the presence of highly genetically divergent lineages, implying that this taxon constitutes a cryptic species complex. The relationships among the lineages found in these two different regions and in the rest of the distribution, however, remain unknown. To better understand the phylogeographic patterns of E. braziliensis, we conducted phylogenetic analyses of specimens from much of its entire range. We obtained DNA sequences for fragments of four mitochondrial genes (16S rDNA, 12S rDNA, COI, and Cytb) and also used publicly available sequences. We conducted maximum likelihood and Bayesian phylogenetic reconstruction methods. Phylogeographic patterns revealed the following: (1) new highly divergent lineages of E. braziliensis; (2) three instances of Atlantic-Pacific divergences, some of which appear to predate the closure of the Isthmus of Panama; (3) the distributional limit of highly divergent lineages found in Brazil coincides with the boundary between two major marine coastal provinces; (4) evidence of recent long-distance dispersal in the Caribbean; and (5) populations in the Gulf of California have closer affinities with lineages further south in the Pacific, which contrasts with the closer affinity with the Caribbean reported for other intertidal organisms. The high levels of cryptic diversity detected also bring about challenges for the conservation of this isopod and its fragile environment, the sandy shores. Our findings underscore the importance of comprehensive geographic sampling for phylogeographic and taxonomical studies of broadly distributed putative species harboring extensive cryptic diversity.


Cholesterol promotes hemifusion and pore widening in membrane fusion induced by influenza hemagglutinin.
Cholesterol-specific interactions that affect membrane fusion were tested for using insect cells; cells that have naturally low cholesterol levels (< 4 mol %). Sf9 cells were engineered (HAS cells) to express the hemagglutinin (HA) of the influenza virus X-31 strain. Enrichment of HAS cells with cholesterol reduced the delay between triggering and lipid dye transfer between HAS cells and human red blood cells (RBC), indicating that cholesterol facilitates membrane lipid mixing prior to fusion pore opening. Increased cholesterol also increased aqueous content transfer between HAS cells and RBC over a broad range of HA expression levels, suggesting that cholesterol also favors fusion pore expansion. This interpretation was tested using both trans-cell dye diffusion and fusion pore conductivity measurements in cholesterol-enriched cells. The results of this study support the hypothesis that host cell cholesterol acts at two stages in membrane fusion: (1) early, prior to fusion pore opening, and (2) late, during fusion pore expansion.


Reciprocal inhibition in the forearm during voluntary contraction and thinking about movement.
The regulatory mechanism for the interaction of agonist and antagonist muscles is not still clear. In these studies we investigated reciprocal inhibition during voluntary contraction and thinking about movement by using transcranial magnetic stimulation (TMS) and the H reflex technique. Magnetic stimuli, with intensities 10% above threshold, were delivered at the scalp under 3 experimental conditions; at rest, during voluntary isometric weak contraction and during thinking about movement. Surface EMG responses were taken from extensor carpi radialis (ECR) and flexor carpi radialis (FCR) muscles in 14 normal subjects. Inhibitions of antagonist muscles induced by thinking about agonist movement occurred in 8 ECR and 10 FCR muscles, but most of them were not statistically significant. H reflexes were obtained from the FCR muscles in 10 normal subjects under the same conditions. Voluntary wrist extension and thinking about wrist extension produced inhibition of the H reflex in the FCR muscles. Reciprocal inhibition of antagonist muscles may occur at the cortical and spinal cord levels. The excitability of motoneurons has effects on the H reflex amplitude. Motor evoked potential (MEP) response to TMS might be affected by the combination of input from the motor cortex and the spinal cord. TMS may evoke simultaneous facilitatory and inhibitory volleys to antagonist muscles. This may explain why the inhibitions in the TMS study were weaker than those in the H reflex study.


What did we learn about national organizational collaboration at the advisory committee level?
This article provides an overview of the typical roles and tasks of advisory groups in general, followed by a discussion of the roles and tasks the Interdisciplinary Generalist Curriculum (IGC) Project Advisory Committee was asked to assume and how these were fulfilled. It analyzes the lessons learned about advisory committees as a result of the IGC Project experience. Key elements of success in fulfilling advisory committee obligations include well-defined expectations, periodic evaluations, and clear communication between project leadership and the members of the advisory committee. In the spirit of lessons learned from the IGC Project, this critique identifies several philosophical and logistic issues that might be considered in the design and implementation of future projects, such as the need to choose committed, high-energy advisory committee members who are willing to perform many complex, time-consuming tasks.


[The new biomaterials of embolization: the shift to degradable].
During the last decade, the interest of degradable embolics has considerably grown as alternatives for the currently-used permanent embolics: no permanent foreign body, recanalisation for repeat embolization, complete drug delivery in chemoembolization. Their design is not trivial since a lot of requirements need to be satisfied. The degradable embolics should be easily suspended in physiological solutions and contrast media, injectable in catheters with small internal lumen and they have to recover their size and shape after injection, as for any embolic. Moreover, they need to be loadable with various drugs, and their degradation has to be achieved in a given time before the onset of a chronic inflammatory response and vessel wall remodeling. Various approaches have been tested on diverse materials over the last years and have produced interesting results that make it possible to claim that the revolution of the degradable embolic has begun.


[The determination of molecular sulphur in Matsesta mineral water and its analog Novonukutskaya mineral water].
The method of microcolumn reversed phase high performance liquid chromatography (rp-HPLC) was employed to determine the content of elemental sulphur in mineral waters. The study envisaged the analysis of the samples of sulphide-containing mineral waters Novonukutskaya and Matsesta obtained by the solid phase extraction technique. Based on these data, the authors discuss the origin and the circulation of sulphur in the hydrogen sulphide sources. The elution conditions selected in this study ensured the high-resolution separation of the octasulphur peak from the peaks of allotropic components of the extract whereas the two-wave detection technique allowed to identify the peaks of molecular sulphur.


Novel targeted therapies for the management of liver fibrosis.
Introduction: Prolonged liver injury results in tissue damage and replacement by extracellular matrix and fibrosis. Cirrhosis represents a leading cause of mortality worldwide and imposes a major financial burden on health-care systems. Fortunately, fibrogenesis has proven to be reversible if halted early, encouraging the development of novel anti-fibrotic agents that may accelerate histological restoration. Preclinical data have elucidated numerous potential therapeutic targets and many anti-fibrotic agents are currently at various stages of clinical research.Areas covered: The present review summarizes recent clinical data regarding anti-fibrotic drugs including monoclonal antibodies, targeted conjugates, and small molecule agents.Expert opinion: Although undeniable progress has been made in the development of anti-fibrotic agents in recent years, most data currently available are derived from preclinical and early clinical studies. The efficacy and safety of these agents will need to be corroborated by larger clinical trials, some of which are ongoing with results expected in the upcoming years. Combination therapy with agents targeting different pathways of fibrogenesis will also be of great interest for the future and will need to be explored in clinical trials.


[Effect of immunization with neurospecific proteins and tubulin on learning in the rat].
The paper deals with the effect of anticerebral antibodies on rats learning in T-shaped maze with food reinforcement. The rats of August line were immunized by neurospecific proteins 10-40-4 from the human and rat's brain, 14-3-2 and cerebral tubuline. Rats immunized by bovine serum albumine (BSA) served as a control of the influence of antibodies to noncerebral protein. Rats injected with Freund adjuvant and saline, served as control of Freund adjuvant action. At the time of antibodies formation (from the 7-th day after the last injection) the rats were trained in a T-shaped maze during 4 days. The acquisition of conditioned reactions was inhibited in rats immunized with neurospecific proteins 10-40-4 from the rats brain, 14-3-2 and cerebral tubuline. Antibodies to neurospecific protein 10-40-4 from the rats brain produced the greatest effect on the process of learning. They elicited a significant decrease of conditioning and an increase of the number of errors. Antibodies to the neurospecific protein 10-40-4 from the human brain and to the BSA, i.e. to proteins which are not present in the nervous tissue of the rats, did not affect the learning. The obtained results indirectly confirm the permeability for the antibodies of the blood-brain barrier in immunized animals.


Behavioral factors associated with SARS-CoV-2 infection in Japan.
The relative burden of COVID-19 has been less severe in Japan. One reason for this may be the uniquely strict restrictions imposed upon bars/restaurants. To assess if this approach was appropriately targeting high-risk individuals, we examined behavioral factors associated with SARS-CoV-2 infection in the community. This multicenter case-control study involved individuals receiving SARS-CoV-2 testing in June-August 2021. Behavioral exposures in the past 2 weeks were collected via questionnaire. SARS-CoV-2 PCR-positive individuals were cases, while PCR-negative individuals were controls. The analysis included 778 individuals (266 [34.2%] positives; median age [interquartile range] 33 [27-43] years). Attending three or more social gatherings was associated with SARS-CoV-2 infection (adjusted odds ratio [aOR] 2.00 [95% CI 1.31-3.05]). Attending gatherings with alcohol (aOR 2.29 [1.53-3.42]), at bars/restaurants (aOR 1.55 [1.04-2.30]), outdoors/at parks (aOR 2.87 [1.01-8.13]), at night (aOR 2.07 [1.40-3.04]), five or more people (aOR 1.81 [1.00-3.30]), 2 hours or longer (aOR 1.76 [1.14-2.71]), not wearing a mask during gatherings (aOR 4.18 [2.29-7.64]), and cloth mask use (aOR 1.77 [1.11-2.83]) were associated with infection. Going to karaoke (aOR 2.53 [1.25-5.09]) and to a gym (aOR 1.87 [1.11-3.16]) were also associated with infection. Factors not associated with infection included visiting a cafe with others, ordering takeout, using food delivery services, eating out by oneself, and work/school/travel-related exposures including teleworking. We identified multiple behavioral factors associated with SARS-CoV-2 infection, many of which were in line with the policy/risk communication implemented in Japan. Rapid assessment of risk factors can inform decision making.


Exonuclease-catalyzed recycling and annular four-footed DNA walking amplification-assisted "on-off-super on" signal transitions for photoelectrochemical biosensing of kanamycin.
Photoelectrochemical (PEC) biosensors have exhibited a promising potential for assays of a large variety of analytes; however, how to realize their low background-based "super on" signal output is still a great challenge. Herein, we report a novel multiple nucleic acid amplification-assisted "on-off-super on" signal transition mechanism for the PEC biosensing of kanamycin antibiotics. The biosensing platform was constructed on a perylene-3,4,9,10-tetracarboxylic dianhydride-based photoelectrode, and its strong photocurrent could be well inhibited by an anchored ferrocene (Fc)-labeled hairpin DNA to produce a low background signal. Two target biorecognition-triggered exonuclease III-catalytic reactions were adopted to produce an annular four-footed DNA walker (AFW) and a methylene blue (MB)-labeled DNA strand. By using their synergistic effect to release Fc quenchers and simultaneously capture MB sensitizers, a "super on" signal output was realized. As a result, a very wide linear range from 10 fg mL-1 to 10 ng mL-1 and an ultra-low detection limit of 7.8 fg mL-1 were obtained. Meanwhile, the aptamer recognition-based homogeneous reaction and AFW-based multiple nucleic acid amplification effectively simplified the assay manipulation and well ensured the repeatability of the method. The satisfactory sample experiment results indicated its good reliability and accuracy for the antibiotic residue analysis application.


Predictive value of atrial fibrillation during the postradiofrequency ablation blanking period.
Recurrent arrhythmia following catheter ablation of atrial fibrillation (AF) may present early, during a standard 3-month blanking period. Early recurrence has been correlated to late recurrence, but the degree to which its absence predicts longer-term success has not been quantified. The purpose of this study was to explore and quantify the relationship between early and late arrhythmia recurrence, specifically the negative predictive value, that is, the degree to which absence of blanking period recurrence predicts absence of late recurrence. A systematic literature review and meta-analysis were conducted using statistical methods of a diagnostic test accuracy review. Studies of AF ablation using point-by-point radiofrequency, with repeated monitoring of arrhythmia recurrence including asymptomatic recurrence, and with separate data by AF type, were eligible. Nine studies met the prespecified eligibility criteria. For paroxysmal AF, 89% (confidence interval [CI] 82%-94%) of patients free from early recurrence remained free from late recurrence. The estimate for persistent AF was similar (91%; CI 75%-97%). This finding was robust in sensitivity analyses. Patients with early recurrence had a wider range of likely outcomes with longer-term follow-up. Freedom from AF recurrence during the blanking period is highly predictive of longer-term success in catheter ablation. Clinical trials in this area may be able to leverage these findings to more quickly assess the potential utility of new ablation technologies and methods, for example, by using early surrogate measures of success.


Vemurafenib Treatment of Pleomorphic Xanthoastrocytoma in a Child With Down Syndrome.
Brain tumors are the most common solid neoplasms of childhood, but they are very rarely reported in children with Down Syndrome (DS), who develop more commonly different types of malignancies. In particular, we hereby report the case of an 8-years-old child with DS that presented to our attention for neurological and endocrinological issues. Brain imaging revealed the presence of a mass that was partially resected revealing a histological diagnosis of Pleomorphic Xanthoastrocytoma (PXA), a rare WHO grade II tumor extending from the diencephalic region into the surrounding brain tissue. These tumors can harbor the BRAF mutation p.V600E, targetable by the specific inhibitor Vemurafenib. After confirming the presence of the mutation in the tumor, the patient was treated with Vemurafenib. The treatment proved to be effective, leading to a partial response and a stabilization of the disease. Usually, in patients with DS a reduction of the dose of chemotherapeutic drugs is necessary. Vemurafenib was instead well-tolerated as the only observed adverse effect was grade I skin toxicity. This is, to our knowledge, the first case of a PXA reported in a child with DS and the first DS patient treated with Vemurafenib.


Modelling the dynamics of signalling pathways.
In the present chapter we discuss methodologies for the modelling, calibration and validation of cellular signalling pathway dynamics. The discussion begins with the typical range of techniques for modelling that might be employed to go from the chemical kinetics to a mathematical model of biochemical pathways. In particular, we consider the decision-making processes involved in selecting the right mechanism and level of detail of representation of the biochemical interactions. These include the choice between (i) deterministic and stochastic chemical kinetics representations, (ii) discrete and continuous time models and (iii) representing continuous and discrete state processes. We then discuss the task of calibrating the models using information available in web-based databases. For situations in which the data are not available from existing sources we discuss model calibration based upon measured data and system identification methods. Such methods, together with mathematical modelling databases and computational tools, are often available in standard packages. We therefore make explicit mention of a range of popular and useful sites. As an example of the whole modelling and calibration process, we discuss a study of the cross-talk between the IL-1 (interleukin-1)-stimulated NF-kappaB (nuclear factor kappaB) pathway and the TGF-beta (transforming growth factor beta)-stimulated Smad2 pathway.


Antibody-Mediated Immune Subset Depletion Modulates the Immune Response in a Rabbit (Oryctolagus cuniculus) Model of Epstein-Barr Virus Infection.
Epstein-Barr Virus (EBV) is a γ-herpesvirus which infects over 90% of the adult human population. Most notably, this virus causes infectious mononucleosis but it is also associated with cancers such as Hodgkin and Burkitt lymphoma. EBV is a species-specific virus and has been studied in many animal models, including nonhuman primates, guinea pigs, humanized mice, and tree shrews. However, none of these animal models are considered the "gold standard" for EBV research. Recently, rabbits have emerged as a viable alternative model, as they are susceptible to EBV infection. In addition, the EBV infection progresses after immune suppression with cyclosporine A (CsA), modeling the reactivation of EBV after latency. We sought to refine this model for acute or active EBV infections by performing antibody-mediated depletion of certain immune subsets in rabbits. Fourteen 16 to 20-wk old, NZW rabbits were intravenously inoculated with EBV and concurrently treated with either anti-CD4 T-cell antibody, anti-pan-T-cell antibody (anti CD45), CSA, or, as a control, anti-HPV antibody. Rabbits that received the depleting antibodies were treated with CsA 3 times at a dose of 15 mg/kg SC once per day for 4 d starting at the time of EBV inoculation then the dose was increased to 20 mg/kg SC twice weekly for 2 wk. Weights, temperatures, and clinical signs were monitored, and rabbits were anesthetized once weekly for blood collection. When compared with the control group, anti-CD4-treated rabbits had fewer clinical signs and displayed higher levels of viral DNA via qPCR in splenocytes; however, flow cytometry results showed only a partial depletion of CD4 T-cells. Treatment with anti-pan-T-cell antibody did not result in noticeable T-cell depletion. These data suggest the EBV-infected rabbit is a promising model for testing antiviral medications and prophylactic vaccines for EBV.


Tryptophan residues promote membrane association for a plant lipid glycosyltransferase involved in phosphate stress.
Chloroplast membranes contain a substantial excess of the nonbilayer-prone monogalactosyldiacylglycerol (GalDAG) over the biosynthetically consecutive, bilayer-forming digalactosyldiacylglycerol (GalGalDAG), yielding a high membrane curvature stress. During phosphate shortage, plants replace phospholipids with GalGalDAG to rescue phosphate while maintaining membrane homeostasis. Here we investigate how the activity of the corresponding glycosyltransferase (GT) in Arabidopsis thaliana (atDGD2) depends on local bilayer properties by analyzing structural and activity features of recombinant protein. Fold recognition and sequence analyses revealed a two-domain GT-B monotopic structure, present in other plant and bacterial glycolipid GTs, such as the major chloroplast GalGalDAG GT atDGD1. Modeling led to the identification of catalytically important residues in the active site of atDGD2 by site-directed mutagenesis. The DGD synthases share unique bilayer interface segments containing conserved tryptophan residues that are crucial for activity and for membrane association. More detailed localization studies and liposome binding analyses indicate differentiated anchor and substrate-binding functions for these separated enzyme interface regions. Anionic phospholipids, but not curvature-increasing nonbilayer lipids, strongly stimulate enzyme activity. From our studies, we propose a model for bilayer "control" of enzyme activity, where two tryptophan segments act as interface anchor points to keep the substrate region close to the membrane surface. Binding of the acceptor substrate is achieved by interaction of positive charges in a surface cluster of lysines, arginines, and histidines with the surrounding anionic phospholipids. The diminishing phospholipid fraction during phosphate shortage stress will then set the new GalGalDAG/phospholipid balance by decreasing stimulation of atDGD2.


General mapping between random walks and thermal vibrations in elastic networks: fractal networks as a case study.
We present an approach to mapping between random walks and vibrational dynamics on general networks. Random walk occupation probabilities, first passage time distributions and passage probabilities between nodes are expressed in terms of thermal vibrational correlation functions. Recurrence is demonstrated equivalent to the Landau-Peierls instability. Fractal networks are analyzed as a case study. In particular, we show that the spectral dimension governs whether or not the first passage time distribution is well represented by its mean. We discuss relevance to universal features arising in protein vibrational dynamics.


Cost-Effectiveness of Pharmacomechanical Catheter-Directed Thrombolysis Versus Standard Anticoagulation in Patients With Proximal Deep Vein Thrombosis: Results From the ATTRACT Trial.
In patients with acute deep vein thrombosis (DVT), pharmacomechanical catheter-directed thrombolysis (PCDT) in conjunction with anticoagulation therapy is increasingly used with the goal of preventing postthrombotic syndrome. Long-term costs and cost-effectiveness of these 2 treatment strategies from the perspective of the US healthcare system have not been compared. Between 2009 and 2014, the ATTRACT trial (Acute Venous Thrombosis: Thrombus Removal With Adjunctive Catheter-Directed Thrombolysis) randomized 692 patients with acute proximal DVT to PCDT plus anticoagulation (n=337) or standard treatment with anticoagulation alone (n=355). Costs (2017 US dollars) were assessed over a 24-month follow-up period using a combination of resource-based costing, hospital bills, Medicare reimbursement rates, and the Drug Topics Red Book. Health state utilities were obtained from the Short Form-36. In-trial results and US life tables were used to develop a Markov cohort model to evaluate lifetime cost-effectiveness. For the PCDT group, mean costs of the initial procedure were $13 600; per-patient costs associated with the index hospitalization were $21 509 for PCDT and $3877 for standard care (difference=$17 632; 95% CI, $16 117-$19 243). The 24-month difference in costs was $20 045 (95% CI, $16 093-$24 120). Utility scores increased significantly between baseline and 6 months for both groups, with no significant differences between groups at any follow-up time point. Projected differences in lifetime costs of $16 740 and quality-adjusted life years (QALYs) of 0.08, yield an incremental cost-effectiveness ratio for PCDT of $222 041/QALY gained. In probabilistic sensitivity analysis, the probability that PCDT would achieve a lifetime incremental cost-effectiveness ratio <$50 000/QALY or <$150 000/QALY was 1% and 25%, respectively. For iliofemoral DVT, QALY gains with PCDT were greater, yielding an incremental cost-effectiveness ratio of $137 526/QALY; for femoral-popliteal DVT, standard therapy was an economically dominant strategy. With an incremental cost-effectiveness ratio >$200 000/QALY gained, PCDT is not an economically attractive treatment for proximal DVT. PCDT may be of intermediate value in patients with iliofemoral DVT. Clinical Trial Registration URL: https://www.clinicaltrials.gov. Unique identifier: NCT00790335.


[Use of high doses of glucocorticoids in patients with rheumatic diseases].
Short-term application of high doses of glucocorticoids removes the acuity of the generalized patterns of systemic rheumatic diseases. This is documented by the reported data and analysis of the authors' own 15 observations. The dose of glucocorticoids, duration of their application and administration routes are determined by the degree of the pathological process activity and by the organs concerned (kidneys, central nervous system). If the effect attained is doubtful, repeated courses of intensive treatment with glucocorticoids are indicated in some cases.


Follow-up study of pulmonary artery configuration in hypoplastic left heart syndrome.
Pulmonary artery (PA) distortion significantly compromises the outcome of the staged approach to the Fontan operation in patients with hypoplastic left heart syndrome (HLHS). This retrospective study was designed to investigate the influence of the initial operation on postoperative PA anatomy. Forty-nine patients with HLHS and its variant were enrolled in this study. As an initial palliation, the Norwood operation with a modified Blalock-Taussig (BT) shunt was performed in 12, the Norwood operation with a right ventricle to pulmonary artery (RV-PA) shunt in 31, and bilateral PA banding in 6. The incidence and risk factors of postoperative central pulmonary artery stenosis (PS) were investigated, and the PA configuration was followed up until post-Fontan status. Twenty-two patients (51.2%) had developed central PS after the Norwood operation (33.3% with a BT shunt vs. 58.1% with a RV-PA shunt). The RV-PA shunt with a polytetrafluoroethylene (PTFE) patch at the distal pulmonary stump significantly decreased the central PS (P = 0.035). The PA index after the Norwood operation was not statistically different between the BT and RV-PA shunt groups, although in the RV-PA group it was significantly higher in patients with a PTFE patch on the distal PA stump. PA plasty was performed in 16 patients in the second-stage palliation and in 15 with the Fontan completion. Freedom from PA plasty was significantly lower in the RV-PA shunt group than in the BT shunt group (63.5% vs. 31.1% at 5 years, P = 0.034). Six patients initially palliated with bilateral PA banding had no stenosis at the banding site in the Norwood + Glenn operation, and one patient required stent placement for left PS in the Fontan completion. Post-Fontan catheterization (n = 31) showed central venous pressure of 11.5 +/- 2.6 mmHg, cardiac index of 3.6 +/- 0.8 l/kg/min, and PA index of 194.0 +/- 58.4 mm(2)/m(2); there was no difference between the groups. The incidence of central PS after the Norwood operation was significant, and the shunt type and procedure for the distal PA stump influenced the postoperative configuration of the central PA. With an aggressive surgical approach to central PS, PA anatomy was satisfactory with good hemodynamic variables after Fontan completion. Bilateral PA banding did not cause later vascular deformity.


Nursing students' work, a risk factor for the consumption of alcohol and other drugs.
This descriptive, qualitative study aims to describe risk factors, in the work environment, for the consumption of alcohol and other drugs, and also to confirm the importance of protective factors. Interview technique was used with 51 nursing students at the Rio de Janeiro State University, who work in the city's health services. students related work as a risk factor for the consumption of drugs such as alcohol, cigarettes and anxiolytics. We conclude that working with participative methodologies, valuing student's former experiences, family and group life, and individual characteristics are essential in the preventive approach. Attention to protective factors is recommended.


[Diagnostic circumstances, therapeutic measures and long-term surveillance of medullary cancers of the thyroid].
Diagnosis of medullary thyroid carcinoma is often delayed as a consequence of its rarity and its recent discovery. Fine needle aspiration biopsies of all the cold thyroid nodules and plasma calcitonin (CT) measurement in case of suggestive clinical features allow an accurate pre-operative diagnosis and the best therapeutic conditions. A cervico-mediastinal check-up, a screening for a Multiple Endocrine Neoplasia type 2a or 2b, the removal of all tumoral cervico-thoracic tissue by total thyroidectomy, a careful node excision and the pathological examination by a skillful pathologist using immunohistochemistry are the main therapeutic methods. Additional treatment such as external radiotherapy, chemotherapy and radiopharmaceutics are indicated when surgery is incomplete, when the tumor is inoperable and when there are extensive distant metastases. Basal and pentagastrin stimulated CT is the best tool in post surgical follow-up. Disappearance may indicate a long remission, even a cure. Persistence of an abnormal level necessitates the search for secreting tumoral site with non invasive methods such as ultrasonography, computed tomography and scintigraphy, avoiding blind aggressive treatments. In case of recurrence or distant metastases, surgical excision has to be considered first.


Virus transmission via food.
Viruses are transmitted to humans via foods as a result of direct or indirect contamination of the foods with human faeces. Viruses transmitted by a faecal-oral route are not strongly dependent on foods as vehicles of transmission, but viruses are important among agents of foodborne disease. Vehicles are most often molluscs from contaminated waters, but many other foods are contaminated directly by infected persons. The viruses most often foodborne are the hepatitis A virus and the Norwalk-like gastroenteritis viruses. Detection methods for these viruses in foods are very difficult and costly; the methods are not routine. Indicators that would rapidly and reliably suggest the presence of viral contamination of foods are still being sought. Contamination can be prevented by keeping faeces out of food or by treating vehicles such as water in order to inactivate virus that might be carried to food in this way. Virus cannot multiply in food, but can usually be inactivated by adequate heating. Other methods of inactivating viruses within a food are relatively unreliable, but viruses in water and on exposed surfaces can be inactivated with ultraviolet light or with strong oxidizing agents.


Defecation patterns of the infants mainly breastfed from birth till the 12th month: Prospective cohort study.
Studies about defecation patterns have been mostly conducted on infants who were breastfed in a short term but were fed predominantly with formula. In this study, defecation patterns of 125 infants, most of them being breastfed during 12th month were evaluated. Frequency, consistency and color of the stool were analyzed in relation to the feeding pattern at the 15th day and at the 1st, 2nd, 3rd, 4th, 5th, 6th and 12th months. Frequency of defecation was highest in 15th day with a median of 6/day. It decreased with age (p=0.0001), being 4/day and 3/day in 1st and 2nd months respectively, and 2/day between 3rd-12th months. During first 5 months, the stool frequency was higher in infants who were exclusively breastfed compared to those being fed with breastfed and formula (p<0.05). The effect of the feeding pattern on stool frequency disappeared in the following months (p>0.05). Stool frequency decreased by half at the 2nd month when the rate of less than once/day attained its highest value (24.8%). In those who had a rate of less than once/day, stool frequency stayed low until the 6th month and exclusively breastfed rate was also found lower (p<0.05). Besides the age, exclusive breastfeeding was also effective on stool frequency. It points out that infants who are fed with formula in addition to breastfeeding may defecate less than once per day hence should not be diagnosed as having constipation depending solely on defecation frequency and should not receive unneccesary treatments.


Preclinical modeling of EGFR inhibitor resistance in head and neck cancer.
The epidermal growth factor receptor (EGFR) is widely expressed in head and neck squamous cell carcinomas (HNSCC) and can activate many growth and survival pathways within tumor cells. Despite ubiquitous EGFR expression, therapies targeting the receptor are only modestly effective in the treatment of HNSCC. A consistent mechanism of resistance to EGFR targeting agents has not yet been identified in HNSCC likely due, in part, to the paucity of preclinical models. We assessed the in vitro and in vivo responses of a panel of 10 genotypically validated HNSCC cell lines to the EGFR inhibitors erlotinib and cetuximab to determine their validity as models of resistance to these agents. We defined a narrow range of response to erlotinib in HNSCC cells in vitro and found a positive correlation between EGFR protein expression and erlotinib response. We observed cross-sensitivity in one HNSCC cell line, 686LN, between erlotinib and cetuximab in vivo. We attempted to generate models of cetuximab resistance in HNSCC cell line-derived xenografts and heterotopic tumorgrafts generated directly from primary patient tumors. While all 10 HNSCC cell line xenografts tested were sensitive to cetuximab in vivo, heterotopic patient tumorgrafts varied in response to cetuximab indicating that these models may be more representative of clinical responses. These studies demonstrate the limitations of using HNSCC cell lines to reflect the heterogeneous clinical responses to erlotinib and cetuximab, and suggest that different approaches including heterotopic tumorgrafts may prove more valuable to elucidate mechanisms of clinical resistance to EGFR inhibitors in HNSCC.


Induction of morphogenesis by methionine starvation in Myxococcus xanthus: polyamine control.
The induction of mycrocyst formation by methionine starvation was demonstrated in Myxococcus xanthus by several methods. Growing in a defined medium (M(1)), M. xanthus had a doubling time of 6.5 hr. Four amino acids-leucine, isoleucine, valine, and glycine-were required for growth under these conditions. When the concentration of several amino acids in the medium was reduced (M(2)), the doubling time increased to 10 to 12 hr, and a requirement for methionine was observed. Methionine starvation led to a slow conversion of the population to microcysts. Under conditions of methionine prototrophy (M(1)), microcyst formation could still be triggered in exponentially growing cells by the addition of either 5 mm ethionine or 0.1 m isoleucine plus 0.1 m threonine, feedback inhibitors of methionine biosynthesis. Vegetative growth in the absence of methionine was obtained in medium M(2) if the leucine concentration was raised to its level in medium M(1). Thus, methionine biosynthesis is controlled by the exogenous concentration of the required amino acid, leucine. During an examination of the effects of methionine metabolites on microcyst formation, the involvement of polyamines in morphogenesis was uncovered. Putrescine (0.05 m) induced the formation of microcysts; spermidine (2 to 5 mm) inhibited induction by methionine starvation, ethionine, or high isoleucine-threonine. Spermidine was the only polyamine detected in M. xanthus (16.0 mug/10(9) cells). Its concentration decreased by more than 50% shortly after microcyst induction by high isoleucine-threonine. It is postulated that spermidine is an inhibitor of microcyst induction; when spermidine formation is blocked by methionine starvation, morphogenesis is induced.


Getting bacterial cells into shape.
The conformational state of a structural protein in bacteria can vary, depending on the concentration level of potassium ions or the nucleotide bound to it.


Prevalence of deep venous reflux in patients with primary superficial vein incompetence.
This prospective study was designed to determine the prevalence of deep reflux and the conditions under which it may occur in patients with primary superficial venous reflux and absence of deep venous thrombosis (DVT). We studied 152 limbs in 120 consecutive patients in the standing position who had superficial venous reflux with color flow duplex scanning. Limbs with documented evidence of DVT or post-thrombotic vein wall changes during the examination were studied but not included in the analysis. Limbs were divided into those that had at least reflux in the saphenofemoral, the saphenopopliteal, or the gastropopliteal junction and into those with nonjunctional reflux in the superficial and gastrocnemial veins. Peak velocity and duration of reflux were measured. To examine the recirculation theory, we tested the deep veins by occluding and refluxing saphenous veins 10 cm below the sampling site. Thirteen limbs in 11 patients (9%) were excluded because of previous DVT. Of the remaining 139 limbs, 106 (76%) had junctional reflux. Saphenofemoral junction was involved in 89 limbs (84%), saphenopopliteal junction in 18 (17%), and gastropopliteal junction in 7 (4%). In 33 limbs (24%), reflux was detected in the main trunk or tributaries of the saphenous veins alone with no junctional incompetence. Femoral or popliteal reflux was present in 31 limbs (22%). This reflux was segmental in 27 limbs, and it was limited in the junction in 24 limbs. The mean duration of deep venous reflux was 0.9 seconds, it ranged from 0.6 to 3.7 seconds, and it was significantly shorter than that in the superficial veins (2.6 seconds; P <.0001). In the absence of junctional reflux, the prevalence of deep venous insufficiency (DVI) was significantly lower compared with that in limbs with junctional involvement (2 of 33 vs 29 of 106; P =.038). The mean duration of deep venous reflux in these groups was comparable (0.85 seconds vs 0. 91 seconds; P =.44). Occlusion of the incompetent superficial veins reduced somewhat the duration of the deep venous reflux but did not abolish it (0.88 seconds vs 0.82 seconds; P =.072). The presence of DVI was associated with junctional reflux of high peak velocity and long duration. The prevalence of DVI in patients with primary superficial venous reflux and without history of DVT is 22%. However, this reflux is segmental, mainly in the common femoral vein, and is of short duration. It is associated with the presence of junctional incompetence that has a high peak velocity and long duration. These findings may explain why surgical correction of superficial reflux abolishes DVI.


Investigation of Lupeol as Anti-Melanoma Agent: An In Vitro-In Ovo Perspective.
Malignant melanoma (MM) represents the most life-threatening skin cancer worldwide, with a narrow and inefficient chemotherapeutic arsenal available in advanced disease stages. Lupeol (LUP) is a triterpenoid-type phytochemical possessing a broad spectrum of pharmacological properties, including a potent anticancer effect against several neoplasms (e.g., colorectal, lung, and liver). However, its potential as an anti-melanoma agent has been investigated to a lesser extent. The current study focused on exploring the impact of LUP against two human MM cell lines (A375 and RPMI-7951) in terms of cell viability, confluence, morphology, cytoskeletal distribution, nuclear aspect, and migration. Additionally, the in ovo antiangiogenic effect has been also examined. The in vitro results indicated concentration-dependent and selective cytotoxicity against both MM cell lines, with estimated IC50 values of 66.59 ± 2.20 for A375, and 45.54 ± 1.48 for RPMI-7951, respectively, accompanied by a reduced cell confluence, apoptosis-specific nuclear features, reorganization of cytoskeletal components, and inhibited cell migration. In ovo, LUP interfered with the process of angiogenesis by reducing the formation of neovascularization. Despite the potential anti-melanoma effect illustrated in our in vitro-in ovo study, further investigations are required to elucidate the underlying LUP-induced effects in A375 and RPMI-7951 MM cells.


C4d deposition without rejection correlates with reduced early scarring in ABO-incompatible renal allografts.
C4d deposition in peritubular capillaries is a specific marker for the presence of antidonor antibodies in renal transplant recipients and is usually associated with antibody-mediated rejection (AMR) in conventional allografts. In ABO-incompatible grafts, however, peritubular capillary C4d is often present on protocol biopsies lacking histologic features of AMR; the significance of C4d in this setting remains unclear. For addressing this, data from 33 patients who received ABO-incompatible renal allografts (after desensitization) were retrospectively reviewed. Protocol biopsies were performed at 1 and/or 3 and 6 mo after transplantation in each recipient and at 12 mo in 28 recipients. Twenty-one patients (group A) had strong, diffuse peritubular capillary C4d staining without histologic evidence of AMR or cellular rejection on their initial protocol biopsies. The remaining 12 patients (group B) had negative or weak, focal peritubular capillary C4d staining. Three grafts (two in group B) were lost but not as a result of AMR. Excluding these three patients, serum creatinine levels were similar in the two groups at 6 and 12 mo after transplantation and at last follow-up; however, recipients in group A developed significantly fewer overall chronic changes, as scored by the sum of Banff chronic indices, than group B during the first year after transplantation. These results suggest that diffuse peritubular capillary C4d deposition without rejection is associated with a lower risk for scarring in ABO-incompatible renal allografts; the generalizability of these results to conventional allografts remains unknown.