Science.gov

Sample records for adult tissue samples

  1. General Information about Adult Soft Tissue Sarcoma

    MedlinePlus

    ... Soft Tissue Sarcoma Treatment (PDQ®)–Patient Version General Information About Adult Soft Tissue Sarcoma Go to Health ... the PDQ Adult Treatment Editorial Board . Clinical Trial Information A clinical trial is a study to answer ...

  2. Microfluidic sampling system for tissue analytics

    PubMed Central

    Hokkanen, A.; Stuns, I.; Schmid, P.; Kokkonen, A.; Gao, F.; Steinecker, A.; Budczies, J.; Heimala, P.; Hakalahti, L.

    2015-01-01

    We have developed a microfluidics based sampling system for tissue analytics. The proof-of-concept of the sampling system was demonstrated by extracting lipid samples from tissue biopsies. The sample collection system consists of a disposable silicon based multiport microneedle integrated with polymer microfluidics. The polymethyl methacrylate polymer microfluidic chip has a 10 μl sample reservoir and actuation membranes for liquid pumping. A special automated robotic system was developed to control the positioning of the needle and the sampling procedure on preselected spots on the tissue. Real breast cancer tissue samples were used to test the feasibility of the sampling system. We successfully measured indicative cancer biomarkers from the tissue surface. Phosphatidylcholine and phosphoethanolamine were extracted from the tissue membrane with methyl tert-butyl ether solvent and detected by mass spectrometry. In the future, this tool could be used in characterization of preoperative biopsies and tumour tissues removed during surgery. PMID:26421088

  3. Distribution of 3-hydroxy iC17:0 in subgingival plaque and gingival tissue samples: relationship to adult periodontitis.

    PubMed

    Nichols, F C

    1994-09-01

    Gram-negative organisms incorporate hydroxy fatty acids into the lipid A moiety of lipopolysaccharide (LPS), and in the case of some members of the family Enterobacteriaceae, hydroxy fatty acids are incorporated exclusively into lipid A. However, a limited number of Bacteroides species have been shown to incorporate several classes of 3-hydroxy fatty acids, particularly 3-hydroxy iC17:0, into constitutive lipids as well as LPS. The present study examined the distribution of hydroxy fatty acids in two periodontal pathogens, Prevotella intermedia and Porphyromonas gingivalis, by employing a phospholipid extraction procedure (E. G. Bligh and W. J. Dyer, Can. J. Biochem. Physiol. 37:911-917, 1959) which partitioned constitutive lipids into the organic solvent phase and LPS into the aqueous phase. The distribution of hydroxy fatty acids within organic solvent and aqueous extracts of these bacterial species was then compared with the distribution in subgingival plaque samples isolated from either gingivitis or severe periodontitis sites as well as the distribution in gingival tissue samples. The organic solvent and aqueous extracts were hydrolyzed under strong alkaline conditions, and the free fatty acids were treated to form pentafluorobenzyl-ester, trimethylsilyl-ether derivatives. Hydroxy fatty acid levels were quantified by using gas chromatography-negative-ion chemical ionization-mass spectrometry. By using this approach, the mean values of the 3-hydroxy iC17:0 recovered within organic solvent extracts of P. gingivalis strains ranged from 56 to 63% of total 3-hydroxy iC17:0. Substantially less 3-hydroxy iC17:0 (< 5%) was recovered in organic solvent extracts of P. intermedia. By comparison, 75% of the 3-hydroxy iC17:0 in periodontitis subgingival plaque samples was recovered in organic solvent extracts, while only 43% of the 3-hydroxy iC17:0 in gingivitis plaque samples from the same patients was recovered in organic solvent extracts. However, 3-hydroxy iC17:0 was

  4. Adult stem cells and tissue repair.

    PubMed

    Körbling, M; Estrov, Z; Champlin, R

    2003-08-01

    Recently, adult stem cells originating from bone marrow or peripheral blood have been suggested to contribute to repair and genesis of cells specific for liver, cardiac and skeletal muscle, gut, and brain tissue. The mechanism involved has been termed transdifferentiation, although other explanations including cell fusion have been postulated. Using adult stem cells to generate or repair solid organ tissue obviates the immunologic, ethical, and teratogenic issues that accompany embryonic stem cells.

  5. The landscape of genomic imprinting across diverse adult human tissues

    PubMed Central

    Baran, Yael; Subramaniam, Meena; Biton, Anne; Tukiainen, Taru; Tsang, Emily K.; Rivas, Manuel A.; Pirinen, Matti; Gutierrez-Arcelus, Maria; Smith, Kevin S.; Kukurba, Kim R.; Zhang, Rui; Eng, Celeste; Torgerson, Dara G.; Urbanek, Cydney; Li, Jin Billy; Rodriguez-Santana, Jose R.; Burchard, Esteban G.; Seibold, Max A.; MacArthur, Daniel G.; Montgomery, Stephen B.; Zaitlen, Noah A.; Lappalainen, Tuuli

    2015-01-01

    Genomic imprinting is an important regulatory mechanism that silences one of the parental copies of a gene. To systematically characterize this phenomenon, we analyze tissue specificity of imprinting from allelic expression data in 1582 primary tissue samples from 178 individuals from the Genotype-Tissue Expression (GTEx) project. We characterize imprinting in 42 genes, including both novel and previously identified genes. Tissue specificity of imprinting is widespread, and gender-specific effects are revealed in a small number of genes in muscle with stronger imprinting in males. IGF2 shows maternal expression in the brain instead of the canonical paternal expression elsewhere. Imprinting appears to have only a subtle impact on tissue-specific expression levels, with genes lacking a systematic expression difference between tissues with imprinted and biallelic expression. In summary, our systematic characterization of imprinting in adult tissues highlights variation in imprinting between genes, individuals, and tissues. PMID:25953952

  6. Tissue Sampling Guides for Porcine Biomedical Models.

    PubMed

    Albl, Barbara; Haesner, Serena; Braun-Reichhart, Christina; Streckel, Elisabeth; Renner, Simone; Seeliger, Frank; Wolf, Eckhard; Wanke, Rüdiger; Blutke, Andreas

    2016-04-01

    This article provides guidelines for organ and tissue sampling adapted to porcine animal models in translational medical research. Detailed protocols for the determination of sampling locations and numbers as well as recommendations on the orientation, size, and trimming direction of samples from ∼50 different porcine organs and tissues are provided in the Supplementary Material. The proposed sampling protocols include the generation of samples suitable for subsequent qualitative and quantitative analyses, including cryohistology, paraffin, and plastic histology; immunohistochemistry;in situhybridization; electron microscopy; and quantitative stereology as well as molecular analyses of DNA, RNA, proteins, metabolites, and electrolytes. With regard to the planned extent of sampling efforts, time, and personnel expenses, and dependent upon the scheduled analyses, different protocols are provided. These protocols are adjusted for (I) routine screenings, as used in general toxicity studies or in analyses of gene expression patterns or histopathological organ alterations, (II) advanced analyses of single organs/tissues, and (III) large-scale sampling procedures to be applied in biobank projects. Providing a robust reference for studies of porcine models, the described protocols will ensure the efficiency of sampling, the systematic recovery of high-quality samples representing the entire organ or tissue as well as the intra-/interstudy comparability and reproducibility of results.

  7. Flexible adult flatfoot: soft tissue procedures.

    PubMed

    Walters, Jeremy L; Mendicino, Samuel S

    2014-07-01

    Classically, adult posterior tibial tendon dysfunction (PTTD) was considered primarily a tendon rupture and was treated as such with soft tissue repair alone. The understanding that PTTD involves more than simply an inflammatory condition or tendon rupture but also a muscle imbalance, leading to a flatfoot, osteoarthritis, and peritalar subluxation, led to surgeons advocating osseous procedures as well. The advancements in knowledge of the pathomechanics of the deformity have modified the role that soft tissue repair plays in surgical treatment, but the importance of soft tissue restoration in flatfoot repair should not be overlooked.

  8. Flexible adult flatfoot: soft tissue procedures.

    PubMed

    Walters, Jeremy L; Mendicino, Samuel S

    2014-07-01

    Classically, adult posterior tibial tendon dysfunction (PTTD) was considered primarily a tendon rupture and was treated as such with soft tissue repair alone. The understanding that PTTD involves more than simply an inflammatory condition or tendon rupture but also a muscle imbalance, leading to a flatfoot, osteoarthritis, and peritalar subluxation, led to surgeons advocating osseous procedures as well. The advancements in knowledge of the pathomechanics of the deformity have modified the role that soft tissue repair plays in surgical treatment, but the importance of soft tissue restoration in flatfoot repair should not be overlooked. PMID:24980925

  9. Soft tissue problems in older adults.

    PubMed

    Holland, N W; Gonzalez, E B

    1998-08-01

    This article describes common soft tissue problems encountered in older adults, including fibromyalgia, selected bursitis/tendinitis syndromes, nerve entrapment syndromes, and miscellaneous topics such as Dupuytren's contractures, trigger fingers, palmar fasciitis, and reflex-sympathetic dystrophy. Clinical presentations, diagnosis, and treatment are emphasized. These are conditions that are frequently encountered but are generally diagnosed as arthritis or normal age-related problems. This article will hopefully enlighten the reader in distinguishing between these conditions.

  10. Calibration of redox scanning for tissue samples

    NASA Astrophysics Data System (ADS)

    Xu, He N.; Wu, Baohua; Nioka, Shoko; Chance, Britton; Li, Lin Z.

    2009-02-01

    The fluorescence properties of reduced nicotinamide adenine dinucleotide (NADH) and oxidized flavoproteins (Fp) such as flavin adenine dinucleotide (FAD) in the respiratory chain are sensitive indicators of intracellular redox states and have been applied to the studies of mitochondrial function with energy-linked processes. The redox scanner, a threedimensional (3D) redox cryo-imager previously developed by Chance et al., can quantitatively determine the metabolic properties of tissue samples by acquiring the fluorescence images of NADH and Fp. The redox ratios, i.e., Fp/(Fp+NADH) and NADH/(Fp+NADH), obtained on the basis of relative signal intensity ratios, provide a sensitive index of steady-state of the mitochondrial metabolism that has been determined for a variety of biological tissues. This paper presents the further development of the instrument by establishing a calibration method to quantify the concentration of the fluorophores and facilitate the comparison of redox images obtained at different time or with different instrument functions. Calibration curves of both NADH and Fp have been obtained using snap-frozen standard references with NADH concentration ranging from 150-1400 μM and Fp from 80-720 μM. Snap-freeze tissue samples such as human breast tumors xenografted in mice, normal mouse pancreases and spleens were imaged. The NADH and Fp concentrations as well as the redox ratios in the tissue samples were quantified based on the adjacent solution standards of NADH and Fp. The obtained multi-slice redox images revealed high heterogeneity of the tissue samples which can be quantitatively interpreted.

  11. Collecting and Storing Tissue, Blood, and Bone Marrow Samples From Patients With Rhabdomyosarcoma or Other Soft Tissue Sarcoma

    ClinicalTrials.gov

    2016-09-23

    Adult Rhabdomyosarcoma; Childhood Desmoplastic Small Round Cell Tumor; Chordoma; Desmoid Tumor; Metastatic Childhood Soft Tissue Sarcoma; Nonmetastatic Childhood Soft Tissue Sarcoma; Previously Treated Childhood Rhabdomyosarcoma; Previously Untreated Childhood Rhabdomyosarcoma; Recurrent Adult Soft Tissue Sarcoma; Recurrent Childhood Rhabdomyosarcoma; Recurrent Childhood Soft Tissue Sarcoma; Stage I Adult Soft Tissue Sarcoma; Stage II Adult Soft Tissue Sarcoma; Stage III Adult Soft Tissue Sarcoma; Stage IV Adult Soft Tissue Sarcoma

  12. Competency-Based Adult Education Sample Activities.

    ERIC Educational Resources Information Center

    Dauzat, Sam V.; Bryant, Nerissa

    For use by adult basic education teachers, this curriculum guide is intended as a frame of reference for building a total instructional program designed around a competency-based and skill-based curriculum. The 104 sample activities are categorized under subheadings of the following topics: Occupational Knowledge, Health, Government and Law,…

  13. Treatment Options for Adult Soft Tissue Sarcoma

    MedlinePlus

    ... superficial (in subcutaneous tissue with no spread into connective tissue or muscle below) or deep (in the muscle ... superficial (in subcutaneous tissue with no spread into connective tissue or muscle below) or deep (in the muscle ...

  14. Treatment Option Overview (Adult Soft Tissue Sarcoma)

    MedlinePlus

    ... superficial (in subcutaneous tissue with no spread into connective tissue or muscle below) or deep (in the muscle ... superficial (in subcutaneous tissue with no spread into connective tissue or muscle below) or deep (in the muscle ...

  15. Stages of Adult Soft Tissue Sarcoma

    MedlinePlus

    ... superficial (in subcutaneous tissue with no spread into connective tissue or muscle below) or deep (in the muscle ... superficial (in subcutaneous tissue with no spread into connective tissue or muscle below) or deep (in the muscle ...

  16. Detection of Quiescent Infections with Multiple Elephant Endotheliotropic Herpesviruses (EEHVs), Including EEHV2, EEHV3, EEHV6, and EEHV7, within Lymphoid Lung Nodules or Lung and Spleen Tissue Samples from Five Asymptomatic Adult African Elephants

    PubMed Central

    Zong, Jian-Chao; Heaggans, Sarah Y.; Long, Simon Y.; Latimer, Erin M.; Nofs, Sally A.; Bronson, Ellen; Casares, Miguel; Fouraker, Michael D.; Pearson, Virginia R.; Richman, Laura K.

    2015-01-01

    ABSTRACT More than 80 cases of lethal hemorrhagic disease associated with elephant endotheliotropic herpesviruses (EEHVs) have been identified in young Asian elephants worldwide. Diagnostic PCR tests detected six types of EEHV in blood of elephants with acute disease, although EEHV1A is the predominant pathogenic type. Previously, the presence of herpesvirus virions within benign lung and skin nodules from healthy African elephants led to suggestions that African elephants may be the source of EEHV disease in Asian elephants. Here, we used direct PCR-based DNA sequencing to detect EEHV genomes in necropsy tissue from five healthy adult African elephants. Two large lung nodules collected from culled wild South African elephants contained high levels of either EEHV3 alone or both EEHV2 and EEHV3. Similarly, a euthanized U.S. elephant proved to harbor multiple EEHV types distributed nonuniformly across four small lung nodules, including high levels of EEHV6, lower levels of EEHV3 and EEHV2, and a new GC-rich branch type, EEHV7. Several of the same EEHV types were also detected in random lung and spleen samples from two other elephants. Sanger PCR DNA sequence data comprising 100 kb were obtained from a total of 15 different strains identified, with (except for a few hypervariable genes) the EEHV2, EEHV3, and EEHV6 strains all being closely related to known genotypes from cases of acute disease, whereas the seven loci (4.0 kb) obtained from EEHV7 averaged 18% divergence from their nearest relative, EEHV3. Overall, we conclude that these four EEHV species, but probably not EEHV1, occur commonly as quiescent infections in African elephants. IMPORTANCE Acute hemorrhagic disease characterized by high-level viremia due to infection by members of the Proboscivirus genus threatens the future breeding success of endangered Asian elephants worldwide. Although the genomes of six EEHV types from acute cases have been partially or fully characterized, lethal disease predominantly

  17. [Radiotherapy of adult soft tissue sarcoma].

    PubMed

    Le Péchoux, C; Moureau-Zabotto, L; Llacer, C; Ducassou, A; Sargos, P; Sunyach, M P; Thariat, J

    2016-09-01

    Incidence of soft tissue sarcoma is low and requires multidisciplinary treatment in specialized centers. The objective of this paper is to report the state of the art regarding indications and treatment techniques of main soft tissue sarcoma localisations.

  18. [Radiotherapy of adult soft tissue sarcoma].

    PubMed

    Le Péchoux, C; Moureau-Zabotto, L; Llacer, C; Ducassou, A; Sargos, P; Sunyach, M P; Thariat, J

    2016-09-01

    Incidence of soft tissue sarcoma is low and requires multidisciplinary treatment in specialized centers. The objective of this paper is to report the state of the art regarding indications and treatment techniques of main soft tissue sarcoma localisations. PMID:27523415

  19. Epimorphic regeneration approach to tissue replacement in adult mammals

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Urodeles and fetal mammals are capable of impressive epimorphic regeneration in a variety of tissues, whereas the typical default response to injury in adult mammals consists of inflammation and scar tissue formation. One component of epimorphic regeneration is the recruitment of resident progenitor...

  20. Adult stem cells applied to tissue engineering and regenerative medicine.

    PubMed

    Cuenca-López, M D; Zamora-Navas, P; García-Herrera, J M; Godino, M; López-Puertas, J M; Guerado, E; Becerra, J; Andrades, J A

    2008-01-01

    Regeneration takes place in the body at a moment or another throughout life. Bone, cartilage, and tendons (the key components of the structure and articulation in the body) have a limited capacity for self-repair and, after traumatic injury or disease, the regenerative power of adult tissue is often insufficient. When organs or tissues are irreparably damaged, they may be replaced by an artificial device or by a donor organ. However, the number of available donor organs is considerably limited. Generation of tissue-engineered replacement organs by extracting stem cells from the patient, growing them and modifying them in clinical conditions after re-introduction in the body represents an ideal source for corrective treatment. Mesenchymal stem cells (MSCs) are the multipotential progenitors that give rise to skeletal cells, vascular smooth muscle cells, muscle (skeletal and cardiac muscle), adipocytes (fat tissue) and hematopoietic (blood)-supportive stromal cells. MSCs are found in multiple connective tissues, in adult bone marrow, skeletal muscles and fat pads. The wide representation in adult tissues may be related to the existence of a circulating blood pool or that MSCs are associated to the vascular system.

  1. Technical note: Alternatives to reduce adipose tissue sampling bias.

    PubMed

    Cruz, G D; Wang, Y; Fadel, J G

    2014-10-01

    Understanding the mechanisms by which nutritional and pharmaceutical factors can manipulate adipose tissue growth and development in production animals has direct and indirect effects in the profitability of an enterprise. Adipocyte cellularity (number and size) is a key biological response that is commonly measured in animal science research. The variability and sampling of adipocyte cellularity within a muscle has been addressed in previous studies, but no attempt to critically investigate these issues has been proposed in the literature. The present study evaluated 2 sampling techniques (random and systematic) in an attempt to minimize sampling bias and to determine the minimum number of samples from 1 to 15 needed to represent the overall adipose tissue in the muscle. Both sampling procedures were applied on adipose tissue samples dissected from 30 longissimus muscles from cattle finished either on grass or grain. Briefly, adipose tissue samples were fixed with osmium tetroxide, and size and number of adipocytes were determined by a Coulter Counter. These results were then fit in a finite mixture model to obtain distribution parameters of each sample. To evaluate the benefits of increasing number of samples and the advantage of the new sampling technique, the concept of acceptance ratio was used; simply stated, the higher the acceptance ratio, the better the representation of the overall population. As expected, a great improvement on the estimation of the overall adipocyte cellularity parameters was observed using both sampling techniques when sample size number increased from 1 to 15 samples, considering both techniques' acceptance ratio increased from approximately 3 to 25%. When comparing sampling techniques, the systematic procedure slightly improved parameters estimation. The results suggest that more detailed research using other sampling techniques may provide better estimates for minimum sampling.

  2. Specimen Sample Preservation for Cell and Tissue Cultures

    NASA Technical Reports Server (NTRS)

    Meeker, Gabrielle; Ronzana, Karolyn; Schibner, Karen; Evans, Robert

    1996-01-01

    The era of the International Space Station with its longer duration missions will pose unique challenges to microgravity life sciences research. The Space Station Biological Research Project (SSBRP) is responsible for addressing these challenges and defining the science requirements necessary to conduct life science research on-board the International Space Station. Space Station will support a wide range of cell and tissue culture experiments for durations of 1 to 30 days. Space Shuttle flights to bring experimental samples back to Earth for analyses will only occur every 90 days. Therefore, samples may have to be retained for periods up to 60 days. This presents a new challenge in fresh specimen sample storage for cell biology. Fresh specimen samples are defined as samples that are preserved by means other than fixation and cryopreservation. The challenge of long-term storage of fresh specimen samples includes the need to suspend or inhibit proliferation and metabolism pending return to Earth-based laboratories. With this challenge being unique to space research, there have not been any ground based studies performed to address this issue. It was decided hy SSBRP that experiment support studies to address the following issues were needed: Fixative Solution Management; Media Storage Conditions; Fresh Specimen Sample Storage of Mammalian Cell/Tissue Cultures; Fresh Specimen Sample Storage of Plant Cell/Tissue Cultures; Fresh Specimen Sample Storage of Aquatic Cell/Tissue Cultures; and Fresh Specimen Sample Storage of Microbial Cell/Tissue Cultures. The objective of these studies was to derive a set of conditions and recommendations that can be used in a long duration microgravity environment such as Space Station that will permit extended storage of cell and tissue culture specimens in a state consistent with zero or minimal growth, while at the same time maintaining their stability and viability.

  3. Alkaline diets favor lean tissue mass in older adults

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Maintaining muscle mass in aging is important to prevent falls and fractures. The net acid load from diets that are rich in acidogenic protein and cereal grains relative to their content of alkalinogenic fruits and vegetables may contribute to reduced lean tissue mass in older adults. This analysis ...

  4. Tissue adaptations to gravitational stress - Newborn versus adult giraffes

    NASA Technical Reports Server (NTRS)

    Hargens, Alan R; Gershuni, David H.; Danzig, Larry A.; Millard, Ronald W.; Pettersson, Knut

    1988-01-01

    Preliminary results on developmental alterations in load-bearing tissues of newborn and adult giraffes are presented. Attention is focused on vascular wall thickness in relation to local blood pressure, and on meniscal adaptations to increased load bearing in the developing giraffe. It is believed that the developing giraffe provides an excellent model for investigations of adaptive mechanisms of increased weight bearing.

  5. Adult mesenchymal stem cells for tissue engineering versus regenerative medicine.

    PubMed

    Caplan, Arnold I

    2007-11-01

    Adult mesenchymal stem cells (MSCs) can be isolated from bone marrow or marrow aspirates and because they are culture-dish adherent, they can be expanded in culture while maintaining their multipotency. The MSCs have been used in preclinical models for tissue engineering of bone, cartilage, muscle, marrow stroma, tendon, fat, and other connective tissues. These tissue-engineered materials show considerable promise for use in rebuilding damaged or diseased mesenchymal tissues. Unanticipated is the realization that the MSCs secrete a large spectrum of bioactive molecules. These molecules are immunosuppressive, especially for T-cells and, thus, allogeneic MSCs can be considered for therapeutic use. In this context, the secreted bioactive molecules provide a regenerative microenvironment for a variety of injured adult tissues to limit the area of damage and to mount a self-regulated regenerative response. This regenerative microenvironment is referred to as trophic activity and, therefore, MSCs appear to be valuable mediators for tissue repair and regeneration. The natural titers of MSCs that are drawn to sites of tissue injury can be augmented by allogeneic MSCs delivered via the bloodstream. Indeed, human clinical trials are now under way to use allogeneic MSCs for treatment of myocardial infarcts, graft-versus-host disease, Crohn's Disease, cartilage and meniscus repair, stroke, and spinal cord injury. This review summarizes the biological basis for the in vivo functioning of MSCs through development and aging. PMID:17620285

  6. Analysis of chemical components from plant tissue samples

    NASA Technical Reports Server (NTRS)

    Laseter, J. L.

    1972-01-01

    Information is given on the type and concentration of sterols, free fatty acids, and total fatty acids in plant tissue samples. All samples were analyzed by gas chromatography and then by gas chromatography-mass spectrometry combination. In each case the mass spectral data was accumulated as a computer printout and plot. Typical gas chromatograms are included as well as tables describing test results.

  7. Multivariate classification of infrared spectra of cell and tissue samples

    DOEpatents

    Haaland, David M.; Jones, Howland D. T.; Thomas, Edward V.

    1997-01-01

    Multivariate classification techniques are applied to spectra from cell and tissue samples irradiated with infrared radiation to determine if the samples are normal or abnormal (cancerous). Mid and near infrared radiation can be used for in vivo and in vitro classifications using at least different wavelengths.

  8. Solid phase microextraction fills the gap in tissue sampling protocols.

    PubMed

    Bojko, Barbara; Gorynski, Krzysztof; Gomez-Rios, German Augusto; Knaak, Jan Matthias; Machuca, Tiago; Spetzler, Vinzent Nikolaus; Cudjoe, Erasmus; Hsin, Michael; Cypel, Marcelo; Selzner, Markus; Liu, Mingyao; Keshavjee, Shaf; Pawliszyn, Janusz

    2013-11-25

    Metabolomics and biomarkers discovery are an integral part of bioanalysis. However, untargeted tissue analysis remains as the bottleneck of such studies due to the invasiveness of sample collection, as well as the laborious and time-consuming sample preparation protocols. In the current study, technology integrating in vivo sampling, sample preparation and global extraction of metabolites--solid phase microextraction was presented and evaluated during liver and lung transplantation in pig model. Sampling approaches, including selection of the probe, transportation, storage conditions and analyte coverage were discussed. The applicability of the method for metabolomics studies was demonstrated during lung transplantation experiments. PMID:24216199

  9. Tissue tropism of recombinant coxsackieviruses in an adult mouse model.

    PubMed

    Harvala, Heli; Kalimo, Hannu; Bergelson, Jeffrey; Stanway, Glyn; Hyypiä, Timo

    2005-07-01

    Recombinant viruses, constructed by exchanging the 5' non-coding region (5'NCR), structural and non-structural protein coding sequences were used to investigate determinants responsible for differences between coxsackievirus A9 (CAV9) and coxsackievirus B3 (CBV3) infections in adult mice and two cell lines. Plaque assay titration of recombinant and parental viruses from different tissues from adult BALB/c mice demonstrated that the structural region of CBV3 determined tropism to the liver tissue due to receptor recognition, and the 5'NCR of CBV3 enhanced viral multiplication in the mouse pancreas. Infection with a chimeric virus, containing the structural region from CBV3 and the rest of the genome from CAV9, and the parental CBV3 strain, caused high levels of viraemia in adult mice. The ability of these viruses to infect the central nervous system suggested that neurotropism is associated with high replication levels and the presence of the CBV3 capsid proteins, which also enhanced formation of neutralizing antibodies. Moreover, the appearance of neutralizing antibodies correlated directly with the clearance of the viruses from the tissues. These results demonstrate potential pathogenicity of intraspecies recombinant coxsackieviruses, and the complexity of the genetic determinants underlying tissue tropism.

  10. Using tissue samples for proteomic studies-critical considerations.

    PubMed

    Becker, Karl-Friedrich

    2015-04-01

    From my experience of 22 years working in a pathology research laboratory and overseeing dozens of collaborations with research groups from basic sciences and industry, I have the impression that researchers are rarely aware of the special issues related to acquisition and processing of frozen or formalin-fixed tissue samples for proteomic analysis. While challenges are expected for formalin-fixed tissues because of the cross-linking activities of formaldehyde, researchers believe when using frozen tissue samples they are safe and always have excellent material to analyze-but this is not always the case. It is alarming that many researchers do not question the quality of the tissue samples they are analyzing and focus only on their analytical technique. Standardization of the entire workflow from test ordering to the report of the proteomic assay, with special emphasis on the preanalytical phase, is crucial for successful integration of proteomic studies in the clinic as protein profiles may change due to sample processing before the proteomic analysis is performed. The aim of this review is to discuss the progress of proteomic studies with human tissues and to highlight the challenges that must be understood and addressed for successful translation of proteomic methods to clinical practice.

  11. Preparation of tissue samples for X-ray fluorescence microscopy

    NASA Astrophysics Data System (ADS)

    Chwiej, Joanna; Szczerbowska-Boruchowska, Magdalena; Lankosz, Marek; Wojcik, Slawomir; Falkenberg, Gerald; Stegowski, Zdzislaw; Setkowicz, Zuzanna

    2005-12-01

    As is well-known, trace elements, especially metals, play an important role in the pathogenesis of many disorders. The topographic and quantitative elemental analysis of pathologically changed tissues may shed some new light on processes leading to the degeneration of cells in the case of selected diseases. An ideal and powerful tool for such purpose is the Synchrotron Microbeam X-ray Fluorescence technique. It enables the carrying out of investigations of the elemental composition of tissues even at the single cell level. The tissue samples for histopathological investigations are routinely fixed and embedded in paraffin. The authors try to verify the usefulness of such prepared tissue sections for elemental analysis with the use of X-ray fluorescence microscopy. Studies were performed on rat brain samples. Changes in elemental composition caused by fixation in formalin or paraformaldehyde and embedding in paraffin were examined. Measurements were carried out at the bending magnet beamline L of the Hamburger Synchrotronstrahlungslabor HASYLAB in Hamburg. The decrease in mass per unit area of K, Br and the increase in P, S, Fe, Cu and Zn in the tissue were observed as a result of the fixation. For the samples embedded in paraffin, a lower level of most elements was observed. Additionally, for these samples, changes in the composition of some elements were not uniform for different analyzed areas of rat brain.

  12. Tissue sampling methods and standards for vertebrate genomics

    PubMed Central

    2012-01-01

    The recent rise in speed and efficiency of new sequencing technologies have facilitated high-throughput sequencing, assembly and analyses of genomes, advancing ongoing efforts to analyze genetic sequences across major vertebrate groups. Standardized procedures in acquiring high quality DNA and RNA and establishing cell lines from target species will facilitate these initiatives. We provide a legal and methodological guide according to four standards of acquiring and storing tissue for the Genome 10K Project and similar initiatives as follows: four-star (banked tissue/cell cultures, RNA from multiple types of tissue for transcriptomes, and sufficient flash-frozen tissue for 1 mg of DNA, all from a single individual); three-star (RNA as above and frozen tissue for 1 mg of DNA); two-star (frozen tissue for at least 700 μg of DNA); and one-star (ethanol-preserved tissue for 700 μg of DNA or less of mixed quality). At a minimum, all tissues collected for the Genome 10K and other genomic projects should consider each species’ natural history and follow institutional and legal requirements. Associated documentation should detail as much information as possible about provenance to ensure representative sampling and subsequent sequencing. Hopefully, the procedures outlined here will not only encourage success in the Genome 10K Project but also inspire the adaptation of standards by other genomic projects, including those involving other biota. PMID:23587255

  13. Magnetic resonance imaging of benign soft tissue neoplasms in adults.

    PubMed

    Walker, Eric A; Fenton, Michael E; Salesky, Joel S; Murphey, Mark D

    2011-11-01

    This article reviews a spectrum of benign soft tissue tumors found in adults. Rather than presenting a complete review, the focus of this article is on benign tumors for which the diagnosis may be confidently made or strongly suggested on the basis of imaging. Diagnoses presented include nodular fasciitis, superficial and deep fibromatosis, elastofibroma, lipomatous lesions, giant cell tumor of the tendon sheath, pigmented villonodular synovitis, peripheral nerve sheath tumors, Morton neuroma, hemangioma, and myxoma.

  14. Translational research in pediatrics: tissue sampling and biobanking.

    PubMed

    Brisson, Alayne R; Matsui, Doreen; Rieder, Michael J; Fraser, Douglas D

    2012-01-01

    Translational research is expanding and has become a focus of National Research funding agencies, touted as the primary avenue to improve health care practice. The use of human tissues for research on disease etiology is a pillar of translational research, particularly with innovations in research technologies to investigate the building blocks of disease. In pediatrics, translational research using human tissues has been hindered by the many practical and ethical considerations associated with tissue procurement from children and also by a limited population base for study, by the increasing complexities in conducting clinical research, and by a lack of dedicated child-health research funding. Given these obstacles, pediatric translational research can be enhanced by developing strategic and efficient biobanks that will provide scientists with quality tissue specimens to render accurate and reproducible research results. Indeed, tissue sampling and biobanking within pediatric academic settings has potential to impact child health by promoting bidirectional interaction between clinicians and scientists, helping to maximize research productivity, and providing a competitive edge for attracting and maintaining high-quality personnel. The authors of this review outline key issues and practical solutions to optimize pediatric tissue sampling and biobanking for translational research, activities that will ultimately reduce the burden of childhood disease. PMID:22144705

  15. Lack of tissue renewal in human adult Achilles tendon is revealed by nuclear bomb 14C

    PubMed Central

    Heinemeier, Katja Maria; Schjerling, Peter; Heinemeier, Jan; Magnusson, Stig Peter; Kjaer, Michael

    2013-01-01

    Tendons are often injured and heal poorly. Whether this is caused by a slow tissue turnover is unknown, since existing data provide diverging estimates of tendon protein half-life that range from 2 mo to 200 yr. With the purpose of determining life-long turnover of human tendon tissue, we used the 14C bomb-pulse method. This method takes advantage of the dramatic increase in atmospheric levels of 14C, produced by nuclear bomb tests in 1955–1963, which is reflected in all living organisms. Levels of 14C were measured in 28 forensic samples of Achilles tendon core and 4 skeletal muscle samples (donor birth years 1945–1983) with accelerator mass spectrometry (AMS) and compared to known atmospheric levels to estimate tissue turnover. We found that Achilles tendon tissue retained levels of 14C corresponding to atmospheric levels several decades before tissue sampling, demonstrating a very limited tissue turnover. The tendon concentrations of 14C approximately reflected the atmospheric levels present during the first 17 yr of life, indicating that the tendon core is formed during height growth and is essentially not renewed thereafter. In contrast, 14C levels in muscle indicated continuous turnover. Our observation provides a fundamental premise for understanding tendon function and pathology, and likely explains the poor regenerative capacity of tendon tissue.—Heinemeier, K. M., Schjerling, P., Heinemeier, J., Magnusson, S. P., Kjaer, M. Lack of tissue renewal in human adult Achilles tendon is revealed by nuclear bomb 14C. PMID:23401563

  16. Immunohistochemical Study of Expression of Sohlh1 and Sohlh2 in Normal Adult Human Tissues

    PubMed Central

    Zhang, Xiaoli; Liu, Ruihua; Su, Zhongxue; Zhang, Yuecun; Zhang, Wenfang; Liu, Xinyu; Wang, Fuwu; Guo, Yuji; Li, Chuangang; Hao, Jing

    2015-01-01

    The expression pattern of Sohlh1 (spermatogenesis and oogenesis specific basic helix-loop-helix 1) and Sohlh2 in mice has been reported in previous studies. Sohlh1 and Sohlh2 are specifically expressed in spermatogonia, prespermatogonia in male mice and oocytes of primordial and primary follicles in female mice. In this report, we studied the expression pattern of Sohlh1 and Sohlh2 in human adult tissues. Immunohistochemical staining of Sohlh1 and Sohlh2 was performed in 5 samples of normal ovaries and testes, respectively. The results revealed that Sohlh genes are not only expressed in oocytes and spermatogonia, but also in granular cells, theca cells, Sertoli cells and Leydig cells, and in smooth muscles of blood vessel walls. To further investigate the expression of Sohlh genes in other adult human tissues, we collected representative normal adult tissues developed from three embryonic germ layers. Compared with the expression in mice, Sohlhs exhibited a much more extensive expression pattern in human tissues. Sohlhs were detected in testis, ovary and epithelia developed from embryonic endoderm, ectoderm and tissues developed from embryonic mesoderm. Sohlh signals were found in spermatogonia, Sertoli cells and also Leydig cells in testis, while in ovary, the expression was mainly in oocytes of primordial and primary follicles, granular cells and theca cells of secondary follicles. Compared with Sohlh2, the expression of Sohlh1 was stronger and more extensive. Our study explored the expression of Sohlh genes in human tissues and might provide insights for functional studies of Sohlh genes. PMID:26375665

  17. [Features of glutamate dehydrogenase in fetal and adult rumen tissue].

    PubMed

    Kalachniuk, H I; Fomenko, I S; Kalachniuk, L H; Kavai, Sh; Marounek, M; Savka, O H

    2001-01-01

    Glutamate dehydrogenase (GDH) from rumen mucosa of cow fetus, liver and two forms from mucosa (bacterial and tissue) of the adult animal were partly purified and characterized. The activity of the bacterial glutamate dehydrogenase was shown to depend on qualities of a biomass of microbes, adhered on surface of rumen mucosa. All enzymes from tissues (GDHTRF, TRC, TLC), revealed the hypersensibility to increase in the concentration medium of Zn2+, guanosine triphosphate (GTP), acting here in a role of negative modulators, and also adenosine monophosphate (AMP) and leucine, which acted as activators. However, in the same concentrations these effectors do not influence the activity of the bacterial glutamate dehydrogenase. And if all tissues enzymes are highly specific to coenzyme NADH, the bacterial ones almost in 3 times is more active at NADPH use. PMID:11642036

  18. Epimorphic regeneration approach to tissue replacement in adult mammals

    PubMed Central

    Agrawal, Vineet; Johnson, Scott A.; Reing, Janet; Zhang, Li; Tottey, Stephen; Wang, Gang; Hirschi, Karen K.; Braunhut, Susan; Gudas, Lorraine J.; Badylak, Stephen F.

    2009-01-01

    Urodeles and fetal mammals are capable of impressive epimorphic regeneration in a variety of tissues, whereas the typical default response to injury in adult mammals consists of inflammation and scar tissue formation. One component of epimorphic regeneration is the recruitment of resident progenitor and stem cells to a site of injury. Bioactive molecules resulting from degradation of extracellular matrix (ECM) have been shown to recruit a variety of progenitor and stem cells in vitro in adult mammals. The ability to recruit multipotential cells to the site of injury by in vivo administration of chemotactic ECM degradation products in a mammalian model of digit amputation was investigated in the present study. Adult, 6- to 8-week-old C57/BL6 mice were subjected to midsecond phalanx amputation of the third digit of the right hind foot and either treated with chemotactic ECM degradation products or left untreated. At 14 days after amputation, mice treated with ECM degradation products showed an accumulation of heterogeneous cells that expressed markers of multipotency, including Sox2, Sca1, and Rex1 (Zfp42). Cells isolated from the site of amputation were capable of differentiation along neuroectodermal and mesodermal lineages, whereas cells isolated from control mice were capable of differentiation along only mesodermal lineages. The present findings demonstrate the recruitment of endogenous stem cells to a site of injury, and/or their generation/proliferation therein, in response to ECM degradation products. PMID:19966310

  19. Strategies to Optimize Adult Stem Cell Therapy for Tissue Regeneration

    PubMed Central

    Liu, Shan; Zhou, Jingli; Zhang, Xuan; Liu, Yang; Chen, Jin; Hu, Bo; Song, Jinlin; Zhang, Yuanyuan

    2016-01-01

    Stem cell therapy aims to replace damaged or aged cells with healthy functioning cells in congenital defects, tissue injuries, autoimmune disorders, and neurogenic degenerative diseases. Among various types of stem cells, adult stem cells (i.e., tissue-specific stem cells) commit to becoming the functional cells from their tissue of origin. These cells are the most commonly used in cell-based therapy since they do not confer risk of teratomas, do not require fetal stem cell maneuvers and thus are free of ethical concerns, and they confer low immunogenicity (even if allogenous). The goal of this review is to summarize the current state of the art and advances in using stem cell therapy for tissue repair in solid organs. Here we address key factors in cell preparation, such as the source of adult stem cells, optimal cell types for implantation (universal mesenchymal stem cells vs. tissue-specific stem cells, or induced vs. non-induced stem cells), early or late passages of stem cells, stem cells with endogenous or exogenous growth factors, preconditioning of stem cells (hypoxia, growth factors, or conditioned medium), using various controlled release systems to deliver growth factors with hydrogels or microspheres to provide apposite interactions of stem cells and their niche. We also review several approaches of cell delivery that affect the outcomes of cell therapy, including the appropriate routes of cell administration (systemic, intravenous, or intraperitoneal vs. local administration), timing for cell therapy (immediate vs. a few days after injury), single injection of a large number of cells vs. multiple smaller injections, a single site for injection vs. multiple sites and use of rodents vs. larger animal models. Future directions of stem cell-based therapies are also discussed to guide potential clinical applications. PMID:27338364

  20. Strategies to Optimize Adult Stem Cell Therapy for Tissue Regeneration.

    PubMed

    Liu, Shan; Zhou, Jingli; Zhang, Xuan; Liu, Yang; Chen, Jin; Hu, Bo; Song, Jinlin; Zhang, Yuanyuan

    2016-06-21

    Stem cell therapy aims to replace damaged or aged cells with healthy functioning cells in congenital defects, tissue injuries, autoimmune disorders, and neurogenic degenerative diseases. Among various types of stem cells, adult stem cells (i.e., tissue-specific stem cells) commit to becoming the functional cells from their tissue of origin. These cells are the most commonly used in cell-based therapy since they do not confer risk of teratomas, do not require fetal stem cell maneuvers and thus are free of ethical concerns, and they confer low immunogenicity (even if allogenous). The goal of this review is to summarize the current state of the art and advances in using stem cell therapy for tissue repair in solid organs. Here we address key factors in cell preparation, such as the source of adult stem cells, optimal cell types for implantation (universal mesenchymal stem cells vs. tissue-specific stem cells, or induced vs. non-induced stem cells), early or late passages of stem cells, stem cells with endogenous or exogenous growth factors, preconditioning of stem cells (hypoxia, growth factors, or conditioned medium), using various controlled release systems to deliver growth factors with hydrogels or microspheres to provide apposite interactions of stem cells and their niche. We also review several approaches of cell delivery that affect the outcomes of cell therapy, including the appropriate routes of cell administration (systemic, intravenous, or intraperitoneal vs. local administration), timing for cell therapy (immediate vs. a few days after injury), single injection of a large number of cells vs. multiple smaller injections, a single site for injection vs. multiple sites and use of rodents vs. larger animal models. Future directions of stem cell-based therapies are also discussed to guide potential clinical applications.

  1. Multipotent progenitor cells isolated from adult human pancreatic tissue.

    PubMed

    Todorov, I; Nair, I; Ferreri, K; Rawson, J; Kuroda, A; Pascual, M; Omori, K; Valiente, L; Orr, C; Al-Abdullah, I; Riggs, A; Kandeel, F; Mullen, Y

    2005-10-01

    The supply of islet cells is a limiting factor for the widespread application of islet transplantation of type-1 diabetes. Islets constitute 1% to 2% of pancreatic tissue, leaving approximately 98% as discard after islet isolation and purification. In this report we present our data on the isolation of multipotent progenitor cells from discarded adult human pancreatic tissue. The collected cells from discarded nonislet fractions, after enzymatic digestion and gradient purification of islets, were dissociated for suspension culture in a serum-free medium. The cell clusters grown to a size of 100 to 150 mum contained cells staining for stage-specific embryonic antigens, but not insulin or C-peptide. To direct cell differentiation toward islets, clusters were recultured in a pancreatic differentiation medium. Insulin and C-peptide-positive cells by immunocytochemistry appeared within a week, reaching over 10% of the cell population. Glucagon and somatostatin-positive cells were also detected. The cell clusters were found to secrete insulin in response to glucose stimulation. Cells from the same clusters also had the capacity for differentiation into neural cells, as documented by staining for neural and glial cell markers when cultured as monolayers in media containing neurotrophic factors. These data suggest that multipotent pancreatic progenitor cells exist within the human pancreatic tissue that is typically discarded during islet isolation procedures. These adult progenitor cells can be successfully differentiated into insulin-producing cells, and thus they have the potential for treatment of type-1 diabetes mellitus. PMID:16298614

  2. Semiautomated Device for Batch Extraction of Metabolites from Tissue Samples

    PubMed Central

    2012-01-01

    Metabolomics has become a mainstream analytical strategy for investigating metabolism. The quality of data derived from these studies is proportional to the consistency of the sample preparation. Although considerable research has been devoted to finding optimal extraction protocols, most of the established methods require extensive sample handling. Manual sample preparation can be highly effective in the hands of skilled technicians, but an automated tool for purifying metabolites from complex biological tissues would be of obvious utility to the field. Here, we introduce the semiautomated metabolite batch extraction device (SAMBED), a new tool designed to simplify metabolomics sample preparation. We discuss SAMBED’s design and show that SAMBED-based extractions are of comparable quality to extracts produced through traditional methods (13% mean coefficient of variation from SAMBED versus 16% from manual extractions). Moreover, we show that aqueous SAMBED-based methods can be completed in less than a quarter of the time required for manual extractions. PMID:22292466

  3. Switching roles: the functional plasticity of adult tissue stem cells

    PubMed Central

    Wabik, Agnieszka; Jones, Philip H

    2015-01-01

    Adult organisms have to adapt to survive, and the same is true for their tissues. Rates and types of cell production must be rapidly and reversibly adjusted to meet tissue demands in response to both local and systemic challenges. Recent work reveals how stem cell (SC) populations meet these requirements by switching between functional states tuned to homoeostasis or regeneration. This plasticity extends to differentiating cells, which are capable of reverting to SCs after injury. The concept of the niche, the micro-environment that sustains and regulates stem cells, is broadening, with a new appreciation of the role of physical factors and hormonal signals. Here, we review different functions of SCs, the cellular mechanisms that underlie them and the signals that bias the fate of SCs as they switch between roles. PMID:25812989

  4. Adult stem cell plasticity: will engineered tissues be rejected?

    PubMed Central

    Fang, Te-Chao; Alison, Malcolm R; Wright, Nicholas A; Poulsom, Richard

    2004-01-01

    The dogma that adult tissue-specific stem cells remain committed to supporting only their own tissue has been challenged; a new hypothesis, that adult stem cells demonstrate plasticity in their repertoires, is being tested. This is important because it seems possible that haematopoietic stem cells, for example, could be exploited to generate and perhaps deliver cell-based therapies deep within existing nonhaematopoietic organs. Much of the evidence for plasticity derives from histological studies of tissues from patients or animals that have received grafts of cells or whole organs, from a donor bearing (or lacking) a definitive marker. Detection in the recipient of appropriately differentiated cells bearing the donor marker is indicative of a switch in phenotype of a stem cell or a member of a transit amplifying population or of a differentiated cell. In this review, we discuss evidence for these changes occurring but do not consider the molecular basis of cell commitment. In general, the extent of engraftment is low but may be increased if tissues are damaged. In model systems of liver regeneration, the repeated application of a selection pressure increases levels of engraftment considerably; how this occurs is unclear. Cell fusion plays a part in regeneration and remodelling of the liver, skeletal muscle and even regions of the brain. Genetic disease may be amenable to some forms of cell therapy, yet immune rejection will present challenges. Graft-vs.-host disease will continue to present problems, although this may be avoided if the cells were derived from the recipient or they were tolerized. Despite great expectations for cellular therapies, there are indications that attempts to replace missing proteins could be confounded simply by the development of specific immunity that rejects the new phenotype. PMID:15255965

  5. An adapted tissue microarray for the development of a matrix arrangement of tissue samples.

    PubMed

    Gurgel, Daniel C; Dornelas, Conceição A; Lima-Júnior, Roberto C P; Ribeiro, Ronaldo A; Almeida, Paulo R C

    2012-03-15

    The arrangement of tissue samples in a matrix, known as the tissue microarray (TMA) method, is a well-recognized method worldwide. This technique makes it possible to assess the expression of molecular markers on a large scale with high yields in terms of time, costs, and archived material. Some researchers are trying to adapt the technique to expand the research possibilities. This study proposes an adaptive simplification of low-cost instruments for obtaining samples that will be used in the construction of the TMA. The use of a manual leather puncher, which has a very low cost and a long expected life and eliminates the need to use a press machine, is a simple and effective alternative to building blocks of tissue microarrays.

  6. Laser microdissection of small tissue samples--application to chronic pancreatitis tissues.

    PubMed

    Heinmöller, Ernst; Bockholt, Anke; Werther, Meike; Ziemer, Maria; Müller, Annegret; Ghadimi, B Michael; Rüschoff, Josef

    2003-01-01

    Laser microdissection is considered to be the gold standard of tissue sampling, especially if a defined small tissue area consisting of single or few cells within a heterogeneous tissue compartment is of interest. This sophisticated technique offers the opportunity of rapid and contamination-free tissue sampling for RNA- or DNA-based molecular genetic studies. We have applied laser microdissection to a molecular genetic study of pancreatic intraductal lesions (PanINs) in tissues of chronic pancreatitis, where an exact microdissection of small ducts within a dense fibrous tissue is of paramount importance for following analysis. From nine patients suffering from chronic pancreatitis, formalin-fixed, paraffin-embedded tissue specimens were laser microdissected, and a total of 202 normal ducts and PanINs of grade PanIN-1A to grade PanIN-2 were harvested. After whole genome amplification by improved primer extension and preamplification PCR (I-PEP-PCR), microsatellite-PCR based loss of heterozygosity analysis (LOH) of the tumor suppressor gene loci TP53, p16INK4, and DPC4 was performed. One of 85 informative duct lesions (1.2%) had LOH of TP53, 1 of 76 duct lesions (1.3%) had LOH of DPC4, and 2/29 duct lesions (6.9%) showed LOH of p16INK4. Microsatellite instability (MSI) was seen in 2 of 178 duct lesions (1.1%). Immunohistochemical staining of p53 protein and DPC4 protein revealed no aberrant expression. These preliminary data indicate that LOH of tumor suppressor genes, important in pancreatic cancer genesis or MSI, can be found in chronic pancreatitis tissues, but their incidence is low. PMID:12924436

  7. Immunoelectron microscopic localization of elastic tissue components in archival tissue samples.

    PubMed

    Fanning, J C; White, J F; Polewski, R; Cleary, E G

    1991-06-01

    Tissue samples that have been stored for many years, in different media and under a variety of conditions, have been examined by modern techniques of immunoelectron microscopy, using antibodies against elastic tissue components. A range of postembedding restorative procedures has been identified, which will allow reliable immunolocalization of antibodies against the elastic tissue component of such specimens. These methods have been applied successfully to autopsy-derived material, fixed in buffered formaldehyde, to archival material stored frozen at -70 or -20 degrees C, to specimens fixed for electron microscopy and stored for many years in buffer, and even to archival material from formaldehyde-fixed, paraffin-embedded blocks, reprocessed for electron microscopic examination. The successful restorative methods included pre-treatment of the sections with 6 M guanidine hydrochloride, or 1 M Tris/saline, each containing 100 mM dithiothreitol (a reducing agent) followed by alkylation with 220 mM iodoacetamide. The application of these techniques allowed reliable study of elastic tissue antibody distributions in archival tissues that could not be obtained again, as well as comparative studies with tissues processed many years previously.

  8. Field sampling of unfed adults of Amblyomma hebraeum Koch.

    PubMed

    Norval, R A; Yunker, C E; Butler, J F

    1987-08-01

    No methods have previously been available for the field sampling of unfed adults of Amblyomma hebraeum. We released 2000 unfed adults into a 5-m2 area in mopane woodland in southeastern Zimbabwe. The ticks sought shelter beneath the debris on the soil surface but emerged and became active in host seeking when stimulated. Effective stimuli were the odours of cattle and sheep and high concentrations of carbon dioxide from dry ice. The ticks did not respond to low concentrations of carbon dioxide, vibrations generated by stamping the ground with poles, or two volatile components of ox breath known to attract tsetse flies. The presence of humans in the release area also had little or no effect in stimulating the ticks. As adults of A. hebraeum are large and conspicuous they can easily be collected from the soil surface, either manually or using a vacuum device, after stimulation by cattle or the release of high concentrations of carbon dioxide.

  9. Imaging of Proteins in Tissue Samples Using Nanospray Desorption Electrospray Ionization Mass Spectrometry.

    PubMed

    Hsu, Cheng-Chih; Chou, Pi-Tai; Zare, Richard N

    2015-11-17

    Chemical maps of tissue samples provide important information on biological processes therein. Recently, advances in tissue imaging have been achieved using ambient ionization techniques, such as desorption electrospray ionization mass spectrometry (DESI-MS), but such techniques have been almost exclusively confined to the mapping of lipids and metabolites. We report here the use of nanospray desorption electrospray ionization (nanoDESI) that allows us to image proteins in tissue samples in a label-free manner at atmospheric pressure with only minimum sample preparation. Multiply charged proteins with masses up to 15 kDa were successfully detected by nanoDESI using an LTQ Orbitrap mass spectrometer. In an adult mice brain section, expression of proteins including ubiquitin, β-thymosin, myelin basic protein, and hemoglobin were spatially mapped and characterized. We also determined the location of methylation on myelin basic protein. This imaging modality was further implemented to MYC-induced lymphomas. We observed an array of truncated proteins in the region where normal thymus cells were infiltrated by tumor cells, in contrast to healthy tissue.

  10. Weirs: Counting and sampling adult salmonids in streams and rivers

    USGS Publications Warehouse

    Zimmerman, Christian E.; Zabkar, Laura M.; Johnson, David H.; Shrier, Brianna M.; O'Neal, Jennifer S.; Knutzen, John A.; Augerot, Xanthippe; O'Neal, Thomas A.; Pearsons, Todd N.

    2007-01-01

    generally regarded as the most accurate technique available to quantify escapement as the result is supposedly an absolute count (Cousens et al. 1982). Weirs also provide the opportunity to capture fish for observation and sampling of biological characteristics and tissues; they may also serve as recapture sites for basin-wide, mark–recapture population estimates. Temporary weirs are useful in monitoring wild populations of salmonids as well as for capturing broodstock for artificial propagation.

  11. Swine infectious agents in Tayassu pecari and Pecari tajacu tissue samples from Brazil.

    PubMed

    de Castro, Alessandra Marnie Martins Gomes; Brombila, Talita; Bersano, Josete Garcia; Soares, Herbert Sousa; Silva, Sheila Oliveira de Souza; Minervino, Antonio Humberto Hamad; Ogata, Renato Akio; Gennari, Solange Maria; Richtzenhain, Leonardo Jose

    2014-04-01

    Peccaries and pigs, Tayassuidae and Suidae respectively, diverged approximately one million years ago from a common ancestor. Because these families share some pathogens, peccaries can act as reservoirs of infectious pathogens for domestic and wild swine. We evaluated the presence of swine infectious agents in the spleen and lung tissues of white-lipped peccaries (WLP; Tayassu pecari) and collared peccaries (CP; Pecari tajacu) in Brazil. Samples from 10 adult CP and three WLP, which had been hunted by locals or hit by motor vehicles, were obtained from two free-ranging Brazilian populations. The samples were tested by PCR for Mycoplasma hyopneumoniae, Bordetella bronchiseptica, Pasteurella multocida, porcine circovirus 2 (PCV2), Suid herpesvirus 1 (SuHV-1), and porcine parvovirus (PPV). Positive samples were sequenced. Both species were negative for PPV and B. bronchiseptica and positive for PCV2 and SuHV-1. The lungs of two animals were positive for M. hyopneumoniae and P. multocida. This report is the first demonstration of PCV2 and SuHV-1 swine viruses and of M. hyopneumoniae and P. multocida bacteria in peccaries. One factor contributing to this detection was access to tissue samples, which is uncommon. The role of these infectious agents in peccaries is unknown and further epidemiologic studies should be performed. This study identified several infectious agents in peccaries and highlighted the importance of the tissue type used to detect pathogens.

  12. A Digital Gene Expression-Based Bovine Gene Atlas Evaluating 92 Adult, Juvenile and Fetal Cattle Tissues

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A comprehensive transcriptome survey, or “Gene Atlas,” provides information essential for a complete understanding of the genomic biology of an organism. Using a digital gene expression approach, we developed a Gene Atlas of RNA abundance in 92 adult, juvenile and fetal cattle tissues. The samples...

  13. Measurement of phthalates in small samples of mammalian tissue

    SciTech Connect

    Acott, P.D.; Murphy, M.G.; Ogborn, M.R.; Crocker, J.F.S.

    1987-03-01

    Di-(2-ethylhexyl)-phthalate (DEHP) is a phthalic acid ester that is used as a plasticizer in polyvinyl chloride products, many of which have widespread medical application. DEHP has been shown to be leached from products used for storage and delivery of blood transfusions during procedures such as plasmaphoresis, hemodialysis and open heart surgery. Results of studies in this laboratory have suggested that there is an association between the absorption and deposition of DEHP (and/or related chemicals) in the kidney and the acquired renal cystic disease (ACD) frequently seen in patients who have undergone prolonged dialysis treatment. In order to determine the relationship between the two, it has been necessary to establish a method for extracting and accurately quantitating minute amounts of these chemicals in small tissue samples. The authors have now established such a method using kidneys from normal rats and from a rat model for ACD.

  14. Quantitative laser-induced breakdown spectroscopy analysis of calcified tissue samples

    NASA Astrophysics Data System (ADS)

    Samek, O.; Beddows, D. C. S.; Telle, H. H.; Kaiser, J.; Liška, M.; Cáceres, J. O.; Gonzáles Ureña, A.

    2001-06-01

    We report on the application of laser-induced breakdown spectroscopy (LIBS) to the analysis of important minerals and the accumulation of potentially toxic elements in calcified tissue, to trace e.g. the influence of environmental exposure, and other medical or biological factors. This theme was exemplified for quantitative detection and mapping of Al, Pb and Sr in representative samples, including teeth (first teeth of infants, second teeth of children and teeth of adults) and bones (tibia and femur). In addition to identifying and quantifying major and trace elements in the tissues, one- and two-dimensional profiles and maps were generated. Such maps (a) provide time/concentration relations, (b) allow to follow mineralisation of the hydroxyapatite matrix and the migration of the elements within it and (c) enable to identify disease states, such as caries in teeth. In order to obtain quantitative calibration, reference samples in the form of pressed pellets with calcified tissue-equivalent material (majority compound of pellets is CaCO 3) were used whose physical properties closely resembled hydroxyapatite. Compounds of Al, Sr and Pb were added to the pellets, containing atomic concentrations in the range 100-10 000 ppm relative to the Ca content of the matrix. Analytical results based on this calibration against artificial samples for the trace elements under investigation agree with literature values, and with our atomic absorption spectroscopy (AAS) cross-validation measurements.

  15. Optimization of Evans blue quantitation in limited rat tissue samples

    NASA Astrophysics Data System (ADS)

    Wang, Hwai-Lee; Lai, Ted Weita

    2014-10-01

    Evans blue dye (EBD) is an inert tracer that measures plasma volume in human subjects and vascular permeability in animal models. Quantitation of EBD can be difficult when dye concentration in the sample is limited, such as when extravasated dye is measured in the blood-brain barrier (BBB) intact brain. The procedure described here used a very small volume (30 µl) per sample replicate, which enabled high-throughput measurements of the EBD concentration based on a standard 96-well plate reader. First, ethanol ensured a consistent optic path length in each well and substantially enhanced the sensitivity of EBD fluorescence spectroscopy. Second, trichloroacetic acid (TCA) removed false-positive EBD measurements as a result of biological solutes and partially extracted EBD into the supernatant. Moreover, a 1:2 volume ratio of 50% TCA ([TCA final] = 33.3%) optimally extracted EBD from the rat plasma protein-EBD complex in vitro and in vivo, and 1:2 and 1:3 weight-volume ratios of 50% TCA optimally extracted extravasated EBD from the rat brain and liver, respectively, in vivo. This procedure is particularly useful in the detection of EBD extravasation into the BBB-intact brain, but it can also be applied to detect dye extravasation into tissues where vascular permeability is less limiting.

  16. Adult human adipose tissue contains several types of multipotent cells.

    PubMed

    Tallone, Tiziano; Realini, Claudio; Böhmler, Andreas; Kornfeld, Christopher; Vassalli, Giuseppe; Moccetti, Tiziano; Bardelli, Silvana; Soldati, Gianni

    2011-04-01

    Multipotent mesenchymal stromal cells (MSCs) are a type of adult stem cells that can be easily isolated from various tissues and expanded in vitro. Many reports on their pluripotency and possible clinical applications have raised hopes and interest in MSCs. In an attempt to unify the terminology and the criteria to label a cell as MSC, in 2006 the International Society for Cellular Therapy (ISCT) proposed a standard set of rules to define the identity of these cells. However, MSCs are still extracted from different tissues, by diverse isolation protocols, are cultured and expanded in different media and conditions. All these variables may have profound effects on the selection of cell types and the composition of heterogeneous subpopulations, on the selective expansion of specific cell populations with totally different potentials and ergo, on the long-term fate of the cells upon in vitro culture. Therefore, specific molecular and cellular markers that identify MSCs subsets as well as standardization of expansion protocols for these cells are urgently needed. Here, we briefly discuss new useful markers and recent data supporting the rapidly emerging concept that many different types of progenitor cells are found in close association with blood vessels. This knowledge may promote the necessary technical improvements required to reduce variability and promote higher efficacy and safety when isolating and expanding these cells for therapeutic use. In the light of the discussed data, particularly the identification of new markers, and advances in the understanding of fundamental MSC biology, we also suggest a revision of the 2006 ISCT criteria.

  17. Long-term room temperature preservation of corpse soft tissue: an approach for tissue sample storage

    PubMed Central

    2011-01-01

    Background Disaster victim identification (DVI) represents one of the most difficult challenges in forensic sciences, and subsequent DNA typing is essential. Collected samples for DNA-based human identification are usually stored at low temperature to halt the degradation processes of human remains. We have developed a simple and reliable procedure for soft tissue storage and preservation for DNA extraction. It ensures high quality DNA suitable for PCR-based DNA typing after at least 1 year of room temperature storage. Methods Fragments of human psoas muscle were exposed to three different environmental conditions for diverse time periods at room temperature. Storage conditions included: (a) a preserving medium consisting of solid sodium chloride (salt), (b) no additional substances and (c) garden soil. DNA was extracted with proteinase K/SDS followed by organic solvent treatment and concentration by centrifugal filter devices. Quantification was carried out by real-time PCR using commercial kits. Short tandem repeat (STR) typing profiles were analysed with 'expert software'. Results DNA quantities recovered from samples stored in salt were similar up to the complete storage time and underscored the effectiveness of the preservation method. It was possible to reliably and accurately type different genetic systems including autosomal STRs and mitochondrial and Y-chromosome haplogroups. Autosomal STR typing quality was evaluated by expert software, denoting high quality profiles from DNA samples obtained from corpse tissue stored in salt for up to 365 days. Conclusions The procedure proposed herein is a cost efficient alternative for storage of human remains in challenging environmental areas, such as mass disaster locations, mass graves and exhumations. This technique should be considered as an additional method for sample storage when preservation of DNA integrity is required for PCR-based DNA typing. PMID:21846338

  18. Gene expression analysis distinguishes tissue-specific and gender-related functions among adult Ascaris suum tissues.

    PubMed

    Wang, Zhengyuan; Gao, Xin; Martin, John; Yin, Yong; Abubucker, Sahar; Rash, Amy C; Li, Ben-Wen; Nash, Bill; Hallsworth-Pepin, Kym; Jasmer, Douglas P; Mitreva, Makedonka

    2013-06-01

    Over a billion people are infected by Ascaris spp. intestinal parasites. To clarify functional differences among tissues of adult A. suum, we compared gene expression by various tissues of these worms by expression microarray methods. The A. suum genome was sequenced and assembled to allow generation of microarray elements. Expression of over 40,000 60-mer elements was investigated in a variety of tissues from both male and female adult worms. Nearly 50 percent of the elements for which signal was detected exhibited differential expression among different tissues. The unique profile of transcripts identified for each tissue clarified functional distinctions among tissues, such as chitin binding in the ovary and peptidase activity in the intestines. Interestingly, hundreds of gender-specific elements were characterized in multiple non-reproductive tissues of female or male worms, with most prominence of gender differences in intestinal tissue. A. suum genes from the same family were frequently expressed differently among tissues. Transcript abundance for genes specific to A. suum, by comparison to Caenorhabditis elegans, varied to a greater extent among tissues than for genes conserved between A. suum and C. elegans. Analysis using C. elegans protein interaction data identified functional modules conserved between these two nematodes, resulting in identification of functional predictions of essential subnetworks of protein interactions and how these networks may vary among nematode tissues. A notable finding was very high module similarity between adult reproductive tissues and intestine. Our results provide the most comprehensive assessment of gene expression among tissues of a parasitic nematode to date. PMID:23572074

  19. Variation in glycogen concentrations within mantle and foot tissue in Amblema plicata plicata: Implications for tissue biopsy sampling

    USGS Publications Warehouse

    Naimo, T.J.; Monroe, E.M.

    1999-01-01

    With the development of techniques to non-lethally biopsy tissue from unionids, a new method is available to measure changes in biochemical, contaminant, and genetic constituents in this imperiled faunal group. However, before its widespread application, information on the variability of biochemical components within and among tissues needs to be evaluated. We measured glycogen concentrations in foot and mantle tissue in Amblema plicata plicata (Say, 1817) to determine if glycogen was evenly distributed within and between tissues and to determine which tissue might be more responsive to the stress associated with relocating mussels. Glycogen was measured in two groups of mussels: those sampled from their native environment (undisturbed mussels) and quickly frozen for analysis and those relocated into an artificial pond (relocated mussels) for 24 months before analysis. In both undisturbed and relocated mussels, glycogen concentrations were evenly distributed within foot, but not within mantle tissue. In mantle tissue, concentrations of glycogen varied about 2-fold among sections. In addition, glycogen varied significantly between tissues in undisturbed mussels, but not in relocated mussels. Twenty-four months after relocation, glycogen concentrations had declined by 80% in mantle tissue and by 56% in foot tissue relative to the undisturbed mussels. These data indicate that representative biopsy samples can be obtained from foot tissue, but not mantle tissue. We hypothesize that mantle tissue could be more responsive to the stress of relocation due to its high metabolic activity associated with shell formation.

  20. Persistent synthetic chlorinated hydrocarbons in albatross tissue samples from Midway Atoll

    SciTech Connect

    Jones, P.D.; Hannah, D.J.; Buckland, S.J.

    1996-10-01

    Anthropogenic organic contaminants have been found in even the most remote locations. To assess the global distribution and possible effects of such contaminants, the authors examined the tissues of two species of albatross collected from Midway Atoll in the central North Pacific Ocean. These birds have an extensive feeding range covering much of the subtropical and northern Pacific Ocean. Anthropogenic contaminants were found at relatively great concentrations in these birds. The sum of 19 polychlorinated biphenyl (PCB) congeners ranged from 177 ng/g wet weight in eggs to 2,750 ng/g wet weight in adult fat. Total toxic equivalents (TEQs) derived from polychlorinated dibenzo-p-dioxins (PCDDs) and dibenzofurans (PCDFs) ranged from 17.2 to 297 pg/g wet weight in the same tissues, while the inclusion of TEQs from PCBs increased these values to 48.4 and 769 pg/g wet weight, respectively. While contaminant concentrations varied between species and tissues, the contaminant profile was relatively uniform. The profile of contaminants detected was unusual in that much of the TEQs was contributed by two pentachlorinated congeners (2,3,4,7,8-pentachlorinated dibenzo-p-dioxin), and the profiles of PCB congeners did not match known sources. When compared to other studies the concentrations detected in the Midway Atoll samples were near or above the thresholds known to cause adverse effects in other fish-eating bird species.

  1. Defining Adult Experiences: Perspectives of a Diverse Sample of Young Adults.

    PubMed

    Lowe, Sarah R; Dillon, Colleen O; Rhodes, Jean E; Zwiebach, Liza

    2013-01-01

    This study explored the roles and psychological experiences identified as defining adult moments using mixed methods with a racially, ethnically, and socioeconomically diverse sample of young adults both enrolled and not enrolled in college (N = 726; ages 18-35). First, we evaluated results from a single survey item that asked participants to rate how adult they feel. Consistent with previous research, the majority of participants (56.9%) reported feeling "somewhat like an adult," and older participants had significantly higher subjective adulthood, controlling for other demographic variables. Next, we analyzed responses from an open-ended question asking participants to describe instances in which they felt like an adult. Responses covered both traditional roles (e.g., marriage, childbearing; 36.1%) and nontraditional social roles and experiences (e.g., moving out of parent's home, cohabitation; 55.6%). Although we found no differences by age and college status in the likelihood of citing a traditional or nontraditional role, participants who had achieved more traditional roles were more likely to cite them in their responses. In addition, responses were coded for psychological experiences, including responsibility for self (19.0%), responsibility for others (15.3%), self-regulation (31.1%), and reflected appraisals (5.1%). Older participants were significantly more likely to include self-regulation and reflected appraisals, whereas younger participants were more likely to include responsibility for self. College students were more likely than noncollege students to include self-regulation and reflected appraisals. Implications for research and practice are discussed. PMID:23554545

  2. Defining Adult Experiences: Perspectives of a Diverse Sample of Young Adults

    PubMed Central

    Lowe, Sarah R.; Dillon, Colleen O.; Rhodes, Jean E.; Zwiebach, Liza

    2013-01-01

    This study explored the roles and psychological experiences identified as defining adult moments using mixed methods with a racially, ethnically, and socioeconomically diverse sample of young adults both enrolled and not enrolled in college (N = 726; ages 18-35). First, we evaluated results from a single survey item that asked participants to rate how adult they feel. Consistent with previous research, the majority of participants (56.9%) reported feeling “somewhat like an adult,” and older participants had significantly higher subjective adulthood, controlling for other demographic variables. Next, we analyzed responses from an open-ended question asking participants to describe instances in which they felt like an adult. Responses covered both traditional roles (e.g., marriage, childbearing; 36.1%) and nontraditional social roles and experiences (e.g., moving out of parent’s home, cohabitation; 55.6%). Although we found no differences by age and college status in the likelihood of citing a traditional or nontraditional role, participants who had achieved more traditional roles were more likely to cite them in their responses. In addition, responses were coded for psychological experiences, including responsibility for self (19.0%), responsibility for others (15.3%), self-regulation (31.1%), and reflected appraisals (5.1%). Older participants were significantly more likely to include self-regulation and reflected appraisals, whereas younger participants were more likely to include responsibility for self. College students were more likely than noncollege students to include self-regulation and reflected appraisals. Implications for research and practice are discussed. PMID:23554545

  3. Automated Classification of Glandular Tissue by Statistical Proximity Sampling

    PubMed Central

    Azar, Jimmy C.; Simonsson, Martin

    2015-01-01

    Due to the complexity of biological tissue and variations in staining procedures, features that are based on the explicit extraction of properties from subglandular structures in tissue images may have difficulty generalizing well over an unrestricted set of images and staining variations. We circumvent this problem by an implicit representation that is both robust and highly descriptive, especially when combined with a multiple instance learning approach to image classification. The new feature method is able to describe tissue architecture based on glandular structure. It is based on statistically representing the relative distribution of tissue components around lumen regions, while preserving spatial and quantitative information, as a basis for diagnosing and analyzing different areas within an image. We demonstrate the efficacy of the method in extracting discriminative features for obtaining high classification rates for tubular formation in both healthy and cancerous tissue, which is an important component in Gleason and tubule-based Elston grading. The proposed method may be used for glandular classification, also in other tissue types, in addition to general applicability as a region-based feature descriptor in image analysis where the image represents a bag with a certain label (or grade) and the region-based feature vectors represent instances. PMID:25685143

  4. Studying Genes in Tissue Samples From Younger and Adolescent Patients With Soft Tissue Sarcomas

    ClinicalTrials.gov

    2016-05-13

    Childhood Alveolar Soft-part Sarcoma; Childhood Angiosarcoma; Childhood Desmoplastic Small Round Cell Tumor; Childhood Epithelioid Sarcoma; Childhood Fibrosarcoma; Childhood Leiomyosarcoma; Childhood Liposarcoma; Childhood Malignant Mesenchymoma; Childhood Neurofibrosarcoma; Childhood Synovial Sarcoma; Chordoma; Desmoid Tumor; Metastatic Childhood Soft Tissue Sarcoma; Nonmetastatic Childhood Soft Tissue Sarcoma; Recurrent Childhood Soft Tissue Sarcoma

  5. The Resilience Scale for Adults: Construct Validity and Measurement in a Belgian Sample

    ERIC Educational Resources Information Center

    Hjemdal, Odin; Friborg, Oddgeir; Braun, Stephanie; Kempenaers, Chantal; Linkowski, Paul; Fossion, Pierre

    2011-01-01

    The Resilience Scale for Adults (RSA) was developed and has been extensively validated in Norwegian samples. The purpose of this study was to explore the construct validity of the Resilience Scale for Adults in a French-speaking Belgian sample and test measurement invariance between the Belgian and a Norwegian sample. A Belgian student sample (N =…

  6. Robust and artifact-free mounting of tissue samples for atomic force microscopy.

    PubMed

    Morgan, Joshua T; Raghunathan, Vijay Krishna; Thomasy, Sara M; Murphy, Christopher J; Russell, Paul

    2014-01-01

    Immobilization of tissue-samples for atomic for microscopy (AFM) is typically done using either semi-dry tissue or by gluing the tissue sample down, both of which can introduce artifacts. Here, we describe the design of a Soft- Clamping Immobilizing Retainer of Tissue (SCIRT) for consistent and nondestructive immobilization of tissues for AFM analysis. We compare the performance of our SCIRT method with glue-immobilization for two difficult to handle tissue types: human trabecular meshwork (HTM) and rabbit cornea (RC). Our results demonstrate that the SCIRT method has several advantages, including: (i) allowing for small sample sizes, (ii) enabling continuous hydration, (iii) eliminating contact with glue or associated solvents, (iv) permitting sample recovery following measurement, and (v) ease of use. In conclusion, the SCIRT method is a simple and effective means of immobilizing soft, hydrated tissue samples consistently and without artifacts.

  7. Robust and artifact-free mounting of tissue samples for atomic force microscopy

    PubMed Central

    Morgan, Joshua T.; Raghunathan, Vijay Krishna; Thomasy, Sara M.; Murphy, Christopher J.; Russell, Paul

    2015-01-01

    Immobilization of tissue-samples for atomic for microscopy (AFM) is typically done using either semi-dry tissue or by gluing the tissue sample down, both of which can introduce artifacts. Here, we describe the design of a Soft-Clamping Immobilizing Retainer of Tissue (SCIRT) for consistent and nondestructive immobilization of tissues for AFM analysis. We compare the performance of our SCIRT method with glue-immobilization for two difficult to handle tissue types: human trabecular meshwork (HTM) and rabbit cornea (RC). Our results demonstrate that the SCIRT method has several advantages, including: (i) allowing for small sample sizes, (ii) enabling continuous hydration, (iii) eliminating contact with glue or associated solvents, (iv) permitting sample recovery following measurement, and (v) ease of use. In conclusion, the SCIRT method is a simple and effective means of immobilizing soft, hydrated tissue samples consistently and without artifacts. PMID:24447138

  8. Functional relationships among selenium concentrations in the diet, target tissues, and nondestructive tissue samples of two species of snakes.

    PubMed

    Hopkins, William A; Snodgrass, Joel W; Baionno, Jennifer A; Roe, John H; Staub, Brandon P; Jackson, Brian P

    2005-02-01

    Nondestructive sampling methods, such as removal of feathers for contaminant analysis, are desirable in ecological monitoring programs that seek to minimize the impacts of harvesting organisms. Although many reptiles are declining worldwide, nondestructive sampling techniques seldom have been employed for assessing contaminant exposure in these organisms. In this study, we examined the utility of nondestructive tissue sampling for assessing Se exposure in reptiles. We describe the functional relationships among dietary Se concentrations, target tissue Se concentrations, and Se concentrations in nondestructive tissue samples (blood and tail tissue biopsy) in two species of snakes that had been exposed to Se under very different experimental protocols. Using nonlinear regression, we found strong positive correlations (r2 > 0.92) in all comparisons among Se concentrations in nondestructive tissues, diet, and target tissues. Moreover, equations describing these relationships can be used to estimate concentrations of Se in diet and target organs, from known concentrations of Se in nondestructive tissue samples. Although the current paucity of toxicity data on reptiles precludes tests of our models, we demonstrate how the equations describing these relationships might be used to make predictions about Se accumulation in target organs for risk assessment. Future studies on reptiles that examine these relationships under different Se exposure conditions, and those that document physiological responses of reptiles to various concentrations of Se, will help to refine our models and test their efficacy for predicting health risk.

  9. Histological and Transcriptomic Analysis of Adult Japanese Medaka Sampled Onboard the International Space Station

    PubMed Central

    Murata, Yasuhiko; Yasuda, Takako; Watanabe-Asaka, Tomomi; Oda, Shoji; Mantoku, Akiko; Takeyama, Kazuhiro; Chatani, Masahiro; Kudo, Akira; Uchida, Satoko; Suzuki, Hiromi; Tanigaki, Fumiaki; Shirakawa, Masaki; Fujisawa, Koichi; Hamamoto, Yoshihiko; Terai, Shuji; Mitani, Hiroshi

    2015-01-01

    To understand how humans adapt to the space environment, many experiments can be conducted on astronauts as they work aboard the Space Shuttle or the International Space Station (ISS). We also need animal experiments that can apply to human models and help prevent or solve the health issues we face in space travel. The Japanese medaka (Oryzias latipes) is a suitable model fish for studying space adaptation as evidenced by adults of the species having mated successfully in space during 15 days of flight during the second International Microgravity Laboratory mission in 1994. The eggs laid by the fish developed normally and hatched as juveniles in space. In 2012, another space experiment (“Medaka Osteoclast”) was conducted. Six-week-old male and female Japanese medaka (Cab strain osteoblast transgenic fish) were maintained in the Aquatic Habitat system for two months in the ISS. Fish of the same strain and age were used as the ground controls. Six fish were fixed with paraformaldehyde or kept in RNA stabilization reagent (n = 4) and dissected for tissue sampling after being returned to the ground, so that several principal investigators working on the project could share samples. Histology indicated no significant changes except in the ovary. However, the RNA-seq analysis of 5345 genes from six tissues revealed highly tissue-specific space responsiveness after a two-month stay in the ISS. Similar responsiveness was observed among the brain and eye, ovary and testis, and the liver and intestine. Among these six tissues, the intestine showed the highest space response with 10 genes categorized as oxidation–reduction processes (gene ontogeny term GO:0055114), and the expression levels of choriogenin precursor genes were suppressed in the ovary. Eleven genes including klf9, klf13, odc1, hsp70 and hif3a were upregulated in more than four of the tissues examined, thus suggesting common immunoregulatory and stress responses during space adaptation. PMID:26427061

  10. Histological and Transcriptomic Analysis of Adult Japanese Medaka Sampled Onboard the International Space Station.

    PubMed

    Murata, Yasuhiko; Yasuda, Takako; Watanabe-Asaka, Tomomi; Oda, Shoji; Mantoku, Akiko; Takeyama, Kazuhiro; Chatani, Masahiro; Kudo, Akira; Uchida, Satoko; Suzuki, Hiromi; Tanigaki, Fumiaki; Shirakawa, Masaki; Fujisawa, Koichi; Hamamoto, Yoshihiko; Terai, Shuji; Mitani, Hiroshi

    2015-01-01

    To understand how humans adapt to the space environment, many experiments can be conducted on astronauts as they work aboard the Space Shuttle or the International Space Station (ISS). We also need animal experiments that can apply to human models and help prevent or solve the health issues we face in space travel. The Japanese medaka (Oryzias latipes) is a suitable model fish for studying space adaptation as evidenced by adults of the species having mated successfully in space during 15 days of flight during the second International Microgravity Laboratory mission in 1994. The eggs laid by the fish developed normally and hatched as juveniles in space. In 2012, another space experiment ("Medaka Osteoclast") was conducted. Six-week-old male and female Japanese medaka (Cab strain osteoblast transgenic fish) were maintained in the Aquatic Habitat system for two months in the ISS. Fish of the same strain and age were used as the ground controls. Six fish were fixed with paraformaldehyde or kept in RNA stabilization reagent (n = 4) and dissected for tissue sampling after being returned to the ground, so that several principal investigators working on the project could share samples. Histology indicated no significant changes except in the ovary. However, the RNA-seq analysis of 5345 genes from six tissues revealed highly tissue-specific space responsiveness after a two-month stay in the ISS. Similar responsiveness was observed among the brain and eye, ovary and testis, and the liver and intestine. Among these six tissues, the intestine showed the highest space response with 10 genes categorized as oxidation-reduction processes (gene ontogeny term GO:0055114), and the expression levels of choriogenin precursor genes were suppressed in the ovary. Eleven genes including klf9, klf13, odc1, hsp70 and hif3a were upregulated in more than four of the tissues examined, thus suggesting common immunoregulatory and stress responses during space adaptation.

  11. Histological and Transcriptomic Analysis of Adult Japanese Medaka Sampled Onboard the International Space Station.

    PubMed

    Murata, Yasuhiko; Yasuda, Takako; Watanabe-Asaka, Tomomi; Oda, Shoji; Mantoku, Akiko; Takeyama, Kazuhiro; Chatani, Masahiro; Kudo, Akira; Uchida, Satoko; Suzuki, Hiromi; Tanigaki, Fumiaki; Shirakawa, Masaki; Fujisawa, Koichi; Hamamoto, Yoshihiko; Terai, Shuji; Mitani, Hiroshi

    2015-01-01

    To understand how humans adapt to the space environment, many experiments can be conducted on astronauts as they work aboard the Space Shuttle or the International Space Station (ISS). We also need animal experiments that can apply to human models and help prevent or solve the health issues we face in space travel. The Japanese medaka (Oryzias latipes) is a suitable model fish for studying space adaptation as evidenced by adults of the species having mated successfully in space during 15 days of flight during the second International Microgravity Laboratory mission in 1994. The eggs laid by the fish developed normally and hatched as juveniles in space. In 2012, another space experiment ("Medaka Osteoclast") was conducted. Six-week-old male and female Japanese medaka (Cab strain osteoblast transgenic fish) were maintained in the Aquatic Habitat system for two months in the ISS. Fish of the same strain and age were used as the ground controls. Six fish were fixed with paraformaldehyde or kept in RNA stabilization reagent (n = 4) and dissected for tissue sampling after being returned to the ground, so that several principal investigators working on the project could share samples. Histology indicated no significant changes except in the ovary. However, the RNA-seq analysis of 5345 genes from six tissues revealed highly tissue-specific space responsiveness after a two-month stay in the ISS. Similar responsiveness was observed among the brain and eye, ovary and testis, and the liver and intestine. Among these six tissues, the intestine showed the highest space response with 10 genes categorized as oxidation-reduction processes (gene ontogeny term GO:0055114), and the expression levels of choriogenin precursor genes were suppressed in the ovary. Eleven genes including klf9, klf13, odc1, hsp70 and hif3a were upregulated in more than four of the tissues examined, thus suggesting common immunoregulatory and stress responses during space adaptation. PMID:26427061

  12. [PREVALENCE OF NON-CARIOUS CERVICAL LESIONS AND ABFRACTIONS OF DENTAL HARD TISSUES IN AN ADULT IN DIFFERENT AGES].

    PubMed

    Iordanishvili, A K; Chernyj, D A; Jankovskij, V V; Orlov, A K; Drobkova, K O

    2015-01-01

    The article is devoted to gerontostomatological and gender-specific prevalence of non-carious lesions of the hard tissue of teeth in adults. The paper presents data of epidemiological study on prevalence of non-carious lesions of dental hard tissues (high abrasion, erosion, wedge-shaped defects, hyperesthesia). Allocated to four age groups: young adults surveyed--from 22 to 39 years; middle ages--from 40 to 59 years; older--from 60 to 74 years of age; senile age--from 75 to 87 years. To determine the frequency of occurrence of different forms of non-carious lesions of the hard tissue of teeth we have used the following: general scientific and special methods: poll, dental examination, groupings, statistical and mathematical methods of processing sample. We have ranked low incidence of non-carious lesions of the hard tissue of the teeth in the sample surveyed: high abrasion, erosion, wedge-shaped defects of solid tissues, hyperesthesia. The features of clinical course of non-carious lesions have been determined. In particular a rare combined lesion of the teeth with advanced erasibility, wedge defects and erosion has been noted. Significant combination of the pathological processes of the hard tissue of teeth with their hyperesthesia has been found. Features of different forms of non-carious lesions of the hard tissue of teeth in different age periods of life have been determined. Noted that older people, due to non-carious esions of the hard tissue of teeth were more likely to require medical intervention aimed at addressing the ncreased sensitivity and loss of hard tissue of teeth by dental therapeutic activities or dental prosthetics. PMID:26856106

  13. Ethical use of tissue samples in genetic research.

    PubMed

    Azarow, Kenneth S; Olmstead, Francis L; Hume, Roderick F; Myers, Jerome; Calhoun, Bryon C; Martin, Laura S

    2003-06-01

    Many centrally based cancer protocols have begun to address the ethical issues concerning tissue banking for genetic research. A multidisciplinary subcommittee of the Madigan Army Medical Center Institutional Review Board was established to determine the scope of the problem and offer a concise, user-friendly policy with guidelines on how to control and monitor the use of stored tissue for future genetic and molecular research. Our institution participates in 69 Southern Oncology Group or National Surgical Adjuvant Breast and Bowel Project protocols and 47 Children's Oncology Group protocols. Of these protocols, 22 of 69 and 36 of 47, respectively, asked for tissue to be stored for future biologic study. Only 4 of 69 and 3 of 47, respectively, deal with specific consent for future genetic/biologic research. The multidisciplinary committee developed a policy that dealt with the following areas: exempt status, waived consent, informed consent, deceased status, family studies, and information flow. An algorithm was created to establish a system of checks and balances concerning privacy, protection and an appeals process.

  14. Adults' Views on Mathematics Education: A Midwest Sample

    ERIC Educational Resources Information Center

    Brez, Caitlin C.; Allen, Jessica J.

    2016-01-01

    Currently, few studies have addressed public opinions regarding math education. The current study surveyed adults in a Midwestern town in the United States to assess opinions regarding math and math education. Overall, we found that adults believe that math is useful and that math education is important. We found that parents who currently have a…

  15. Concise Review: Quiescence in Adult Stem Cells: Biological Significance and Relevance to Tissue Regeneration.

    PubMed

    Rumman, Mohammad; Dhawan, Jyotsna; Kassem, Moustapha

    2015-10-01

    Adult stem cells (ASCs) are tissue resident stem cells responsible for tissue homeostasis and regeneration following injury. In uninjured tissues, ASCs exist in a nonproliferating, reversibly cell cycle-arrested state known as quiescence or G0. A key function of the quiescent state is to preserve stemness in ASCs by preventing precocious differentiation, and thus maintaining a pool of undifferentiated ASCs. Recent evidences suggest that quiescence is an actively maintained state and that excessive or defective quiescence may lead to compromised tissue regeneration or tumorigenesis. The aim of this review is to provide an update regarding the biological mechanisms of ASC quiescence and their role in tissue regeneration.

  16. Final LDRD report : development of sample preparation methods for ChIPMA-based imaging mass spectrometry of tissue samples.

    SciTech Connect

    Maharrey, Sean P.; Highley, Aaron M.; Behrens, Richard, Jr.; Wiese-Smith, Deneille

    2007-12-01

    The objective of this short-term LDRD project was to acquire the tools needed to use our chemical imaging precision mass analyzer (ChIPMA) instrument to analyze tissue samples. This effort was an outgrowth of discussions with oncologists on the need to find the cellular origin of signals in mass spectra of serum samples, which provide biomarkers for ovarian cancer. The ultimate goal would be to collect chemical images of biopsy samples allowing the chemical images of diseased and nondiseased sections of a sample to be compared. The equipment needed to prepare tissue samples have been acquired and built. This equipment includes an cyro-ultramicrotome for preparing thin sections of samples and a coating unit. The coating unit uses an electrospray system to deposit small droplets of a UV-photo absorbing compound on the surface of the tissue samples. Both units are operational. The tissue sample must be coated with the organic compound to enable matrix assisted laser desorption/ionization (MALDI) and matrix enhanced secondary ion mass spectrometry (ME-SIMS) measurements with the ChIPMA instrument Initial plans to test the sample preparation using human tissue samples required development of administrative procedures beyond the scope of this LDRD. Hence, it was decided to make two types of measurements: (1) Testing the spatial resolution of ME-SIMS by preparing a substrate coated with a mixture of an organic matrix and a bio standard and etching a defined pattern in the coating using a liquid metal ion beam, and (2) preparing and imaging C. elegans worms. Difficulties arose in sectioning the C. elegans for analysis and funds and time to overcome these difficulties were not available in this project. The facilities are now available for preparing biological samples for analysis with the ChIPMA instrument. Some further investment of time and resources in sample preparation should make this a useful tool for chemical imaging applications.

  17. Facial soft tissue depths in craniofacial identification (part II): An analytical review of the published sub-adult data.

    PubMed

    Stephan, Carl N; Simpson, Ellie K

    2008-11-01

    Prior research indicates that while statistically significant differences exist between subcategories of the adult soft tissue depth data, magnitudes of difference are small and possess little practical meaning when measurement errors and variations between measurement methods are considered. These findings raise questions as to what variables may or may not hold meaning for the sub-adult data. Of primary interest is the effect of age, as these differences have the potential to surpass the magnitude of measurement error. Data from the five studies in the literature on sub-adults which describe values for single integer age groups were pooled and differences across the ages examined. From 1 to 18 years, most soft tissue depth measurements increased by less than 3 mm. These results suggest that dividing the data for children into more than two age groups is unlikely to hold many advantages. Data were therefore split into two groups with the division point corresponding to the mid-point of the observed trends and main data density (0-11 and 12-18 years; division point = 11.5 years). Published sub-adult data for seven further studies which reported broader age groups were pooled with the data above to produce the final tallied soft tissue depth tables. These tables hold the advantages of increased sample sizes (pogonion has greater than 1770 individuals for either age group) and increased levels of certainty (as random and opposing systematic errors specific to each independent study should average out when the data are combined).

  18. Micro-organisms isolated from cadaveric samples of allograft musculoskeletal tissue.

    PubMed

    Varettas, Kerry

    2013-12-01

    Allograft musculoskeletal tissue is commonly used in orthopaedic surgical procedures. Cadaveric donors of musculoskeletal tissue supply multiple allografts such as tendons, ligaments and bone. The microbiology laboratory of the South Eastern Area Laboratory Services (SEALS, Australia) has cultured cadaveric allograft musculoskeletal tissue samples for bacterial and fungal isolates since 2006. This study will retrospectively review the micro-organisms isolated over a 6-year period, 2006-2011. Swab and tissue samples were received for bioburden testing and were inoculated onto agar and/or broth culture media. Growth was obtained from 25.1 % of cadaveric allograft musculoskeletal tissue samples received. The predominant organisms isolated were coagulase-negative staphylococci and coliforms, with the heaviest bioburden recovered from the hemipelvis. The rate of bacterial and fungal isolates from cadaveric allograft musculoskeletal tissue samples is higher than that from living donors. The type of organism isolated may influence the suitability of the allograft for transplant.

  19. Resilience to Adult Psychopathology Following Childhood Maltreatment: Evidence from a Community Sample

    ERIC Educational Resources Information Center

    Collishaw, Stephan; Pickles, Andrew; Messer, Julie; Rutter, Michael; Shearer, Christina; Maughan, Barbara

    2007-01-01

    Objective: Child abuse is an important risk for adult psychiatric morbidity. However, not all maltreated children experience mental health problems as adults. The aims of the present study were to address the extent of resilience to adult psychopathology in a representative community sample, and to explore predictors of a good prognosis. Methods:…

  20. Elemental distribution and sample integrity comparison of freeze-dried and frozen-hydrated biological tissue samples with nuclear microprobe

    NASA Astrophysics Data System (ADS)

    Vavpetič, P.; Vogel-Mikuš, K.; Jeromel, L.; Ogrinc Potočnik, N.; Pongrac, P.; Drobne, D.; Pipan Tkalec, Ž.; Novak, S.; Kos, M.; Koren, Š.; Regvar, M.; Pelicon, P.

    2015-04-01

    The analysis of biological samples in frozen-hydrated state with micro-PIXE technique at Jožef Stefan Institute (JSI) nuclear microprobe has matured to a point that enables us to measure and examine frozen tissue samples routinely as a standard research method. Cryotome-cut slice of frozen-hydrated biological sample is mounted between two thin foils and positioned on the sample holder. The temperature of the cold stage in the measuring chamber is kept below 130 K throughout the insertion of the samples and the proton beam exposure. Matrix composition of frozen-hydrated tissue is consisted mostly of ice. Sample deterioration during proton beam exposure is monitored during the experiment, as both Elastic Backscattering Spectrometry (EBS) and Scanning Transmission Ion Microscopy (STIM) in on-off axis geometry are recorded together with the events in two PIXE detectors and backscattered ions from the chopper in a single list-mode file. The aim of this experiment was to determine differences and similarities between two kinds of biological sample preparation techniques for micro-PIXE analysis, namely freeze-drying and frozen-hydrated sample preparation in order to evaluate the improvements in the elemental localisation of the latter technique if any. In the presented work, a standard micro-PIXE configuration for tissue mapping at JSI was used with five detection systems operating in parallel, with proton beam cross section of 1.0 × 1.0 μm2 and a beam current of 100 pA. The comparison of the resulting elemental distributions measured at the biological tissue prepared in the frozen-hydrated and in the freeze-dried state revealed differences in elemental distribution of particular elements at the cellular level due to the morphology alteration in particular tissue compartments induced either by water removal in the lyophilisation process or by unsatisfactory preparation of samples for cutting and mounting during the shock-freezing phase of sample preparation.

  1. Monitoring the marine environment using marine mammal tissue samples

    SciTech Connect

    Jones, P.D.; Hannah, D.J.; Day, P.J.

    1995-12-31

    Marine environments, both inshore and open ocean, receive numerous inputs of anthropogenic chemicals. Cetaceans provide a valuable resource for monitoring the low level contamination of marine environments with persistent organic contaminants. Comparative studies using inshore and offshore southern ocean cetaceans have revealed significant differences in the types of contamination in these two environments. The polychlorinated biphenyls (PCBs) deposited in the southern oceans are characterized by an abundance of lower chlorinated congeners. Polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/F) are not present at significant concentrations in cetaceans from the open southern ocean. In contrast significant concentrations of PCDD/F congeners are detected in the blubber of the inshore living Hector`s dolphin. This species lives close to the shore and has a very small home range (approximately 30 km) for a cetacean. Analysis of tissue PCDD/F and PCB profiles from different populations and their food sources will be presented. The data are being used to determine if there are local variations in the contamination of the New Zealand inshore marine environment.

  2. Quantitation of two endogenous lactose-inhibitable lectins in embryonic and adult chicken tissues

    SciTech Connect

    Beyer, E.C.; Barondes, S.H.

    1982-01-01

    Two lactose-binding lectins from chicken tissues, chicken-lactose-lectin-I (CLL-I) and chicken-lactose-lectin-II (CLL-II) were quantified with a radioimmunoassay in extracts of a number of developing and adult chicken tissues. Both lectins could be measured in the same extract without separation, because they showed no significant immunological cross- reactivity. Many embryonic and adult tissues, including brain, heart, intestine, kidney, liver, lung, muscle, pancreas, and spleen, contained one or both lectins, although their concentrations differed markedly. For example, embryonic muscle, the richest source of CLL-I contained only traces of CLL-II whereas embryonic kidney, a very rich source of CLL-II contained substantial CLL-I. In both muscle and kidney, lectin levels in adulthood were much lower than in the embryonic state. In contrast, CLL-I in liver and CLL-II in intestine were 10-fold to 30-fold more concentrated in the adult than in the 15-d embryo. CLL-I and CLL-II from several tissues were purified by affinity chromatography and their identity in the various tissues was confirmed by polyacrylamide gel electrophoresis, isoelectric focusing, and peptide mapping. The results suggest that these lectins might have different functions in the many developing and adult tissues in which they are found.

  3. Adult-Child Incest: A Sample of Substantiated Cases.

    ERIC Educational Resources Information Center

    Vander Mey, Brenda J.; Neff, Ronald L.

    1984-01-01

    Describes adult-child incest involving 16 victims. Cases were analyzed and discussed in relationship to previous studies. Emphasis is placed on replicated and newly identified risk factors including alcohol dependency, spouse abuse, other (nonsexual) child abuse or neglect, inadequate housing, social isolation, birth order, and other family…

  4. Screening for ADHD in an Adult Social Phobia Sample

    ERIC Educational Resources Information Center

    Mortberg, Ewa; Tilfors, Kerstin; Bejerot, Susanne

    2012-01-01

    Objective: Recent studies have suggested a link between a primary anxiety disorder and ADHD. Method: A total of 39 participants with a primary diagnosis of social phobia were compared with 178 patients with ADHD and 88 patients with other psychiatric disorders on measures for childhood and adult ADHD (the Wender Utah Rating Scale and the Adult…

  5. The molecular nature of very small embryonic-like stem cells in adult tissues.

    PubMed

    Kim, YongHwan; Jeong, Jaeho; Kang, Hyunsook; Lim, Jisun; Heo, Jinbeom; Ratajczak, Janina; Ratajczak, Mariusz Z; Shin, Dong-Myung

    2014-11-01

    Pluripotent stem cells (PSCs) have been considered as the most important cells in regenerative medicine as they are able to differentiate into all types of cells in the human body. PSCs have been established from several sources of embryo tissue or by reprogramming of terminally differentiated adult tissue by transduction of so-called Yamanaka factors (Oct4, Sox2, Klf4, and cMyc). Interestingly, accumulating evidence has demonstrated the residence of PSCs in adult tissue and with the ability to differentiate into multiple types of tissue-committed stem cells (TCSCs). We also recently demonstrated that a population of pluripotent Oct4(+) SSEA-1(+)Sca-1(+)Lin(-)CD45(-) very small embryonic-like stem cells (VSELs) resides in the adult murine bone marrow (BM) and in other murine tissue. These very small (∼3-6 μm) cells express pluripotent markers such as Oct4, Nanog, and SSEA-1. VSELs could be specified into several tissue-residing TCSCs in response to tissue/organ injury, and thus suggesting that these cells have a physiological role in the rejuvenation of a pool of TCSCs under steady-state conditions. In this review article, we discuss the molecular nature of the rare population of VSELs which have a crucial role in regulating the pluripotency, proliferation, differentiation, and aging of these cells. PMID:25473442

  6. A resource of ribosomal RNA-depleted RNA-Seq data from different normal adult and fetal human tissues.

    PubMed

    Choy, Jocelyn Y H; Boon, Priscilla L S; Bertin, Nicolas; Fullwood, Melissa J

    2015-01-01

    Gene expression is the most fundamental level at which the genotype leads to the phenotype of the organism. Enabled by ultra-high-throughput next-generation DNA sequencing, RNA-Seq involves shotgun sequencing of fragmented RNA transcripts by next-generation sequencing followed by in silico assembly, and is rapidly becoming the most popular method for gene expression analysis. Poly[A]+ RNA-Seq analyses of normal human adult tissue samples such as Illumina's Human BodyMap 2.0 Project and the RNA-Seq atlas have provided a useful global resource and framework for comparisons with diseased tissues such as cancer. However, these analyses have failed to provide information on poly[A]-RNA, which is abundant in our cells. The most recent advances in RNA-Seq analyses use ribosomal RNA-depletion to provide information on both poly[A]+ and poly[A]-RNA. In this paper, we describe the use of Illumina's HiSeq 2000 to generate high quality rRNA-depleted RNA-Seq datasets from human fetal and adult tissues. The datasets reported here will be useful in understanding the different expression profiles in different tissues.

  7. Brain tissue pressure measurements in perinatal and adult rabbits.

    PubMed

    Hornig, G W; Lorenzo, A V; Zavala, L M; Welch, K

    1987-12-01

    Brain tissue pressure (BTP) in pre- and post-natal anesthetized rabbits, held in a stereotactic head holder, was measured with a fluid filled 23 gauge open-ended cannula connected distally to a pressure transducer. By advancing the cannula step wise through a hole in the cranium it was possible to sequentially measure pressure from the cranial subarachnoid space, cortex, ventricle and basal ganglia. Separate cannulas and transducers were used to measure CSFP from the cisterna magna and arterial and/or venous pressure. Pressure recordings obtained when the tip of the BTP cannula was located in the cranial subarachnoid space or ventricle exhibited respiratory and blood pressure pulsations equivalent to and in phase with CSF pulsations recorded from the cisterna magna. When the tip was advanced into brain parenchymal sites such pulsations were suppressed or non-detectable unless communication with a CSF compartment had been established inadvertently. Although CSF pressures in the three spinal fluid compartments were equivalent, in most animals BTP was higher than CSFP. However, after momentary venting of the system BTP equilibrated at a pressure below that of CSFP. We speculate that venting of the low compliance system (1.20 x 10(-5) ml/mmHg) relieves the isometric pressure build-up due to insertion of the cannula into brain parenchyma. Under these conditions, and at all ages examined, BTP in the rabbit is consistently lower than CSFP and, as with CSFP, it increases as the animal matures.

  8. Tissue Microarray Technology for Molecular Applications: Investigation of Cross-Contamination between Tissue Samples Obtained from the Same Punching Device

    PubMed Central

    Vassella, Erik; Galván, José A.; Zlobec, Inti

    2015-01-01

    Background: Tissue microarray (TMA) technology allows rapid visualization of molecular markers by immunohistochemistry and in situ hybridization. In addition, TMA instrumentation has the potential to assist in other applications: punches taken from donor blocks can be placed directly into tubes and used for nucleic acid analysis by PCR approaches. However, the question of possible cross-contamination between samples punched with the same device has frequently been raised but never addressed. Methods: Two experiments were performed. (1) A block from mycobacterium tuberculosis (TB) positivetissue and a second from an uninfected patient were aligned side-by-side in an automated tissue microarrayer. Four 0.6 mm punches were cored from each sample and placed inside their corresponding tube. Between coring of each donor block, a mechanical cleaning step was performed by insertion of the puncher into a paraffin block. This sequence of coring and cleaning was repeated three times, alternating between positive and negative blocks. A fragment from the 6110 insertion sequence specific for mycobacterium tuberculosis was analyzed; (2) Four 0.6 mm punches were cored from three KRAS mutated colorectal cancer blocks, alternating with three different wild-type tissues using the same TMA instrument (sequence of coring: G12D, WT, G12V, WT, G13D and WT). Mechanical cleaning of the device between each donor block was made. Mutation analysis by pyrosequencing was carried out. This sequence of coring was repeated manually without any cleaning step between blocks. Results/Discussion: In both analyses, all alternating samples showed the expected result (samples 1, 3 and 5: positive or mutated, samples 2, 4 and 6: negative or wild-type). Similar results were obtained without cleaning step. These findings suggest that no cross-contamination of tissue samples occurs when donor blocks are punched using the same device, however a cleaning step is nonetheless recommended. Our result supports

  9. Experimental implementation of coded aperture coherent scatter spectral imaging of cancerous and healthy breast tissue samples

    NASA Astrophysics Data System (ADS)

    Lakshmanan, Manu N.; Greenberg, Joel A.; Samei, Ehsan; Kapadia, Anuj J.

    2015-03-01

    A fast and accurate scatter imaging technique to differentiate cancerous and healthy breast tissue is introduced in this work. Such a technique would have wide-ranging clinical applications from intra-operative margin assessment to breast cancer screening. Coherent Scatter Computed Tomography (CSCT) has been shown to differentiate cancerous from healthy tissue, but the need to raster scan a pencil beam at a series of angles and slices in order to reconstruct 3D images makes it prohibitively time consuming. In this work we apply the coded aperture coherent scatter spectral imaging technique to reconstruct 3D images of breast tissue samples from experimental data taken without the rotation usually required in CSCT. We present our experimental implementation of coded aperture scatter imaging, the reconstructed images of the breast tissue samples and segmentations of the 3D images in order to identify the cancerous and healthy tissue inside of the samples. We find that coded aperture scatter imaging is able to reconstruct images of the samples and identify the distribution of cancerous and healthy tissues (i.e., fibroglandular, adipose, or a mix of the two) inside of them. Coded aperture scatter imaging has the potential to provide scatter images that automatically differentiate cancerous and healthy tissue inside of ex vivo samples within a time on the order of a minute.

  10. Mitochondrial Respiration Chain Enzymatic Activities in the Human Brain: Methodological Implications for Tissue Sampling and Storage.

    PubMed

    Ronsoni, Marcelo Fernando; Remor, Aline Pertile; Lopes, Mark William; Hohl, Alexandre; Troncoso, Iris H Z; Leal, Rodrigo Bainy; Boos, Gustavo Luchi; Kondageski, Charles; Nunes, Jean Costa; Linhares, Marcelo Neves; Lin, Kátia; Latini, Alexandra Susana; Walz, Roger

    2016-04-01

    Mitochondrial respiratory chain complexes enzymatic (MRCCE) activities were successfully evaluated in frozen brain samples. Epilepsy surgery offers an ethical opportunity to study human brain tissue surgically removed to treat drug resistant epilepsies. Epilepsy surgeries are done with hemodynamic and laboratory parameters to maintain physiology, but there are no studies analyzing the association among these parameters and MRCCE activities in the human brain tissue. We determined the intra-operative parameters independently associated with MRCCE activities in middle temporal neocortex (Cx), amygdala (AMY) and head of hippocampus (HIP) samples of patients (n = 23) who underwent temporal lobectomy using multiple linear regressions. MRCCE activities in Cx, AMY and HIP are differentially associated to trans-operative mean arterial blood pressure, O2 saturation, hemoglobin, and anesthesia duration to time of tissue sampling. The time-course between the last seizure occurrence and tissue sampling as well as the sample storage to biochemical assessments were also associated with enzyme activities. Linear regression models including these variables explain 13-17 % of MRCCE activities and show a moderate to strong effect (r = 0.37-0.82). Intraoperative hemodynamic and laboratory parameters as well as the time from last seizure to tissue sampling and storage time are associated with MRCCE activities in human samples from the Cx, AMYG and HIP. Careful control of these parameters is required to minimize confounding biases in studies using human brain samples collected from elective neurosurgery. PMID:26586405

  11. Culture methods of allograft musculoskeletal tissue samples in Australian bacteriology laboratories.

    PubMed

    Varettas, Kerry

    2013-12-01

    Samples of allograft musculoskeletal tissue are cultured by bacteriology laboratories to determine the presence of bacteria and fungi. In Australia, this testing is performed by 6 TGA-licensed clinical bacteriology laboratories with samples received from 10 tissue banks. Culture methods of swab and tissue samples employ a combination of solid agar and/or broth media to enhance micro-organism growth and maximise recovery. All six Australian laboratories receive Amies transport swabs and, except for one laboratory, a corresponding biopsy sample for testing. Three of the 6 laboratories culture at least one allograft sample directly onto solid agar. Only one laboratory did not use a broth culture for any sample received. An international literature review found that a similar combination of musculoskeletal tissue samples were cultured onto solid agar and/or broth media. Although variations of allograft musculoskeletal tissue samples, culture media and methods are used in Australian and international bacteriology laboratories, validation studies and method evaluations have challenged and supported their use in recovering fungi and aerobic and anaerobic bacteria.

  12. Heavy Metal Content in Thoracic Tissue Samples from Patients with and without NSCLC.

    PubMed

    Tran, Jessica Q; Dranikov, Alexandra; Iannucci, Anita; Wagner, Walter P; LoBello, Janine; Allen, Jeffrey; Weiss, Glen J

    2014-01-01

    Objectives. Environmental factors expose an individual to heavy metals that may stimulate cancer growth preclinically including non-small cell lung cancer (NSCLC) cells. Here, we examine the prevalence of four heavy metals present in postsurgical tissues from individuals with and without NSCLC. Materials and Methods. Thoracic tissue samples from two separate sample sets were analyzed for cadmium (Cd), arsenic (As), mercury (Hg), and lead (Pb) content. Results. In the first sample set, there was no significant measurable amount of Pb and Hg found in either NSCLC tissue or nonmalignant lung tissue samples. Cd was the most prevalent heavy metal and As was present in moderate amounts. In the second sample set, Cd was measurable across all tissue types taken from 28 NSCLC patients and significantly higher Cd was measurable in noncancer benign lung (n = 9). In the NSCLC samples, As was measurable in moderate amounts, while Hg and Pb amounts were negligible. Conclusion. Cd and As are present in lung tissues for patients with NSCLC. With existing preclinical evidence of their tumorigenecity, it is plausible that Cd and/or As may have an impact on NSCLC development. Additional studies examining the prevalence and association between smokers and nonsmokers are suggested. PMID:26316947

  13. High prevalence of thymic tissue in adults with human immunodeficiency virus-1 infection.

    PubMed Central

    McCune, J M; Loftus, R; Schmidt, D K; Carroll, P; Webster, D; Swor-Yim, L B; Francis, I R; Gross, B H; Grant, R M

    1998-01-01

    The thymus in adults infected with the HIV-1 is generally thought to be inactive, both because of age-related involution and viral destruction. We have revisited the question of thymic function in adults, using chest-computed tomography (CT) to measure thymic tissue in HIV-1-seropositive (n = 99) or HIV-1-seronegative (n = 32) subjects, and correlating these results with the level of circulating CD4(+) and CD8(+) T cells that are phenotypically described as naive thymic emigrants. Abundant thymic tissue was detectable in many (47/99) HIV-1-seropositive adults, aged 20-59. Independent of age, radiographic demonstration of thymic tissue was significantly associated with both a higher CD4(+) T cell count (P = 0.02) and a higher percentage and absolute number of circulating naive (CD45RA+CD62L+) CD4(+) T cells (P < 0.04). The prevalence of an abundant thymus was especially high in younger HIV-1-seropositive adults ( 40 yr) regardless of CD4 count (P = 0.03). These studies suggest that the thymus is functional in some but not all adults with HIV-1 disease. PMID:9616201

  14. JH Biosynthesis by Reproductive Tissues and Corpora Allata in Adult Longhorned Beetles, Apriona germari

    Technology Transfer Automated Retrieval System (TEKTRAN)

    We report on juvenile hormone (JH) biosynthesis from long-chain intermediates by specific reproductive system tissues and the corpora allata (CA) prepared from adult longhorned beetles, Apriona germari. Testes, male accessory glands (MAGs), ovaries and CA contain the long-chain intermediates in the ...

  15. The osmotic stability of lysosomes from adult and foetal guinea-pig liver tissue.

    PubMed

    Turnbull, J M; Neil, M W

    1969-02-01

    1. Lysosome-rich fractions were obtained from foetal liver tissues as early as 35 days uterine age. Foetal lysosomes showed the same ;structure-linked latency' and acid hydrolytic potentiality characteristic of their adult counterparts. 2. The osmotic stability of lysosome-rich fraction from foetal guinea-pig liver tissue was greater than that of the corresponding adult lysosome fractions, p-nitrophenyl-phosphatase being used as marker enzyme. 3. The observation was confirmed by using beta-glycerophosphatase and phenolphthalein beta-glucuronidase as alternative marker enzymes. p-Nitrophenyl phosphate and beta-glycerophosphate appear to act as substrates for the same enzyme. 4. By using p-nitrophenylphosphatase activity measurements it was shown that the osmotic stability of foetal lysosomal fractions decreased with increasing foetal age, but at no time achieved the degree of osmotic instability characteristic of adult lysosomal fractions. 5. The correlation of these findings with the intracellular environment of lysosomes is discussed.

  16. Wheelchair Use among Community-Dwelling Older Adults: Prevalence and Risk Factors in a National Sample

    ERIC Educational Resources Information Center

    Clarke, Philippa; Colantonio, Angela

    2005-01-01

    Older adults are the largest group of wheelchair users yet there are no peer-reviewed studies on the national profile of older wheelchair users in Canada. We investigated the characteristics of wheelchair users in a national sample of community-dwelling older adults from the Canadian Study of Health and Aging (CSHA-2). Questions on the use of…

  17. Ex vivo confocal microscopy imaging to identify tumor tissue on freshly removed brain sample.

    PubMed

    Forest, Fabien; Cinotti, Elisa; Yvorel, Violaine; Habougit, Cyril; Vassal, François; Nuti, Christophe; Perrot, Jean-Luc; Labeille, Bruno; Péoc'h, Michel

    2015-09-01

    Confocal microscopy is a technique able to realize "optic sections" of a tissue with increasing applications. We wondered if we could apply an ex vivo confocal microscope designed for dermatological purpose in a routine use for the most frequent brain tumors. The aim of this work was to identify tumor tissue and its histopathological hallmarks, and to assess grading criteria used in neuropathological practice without tissue loss on freshly removed brain tissue. Seven infiltrating gliomas, nine meningiomas and three metastases of carcinomas were included. We compared imaging results obtained with the confocal microscope to frozen sections, smears and tissue sections of formalin-fixed tissue. Our results show that ex vivo confocal microscopy imaging can be applied to brain tumors in order to quickly identify tumor tissue without tissue loss. It can differentiate tumors and can assess most of grading criteria. Confocal microscopy could represent a new tool to identify tumor tissue on freshly removed sample and could help in selecting areas for biobanking of tumor tissue.

  18. Efficient and scalable serial extraction of DNA and RNA from frozen tissue samples.

    PubMed

    Mathot, Lucy; Lindman, Monica; Sjöblom, Tobias

    2011-01-01

    Advances in cancer genomics have created a demand for scalable sample processing. We here present a process for serial extraction of nucleic acids from the same frozen tissue sample based on magnetic silica particles. The process is automation friendly with high recoveries of pure DNA and RNA suitable for analysis.

  19. Terahertz spectroscopy and detection of brain tumor in rat fresh-tissue samples

    NASA Astrophysics Data System (ADS)

    Yamaguchi, S.; Fukushi, Y.; Kubota, O.; Itsuji, T.; Yamamoto, S.; Ouchi, T.

    2015-03-01

    Terahertz (THz) spectroscopy and imaging of biomedical samples is expected to be an important application of THz analysis techniques. Identification and localization of tumor tissue, imaging of biological samples, and analysis of DNA by THz spectroscopy have been reported. THz time-domain spectroscopy (TDS) is useful for obtaining the refractive index over a broad frequency range. However, THz-TDS spectra of fresh tissue samples are sensitive to procedures such as sample preparation, and a standardized measurement protocol is required. Therefore, in this work, we establish a protocol for measurements of THz spectra of fresh tissue and demonstrate reliable detection of rat brain tumor tissue. We use a reflection THz-TDS system to measure the refractive index spectra of the samples mounted on a quartz plate. The tissue samples were measured immediately after sectioning to avoid sample denaturalization during storage. Special care was taken in THz data processing to eliminate parasitic reflections and reduce noise. The error level in our refractive index measurements was as low as 0.02 in the frequency range 0.8-1.5 THz. With increasing frequency, the refractive index in the tumor and normal regions monotonically decreased, similarly to water, and it was 0.02 higher in the tumor regions. The spectral data suggest that the tumor regions have higher water content. Hematoxylin-eosin stained images showed that increased cell density was also responsible for the observed spectral features. A set of samples from 10 rats showed consistent results. Our results suggest that reliable tumor detection in fresh tissue without pretreatment is possible with THz spectroscopy measurements. THz spectroscopy has the potential to become a real-time in vivo diagnostic method.

  20. Sample Preparation for Mass Spectrometry Imaging of Plant Tissues: A Review.

    PubMed

    Dong, Yonghui; Li, Bin; Malitsky, Sergey; Rogachev, Ilana; Aharoni, Asaph; Kaftan, Filip; Svatoš, Aleš; Franceschi, Pietro

    2016-01-01

    Mass spectrometry imaging (MSI) is a mass spectrometry based molecular ion imaging technique. It provides the means for ascertaining the spatial distribution of a large variety of analytes directly on tissue sample surfaces without any labeling or staining agents. These advantages make it an attractive molecular histology tool in medical, pharmaceutical, and biological research. Likewise, MSI has started gaining popularity in plant sciences; yet, information regarding sample preparation methods for plant tissues is still limited. Sample preparation is a crucial step that is directly associated with the quality and authenticity of the imaging results, it therefore demands in-depth studies based on the characteristics of plant samples. In this review, a sample preparation pipeline is discussed in detail and illustrated through selected practical examples. In particular, special concerns regarding sample preparation for plant imaging are critically evaluated. Finally, the applications of MSI techniques in plants are reviewed according to different classes of plant metabolites. PMID:26904042

  1. Sample Preparation for Mass Spectrometry Imaging of Plant Tissues: A Review

    PubMed Central

    Dong, Yonghui; Li, Bin; Malitsky, Sergey; Rogachev, Ilana; Aharoni, Asaph; Kaftan, Filip; Svatoš, Aleš; Franceschi, Pietro

    2016-01-01

    Mass spectrometry imaging (MSI) is a mass spectrometry based molecular ion imaging technique. It provides the means for ascertaining the spatial distribution of a large variety of analytes directly on tissue sample surfaces without any labeling or staining agents. These advantages make it an attractive molecular histology tool in medical, pharmaceutical, and biological research. Likewise, MSI has started gaining popularity in plant sciences; yet, information regarding sample preparation methods for plant tissues is still limited. Sample preparation is a crucial step that is directly associated with the quality and authenticity of the imaging results, it therefore demands in-depth studies based on the characteristics of plant samples. In this review, a sample preparation pipeline is discussed in detail and illustrated through selected practical examples. In particular, special concerns regarding sample preparation for plant imaging are critically evaluated. Finally, the applications of MSI techniques in plants are reviewed according to different classes of plant metabolites. PMID:26904042

  2. A tissue fixative that protects macromolecules (DNA, RNA, and protein) and histomorphology in clinical samples.

    PubMed

    Vincek, Vladimir; Nassiri, Mehdi; Nadji, Mehrdad; Morales, Azorides R

    2003-10-01

    Preservation of macromolecules (DNA, RNA, and proteins) in tissue is traditionally achieved by immediate freezing of the sample. Although isolation of PCR-able RNA has been reported from formalin-fixed, paraffin-embedded tissues, the process has not been shown to be reproducible because high molecular weight RNA is usually degraded. We investigated the potential value of a new universal molecular fixative (UMFIX, Sakura Finetek USA, Inc., Torrance, California) in preservation of macromolecules in paraffin-embedded tissue. Mouse and human tissues were fixed in UMFIX from 1 hour to 8 weeks. They were then processed by a rapid tissue processing (RTP) system, embedded in paraffin, and evaluated for routine histology as well as for the quality and quantity of DNA, RNA, and proteins. Formalin-fixed tissues were processed by RTP and evaluated in a similar manner. Fresh-frozen samples were used as controls. The morphology of UMFIX-exposed tissue was comparable to that fixed in formalin. High molecular weight RNA was preserved in tissue that was immediately fixed in UMFIX and stored from 1 hour to 8 weeks at room temperature. There were no significant differences between UMFIX-exposed and frozen tissues on PCR, RT-PCR, real-time PCR, and expression microarrays. Similarly, physical and antigenic preservation of proteins in UMFIX tissue was similar to fresh state. Both RNA and proteins were substantially degraded in formalin-fixed and similarly processed specimens. We concluded that it is now possible to preserve histomorphology and intact macromolecules in the same archival paraffin-embedded tissue through the use of a novel fixative and a rapid processing system.

  3. Declining estimated prevalence of alcohol drinking and smoking among young adults nationally: artifacts of sample undercoverage?

    PubMed

    Delnevo, Cristine D; Gundersen, Daniel A; Hagman, Brett T

    2008-01-01

    A growing concern in public health surveillance surveys that rely on random digit dialing for sampling is the exclusion of adults in cell-phone-only households. The purpose of this study was to examine whether recent increases in wireless substitution have affected estimates of tobacco and alcohol use in the Behavioral Risk Factor Surveillance System (BRFSS) in a subpopulation with notable cell-phone usage (i.e., young adults). BRFSS data from 2001-2005 were examined. Analyses were limited to participants aged 18-24 years, and the sample contained approximately 18,500 persons in each year. Prevalence estimates were generated with SUDAAN software for three health behaviors: cigarette smoking, binge drinking, and heavy alcohol consumption. In addition, the authors examined sample completeness for young adults relative to US Census estimates. Overall, prevalences of all three health behaviors among young adults were fairly stable between 2001 and 2003 but significantly decreased between 2003 and 2005. These trends are not replicated in national surveys that use area probability samples. The authors found a declining trend in the sample completeness ratio for young adults; it declined from 0.32 in 2001 to 0.15 in 2005. Given the high prevalence of wireless substitution among young adults and the declining sample completeness ratio, the authors suspect that the observed decreases in prevalence are artifacts of undercoverage.

  4. Declining estimated prevalence of alcohol drinking and smoking among young adults nationally: artifacts of sample undercoverage?

    PubMed

    Delnevo, Cristine D; Gundersen, Daniel A; Hagman, Brett T

    2008-01-01

    A growing concern in public health surveillance surveys that rely on random digit dialing for sampling is the exclusion of adults in cell-phone-only households. The purpose of this study was to examine whether recent increases in wireless substitution have affected estimates of tobacco and alcohol use in the Behavioral Risk Factor Surveillance System (BRFSS) in a subpopulation with notable cell-phone usage (i.e., young adults). BRFSS data from 2001-2005 were examined. Analyses were limited to participants aged 18-24 years, and the sample contained approximately 18,500 persons in each year. Prevalence estimates were generated with SUDAAN software for three health behaviors: cigarette smoking, binge drinking, and heavy alcohol consumption. In addition, the authors examined sample completeness for young adults relative to US Census estimates. Overall, prevalences of all three health behaviors among young adults were fairly stable between 2001 and 2003 but significantly decreased between 2003 and 2005. These trends are not replicated in national surveys that use area probability samples. The authors found a declining trend in the sample completeness ratio for young adults; it declined from 0.32 in 2001 to 0.15 in 2005. Given the high prevalence of wireless substitution among young adults and the declining sample completeness ratio, the authors suspect that the observed decreases in prevalence are artifacts of undercoverage. PMID:17977896

  5. Unpacking Sleep and Suicide in Older Adults in a Combined Online Sample

    PubMed Central

    Golding, Shea; Nadorff, Michael R.; Winer, E. Samuel; Ward, Kathryn Claire

    2015-01-01

    Objective: Duration of insomnia symptoms and nightmares are related to suicidal risk in young adults independent of current symptoms of insomnia, nightmares, anxiety, depression, and PTSD. However, this relation has yet to be examined among older adults, despite older adults being at higher risk of suicidal behavior. Further, the current study aims to replicate previous research among younger adults showing that insomnia symptoms and nightmares are associated with suicide risk independent of the interpersonal psychological theory of suicide (IPTS). Methods: The present study utilized 167 participants age 55 and older obtained by combining two independent mTurk data collections of adults in the United States. Results: In the current sample, duration of nightmares was associated with suicide risk in older adults independent of symptoms of current insomnia and nightmares, duration of insomnia, and symptoms of PTSD, anhedonia, and the IPTS. Conclusions: Our findings suggest that the duration of nightmares (i.e., how long someone has been experiencing nightmares) predict substantial variance in suicide risk among older adults in addition to the risk factors typically examined. Thus, assessment of sleep dysfunction is important when assessing suicide risk among older adults. Citation: Golding S, Nadorff MR, Winer ES, Ward KC. Unpacking sleep and suicide in older adults in a combined online sample. J Clin Sleep Med 2015;11(12):1385–1392. PMID:26194726

  6. Gender differences in a sample of adults with attention deficit hyperactivity disorder.

    PubMed

    Biederman, J; Faraone, S V; Spencer, T; Wilens, T; Mick, E; Lapey, K A

    1994-07-01

    Although originally conceptualized as a childhood disorder, attention deficit hyperactivity disorder (ADHD) may also be an adult disorder. However, despite increasing media attention to adult ADHD, its validity has only recently been studied in a systematic fashion. The overrepresentation of females in adult samples in comparison to pediatric samples of ADHD raises additional questions about the validity of this disorder in adults. The goal of this article is to explore whether ADHD is a valid clinical entity in female subjects and whether it is expressed differently in male and female adults. To this end, we examined the clinical, cognitive, and functional characteristics of 128 referred adult ADHD cases of both sexes. Each subject had a clinical diagnosis of childhood-onset ADHD confirmed by structured interview. The male and female ADHD adults were similar to one another but more disturbed and impaired than non-ADHD adult control subjects. Compared with normal control females, ADHD women had higher rates of major depression, anxiety disorders, and conduct disorder; and more evidence of school failure and cognitive impairment. The consistency of these findings in both genders further supports the validity of the diagnosis of ADHD in adults. Our results stress the viability and importance of identification of female subjects with ADHD. The underidentification and undertreatment of females with ADHD may have substantial mental health and educational implications, suggesting that research is needed to develop a better understanding of clinical indicators of ADHD in females.

  7. MALDI direct analysis and imaging of frozen versus FFPE tissues: what strategy for which sample?

    PubMed

    Wisztorski, Maxence; Franck, Julien; Salzet, Michel; Fournier, Isabelle

    2010-01-01

    Significant advances have been made in the past decade in the field of mass spectrometry imaging with MALDI ion sources (MALDI-MSI). While MALDI-MSI has high potential in the field of biology and in the clinic, a challenge for MALDI-MSI has been to adapt itself to a greater range of sample types. In particular, much of the biological archived materials for pathology studies are tissue biopsies fixed with paraformaldehyde and embedded in paraffin (FFPE tissues) because of the high stability of such samples. Thus, there has been a need to develop strategies for analyzing FFPE samples as this would allow retrospective studies of past clinical cases on large cohorts of existing samples. Obviously, PAF fixation, by inducing protein cross-linking, causes problems for molecular analysis by MS. We developed on tissue digestion strategies for overcoming these difficulties and allowing molecular data to be retrieved from FFPE samples no matter how long they have been stored. These digestion strategies preserve localization from digested proteins making MALDI-MSI of proteins possible by monitoring the resulting peptides. We present methods and protocols for FFPE samples. These strategies have proven to be valuable for all tested FFPE samples and have opened archived tissues from hospital banks to MALDI-MSI.

  8. Spontaneous myogenic differentiation of Flk-1-positive cells from adult pancreas and other nonmuscle tissues.

    PubMed

    Di Rocco, Giuliana; Tritarelli, Alessandra; Toietta, Gabriele; Gatto, Ilaria; Iachininoto, Maria Grazia; Pagani, Francesca; Mangoni, Antonella; Straino, Stefania; Capogrossi, Maurizio C

    2008-02-01

    At the embryonic or fetal stages, autonomously myogenic cells (AMCs), i.e., cells able to spontaneously differentiate into skeletal myotubes, have been identified from several different sites other than skeletal muscle, including the vascular compartment. However, in the adult animal, AMCs from skeletal muscle-devoid tissues have been described in only two cases. One is represented by thymic myoid cells, a restricted population of committed myogenic progenitors of unknown derivation present in the thymic medulla; the other is represented by a small subset of adipose tissue-associated cells, which we recently identified. In the present study we report, for the first time, the presence of spontaneously differentiating myogenic precursors in the pancreas and in other skeletal muscle-devoid organs such as spleen and stomach, as well as in the periaortic tissue of adult mice. Immunomagnetic selection procedures indicate that AMCs derive from Flk-1(+) progenitors. Individual clones of myogenic cells from nonmuscle organs are morphologically and functionally indistinguishable from skeletal muscle-derived primary myoblasts. Moreover, they can be induced to proliferate in vitro and are able to participate in muscle regeneration in vivo. Thus, we provide evidence that fully competent myogenic progenitors can be derived from the Flk-1(+) compartment of several adult tissues that are embryologically unrelated to skeletal muscle. PMID:18094147

  9. Broth versus solid agar culture of swab samples of cadaveric allograft musculoskeletal tissue.

    PubMed

    Varettas, Kerry

    2013-12-01

    As part of the donor assessment protocol, bioburden assessment must be performed on allograft musculoskeletal tissue samples collected at the time of tissue retrieval. Swab samples of musculoskeletal tissue allografts from cadaveric donors are received at the microbiology department of the South Eastern Area Laboratory Services (Australia) to determine the presence of bacteria and fungi. This study will review the isolation rate of organisms from solid agar and broth culture of swab samples of cadaveric allograft musculoskeletal tissue over a 6-year period, 2006-2011. Swabs were inoculated onto horse blood agar (anaerobic, 35 °C) and chocolate agar (CO2, 35 °C) and then placed into a cooked meat broth (aerobic, 35 °C). A total of 1,912 swabs from 389 donors were received during the study period. 557 (29.1 %) swabs were culture positive with the isolation of 713 organisms, 249 (34.9 %) from solid agar culture and an additional 464 (65.1 %) from broth culture only. This study has shown that the broth culture of cadaveric allograft musculoskeletal swab samples recovered a greater amount of organisms than solid agar culture. Isolates such as Clostridium species and Staphylococcus aureus would not have been isolated from solid agar culture alone. Broth culture is an essential part of the bioburden assessment protocol of swab samples of cadaveric allograft musculoskeletal tissue in this laboratory.

  10. An antigen trapping ELISA for the detection of capripoxvirus in tissue culture supernatant and biopsy samples.

    PubMed

    Carn, V M

    1995-01-01

    A trapping ELISA for the detection of capripoxvirus antigen in tissue culture supernatant and biopsy material was developed, using a guinea-pig polyclonal detector antiserum raised against a recombinant capripoxvirus specific antigen, expressed in Escherichia coli using the plasmid vector pGEX-2T. The ELISA detected antigen in tissue culture samples that on virus titration contained equal to or in excess of 10(2.8) TCID50/ml. Virus isolation and ELISA were compared for the detection of capripoxvirus in skin biopsy samples from sheep, goats and cattle. The ELISA compared well with virus isolation, and has applications as a diagnostic test. This assay reduces the reliance of diagnostic laboratories on tissue culture facilities, and can be used to confirm the presence of capripoxvirus in tissue culture.

  11. Detection of Slit2 promoter hypermethylation in tissue and serum samples from breast cancer patients.

    PubMed

    Kim, Ga-Eon; Lee, Kyung Hwa; Choi, Yoo Duk; Lee, Ji Shin; Lee, Jae Hyuk; Nam, Jong Hee; Choi, Chan; Park, Min Ho; Yoon, Jung Han

    2011-10-01

    Promoter hypermethylation has been shown to be a common mechanism for inactivation of tumor suppressor genes in breast cancer. The aim of this study was to investigate the prevalence of Slit2 promoter hypermethylation in both the tumor and serum samples of breast cancer patients with ductal carcinoma in situ (DCIS) or invasive breast carcinoma (IBC). The methylation status of Slit2 was investigated in 210 tissue samples (15 breast with no pathological findings, 26 DCIS, and 169 IBC samples) and 123 corresponding serum samples (15 breast with no pathological findings, 26 DCIS, and 82 IBC samples) using methylation-specific polymerase chain reaction. Immunohistochemical staining for Slit2 was also performed using tissue microarray blocks to determine whether Slit2 promoter hypermethylation correlated with loss of Slit2 expression. Slit2 promoter hypermethylation was not detected in breast tissue and serum samples from patients with no pathological findings. DCIS or IBC showed a statistically higher frequency of Slit2 promoter hypermethylation compared to breast with no pathological findings in both the tissue and serum samples; however, there were no statistically significant differences between DCIS and IBC samples. Similar Slit2 promoter hypermethylation patterns were seen in the tissue samples and corresponding serum specimens (p < 0.001). Slit2 promoter hypermethylation was associated with loss of Slit2 expression. These results suggest that Slit2 promoter hypermethylation appears to be responsible for functionally silencing Slit2 expression. Slit2 promoter hypermethylation may be considered as a possible serum marker for early detection of breast cancer.

  12. X-ray scattering for the characterization of lyophilized breast tissue samples

    NASA Astrophysics Data System (ADS)

    Elshemey, Wael M.; Mohamed, Fayrouz S.; Khater, Ibrahim M.

    2013-09-01

    This work investigates the possibility of characterizing breast cancer by measuring the X-ray scattering profiles of lyophilized excised breast tissue samples. Since X-ray scattering from water-rich tissue is dominated by scattering from water, the removal of water by lyophilization would enhance the characterization process. In the present study, X-ray scattering profiles of 22 normal, 22 malignant and 10 benign breast tissue samples are measured. The cut-offs of scatter diagrams, sensitivity, specificity and diagnostic accuracy of three characterization parameters (full width at half maximum (FWHM) for the peak at 1.1 nm-1, area under curve (AUC), and ratio of 1st to 2nd scattering peak intensities (I1/I2%)) are calculated and compared to the data from non-lyophilized samples. Results show increased sensitivity (up to 100%) of the present data on lyophilized breast tissue samples compared to previously reported data for non-lyophilized samples while the specificity (up to 95.4%), diagnostic accuracy (up to 95.4%) and receiver operating characteristic (ROC) curve values (up to 0.9979) for both sets of data are comparable. The present study shows significant differences between normal samples and each of malignant and benign samples. Only subtle differences exist between malignant and benign lyophilized breast tissue samples where FWHM=0.7±0.1 and 0.8±0.3, AUC=1.3±0.2 and 1.4±0.2 and I1/I2%=44.9±11.0 and 52.4±7.6 for malignant and benign samples respectively.

  13. Evidence for tissue-resident mesenchymal stem cells in human adult lung from studies of transplanted allografts.

    PubMed

    Lama, Vibha N; Smith, Lisa; Badri, Linda; Flint, Andrew; Andrei, Adin-Cristian; Murray, Susan; Wang, Zhuo; Liao, Hui; Toews, Galen B; Krebsbach, Paul H; Peters-Golden, Marc; Pinsky, David J; Martinez, Fernando J; Thannickal, Victor J

    2007-04-01

    The origin and turnover of connective tissue cells in adult human organs, including the lung, are not well understood. Here, studies of cells derived from human lung allografts demonstrate the presence of a multipotent mesenchymal cell population, which is locally resident in the human adult lung and has extended life span in vivo. Examination of plastic-adherent cell populations in bronchoalveolar lavage samples obtained from 76 human lung transplant recipients revealed clonal proliferation of fibroblast-like cells in 62% (106 of 172) of samples. Immunophenotyping of these isolated cells demonstrated expression of vimentin and prolyl-4-hydroxylase, indicating a mesenchymal phenotype. Multiparametric flow cytometric analyses revealed expression of cell-surface proteins, CD73, CD90, and CD105, commonly found on mesenchymal stem cells (MSCs). Hematopoietic lineage markers CD14, CD34, and CD45 were absent. Multipotency of these cells was demonstrated by their capacity to differentiate into adipocytes, chondrocytes, and osteocytes. Cytogenetic analysis of cells from 7 sex-mismatched lung transplant recipients harvested up to 11 years after transplant revealed that 97.2% +/- 2.1% expressed the sex genotype of the donor. The presence of MSCs of donor sex identity in lung allografts even years after transplantation provides what we believe to be the first evidence for connective tissue cell progenitors that reside locally within a postnatal, nonhematopoietic organ.

  14. Distribution of bisphenol A into tissues of adult, neonatal, and fetal Sprague-Dawley rats

    SciTech Connect

    Doerge, Daniel R.; Twaddle, Nathan C.; Vanlandingham, Michelle; Brown, Ronald P.; Fisher, Jeffrey W.

    2011-09-15

    Bisphenol A (BPA) is an important industrial chemical used in the manufacture of polycarbonate plastic products and epoxy resin-based food can liners. The presence of BPA metabolites in urine of > 90% of Americans aged 6-60 suggests ubiquitous and frequent exposure in the range of 0.02-0.2 {mu}g/kg bw/d (25th-95th percentiles). The current study used LC/MS/MS to measure placental transfer and concentrations of aglycone (receptor-active) and conjugated (inactive) BPA in tissues from Sprague-Dawley rats administered deuterated BPA (100 {mu}g/kg bw) by oral and IV routes. In adult female rat tissues, the tissue/serum concentration ratios for aglycone BPA ranged from 0.7 in liver to 5 in adipose tissue, reflecting differences in tissue perfusion, composition, and metabolic capacity. Following IV administration to dams, placental transfer was observed for aglycone BPA into fetuses at several gestational days (GD), with fetal/maternal serum ratios of 2.7 at GD 12, 1.2 at GD 16, and 0.4 at GD 20; the corresponding ratios for conjugated BPA were 0.43, 0.65, and 3.7. These ratios were within the ranges observed in adult tissues and were not indicative of preferential accumulation of aglycone BPA or hydrolysis of conjugates in fetal tissue in vivo. Concentrations of aglycone BPA in GD 20 fetal brain were higher than in liver or serum. Oral administration of the same dose did not produce measurable levels of aglycone BPA in fetal tissues. Amniotic fluid consistently contained levels of BPA at or below those in maternal serum. Concentrations of aglycone BPA in tissues of neonatal rats decreased with age in a manner consistent with the corresponding circulating levels. Phase II metabolism of BPA increased with fetal age such that near-term fetus was similar to early post-natal rats. These results show that concentrations of aglycone BPA in fetal tissues are similar to those in other maternal and neonatal tissues and that maternal Phase II metabolism, especially following oral

  15. Effects of formalin fixation on tissue optical properties of in-vitro brain samples

    NASA Astrophysics Data System (ADS)

    Anand, Suresh; Cicchi, Riccardo; Martelli, Fabrizio; Giordano, Flavio; Buccoliero, Anna Maria; Guerrini, Renzo; Pavone, Francesco S.

    2015-03-01

    Application of light spectroscopy based techniques for the detection of cancers have emerged as a promising approach for tumor diagnostics. In-vivo or freshly excised samples are normally used for point spectroscopic studies. However, ethical issues related to in-vivo studies, rapid decay of surgically excised tissues and sample availability puts a limitation on in-vivo and in-vitro studies. There has been a few studies reported on the application of formalin fixed samples with good discrimination capability. Usually formalin fixation is performed to prevent degradation of tissues after surgical resection. Fixing tissues in formalin prevents cell death by forming cross-linkages with proteins. Previous investigations have revealed that washing tissues fixed in formalin using phosphate buffered saline is known to reduce the effects of formalin during spectroscopic measurements. But this could not be the case with reflectance measurements. Hemoglobin is a principal absorbing medium in biological tissues in the visible range. Formalin fixation causes hemoglobin to seep out from red blood cells. Also, there could be alterations in the refractive index of tissues when fixed in formalin. In this study, we propose to investigate the changes in tissue optical properties between freshly excised and formalin fixed brain tissues. The results indicate a complete change in the spectral profile in the visible range where hemoglobin has its maximum absorption peaks. The characteristic bands of oxy-hemoglobin at 540, 580 nm and deoxy-hemoglobin at 555 nm disappear in the case of samples fixed in formalin. In addition, an increased spectral intensity was observed for the wavelengths greater than 650 nm where scattering phenomena are presumed to dominate.

  16. Gene expression profiling of adult female tissues in feeding Rhipicephalus microplus cattle ticks.

    PubMed

    Stutzer, Christian; van Zyl, Willem A; Olivier, Nicholas A; Richards, Sabine; Maritz-Olivier, Christine

    2013-06-01

    The southern cattle tick, Rhipicephalus microplus, is an economically important pest, especially for resource-poor countries, both as a highly adaptive invasive species and prominent vector of disease. The increasing prevalence of resistance to chemical acaricides and variable efficacy of current tick vaccine candidates highlight the need for more effective control methods. In the absence of a fully annotated genome, the wealth of available expressed sequence tag sequence data for this species presents a unique opportunity to study the genes that are expressed in tissues involved in blood meal acquisition, digestion and reproduction during feeding. Utilising a custom oligonucleotide microarray designed from available singletons (BmiGI Version 2.1) and expressed sequence tag sequences of R. microplus, the expression profiles in feeding adult female midgut, salivary glands and ovarian tissues were compared. From 13,456 assembled transcripts, 588 genes expressed in all three tissues were identified from fed adult females 20 days post infestation. The greatest complement of genes relate to translation and protein turnover. Additionally, a number of unique transcripts were identified for each tissue that relate well to their respective physiological/biological function/role(s). These transcripts include secreted anti-hemostatics and defense proteins from the salivary glands for acquisition of a blood meal, proteases as well as enzymes and transporters for digestion and nutrient acquisition from ingested blood in the midgut, and finally proteins and associated factors involved in DNA replication and cell-cycle control for oogenesis in the ovaries. Comparative analyses of adult female tissues during feeding enabled the identification of a catalogue of transcripts that may be essential for successful feeding and reproduction in the cattle tick, R. microplus. Future studies will increase our understanding of basic tick biology, allowing the identification of shared proteins

  17. Comparison of organochlorine residues in human adipose tissue autopsy samples from two Ontario municipalities

    SciTech Connect

    Williams, D.T.; LeBel, G.L.; Junkins, E.

    1984-01-01

    Human adipose tissue samples obtained during autopsies in a Canadian Great Lakes community, Kingston, Ontario, and a second community, Ottawa, Ontario, were analyzed for organochlorine pesticides, polychlorobiphenyls, chlorobenzenes, and chlorophenols. Significantly different levels of Dichlorodiphenyl-dichlorethane, mirex, hexachlorobenzene, and 2,3,4,6-tetrachlorophenol were found in Kingston adipose tissues compared to Ottawa tissues. Residue levels of oxychlordane, mirex, and polychlorinated biphenyls were significantly different in Kingston males versus Kingston females. The means and ranges of residue levels were contrasted with those reported in previous Canadian surveys.

  18. Characteristics of electrically injured skin from human hand tissue samples using Fourier transform infrared microspectroscopy.

    PubMed

    Li, Shi-Ying; Zou, Dong-Hua; Luo, Yi-Wen; Sun, Qi-Ran; Deng, Kai-Fei; Chen, Yi-Jiu; Huang, Ping

    2014-01-01

    This technical note describes a method for distinguishing normal skin tissue samples from those electrically injured by Fourier transform infrared microspectroscopy (FTIR MSP). Furthermore, the infrared spectral features of electrically injured cells and tissues were evaluated to identify molecular changes in epidermal cells. In the present study, 20 human hand tissue samples were evaluated macroscopically and histopathologically. The electrically injured skin samples were subdivided into 2 regions [normal cell regions (NCRs) and polarized cell regions (PCRs)] and 14 major spectral absorption bands were selected. The spectral results showed that the band absorbance at 1080, 1126, 1172, 1242, 1307, 1403, 1456, 1541, 2852, 2925, 2957, 3075, and 3300cm(-1) increased significantly both in the stratum and non-stratum corneum of the PCRs in electrically injured skin tissues samples. No significant difference was found between normal skin and the NCR of the electrically injured skin samples. The band absorbance ratios of A1172/A1126, A1456/A1403, and A2925/A2957 were significantly increased, whereas the A1652/A1541 ratio was decreased in the PCR of the stratum corneum and non-stratum corneum. Baseline changes from 4000 to near 1737cm(-1) were observed in the spectra of the electrically injured skin samples, which were interpreted in terms of the pathological process involved in electrical injury. FTIR-MSP presents a useful method to provide objective spectral markers for the assisted diagnosis of electrical marks.

  19. The connective tissue of the adductor canal--a morphological study in fetal and adult specimens.

    PubMed

    de Oliveira, Flavia; de Vasconcellos Fontes, Ricardo Bragança; da Silva Baptista, Josemberg; Mayer, William Paganini; de Campos Boldrini, Silvia; Liberti, Edson Aparecido

    2009-03-01

    The adductor canal is a conical or pyramid-shaped pathway that contains the femoral vessels, saphenous nerve and a varying amount of fibrous tissue. It is involved in adductor canal syndrome, a claudication syndrome involving young individuals. Our objective was to study modifications induced by aging on the connective tissue and to correlate them to the proposed pathophysiological mechanism. The bilateral adductor canals and femoral vessels of four adult and five fetal specimens were removed en bloc and analyzed. Sections 12 microm thick were obtained and the connective tissue studied with Sirius Red, Verhoeff, Weigert and Azo stains. Scanning electron microscopy (SEM) photomicrographs of the surfaces of each adductor canal were also analyzed. Findings were homogeneous inside each group. The connective tissue of the canal was continuous with the outer layer of the vessels in both groups. The pattern of concentric, thick collagen type I bundles in fetal specimens was replaced by a diffuse network of compact collagen bundles with several transversal fibers and an impressive content of collagen III fibers. Elastic fibers in adults were not concentrated in the thick bundles but dispersed in line with the transversal fiber system. A dynamic compression mechanism with or without an evident constricting fibrous band has been proposed previously for adductor canal syndrome, possibly involving the connective tissue inside the canal. The vessels may not slide freely during movement. These age-related modifications in normal individuals may represent necessary conditions for this syndrome to develop.

  20. The connective tissue of the adductor canal – a morphological study in fetal and adult specimens

    PubMed Central

    de Oliveira, Flavia; de Vasconcellos Fontes, Ricardo Bragança; da Silva Baptista, Josemberg; Mayer, William Paganini; de Campos Boldrini, Silvia; Liberti, Edson Aparecido

    2009-01-01

    The adductor canal is a conical or pyramid-shaped pathway that contains the femoral vessels, saphenous nerve and a varying amount of fibrous tissue. It is involved in adductor canal syndrome, a claudication syndrome involving young individuals. Our objective was to study modifications induced by aging on the connective tissue and to correlate them to the proposed pathophysiological mechanism. The bilateral adductor canals and femoral vessels of four adult and five fetal specimens were removed en bloc and analyzed. Sections 12 µm thick were obtained and the connective tissue studied with Sirius Red, Verhoeff, Weigert and Azo stains. Scanning electron microscopy (SEM) photomicrographs of the surfaces of each adductor canal were also analyzed. Findings were homogeneous inside each group. The connective tissue of the canal was continuous with the outer layer of the vessels in both groups. The pattern of concentric, thick collagen type I bundles in fetal specimens was replaced by a diffuse network of compact collagen bundles with several transversal fibers and an impressive content of collagen III fibers. Elastic fibers in adults were not concentrated in the thick bundles but dispersed in line with the transversal fiber system. A dynamic compression mechanism with or without an evident constricting fibrous band has been proposed previously for adductor canal syndrome, possibly involving the connective tissue inside the canal. The vessels may not slide freely during movement. These age-related modifications in normal individuals may represent necessary conditions for this syndrome to develop. PMID:19245505

  1. Quantitative mapping of collagen fiber alignment in thick tissue samples using transmission polarized-light microscopy.

    PubMed

    Yakovlev, Dmitry D; Shvachkina, Marina E; Sherman, Maria M; Spivak, Andrey V; Pravdin, Alexander B; Yakovlev, Dmitry A

    2016-07-01

    Immersion optical clearing makes it possible to use transmission polarized-light microscopy for characterization of thick (200 to 2000  μm) layers of biological tissues. We discuss polarization properties of thick samples in the context of the problem of characterization of collagen fiber alignment in connective tissues such as sclera and dermis. Optical chirality caused by azimuthal variations of the macroscopic (effective) optic axis of the medium across the sample thickness should be considered in polarization mapping of thick samples of these tissues. We experimentally evaluate to what extent the optical chirality affects the measurement results in typical situations and show under what conditions it can be easily taken into account and does not hinder, but rather helps, in characterization of collagen fiber alignment.

  2. Quantitative mapping of collagen fiber alignment in thick tissue samples using transmission polarized-light microscopy

    NASA Astrophysics Data System (ADS)

    Yakovlev, Dmitry D.; Shvachkina, Marina E.; Sherman, Maria M.; Spivak, Andrey V.; Pravdin, Alexander B.; Yakovlev, Dmitry A.

    2016-07-01

    Immersion optical clearing makes it possible to use transmission polarized-light microscopy for characterization of thick (200 to 2000 μm) layers of biological tissues. We discuss polarization properties of thick samples in the context of the problem of characterization of collagen fiber alignment in connective tissues such as sclera and dermis. Optical chirality caused by azimuthal variations of the macroscopic (effective) optic axis of the medium across the sample thickness should be considered in polarization mapping of thick samples of these tissues. We experimentally evaluate to what extent the optical chirality affects the measurement results in typical situations and show under what conditions it can be easily taken into account and does not hinder, but rather helps, in characterization of collagen fiber alignment.

  3. Isolation of high quality protein samples from punches of formalin fixed and paraffin embedded tissue blocks.

    PubMed

    Kroll, J; Becker, K F; Kuphal, S; Hein, R; Hofstädter, F; Bosserhoff, A K

    2008-04-01

    In general, it is believed that the extraction of proteins from formalin-fixed paraffin embedded samples is not feasible. However, recently a new technique was developed, presenting the extraction of non-degraded, full length proteins from formalin fixed tissues, usable for western blotting and protein arrays. In the study presented here, we applied this technique to punch biopsies of formalin fixed tissues embedded in paraffin to reduce heterogeneity of the tissue represented in sections, and to ensure analysing mainly defined cellular material. Successful extraction was achieved even from very small samples (0.7 mm(3)). Additionally, we were able to detect highly glycosylated proteins and protein modification, such as phosphorylation. Interestingly, with this technique it is feasible to extract high quality proteins from 14 year old samples. In summary, the new technique makes a great pool of material now usable for molecular analysis with high throughput tools. PMID:18228195

  4. Method for the detection of desmethylbromethalin in animal tissue samples for the determination of bromethalin exposure.

    PubMed

    Filigenzi, Michael S; Bautista, Adrienne C; Aston, Linda S; Poppenga, Robert H

    2015-06-01

    Bromethalin, a potent neurotoxin, is widely available for use as a rodenticide. As access to other rodenticides is reduced due to regulatory pressure, the use of bromethalin is likely to increase with a concomitant increase in poisonings in nontarget animals. Analytical methods for the detection of bromethalin residues in animals suspected to have been exposed to this rodenticide are needed to support post-mortem diagnosis of toxicosis. This paper describes a novel method for the analysis of desmethylbromethalin (DMB), bromethalin's toxic metabolite, in tissue samples such as liver, brain, and adipose. Samples were extracted with 5% ethanol in ethyl acetate, and an aliquot of the extract was evaporated dry, reconstituted, and analyzed by reverse phase ultrahigh-performance liquid chromatography-mass spectrometry. The mass spectrometer utilized electrospray ionization in negative ion mode with multiple reaction monitoring. This method was qualitatively validated at a level of 1.0 ng/g in liver tissue. The quantitative potential of the method was also evaluated, and a method detection limit of 0.35 ng/g wet weight was determined in fat tissue. DMB was detected in tissue samples from animals suspected to have been poisoned by this compound. To the authors' knowledge, there have been no other methods reported for analysis of DMB in tissue samples using LC-MS/MS. PMID:25688571

  5. Sample processing considerations for detecting copy number changes in formalin-fixed, paraffin-embedded tissues.

    PubMed

    Jacobs, Sharoni

    2012-11-01

    The Whole Genome Sampling Analysis (WGSA) assay in combination with Affymetrix GeneChip Mapping Arrays is used for copy number analysis of high-quality DNA samples (i.e., samples that have been collected from blood, fresh or frozen tissue, or cell lines). Formalin-fixed, paraffin-embedded (FFPE) samples, however, represent the most prevalent form of archived clinical samples, but they provide additional challenges for molecular assays. FFPE processing usually results in the degradation of FFPE DNA and in the contamination and chemical modification of these DNA samples. Because of these issues, FFPE DNA is not suitable for all molecular assays designed for high-quality DNA samples. Strategies recommended for processing FFPE DNA samples through WGSA and to the Mapping arrays are described here. PMID:23118355

  6. Sample processing considerations for detecting copy number changes in formalin-fixed, paraffin-embedded tissues.

    PubMed

    Jacobs, Sharoni

    2012-11-01

    The Whole Genome Sampling Analysis (WGSA) assay in combination with Affymetrix GeneChip Mapping Arrays is used for copy number analysis of high-quality DNA samples (i.e., samples that have been collected from blood, fresh or frozen tissue, or cell lines). Formalin-fixed, paraffin-embedded (FFPE) samples, however, represent the most prevalent form of archived clinical samples, but they provide additional challenges for molecular assays. FFPE processing usually results in the degradation of FFPE DNA and in the contamination and chemical modification of these DNA samples. Because of these issues, FFPE DNA is not suitable for all molecular assays designed for high-quality DNA samples. Strategies recommended for processing FFPE DNA samples through WGSA and to the Mapping arrays are described here.

  7. Identifying proteins in zebrafish embryos using spectral libraries generated from dissected adult organs and tissues.

    PubMed

    van der Plas-Duivesteijn, Suzanne J; Mohammed, Yassene; Dalebout, Hans; Meijer, Annemarie; Botermans, Anouk; Hoogendijk, Jordy L; Henneman, Alex A; Deelder, André M; Spaink, Herman P; Palmblad, Magnus

    2014-03-01

    Spectral libraries provide a sensitive and accurate method for identifying peptides from tandem mass spectra, complementary to searching genome-derived databases or sequencing de novo. Their application requires comprehensive libraries including peptides from low-abundant proteins. Here we describe a method for constructing such libraries using biological differentiation to "fractionate" the proteome by harvesting adult organs and tissues and build comprehensive libraries for identifying proteins in zebrafish (Danio rerio) embryos and larvae (an important and widely used model system). Hierarchical clustering using direct comparison of spectra was used to prioritize organ selection. The resulting and publicly available library covers 14,164 proteins, significantly improved the number of peptide-spectrum matches in zebrafish developmental stages, and can be used on data from different instruments and laboratories. The library contains information on tissue and organ expression of these proteins and is also applicable for adult experiments. The approach itself is not limited to zebrafish but would work for any model system.

  8. Lack of tissue renewal in human adult Achilles tendon is revealed by nuclear bomb (14)C.

    PubMed

    Heinemeier, Katja Maria; Schjerling, Peter; Heinemeier, Jan; Magnusson, Stig Peter; Kjaer, Michael

    2013-05-01

    Tendons are often injured and heal poorly. Whether this is caused by a slow tissue turnover is unknown, since existing data provide diverging estimates of tendon protein half-life that range from 2 mo to 200 yr. With the purpose of determining life-long turnover of human tendon tissue, we used the (14)C bomb-pulse method. This method takes advantage of the dramatic increase in atmospheric levels of (14)C, produced by nuclear bomb tests in 1955-1963, which is reflected in all living organisms. Levels of (14)C were measured in 28 forensic samples of Achilles tendon core and 4 skeletal muscle samples (donor birth years 1945-1983) with accelerator mass spectrometry (AMS) and compared to known atmospheric levels to estimate tissue turnover. We found that Achilles tendon tissue retained levels of (14)C corresponding to atmospheric levels several decades before tissue sampling, demonstrating a very limited tissue turnover. The tendon concentrations of (14)C approximately reflected the atmospheric levels present during the first 17 yr of life, indicating that the tendon core is formed during height growth and is essentially not renewed thereafter. In contrast, (14)C levels in muscle indicated continuous turnover. Our observation provides a fundamental premise for understanding tendon function and pathology, and likely explains the poor regenerative capacity of tendon tissue.

  9. Automated MALDI matrix coating system for multiple tissue samples for imaging mass spectrometry.

    PubMed

    Mounfield, William P; Garrett, Timothy J

    2012-03-01

    Uniform matrix deposition on tissue samples for matrix-assisted laser desorption/ionization (MALDI) is key for reproducible analyte ion signals. Current methods often result in nonhomogenous matrix deposition, and take time and effort to produce acceptable ion signals. Here we describe a fully-automated method for matrix deposition using an enclosed spray chamber and spray nozzle for matrix solution delivery. A commercial air-atomizing spray nozzle was modified and combined with solenoid controlled valves and a Programmable Logic Controller (PLC) to control and deliver the matrix solution. A spray chamber was employed to contain the nozzle, sample, and atomized matrix solution stream, and to prevent any interference from outside conditions as well as allow complete control of the sample environment. A gravity cup was filled with MALDI matrix solutions, including DHB in chloroform/methanol (50:50) at concentrations up to 60 mg/mL. Various samples (including rat brain tissue sections) were prepared using two deposition methods (spray chamber, inkjet). A linear ion trap equipped with an intermediate-pressure MALDI source was used for analyses. Optical microscopic examination showed a uniform coating of matrix crystals across the sample. Overall, the mass spectral images gathered from tissues coated using the spray chamber system were of better quality and more reproducible than from tissue specimens prepared by the inkjet deposition method.

  10. Rapid mass spectrometric conversion of tissue biopsy samples into permanent quantitative digital proteome maps

    PubMed Central

    Guo, Tiannan; Kouvonen, Petri; Koh, Ching Chiek; Gillet, Ludovic C; Wolski, Witold E; Röst, Hannes L; Rosenberger, George; Collins, Ben C; Blum, Lorenz C; Gillessen, Silke; Joerger, Markus; Jochum, Wolfram; Aebersold, Ruedi

    2015-01-01

    Clinical specimens are each inherently unique, limited and non-renewable. As such, small samples such as tissue biopsies are often completely consumed after a limited number of analyses. Here we present a method that enables fast and reproducible conversion of a small amount of tissue (approximating the quantity obtained by a biopsy) into a single, permanent digital file representing the mass spectrometry-measurable proteome of the sample. The method combines pressure cycling technology (PCT) and SWATH mass spectrometry (MS), and the resulting proteome maps can be analyzed, re-analyzed, compared and mined in silico to detect and quantify specific proteins across multiple samples. We used this method to process and convert 18 biopsy samples from 9 renal cell carcinoma patients into SWATH-MS fragment ion maps. From these proteome maps we detected and quantified more than 2,000 proteins with a high degree of reproducibility across all samples. The identified proteins clearly separated tumorous kidney tissues from healthy tissue, and differentiated distinct histomorphological kidney cancer subtypes. PMID:25730263

  11. Automated MALDI Matrix Coating System for Multiple Tissue Samples for Imaging Mass Spectrometry

    NASA Astrophysics Data System (ADS)

    Mounfield, William P.; Garrett, Timothy J.

    2012-03-01

    Uniform matrix deposition on tissue samples for matrix-assisted laser desorption/ionization (MALDI) is key for reproducible analyte ion signals. Current methods often result in nonhomogenous matrix deposition, and take time and effort to produce acceptable ion signals. Here we describe a fully-automated method for matrix deposition using an enclosed spray chamber and spray nozzle for matrix solution delivery. A commercial air-atomizing spray nozzle was modified and combined with solenoid controlled valves and a Programmable Logic Controller (PLC) to control and deliver the matrix solution. A spray chamber was employed to contain the nozzle, sample, and atomized matrix solution stream, and to prevent any interference from outside conditions as well as allow complete control of the sample environment. A gravity cup was filled with MALDI matrix solutions, including DHB in chloroform/methanol (50:50) at concentrations up to 60 mg/mL. Various samples (including rat brain tissue sections) were prepared using two deposition methods (spray chamber, inkjet). A linear ion trap equipped with an intermediate-pressure MALDI source was used for analyses. Optical microscopic examination showed a uniform coating of matrix crystals across the sample. Overall, the mass spectral images gathered from tissues coated using the spray chamber system were of better quality and more reproducible than from tissue specimens prepared by the inkjet deposition method.

  12. Soft Tissue Deformations Contribute to the Mechanics of Walking in Obese Adults

    PubMed Central

    Fu, Xiao-Yu; Zelik, Karl E.; Board, Wayne J.; Browning, Raymond C.; Kuo, Arthur D.

    2014-01-01

    Obesity not only adds to the mass that must be carried during walking, but also changes body composition. Although extra mass causes roughly proportional increases in musculoskeletal loading, less well understood is the effect of relatively soft and mechanically compliant adipose tissue. Purpose To estimate the work performed by soft tissue deformations during walking. The soft tissue would be expected to experience damped oscillations, particularly from high force transients following heel strike, and could potentially change the mechanical work demands for walking. Method We analyzed treadmill walking data at 1.25 m/s for 11 obese (BMI > 30 kg/m2) and 9 non-obese (BMI < 30 kg/m2) adults. The soft tissue work was quantified with a method that compares the work performed by lower extremity joints as derived using assumptions of rigid body segments, with that estimated without rigid body assumptions. Results Relative to body mass, obese and non-obese individuals perform similar amounts of mechanical work. But negative work performed by soft tissues was significantly greater in obese individuals (p= 0.0102), equivalent to about 0.36 J/kg vs. 0.27 J/kg in non-obese individuals. The negative (dissipative) work by soft tissues occurred mainly after heel strike, and for obese individuals was comparable in magnitude to the total negative work from all of the joints combined (0.34 J/kg vs. 0.33 J/kg for obese and non-obese adults, respectively). Although the joints performed a relatively similar amount of work overall, obese individuals performed less negative work actively at the knee. Conclusion The greater proportion of soft tissues in obese individuals results in substantial changes in the amount, location, and timing of work, and may also impact metabolic energy expenditure during walking. PMID:25380475

  13. Enrichment of a bipotent hepatic progenitor cell from naive adult liver tissue

    SciTech Connect

    Wright, Natasha; Samuelson, Lisa; Walkup, Maggie H.; Chandrasekaran, Prakash; Gerber, David A.

    2008-02-08

    Background/Aim: Recent interest in the liver stem cell field has led to the identification and characterization of several hepatic progenitor cell populations from fetal and adult tissues. We isolated a hepatic progenitor cell from naive adult liver and the current studies focus on differentiation and growth. Results: A Sca-1{sup +} hepatic progenitor cell was identified within the liver parenchyma. This cell expresses numerous liver related genes and transcription found in the developing and/or adult liver. It is located in the peri-portal region and expresses markers associated with undifferentiated hepatic cell populations, mature hepatocytes and biliary cells which distinguish it from the Sca-1{sup -} fraction. Conclusion: This hepatic progenitor cell from uninjured liver has features of both hepatocytic and biliary populations and demonstrates proliferative potential. Further studies will focus on sca-HPC subsets and conditions that regulate differentiation towards hepatic or biliary lineages.

  14. Rapid quantification of inflammation in tissue samples using perfluorocarbon emulsion and fluorine-19 nuclear magnetic resonance

    PubMed Central

    Ahrens, Eric T.; Young, Won-Bin; Xu, Hongyan; Pusateri, Lisa K.

    2016-01-01

    Quantification of inflammation in tissue samples can be a time-intensive bottleneck in therapeutic discovery and preclinical endeavors. We describe a versatile and rapid approach to quantitatively assay macrophage burden in intact tissue samples. Perfluorocarbon (PFC) emulsion is injected intravenously, and the emulsion droplets are effectively taken up by monocytes and macrophages. These ‘in situ’ labeled cells participate in inflammatory events in vivo resulting in PFC accumulation at inflammatory loci. Necropsied tissues or intact organs are subjected to conventional fluorine-19 (19F) NMR spectroscopy to quantify the total fluorine content per sample, proportional to the macrophage burden. We applied these methods to a rat model of experimental allergic encephalomyelitis (EAE) exhibiting extensive inflammation and demyelination in the central nervous system (CNS), particularly in the spinal cord. In a cohort of EAE rats, we used 19F NMR to derive an inflammation index (IFI) in intact CNS tissues. Immunohistochemistry was used to confirm intracellular colocalization of the PFC droplets within CNS CD68+ cells having macrophage morphology. The IFI linearly correlated to mRNA levels of CD68 via real-time PCR analysis. This 19F NMR approach can accelerate tissue analysis by at least an order of magnitude compared with histological approaches. PMID:21548906

  15. Spatial pattern and sequential sampling of squash bug (Heteroptera: Coreidae) adults in watermelon.

    PubMed

    Dogramaci, Mahmut; Shrefler, James W; Giles, Kristopher; Edelson, J V

    2006-04-01

    Spatial distribution patterns of adult squash bugs were determined in watermelon, Citrullus lanatus (Thunberg) Matsumura and Nakai, during 2001 and 2002. Results of analysis using Taylor's power law regression model indicated that squash bugs were aggregated in watermelon. Taylor's power law provided a good fit with r2 = 0.94. A fixed precision sequential sampling plan was developed for estimating adult squash bug density at fixed precision levels in watermelon. The plan was tested using a resampling simulation method on nine and 13 independent data sets ranging in density from 0.15 to 2.52 adult squash bugs per plant. Average estimated means obtained in 100 repeated simulation runs were within the 95% CI of the true means for all the data. Average estimated levels of precision were similar to the desired level of precision, particularly when the sampling plan was tested on data having an average mean density of 1.19 adult squash bugs per plant. Also, a sequential sampling for classifying adult squash bug density as below or above economic threshold was developed to assist in the decision-making process. The classification sampling plan is advantageous in that it requires smaller sample sizes to estimate the population status when the population density differs greatly from the action threshold. However, the plan may require excessively large sample sizes when the density is close to the threshold. Therefore, an integrated sequential sampling plan was developed using a combination of a fixed precision and classification sequential sampling plans. The integration of sampling plans can help reduce sampling requirements. PMID:16686160

  16. Semiquantitative determination of polychlorinated biphenyls in tissue samples by thin layer chromatography

    USGS Publications Warehouse

    Mulhern, B.M.; Cromartie, E.; Reichel, W.L.; Belisle, A.A.

    1971-01-01

    A method is described for the analysis of polychlorinated biphenyl (PCB) compounds in tissue samples. Cleanup by hexane-aceto-nitrile partitioning and Florisil column chromatography are performed on samples before oxidative treatment to convert DDE to DCBP. PCB components are then determined semi-quantitatively by TLC. No prior separation of PCB from chlorinated pesticides is required. The lower limit of sensitivity is 0.2 ?g.

  17. Three Dimensional Imaging of Paraffin Embedded Human Lung Tissue Samples by Micro-Computed Tomography

    PubMed Central

    Scott, Anna E.; Vasilescu, Dragos M.; Seal, Katherine A. D.; Keyes, Samuel D.; Mavrogordato, Mark N.; Hogg, James C.; Sinclair, Ian; Warner, Jane A.; Hackett, Tillie-Louise; Lackie, Peter M.

    2015-01-01

    Background Understanding the three-dimensional (3-D) micro-architecture of lung tissue can provide insights into the pathology of lung disease. Micro computed tomography (µCT) has previously been used to elucidate lung 3D histology and morphometry in fixed samples that have been stained with contrast agents or air inflated and dried. However, non-destructive microstructural 3D imaging of formalin-fixed paraffin embedded (FFPE) tissues would facilitate retrospective analysis of extensive tissue archives of lung FFPE lung samples with linked clinical data. Methods FFPE human lung tissue samples (n = 4) were scanned using a Nikon metrology µCT scanner. Semi-automatic techniques were used to segment the 3D structure of airways and blood vessels. Airspace size (mean linear intercept, Lm) was measured on µCT images and on matched histological sections from the same FFPE samples imaged by light microscopy to validate µCT imaging. Results The µCT imaging protocol provided contrast between tissue and paraffin in FFPE samples (15mm x 7mm). Resolution (voxel size 6.7 µm) in the reconstructed images was sufficient for semi-automatic image segmentation of airways and blood vessels as well as quantitative airspace analysis. The scans were also used to scout for regions of interest, enabling time-efficient preparation of conventional histological sections. The Lm measurements from µCT images were not significantly different to those from matched histological sections. Conclusion We demonstrated how non-destructive imaging of routinely prepared FFPE samples by laboratory µCT can be used to visualize and assess the 3D morphology of the lung including by morphometric analysis. PMID:26030902

  18. Aberrant Synaptic Integration in Adult Lamina I Projection Neurons Following Neonatal Tissue Damage

    PubMed Central

    Li, Jie; Kritzer, Elizabeth; Craig, Paige E.

    2015-01-01

    Mounting evidence suggests that neonatal tissue damage evokes alterations in spinal pain reflexes which persist into adulthood. However, less is known about potential concomitant effects on the transmission of nociceptive information to the brain, as the degree to which early injury modulates synaptic integration and membrane excitability in mature spinal projection neurons remains unclear. Here we demonstrate that neonatal surgical injury leads to a significant shift in the balance between synaptic excitation and inhibition onto identified lamina I projection neurons of the adult mouse spinal cord. The strength of direct primary afferent input to mature spino-parabrachial neurons was enhanced following neonatal tissue damage, whereas the efficacy of both GABAergic and glycinergic inhibition onto the same population was compromised. This was accompanied by reorganization in the pattern of sensory input to adult projection neurons, which included a greater prevalence of monosynaptic input from low-threshold A-fibers when preceded by early tissue damage. In addition, neonatal incision resulted in greater primary afferent-evoked action potential discharge in mature projection neurons. Overall, these results demonstrate that tissue damage during early life causes a long-term increase in the gain of spinal nociceptive circuits, and suggest that the prolonged consequences of neonatal trauma may not be restricted to the spinal cord but rather include excessive ascending signaling to supraspinal pain centers. PMID:25673839

  19. Phase-contrast Hounsfield units of fixated and non-fixated soft-tissue samples

    SciTech Connect

    Willner, Marian; Fior, Gabriel; Marschner, Mathias; Birnbacher, Lorenz; Schock, Jonathan; Braun, Christian; Fingerle, Alexander A.; Noël, Peter B.; Rummeny, Ernst J.; Pfeiffer, Franz; Herzen, Julia; Rozhkova, Elena A.

    2015-08-31

    X-ray phase-contrast imaging is a novel technology that achieves high soft-tissue contrast. Although its clinical impact is still under investigation, the technique may potentially improve clinical diagnostics. In conventional attenuation-based X-ray computed tomography, radiological diagnostics are quantified by Hounsfield units. Corresponding Hounsfield units for phase-contrast imaging have been recently introduced, enabling a setup-independent comparison and standardized interpretation of imaging results. Thus far, the experimental values of few tissue types have been reported; these values have been determined from fixated tissue samples. This study presents phase-contrast Hounsfield units for various types of non-fixated human soft tissues. A large variety of tissue specimens ranging from adipose, muscle and connective tissues to liver, kidney and pancreas tissues were imaged by a grating interferometer with a rotating-anode X-ray tube and a photon-counting detector. In addition, we investigated the effects of formalin fixation on the quantitative phase-contrast imaging results.

  20. [The three-dimensional culture of adult mesenchymal stem cells for intervertebral disc tissue engineering].

    PubMed

    Feng, Ganjun; Liu, Hao; Deng, Li; Chen, Xiaohe; Zhao, Xianfeng; Liang, Tao; Li, Xiuqiong

    2009-12-01

    Intervertebral disc (IVD) degeneration is one of the major causes of low back pain. As current clinical treatments are aimed at restoring biomechanical function and providing symptomatic relief, the methods focused on biological repair have aroused interest and several tissue engineering approaches using different cell types have been proposed. Owing to the unsuitable nature of degenerate cells for tissue engineering, attention has been given to the use of mesenchymal stem cells (MSCs). In this connection, we have made a study on the characteristics of MSCs derived from adult bone marrow and on the feasibility of constructing IVD tissue-engineering cell under a Three-Dimensional Pellet Culture System. The human bone marrow MSCs were isolated and purified with density gradient solution and attachment-independent culture system. MSCs isolated using this method are a homogeneous population as indicated by morphology and other criteria. They have the capacity for self-renewal and proliferation, and the multilineage potential to differentiate.

  1. Evaluation of frozen tissue-derived prognostic gene expression signatures in FFPE colorectal cancer samples

    PubMed Central

    Zhu, Jing; Deane, Natasha G.; Lewis, Keeli B.; Padmanabhan, Chandrasekhar; Washington, M. Kay; Ciombor, Kristen K.; Timmers, Cynthia; Goldberg, Richard M.; Beauchamp, R. Daniel; Chen, Xi

    2016-01-01

    Defining molecular features that can predict the recurrence of colorectal cancer (CRC) for stage II-III patients remains challenging in cancer research. Most available clinical samples are Formalin-Fixed, Paraffin-Embedded (FFPE). NanoString nCounter® and Affymetrix GeneChip® Human Transcriptome Array 2.0 (HTA) are the two platforms marketed for high-throughput gene expression profiling for FFPE samples. In this study, to evaluate the gene expression of frozen tissue-derived prognostic signatures in FFPE CRC samples, we evaluated the expression of 516 genes from published frozen tissue-derived prognostic signatures in 42 FFPE CRC samples measured by both platforms. Based on HTA platform-derived data, we identified both gene (99 individual genes, FDR < 0.05) and gene set (four of the six reported multi-gene signatures with sufficient information for evaluation, P < 0.05) expression differences associated with survival outcomes. Using nCounter platform-derived data, one of the six multi-gene signatures (P < 0.05) but no individual gene was associated with survival outcomes. Our study indicated that sufficiently high quality RNA could be obtained from FFPE tumor tissues to detect frozen tissue-derived prognostic gene expression signatures for CRC patients. PMID:27623752

  2. Comparison of Microdialysis Sampling Perfusion Fluid Components on the Foreign Body Reaction in Rat Subcutaneous Tissue

    PubMed Central

    Keeler, Geoffrey D.; Durdik, Jeannine M.; Stenken, Julie A.

    2013-01-01

    Microdialysis sampling is a commonly used technique for collecting solutes from the extracellular space of tissues in laboratory animals and humans. Large molecular weight solutes can be collected using high molecular weight cutoff (MWCO) membranes (100 kDa or greater). High MWCO membranes require addition of high molecular weight dextrans or albumin to the perfusion fluid to prevent fluid loss via ultrafiltration. While these perfusion fluid additives are commonly used during microdialysis sampling, the tissue response to the loss of these compounds across the membrane is poorly understood. Tissue reactions to implanted microdialysis sampling probes containing different microdialysis perfusion fluids were compared over a 7-day time period in rats. The base perfusion fluid was Ringer’s solution supplemented with either bovine serum albumin (BSA), rat serum albumin (RSA), Dextran-70, or Dextran-500. A significant inflammatory response to Dextran-70 was observed. No differences in the tissue response between BSA and RSA were observed. Among these agents, the BSA, RSA, and Dextran-500 produced a significantly reduced inflammatory response compared to the Dextran-70. This work demonstrates that use of Dextran-70 in microdialysis sampling perfusion fluids should be eliminated and replaced with Dextran-500 or other alternatives. PMID:24239995

  3. Evaluation of frozen tissue-derived prognostic gene expression signatures in FFPE colorectal cancer samples.

    PubMed

    Zhu, Jing; Deane, Natasha G; Lewis, Keeli B; Padmanabhan, Chandrasekhar; Washington, M Kay; Ciombor, Kristen K; Timmers, Cynthia; Goldberg, Richard M; Beauchamp, R Daniel; Chen, Xi

    2016-01-01

    Defining molecular features that can predict the recurrence of colorectal cancer (CRC) for stage II-III patients remains challenging in cancer research. Most available clinical samples are Formalin-Fixed, Paraffin-Embedded (FFPE). NanoString nCounter® and Affymetrix GeneChip® Human Transcriptome Array 2.0 (HTA) are the two platforms marketed for high-throughput gene expression profiling for FFPE samples. In this study, to evaluate the gene expression of frozen tissue-derived prognostic signatures in FFPE CRC samples, we evaluated the expression of 516 genes from published frozen tissue-derived prognostic signatures in 42 FFPE CRC samples measured by both platforms. Based on HTA platform-derived data, we identified both gene (99 individual genes, FDR < 0.05) and gene set (four of the six reported multi-gene signatures with sufficient information for evaluation, P < 0.05) expression differences associated with survival outcomes. Using nCounter platform-derived data, one of the six multi-gene signatures (P < 0.05) but no individual gene was associated with survival outcomes. Our study indicated that sufficiently high quality RNA could be obtained from FFPE tumor tissues to detect frozen tissue-derived prognostic gene expression signatures for CRC patients. PMID:27623752

  4. Pathology, physiologic parameters, tissue contaminants, and tissue thiamine in morbid and healthy central Florida adult American alligators (Alligator mississippiensis)

    USGS Publications Warehouse

    Honeyfield, D.C.; Ross, J.P.; Carbonneau, D.A.; Terrell, S.P.; Woodward, A.R.; Schoeb, T.R.; Perceval, H.F.; Hinterkopf, J.P.

    2008-01-01

    An investigation of adult alligator (Alligator mississippiensis) mortalities in Lake Griffin, central Florida, was conducted from 1998-2004. Alligator mortality was highest in the months of April and May and annual death count peaked in 2000. Bacterial pathogens, heavy metals, and pesticides were not linked with the mortalities. Blood chemistry did not point to any clinical diagnosis, although differences between impaired and normal animals were noted. Captured alligators with signs of neurologic impairment displayed unresponsive and uncoordinated behavior. Three of 21 impaired Lake Griffin alligators were found to have neural lesions characteristic of thiamine deficiency in the telencephalon, particularly the dorsal ventricular ridge. In some cases, lesions were found in the thalamus, and parts of the midbrain. Liver and muscle tissue concentrations of thiamine (vitamin B"1) were lowest in impaired Lake Griffin alligators when compared to unimpaired alligators or to alligators from Lake Woodruff. The consumption of thiaminase-positive gizzard shad (Dorosoma cepedianum) is thought to have been the cause of the low tissue thiamine and resulting mortalities. ?? Wildlife Disease Association 2008.

  5. Tissue sampling from live blue mussels, Mytilus edulis. A field study from the Swedish west coast

    NASA Astrophysics Data System (ADS)

    Svärdh, Lillemor

    2003-05-01

    Histological techniques are often used to study environmental effects on mussels, but since these techniques include killing of the individuals, rare or endangered populations cannot be studied using conventional tissue sampling. This study is an attempt to find a method that can be used repeatedly with the same mussel individual and which does not affect growth and survival. From 200 mussels, Mytilus edulis, tissue was sampled in different ways, such as drilling a hole in the shell or prising apart the shell valves. Two kind of instruments were used, an injection needle and surgery forceps. Some of the drilled mussels had their holes sealed again with cement. Drilling a hole in the shell, removing tissue sample with surgical forceps and then leaving the holes open did not seriously harm the mussels during the two months the experiment lasted. But if the holes were sealed with cement, both length and weight growth were negatively affected (35% lower length growth and 36% lower weight growth compared to the control mussels). Mortality was highest among the drilled and sealed mussels (80% higher than among the other treatments). The vulnerability of the population, the aim of the study and the duration of the experiment should decide what method to use for tissue sampling. For long-term experiments and repeated sampling, opening the mussels by prizing apart the valves is a better alternative than drilling holes in the shells, but depending on the morphology of the species it could be difficult to sample the anterior part of the mussel body. For a short experiment and to sample anterior parts, drilling the shells, leaving the holes open and using surgical forceps, seems to be an acceptable compromise between the different treatments used.

  6. The Novel Application of Non-Lethal Citizen Science Tissue Sampling in Recreational Fisheries.

    PubMed

    Williams, Samuel M; Holmes, Bonnie J; Pepperell, Julian G

    2015-01-01

    Increasing fishing pressure and uncertainty surrounding recreational fishing catch and effort data promoted the development of alternative methods for conducting fisheries research. A pilot investigation was undertaken to engage the Australian game fishing community and promote the non-lethal collection of tissue samples from the black marlin Istiompax indica, a valuable recreational-only species in Australian waters, for the purpose of future genetic research. Recruitment of recreational anglers was achieved by publicizing the project in magazines, local newspapers, social media, blogs, websites and direct communication workshops at game fishing tournaments. The Game Fishing Association of Australia and the Queensland Game Fishing Association were also engaged to advertise the project and recruit participants with a focus on those anglers already involved in the tag-and-release of marlin. Participants of the program took small tissue samples using non-lethal methods which were stored for future genetic analysis. The program resulted in 165 samples from 49 participants across the known distribution of I. indica within Australian waters which was a sufficient number to facilitate a downstream population genetic analysis. The project demonstrated the potential for the development of citizen science sampling programs to collect tissue samples using non-lethal methods in order to achieve targeted research objects in recreationally caught species. PMID:26376487

  7. The Novel Application of Non-Lethal Citizen Science Tissue Sampling in Recreational Fisheries.

    PubMed

    Williams, Samuel M; Holmes, Bonnie J; Pepperell, Julian G

    2015-01-01

    Increasing fishing pressure and uncertainty surrounding recreational fishing catch and effort data promoted the development of alternative methods for conducting fisheries research. A pilot investigation was undertaken to engage the Australian game fishing community and promote the non-lethal collection of tissue samples from the black marlin Istiompax indica, a valuable recreational-only species in Australian waters, for the purpose of future genetic research. Recruitment of recreational anglers was achieved by publicizing the project in magazines, local newspapers, social media, blogs, websites and direct communication workshops at game fishing tournaments. The Game Fishing Association of Australia and the Queensland Game Fishing Association were also engaged to advertise the project and recruit participants with a focus on those anglers already involved in the tag-and-release of marlin. Participants of the program took small tissue samples using non-lethal methods which were stored for future genetic analysis. The program resulted in 165 samples from 49 participants across the known distribution of I. indica within Australian waters which was a sufficient number to facilitate a downstream population genetic analysis. The project demonstrated the potential for the development of citizen science sampling programs to collect tissue samples using non-lethal methods in order to achieve targeted research objects in recreationally caught species.

  8. The Novel Application of Non-Lethal Citizen Science Tissue Sampling in Recreational Fisheries

    PubMed Central

    Williams, Samuel M.; Holmes, Bonnie J.; Pepperell, Julian G.

    2015-01-01

    Increasing fishing pressure and uncertainty surrounding recreational fishing catch and effort data promoted the development of alternative methods for conducting fisheries research. A pilot investigation was undertaken to engage the Australian game fishing community and promote the non-lethal collection of tissue samples from the black marlin Istiompax indica, a valuable recreational-only species in Australian waters, for the purpose of future genetic research. Recruitment of recreational anglers was achieved by publicizing the project in magazines, local newspapers, social media, blogs, websites and direct communication workshops at game fishing tournaments. The Game Fishing Association of Australia and the Queensland Game Fishing Association were also engaged to advertise the project and recruit participants with a focus on those anglers already involved in the tag-and-release of marlin. Participants of the program took small tissue samples using non-lethal methods which were stored for future genetic analysis. The program resulted in 165 samples from 49 participants across the known distribution of I. indica within Australian waters which was a sufficient number to facilitate a downstream population genetic analysis. The project demonstrated the potential for the development of citizen science sampling programs to collect tissue samples using non-lethal methods in order to achieve targeted research objects in recreationally caught species. PMID:26376487

  9. Non-lethal sampling of walleye for stable isotope analysis: a comparison of three tissues

    USGS Publications Warehouse

    Chipps, Steven R.; VanDeHey, J.A.; Fincel, M.J.

    2012-01-01

    Stable isotope analysis of fishes is often performed using muscle or organ tissues that require sacrificing animals. Non-lethal sampling provides an alternative for evaluating isotopic composition for species of concern or individuals of exceptional value. Stable isotope values of white muscle (lethal) were compared with those from fins and scales (non-lethal) in walleye, Sander vitreus (Mitchill), from multiple systems, size classes and across a range of isotopic values. Isotopic variability was also compared among populations to determine the potential of non-lethal tissues for diet-variability analyses. Muscle-derived isotope values were enriched compared with fins and depleted relative to scales. A split-sample validation technique and linear regression found that isotopic composition of walleye fins and scales was significantly related to that in muscle tissue for both δ13C and δ15N (r2 = 0.79–0.93). However, isotopic variability was significantly different between tissue types in two of six populations for δ15N and three of six populations for δ13C. Although species and population specific, these findings indicate that isotopic measures obtained from non-lethal tissues are indicative of those obtained from muscle.

  10. Phase-contrast Hounsfield units of fixated and non-fixated soft-tissue samples

    DOE PAGES

    Willner, Marian; Fior, Gabriel; Marschner, Mathias; Birnbacher, Lorenz; Schock, Jonathan; Braun, Christian; Fingerle, Alexander A.; Noël, Peter B.; Rummeny, Ernst J.; Pfeiffer, Franz; et al

    2015-08-31

    X-ray phase-contrast imaging is a novel technology that achieves high soft-tissue contrast. Although its clinical impact is still under investigation, the technique may potentially improve clinical diagnostics. In conventional attenuation-based X-ray computed tomography, radiological diagnostics are quantified by Hounsfield units. Corresponding Hounsfield units for phase-contrast imaging have been recently introduced, enabling a setup-independent comparison and standardized interpretation of imaging results. Thus far, the experimental values of few tissue types have been reported; these values have been determined from fixated tissue samples. This study presents phase-contrast Hounsfield units for various types of non-fixated human soft tissues. A large variety of tissuemore » specimens ranging from adipose, muscle and connective tissues to liver, kidney and pancreas tissues were imaged by a grating interferometer with a rotating-anode X-ray tube and a photon-counting detector. In addition, we investigated the effects of formalin fixation on the quantitative phase-contrast imaging results.« less

  11. Tissue-specific mutation accumulation in human adult stem cells during life

    NASA Astrophysics Data System (ADS)

    Blokzijl, Francis; de Ligt, Joep; Jager, Myrthe; Sasselli, Valentina; Roerink, Sophie; Sasaki, Nobuo; Huch, Meritxell; Boymans, Sander; Kuijk, Ewart; Prins, Pjotr; Nijman, Isaac J.; Martincorena, Inigo; Mokry, Michal; Wiegerinck, Caroline L.; Middendorp, Sabine; Sato, Toshiro; Schwank, Gerald; Nieuwenhuis, Edward E. S.; Verstegen, Monique M. A.; van der Laan, Luc J. W.; de Jonge, Jeroen; Ijzermans, Jan N. M.; Vries, Robert G.; van de Wetering, Marc; Stratton, Michael R.; Clevers, Hans; Cuppen, Edwin; van Boxtel, Ruben

    2016-10-01

    The gradual accumulation of genetic mutations in human adult stem cells (ASCs) during life is associated with various age-related diseases, including cancer. Extreme variation in cancer risk across tissues was recently proposed to depend on the lifetime number of ASC divisions, owing to unavoidable random mutations that arise during DNA replication. However, the rates and patterns of mutations in normal ASCs remain unknown. Here we determine genome-wide mutation patterns in ASCs of the small intestine, colon and liver of human donors with ages ranging from 3 to 87 years by sequencing clonal organoid cultures derived from primary multipotent cells. Our results show that mutations accumulate steadily over time in all of the assessed tissue types, at a rate of approximately 40 novel mutations per year, despite the large variation in cancer incidence among these tissues. Liver ASCs, however, have different mutation spectra compared to those of the colon and small intestine. Mutational signature analysis reveals that this difference can be attributed to spontaneous deamination of methylated cytosine residues in the colon and small intestine, probably reflecting their high ASC division rate. In liver, a signature with an as-yet-unknown underlying mechanism is predominant. Mutation spectra of driver genes in cancer show high similarity to the tissue-specific ASC mutation spectra, suggesting that intrinsic mutational processes in ASCs can initiate tumorigenesis. Notably, the inter-individual variation in mutation rate and spectra are low, suggesting tissue-specific activity of common mutational processes throughout life.

  12. Fetal and adult liver stem cells for liver regeneration and tissue engineering.

    PubMed

    Fiegel, H C; Lange, Claudia; Kneser, U; Lambrecht, W; Zander, A R; Rogiers, X; Kluth, D

    2006-01-01

    For the development of innovative cell-based liver directed therapies, e.g. liver tissue engineering, the use of stem cells might be very attractive to overcome the limitation of donor liver tissue. Liver specific differentiation of embryonic, fetal or adult stem cells is currently under investigation. Different types of fetal liver (stem) cells during development were identified, and their advantageous growth potential and bipotential differentiation capacity were shown. However, ethical and legal issues have to be addressed before using fetal cells. Use of adult stem cells is clinically established, e.g. transplantation of hematopoietic stem cells. Other bone marrow derived liver stem cells might be mesenchymal stem cells (MSC). However, the transdifferentiation potential is still in question due to the observation of cellular fusion in several in vivo experiments. In vitro experiments revealed a crucial role of the environment (e.g. growth factors and extracellular matrix) for specific differentiation of stem cells. Co-cultured liver cells also seemed to be important for hepatic gene expression of MSC. For successful liver cell transplantation, a novel approach of tissue engineering by orthotopic transplantation of gel-immobilized cells could be promising, providing optimal environment for the injected cells. Moreover, an orthotopic tissue engineering approach using bipotential stem cells could lead to a repopulation of the recipients liver with healthy liver and biliary cells, thus providing both hepatic functions and biliary excretion. Future studies have to investigate, which stem cell and environmental conditions would be most suitable for the use of stem cells for liver regeneration or tissue engineering approaches.

  13. MicroRNA Stability in FFPE Tissue Samples: Dependence on GC Content

    PubMed Central

    Kakimoto, Yu; Tanaka, Masayuki; Kamiguchi, Hiroshi; Ochiai, Eriko; Osawa, Motoki

    2016-01-01

    MicroRNAs (miRNAs) are small non-coding RNAs responsible for fine-tuning of gene expression at post-transcriptional level. The alterations in miRNA expression levels profoundly affect human health and often lead to the development of severe diseases. Currently, high throughput analyses, such as microarray and deep sequencing, are performed in order to identify miRNA biomarkers, using archival patient tissue samples. MiRNAs are more robust than longer RNAs, and resistant to extreme temperatures, pH, and formalin-fixed paraffin-embedding (FFPE) process. Here, we have compared the stability of miRNAs in FFPE cardiac tissues using next-generation sequencing. The mode read length in FFPE samples was 11 nucleotides (nt), while that in the matched frozen samples was 22 nt. Although the read counts were increased 1.7-fold in FFPE samples, compared with those in the frozen samples, the average miRNA mapping rate decreased from 32.0% to 9.4%. These results indicate that, in addition to the fragmentation of longer RNAs, miRNAs are to some extent degraded in FFPE tissues as well. The expression profiles of total miRNAs in two groups were highly correlated (0.88 tissues instead of miR-1, which was predominant before fixation. Subsequent quantitative PCR (qPCR) analyses revealed that miRNAs with GC content of less than 40% are more degraded than GC-rich miRNAs (p<0.0001). We showed that deep sequencing data obtained using FFPE samples cannot be directly compared with that of fresh frozen samples. The combination of miRNA deep sequencing and other quantitative analyses, such as qPCR, may improve the utility of archival FFPE tissue samples. PMID:27649415

  14. MicroRNA Stability in FFPE Tissue Samples: Dependence on GC Content.

    PubMed

    Kakimoto, Yu; Tanaka, Masayuki; Kamiguchi, Hiroshi; Ochiai, Eriko; Osawa, Motoki

    2016-01-01

    MicroRNAs (miRNAs) are small non-coding RNAs responsible for fine-tuning of gene expression at post-transcriptional level. The alterations in miRNA expression levels profoundly affect human health and often lead to the development of severe diseases. Currently, high throughput analyses, such as microarray and deep sequencing, are performed in order to identify miRNA biomarkers, using archival patient tissue samples. MiRNAs are more robust than longer RNAs, and resistant to extreme temperatures, pH, and formalin-fixed paraffin-embedding (FFPE) process. Here, we have compared the stability of miRNAs in FFPE cardiac tissues using next-generation sequencing. The mode read length in FFPE samples was 11 nucleotides (nt), while that in the matched frozen samples was 22 nt. Although the read counts were increased 1.7-fold in FFPE samples, compared with those in the frozen samples, the average miRNA mapping rate decreased from 32.0% to 9.4%. These results indicate that, in addition to the fragmentation of longer RNAs, miRNAs are to some extent degraded in FFPE tissues as well. The expression profiles of total miRNAs in two groups were highly correlated (0.88 tissues instead of miR-1, which was predominant before fixation. Subsequent quantitative PCR (qPCR) analyses revealed that miRNAs with GC content of less than 40% are more degraded than GC-rich miRNAs (p<0.0001). We showed that deep sequencing data obtained using FFPE samples cannot be directly compared with that of fresh frozen samples. The combination of miRNA deep sequencing and other quantitative analyses, such as qPCR, may improve the utility of archival FFPE tissue samples. PMID:27649415

  15. Tabletop magnetic resonance elastography for the measurement of viscoelastic parameters of small tissue samples

    NASA Astrophysics Data System (ADS)

    Ipek-Ugay, Selcan; Drießle, Toni; Ledwig, Michael; Guo, Jing; Hirsch, Sebastian; Sack, Ingolf; Braun, Jürgen

    2015-02-01

    We demonstrate the feasibility of low-cost tabletop MR elastography (MRE) for quantifying the complex shear modulus G∗ of small soft biological tissue samples as provided by pathologists. The MRE system was developed based on a tabletop MRI scanner equipped with a 0.5 T permanent magnet and a tissue sample holder mounted to a loudspeaker. A spin echo sequence was enhanced with motion-encoding gradients of 250 mT/m amplitude synchronized to acoustic vibration frequencies. Shear wave images suitable for elastography were acquired between vibration frequencies of 0.5 and 1 kHz in agarose, ultrasound gel, porcine liver, porcine skeletal muscle, and bovine heart with a spatial resolution of 234 μm pixel edge length. The measured frequency dependence of G∗ agreed well with previous work based on high-field MR systems. The ratio between loss and storage moduli was highest in liver and ultrasound gel, followed by muscle tissue and agarose gel while ultrasound gel and liver showed similarly low storage moduli compared to the other samples. The shear wave to noise ratio is an important imaging criteria for MRE and was about 4.2 times lower for the preliminary setup of the 0.5 T tabletop system compared to a 7 T animal scanner. In the future, the new tabletop MRE system may serve as a low cost device for preclinical research on the correlation of viscoelastic parameters with histopathology of biological samples.

  16. Screening of Viral Pathogens from Pediatric Ileal Tissue Samples after Vaccination

    DOE PAGES

    Hewitson, Laura; Thissen, James B.; Gardner, Shea N.; McLoughlin, Kevin S.; Glausser, Margaret K.; Jaing, Crystal J.

    2014-01-01

    In 2010, researchers reported that the two US-licensed rotavirus vaccines contained DNA or DNA fragments from porcine circovirus (PCV). Although PCV, a common virus among pigs, is not thought to cause illness in humans, these findings raised several safety concerns. In this study, we sought to determine whether viruses, including PCV, could be detected in ileal tissue samples of children vaccinated with one of the two rotavirus vaccines. A broad spectrum, novel DNA detection technology, the Lawrence Livermore Microbial Detection Array (LLMDA), was utilized, and confirmation of viral pathogens using the polymerase chain reaction (PCR) was conducted. The LLMDAmore » technology was recently used to identify PCV from one rotavirus vaccine. Ileal tissue samples were analyzed from 21 subjects, aged 15–62 months. PCV was not detected in any ileal tissue samples by the LLMDA or PCR. LLMDA identified a human rotavirus A from one of the vaccinated subjects, which is likely due to a recent infection from a wild type rotavirus. LLMDA also identified human parechovirus, a common gastroenteritis viral infection, from two subjects. Additionally, LLMDA detected common gastrointestinal bacterial organisms from the Enterobacteriaceae , Bacteroidaceae , and Streptococcaceae families from several subjects. This study provides a survey of viral and bacterial pathogens from pediatric ileal samples, and may shed light on future studies to identify pathogen associations with pediatric vaccinations.« less

  17. In situ hybridization method for studies of cell wall deficient M. paratuberculosis in tissue samples.

    PubMed

    Hulten, K; Karttunen, T J; El-Zimaity, H M; Naser, S A; Almashhrawi, A; Graham, D Y; El-Zaatari, F A

    2000-12-20

    Cell wall deficient forms of mycobacteria may be important in the pathogenesis of Crohn's disease and sarcoidosis. However, no method has been available to localize this type of organisms in tissue sections. We developed an in situ hybridization method for the demonstration of Mycobacterium paratuberculosis spheroplasts (cell wall deficient forms) in paraffin embedded tissue sections.M. paratuberculosis spheroplasts were prepared by treatment with glycine and lysozyme. Pieces of beef were injected with the prepared spheroplasts. The samples were fixed in buffered formalin and paraffin embedded. A M. paratuberculosis-specific probe derived from the IS900 gene was used. Specificity was controlled by using an irrelevant probe and by hybridizing sections with spheroplasts from other bacteria. Beef samples injected with M. paratuberculosis spheroplasts were the only samples that hybridized with the probe. Beef samples containing acid-fast or spheroplast forms of M. smegmatis and M. tuberculosis as well as the acid-fast forms of M. paratuberculosis did not hybridize with the probe. Unrelated bacterial controls, i.e. Helicobacter pylori and Escherichia coli were also negative in the assay. In situ hybridization with the IS900 probe provides a specific way to localize M. paratuberculosis spheroplasts in tissue sections and may be useful for studies of the connection between M. paratuberculosis and Crohn's disease and sarcoidosis. The assay may also be valuable for studies on Johne's diseased animals. PMID:11118736

  18. The latent structure of attention deficit/hyperactivity disorder in an adult sample.

    PubMed

    Marcus, David K; Norris, Alyssa L; Coccaro, Emil F

    2012-06-01

    The vast majority of studies that have examined the latent structure of attention deficit/hyperactivity disorder (ADHD) in children and adolescents have concluded that ADHD has a dimensional latent structure. In other words, ADHD symptomatology exists along a continuum and there is no natural boundary or qualitative distinction (i.e., taxon) separating youth with ADHD from those with subclinical inattention or hyperactivity/impulsivity problems. Although adult ADHD appears to be less prevalent than ADHD in youth (which could suggest a more severe adult ADHD taxon), researchers have yet to examine the latent structure of ADHD in adults. The present study used a sample (N = 600) of adults who completed a self-report measure of ADHD symptoms. The taxometric analyses revealed a dimensional latent structure for inattention, hyperactivity/impulsivity, and ADHD. These findings are consistent with previous taxometric studies that examined ADHD in children and adolescents, and with contemporary polygenic and multifactorial models of ADHD.

  19. Targeted or whole genome sequencing of formalin fixed tissue samples: potential applications in cancer genomics

    PubMed Central

    Zhao, Yue; Cottrell, Joseph; Klotzle, Brandy; Godwin, Andrew K.; Koestler, Devin; Beyerlein, Peter; Fan, Jian-Bing; Bibikova, Marina; Chien, Jeremy

    2015-01-01

    Current genomic studies are limited by the poor availability of fresh-frozen tissue samples. Although formalin-fixed diagnostic samples are in abundance, they are seldom used in current genomic studies because of the concern of formalin-fixation artifacts. Better characterization of these artifacts will allow the use of archived clinical specimens in translational and clinical research studies. To provide a systematic analysis of formalin-fixation artifacts on Illumina sequencing, we generated 26 DNA sequencing data sets from 13 pairs of matched formalin-fixed paraffin-embedded (FFPE) and fresh-frozen (FF) tissue samples. The results indicate high rate of concordant calls between matched FF/FFPE pairs at reference and variant positions in three commonly used sequencing approaches (whole genome, whole exome, and targeted exon sequencing). Global mismatch rates and C·G > T·A substitutions were comparable between matched FF/FFPE samples, and discordant rates were low (<0.26%) in all samples. Finally, low-pass whole genome sequencing produces similar pattern of copy number alterations between FF/FFPE pairs. The results from our studies suggest the potential use of diagnostic FFPE samples for cancer genomic studies to characterize and catalog variations in cancer genomes. PMID:26305677

  20. Targeted or whole genome sequencing of formalin fixed tissue samples: potential applications in cancer genomics.

    PubMed

    Munchel, Sarah; Hoang, Yen; Zhao, Yue; Cottrell, Joseph; Klotzle, Brandy; Godwin, Andrew K; Koestler, Devin; Beyerlein, Peter; Fan, Jian-Bing; Bibikova, Marina; Chien, Jeremy

    2015-09-22

    Current genomic studies are limited by the poor availability of fresh-frozen tissue samples. Although formalin-fixed diagnostic samples are in abundance, they are seldom used in current genomic studies because of the concern of formalin-fixation artifacts. Better characterization of these artifacts will allow the use of archived clinical specimens in translational and clinical research studies. To provide a systematic analysis of formalin-fixation artifacts on Illumina sequencing, we generated 26 DNA sequencing data sets from 13 pairs of matched formalin-fixed paraffin-embedded (FFPE) and fresh-frozen (FF) tissue samples. The results indicate high rate of concordant calls between matched FF/FFPE pairs at reference and variant positions in three commonly used sequencing approaches (whole genome, whole exome, and targeted exon sequencing). Global mismatch rates and C · G > T · A substitutions were comparable between matched FF/FFPE samples, and discordant rates were low (<0.26%) in all samples. Finally, low-pass whole genome sequencing produces similar pattern of copy number alterations between FF/FFPE pairs. The results from our studies suggest the potential use of diagnostic FFPE samples for cancer genomic studies to characterize and catalog variations in cancer genomes.

  1. Targeted or whole genome sequencing of formalin fixed tissue samples: potential applications in cancer genomics.

    PubMed

    Munchel, Sarah; Hoang, Yen; Zhao, Yue; Cottrell, Joseph; Klotzle, Brandy; Godwin, Andrew K; Koestler, Devin; Beyerlein, Peter; Fan, Jian-Bing; Bibikova, Marina; Chien, Jeremy

    2015-09-22

    Current genomic studies are limited by the poor availability of fresh-frozen tissue samples. Although formalin-fixed diagnostic samples are in abundance, they are seldom used in current genomic studies because of the concern of formalin-fixation artifacts. Better characterization of these artifacts will allow the use of archived clinical specimens in translational and clinical research studies. To provide a systematic analysis of formalin-fixation artifacts on Illumina sequencing, we generated 26 DNA sequencing data sets from 13 pairs of matched formalin-fixed paraffin-embedded (FFPE) and fresh-frozen (FF) tissue samples. The results indicate high rate of concordant calls between matched FF/FFPE pairs at reference and variant positions in three commonly used sequencing approaches (whole genome, whole exome, and targeted exon sequencing). Global mismatch rates and C · G > T · A substitutions were comparable between matched FF/FFPE samples, and discordant rates were low (<0.26%) in all samples. Finally, low-pass whole genome sequencing produces similar pattern of copy number alterations between FF/FFPE pairs. The results from our studies suggest the potential use of diagnostic FFPE samples for cancer genomic studies to characterize and catalog variations in cancer genomes. PMID:26305677

  2. Neonatal tissue injury reduces the intrinsic excitability of adult mouse superficial dorsal horn neurons.

    PubMed

    Li, J; Baccei, M L

    2014-01-01

    Tissue damage during the neonatal period evokes long-lasting changes in nociceptive processing within the adult spinal cord which contribute to persistent alterations in pain sensitivity. However, it remains unclear if the observed modifications in neuronal activity within the mature superficial dorsal horn (SDH) following early injury reflect shifts in the intrinsic membrane properties of these cells. Therefore, the present study was undertaken to identify the effects of neonatal surgical injury on the intrinsic excitability of both GABAergic and presumed glutamatergic neurons within lamina II of the adult SDH using in vitro patch clamp recordings from spinal cord slices prepared from glutamic acid decarboxylase-green fluorescent protein (Gad-GFP) mice. The results demonstrate that hindpaw surgical incision at postnatal day (P) 3 altered the passive membrane properties of both Gad-GFP and adjacent, non-GFP neurons in the mature SDH, as evidenced by decreased membrane resistance and more negative resting potentials in comparison to naïve littermate controls. This was accompanied by a reduction in the prevalence of spontaneous activity within the GABAergic population. Both Gad-GFP and non-GFP neurons displayed a significant elevation in rheobase and decreased instantaneous firing frequency after incision, suggesting that early tissue damage lowers the intrinsic membrane excitability of adult SDH neurons. Isolation of inward-rectifying K(+) (K(ir)) currents revealed that neonatal incision significantly increased K(ir) conductance near physiological membrane potentials in GABAergic, but not glutamatergic, lamina II neurons. Overall, these findings suggest that neonatal tissue injury causes a long-term dampening of intrinsic firing across the general population of lamina II interneurons, but the underlying ionic mechanisms may be cell-type specific.

  3. Simultaneous sampling of tissue oxygenation and oxygen consumption in skeletal muscle.

    PubMed

    Nugent, William H; Song, Bjorn K; Pittman, Roland N; Golub, Aleksander S

    2016-05-01

    Under physiologic conditions, microvascular oxygen delivery appears to be well matched to oxygen consumption in respiring tissues. We present a technique to measure interstitial oxygen tension (PISFO2) and oxygen consumption (VO2) under steady-state conditions, as well as during the transitions from rest to activity and back. Phosphorescence Quenching Microscopy (PQM) was employed with pneumatic compression cycling to achieve 1 to 10 Hz sampling rates of interstitial PO2 and simultaneous recurrent sampling of VO2 (3/min) in the exteriorized rat spinotrapezius muscle. The compression pressure was optimized to 120-130 mmHg without adverse effect on the tissue preparation. A cycle of 5s compression followed by 15s recovery yielded a resting VO2 of 0.98 ± 0.03 ml O2/100 cm(3)min while preserving microvascular oxygen delivery. The measurement system was then used to assess VO2 dependence on PISFO2 at rest and further tested under conditions of isometric muscle contraction to demonstrate a robust ability to monitor the on-kinetics of tissue respiration and the compensatory changes in PISFO2 during contraction and recovery. The temporal and spatial resolution of this approach is well suited to studies seeking to characterize microvascular oxygen supply and demand in thin tissues. PMID:26683232

  4. Cell secretion from the adult lamprey supraneural body tissues possesses cytocidal activity against tumor cells.

    PubMed

    Pang, Yue; Wang, Shiyue; Ba, Wei; Li, Qingwei

    2015-01-01

    The supraneural body was identified in the adult lamprey, and its secretions induced the death of a variety of tumor cells but had no effect on normal cells. The cell secretions from different lamprey tissues were separated, and these secretions killed human tumor cells to varying degrees. The cell secretions induced remarkable cell morphological alterations such as cell blebbing, and the plasma membrane was destroyed by the secretions. In addition, the secretions induced morphological alterations of the mitochondria, cytoskeletal structure, and endoplasmic reticulum, eventually leading to cell death. These observations suggest the presence of a novel protein in the lamprey and the possibility of new applications for the protein in the medical field.

  5. Profiles of Chronic Illness Knowledge in a Community Sample of American Adults

    ERIC Educational Resources Information Center

    Jackson, Todd

    2009-01-01

    The author identified profiles of chronic illness knowledge (i.e., heart disease, cancer, diabetes) in a community sample of American adults and examined the effect of sociodemographic influences on relations of illness knowledge to health practices and well-being. Participants were 181 women and 120 men who completed measures of illness…

  6. Adult Education--Adequacy of Provision. Chapter 7. The Part-Time Tutor Sample Examined

    ERIC Educational Resources Information Center

    Adult Educ (London), 1970

    1970-01-01

    Part of the report of the survey by the National Institute of Adult Education (England and Wales), this chapter provides information gathered from questionnaires distributed to the tutors of classes in the areas from which the student sample was drawn. (NL)

  7. Feasibility of Momentary Sampling Assessment of Cannabis Use in Adolescents and Young Adults

    ERIC Educational Resources Information Center

    Black, Shimrit K.; de Moor, Carl; Kendall, Ashley D.; Shrier, Lydia A.

    2014-01-01

    This study examines the feasibility of recruiting and retaining adolescents and young adults with frequent cannabis use for a 2-week momentary sampling study of cannabis use. Participants responded to random signals on a handheld computer with reports of their use. Participants also initiated reports pre- and post-cannabis use. Participants had…

  8. Factor Structure Invariance of the Kaufman Adolescent and Adult Intelligence Test across Male and Female Samples

    ERIC Educational Resources Information Center

    Immekus, Jason C.; Maller, Susan J.

    2010-01-01

    Multisample confirmatory factor analysis (MCFA) and latent mean structures analysis (LMS) were used to test measurement invariance and latent mean differences on the Kaufman Adolescent and Adult Intelligence Scale[TM] (KAIT) across males and females in the standardization sample. MCFA found that the parameters of the KAIT two-factor model were…

  9. Stress, Social Support, and Outcomes in Two Probability Samples of Homeless Adults

    ERIC Educational Resources Information Center

    Toro, Paul A.; Tulloch, Elizabeth; Ouellette, Nicole

    2008-01-01

    This study investigated the main effects of social support measures and their stress-buffering effects in two samples of homeless adults (Ns =249 and 219) obtained in the same large county (surrounding Detroit) at different points in time over an 8-year period (1992-1994 and 2000-2002). The findings suggest that the construct of social support,…

  10. Periodontal implications of orthodontic treatment in adults with reduced or normal periodontal tissues versus those of adolescents.

    PubMed

    Boyd, R L; Leggott, P J; Quinn, R S; Eakle, W S; Chambers, D

    1989-09-01

    This longitudinal study monitored periodontal status in 20 adults and 20 adolescents undergoing fixed orthodontic treatment. Ten adults had generalized periodontitis and received periodontal treatment, including periodontal surgery, before orthodontic treatment. They also received periodontal maintenance at 3-month intervals during orthodontic treatment. The other 10 adults had normal periodontal tissues. Neither these latter adults nor the adolescents received periodontal maintenance during orthodontic treatment. Periodontal status was determined (1) at six standard sites before fixed appliances were placed (baseline), (2) at 1, 3, 6, 9, 12, and 18 months after appliances had been placed, and (3) 1, 3, 6, and 12 months after appliances had been removed. At each of these visits, these sites were assessed for plaque index, gingival index, bleeding tendency, and pocket depth. Loss of attachment between baseline and 3 months after appliances were removed and tooth loss were also determined. Complete data were obtained for 15 adolescents and 14 adults. During orthodontic treatment the adolescent group showed significantly more (p less than 0.05) periodontal inflammation and supragingival plaque than the adults; after appliances were removed, this pattern was no longer statistically significant. For loss of attachment, there were no significant differences among adolescents, adults with normal periodontal tissues, or adults with reduced but healthy periodontal tissues who had undergone treatment for periodontal disease. For tooth loss, three nonstudy site teeth with pockets deeper than 6 mm and/or furcation involvements were lost because of periodontal abscesses in the adult group treated for periodontal disease. PMID:2773862

  11. Analysis of dissected tissues with digital holographic microscopy: quantification of inflammation mediated tissue alteration, influence of sample preparation, and reliability of numerical autofocusing

    NASA Astrophysics Data System (ADS)

    Kemper, Björn; Lenz, Philipp; Bettenworth, Dominik; Krausewitz, Philipp; Domagk, Dirk; Ketelhut, Steffi

    2015-03-01

    Quantitative phase imaging with digital holographic microscopy (DHM) allows label-free imaging of tissue sections and quantification of the spatial refractive index distribution, which is of interest for applications in digital pathology. We show that DHM allows quantitative imaging of different layers in unstained tissue samples by detection of refractive index changes. In addition, we evaluate the automated refocussing feature of DHM for application on dissected tissues and could achieve highly reproducible holographic autofocusing for unstained and moderately stained samples. Finally, it is demonstrated that in human ulcerative colitis patients the average tissue refractive index is reduced significantly in all parts of the inflamed colonic wall in comparison to patients in remission.

  12. Confidentiality, informed consent, and children's participation in research involving stored tissue samples: interviews with medical professionals from the Middle East.

    PubMed

    Alahmad, Ghiath; Al Jumah, Mohammed; Dierickx, Kris

    2015-01-01

    Ethical issues regarding research biobanks continue to be a topic of intense debate, especially issues of confidentiality, informed consent, and child participation. Although considerable empirical literature concerning research biobank ethics exists, very little information is available regarding the opinions of medical professionals doing genetics research from the Middle East, especially Arabic speaking countries. Ethical guidelines for research biobanks are critically needed as some countries in the Middle East are starting to establish national research biobanks. Islam is the dominant religion in these countries, and it affects people's behavior and influences their positions. Moreover, communities in these countries enjoy a set of customs, traditions and social norms, and have social and familial structures that must be taken into account when developing research policies. We interviewed 12 medical professionals from the Middle East currently working with stored tissue samples to document their opinions. We found general agreement. Participants' primary concerns were similar to the views of researchers internationally. Since children tend to represent a high percentage of Middle Eastern populations, and because children's bodies are not just small adult bodies, the interviewed professionals strongly believed that it is imperative to include children in biobank research. Participants generally believed that protecting confidentiality is socially very important and that informed consent/assent must be obtained from both adult and child participants. This study provides a starting point for additional studies.

  13. Fourier analysis of human soft tissue facial shape: sex differences in normal adults.

    PubMed Central

    Ferrario, V F; Sforza, C; Schmitz, J H; Miani, A; Taroni, G

    1995-01-01

    Sexual dimorphism in human facial form involves both size and shape variations of the soft tissue structures. These variations are conventionally appreciated using linear and angular measurements, as well as ratios, taken from photographs or radiographs. Unfortunately this metric approach provides adequate quantitative information about size only, eluding the problems of shape definition. Mathematical methods such as the Fourier series allow a correct quantitative analysis of shape and of its changes. A method for the reconstruction of outlines starting from selected landmarks and for their Fourier analysis has been developed, and applied to analyse sex differences in shape of the soft tissue facial contour in a group of healthy young adults. When standardised for size, no sex differences were found between both cosine and sine coefficients of the Fourier series expansion. This shape similarity was largely overwhelmed by the very evident size differences and it could be measured only using the proper mathematical methods. PMID:8586558

  14. A pilot study of sampling subcutaneous adipose tissue to examine biomarkers of cancer risk.

    PubMed

    Campbell, Kristin L; Makar, Karen W; Kratz, Mario; Foster-Schubert, Karen E; McTiernan, Anne; Ulrich, Cornelia M

    2009-01-01

    Examination of adipose tissue biology may provide important insight into mechanistic links for the observed association between higher body fat and risk of several types of cancer, in particular colorectal and breast cancer. We tested two different methods of obtaining adipose tissue from healthy individuals. Ten overweight or obese (body mass index, 25-40 kg/m(2)), postmenopausal women were recruited. Two subcutaneous abdominal adipose tissue samples were obtained per individual (i.e., right and left lower abdominal regions) using two distinct methods (method A: 14-gauge needle with incision, versus method B: 16-gauge needle without incision). Gene expression was examined at the mRNA level for leptin, adiponectin, aromatase, interleukin 6 (IL-6), and tumor necrosis factor-alpha (TNF-alpha) in flash-frozen tissue, and at the protein level for leptin, adiponectin, IL-6, and TNF-alpha following short-term culture. Participants preferred biopsy method A and few participants reported any of the usual minor side effects. Gene expression was detectable for leptin, adiponectin, and aromatase, but was below detectable limits for IL-6 and TNF-alpha. For detectable genes, relative gene expression in adipose tissue obtained by methods A and B was similar for adiponectin (r = 0.64, P = 0.06) and leptin (r = 0.80, P = 0.01), but not for aromatase (r = 0.37,P = 0.34). Protein levels in tissue culture supernatant exhibited good intra-assay agreement [coefficient of variation (CV), 1-10%], with less agreement for intraindividual agreement (CV, 17-29%) and reproducibility, following one freeze-thaw cycle (CV, >14%). Subcutaneous adipose tissue biopsies from healthy, overweight individuals provide adequate amounts for RNA extraction, gene expression, and other assays of relevance to cancer prevention research. PMID:19139016

  15. Effects of sample preparation on the optical properties of breast tissue

    NASA Astrophysics Data System (ADS)

    Marks, Fay A.

    1996-04-01

    The optical properties of biological tissue should be determined in vivo whenever possible. However, for those instances when in vivo studies are impractical, too expensive or inappropriate, and when blood flow is not an issue, the ability to perform in vitro studies then becomes invaluable. Optical absorption spectroscopy shows that it may be possible to obtain meaningful information about the optical properties of human breast tissue from in vitro samples if strict preparation and measuring protocols are used. That a strict protocol for storing and handling tissue is critical can be seen from our observations of changes in the optical absorption spectra that occur in response to formalin fixation, the passage of time, application of stains and dyes, and storage in growth medium of the excised tissue. In vivo optical absorption spectroscopy measurements have been made on human breast cancer xenografts and compared with in vitro measurements on breast biopsies prepared according to precise collection and treatment protocols. There is a 'window of opportunity' before time dependent changes in the UV optical absorption spectra of the excised tissue specimens occur. This time window of opportunity widens at longer wavelengths with the least changes occurring in the optical spectra in the NIR.

  16. Transgenic zebrafish reveal tissue-specific differences in estrogen signaling in response to environmental water samples

    USGS Publications Warehouse

    Gorelick, Daniel A.; Iwanowicz, Luke R.; Hung, Alice L.; Blazer, Vicki; Halpern, Marnie E.

    2014-01-01

    Background: Environmental endocrine disruptors (EED) are exogenous chemicals that mimic endogenous hormones, such as estrogens. Previous studies using a zebrafish transgenic reporter demonstrated that the EEDs bisphenol A and genistein preferentially activate estrogen receptors (ER) in the larval heart compared to the liver. However, it was not known whether the transgenic zebrafish reporter was sensitive enough to detect estrogens from environmental samples, whether environmental estrogens would exhibit similar tissue-specific effects as BPA and genistein or why some compounds preferentially target receptors in the heart. Methods: We tested surface water samples using a transgenic zebrafish reporter with tandem estrogen response elements driving green fluorescent protein expression (5xERE:GFP). Reporter activation was colocalized with tissue-specific expression of estrogen receptor genes by RNA in situ hybridization. Results: Selective patterns of ER activation were observed in transgenic fish exposed to river water samples from the Mid-Atlantic United States, with several samples preferentially activating receptors in embryonic and larval heart valves. We discovered that tissue-specificity in ER activation is due to differences in the expression of estrogen receptor subtypes. ERα is expressed in developing heart valves but not in the liver, whereas ERβ2 has the opposite profile. Accordingly, subtype-specific ER agonists activate the reporter in either the heart valves or the liver. Conclusion: The use of 5xERE:GFP transgenic zebrafish has revealed an unexpected tissue-specific difference in the response to environmentally relevant estrogenic compounds. Exposure to estrogenic EEDs in utero is associated with adverse health effects, with the potentially unanticipated consequence of targeting developing heart valves.

  17. Molecular classification of cancer with the 92-gene assay in cytology and limited tissue samples

    PubMed Central

    Brachtel, Elena F.; Operaña, Theresa N.; Sullivan, Peggy S.; Kerr, Sarah E.; Cherkis, Karen A.; Schroeder, Brock E.; Dry, Sarah M.; Schnabel, Catherine A.

    2016-01-01

    Background Detailed molecular evaluation of cytology and limited tissue samples is increasingly becoming the standard for cancer care. Reproducible and accurate diagnostic approaches with reduced demands on cellularity are an ongoing unmet need. This study evaluated the performance of a 92-gene assay for molecular diagnosis of tumor type/subtype in cytology and limited tissue samples. Methods Clinical validation of accuracy for the 92-gene assay in limited tissue samples such as cytology cell blocks, core biopsies and small excisions was conducted in a blinded multi-institutional study (N = 109, 48% metastatic, 53% grade II and III). Analytical success rate and diagnostic utility were evaluated in a consecutive series of 644 cytology cases submitted for clinical testing. Results The 92-gene assay demonstrated 91% sensitivity (95% CI [0.84, 0.95]) for tumor classification, with high accuracy maintained irrespective of specimen type (100%, 92%, and 86% in FNA/cytology cell blocks, core biopsies, and small excisions, respectively; p = 0.26). The assay performed equally well for metastatic versus primary tumors (90% vs 93%, p = 0.73), and across histologic grades (100%, 90%, 89%, in grades I, II, and III, respectively; p = 0.75). In the clinical case series, a molecular diagnosis was reported in 87% of the 644 samples, identifying 23 different tumor types and allowing for additional mutational analysis in selected cases. Conclusions These findings demonstrate high accuracy and analytical success rate of the 92-gene assay, supporting its utility in the molecular diagnosis of cancer for specimens with limited tissue. PMID:27034010

  18. Altered expression of Lewis blood group and related antigens in fetal, normal adult and malignant tissues of the uterine endometrium.

    PubMed

    Inoue, M; Nakayama, M; Tanizawa, O

    1990-01-01

    The expression of the Lewis blood group and its related antigens in fetal, normal adult and malignant tissues of the uterine endometrium was examined immunohistochemically using a panel of mouse monoclonal antibodies with specificities for Lewis-a (La), Sialyl Lewis-a (SLa), Lewis-b (Lb), Lewis-X (LX), Sialyl Lewis-X (SLX) and Lewis-Y (LY) antigens. La, SLa and SLX having one fucose residue were detected in a small number of fetal tissues, while Lb and LY having two fucose residues were found in most cases. In the adult endometrium, expression of Lb and LY was considerably lower than those in fetal tissues, although expression of La and SLa was not different between these two tissues. Expression of LX and SLX was pronounced in adult when compared with fetal tissues. Malignant endometrial glands expressed La, SLa, Lb and LY, extensively, while LX and SLX were expressed less than in normal tissues. Lb and LY can thus be considered oncofetal antigens, extensively expressed in fetal and malignant tissues but not in normal adult tissues. Expression of Lb and LY was greater than that of La and SLA in carcinoma; an increase in the activity of fucose transferase might be associated with malignant transformation in the uterine endometrium.

  19. Cell Competition Modifies Adult Stem Cell and Tissue Population Dynamics in a JAK-STAT-Dependent Manner

    PubMed Central

    Kolahgar, Golnar; Suijkerbuijk, Saskia J.E.; Kucinski, Iwo; Poirier, Enzo Z.; Mansour, Sarah; Simons, Benjamin D.; Piddini, Eugenia

    2015-01-01

    Summary Throughout their lifetime, cells may suffer insults that reduce their fitness and disrupt their function, and it is unclear how these potentially harmful cells are managed in adult tissues. We address this question using the adult Drosophila posterior midgut as a model of homeostatic tissue and ribosomal Minute mutations to reduce fitness in groups of cells. We take a quantitative approach combining lineage tracing and biophysical modeling and address how cell competition affects stem cell and tissue population dynamics. We show that healthy cells induce clonal extinction in weak tissues, targeting both stem and differentiated cells for elimination. We also find that competition induces stem cell proliferation and self-renewal in healthy tissue, promoting selective advantage and tissue colonization. Finally, we show that winner cell proliferation is fueled by the JAK-STAT ligand Unpaired-3, produced by Minute−/+ cells in response to chronic JNK stress signaling. PMID:26212135

  20. Nasopharyngeal versus oropharyngeal sampling for detection of pneumococcal carriage in adults.

    PubMed

    Watt, James P; O'Brien, Katherine L; Katz, Scott; Bronsdon, Melinda A; Elliott, John; Dallas, Jean; Perilla, Mindy J; Reid, Raymond; Murrow, Laurel; Facklam, Richard; Santosham, Mathuram; Whitney, Cynthia G

    2004-11-01

    Several studies have shown that nasopharyngeal sampling is more sensitive than oropharyngeal sampling for the detection of pneumococcal carriage in children. The data for adults are limited and conflicting. This study was part of a larger study of pneumococcal carriage on the Navajo and White Mountain Apache Reservation following a clinical trial of a seven-valent pneumococcal conjugate vaccine. Persons aged 18 years and older living in households with children enrolled in the vaccine trial were eligible. We collected both nasopharyngeal and oropharyngeal specimens by passing a flexible calcium alginate wire swab either nasally to the posterior nasopharynx or orally to the posterior oropharynx. Swabs were placed in skim milk-tryptone-glucose-glycerin medium and frozen at -70 degrees C. Pneumococcal isolation was performed by standard techniques. Analyses were based on specimens collected from 1,994 adults living in 1,054 households. Nasopharyngeal specimens (11.1%; 95% confidence interval [CI], 9.8 and 12.6%) were significantly more likely to grow pneumococci than were oropharyngeal specimens (5.8%; 95% CI, 4.8 to 6.9%) (P < 0.0001). Few persons had pneumococcal growth from both specimens (1.7%). Therefore, both tests together were more likely to identify pneumococcal carriage (15.2%; 95% CI, 13.7 to 16.9%) than either test alone. Although we found that nasopharyngeal sampling was more sensitive than oropharyngeal sampling, nasopharyngeal sampling alone would have underestimated the prevalence of pneumococcal carriage in this adult population. Sampling both sites may give more accurate results than sampling either site alone in studies of pneumococcal carriage in adults. PMID:15528682

  1. Characterising resting-state functional connectivity in a large sample of adults with ADHD.

    PubMed

    Mostert, Jeanette C; Shumskaya, Elena; Mennes, Maarten; Onnink, A Marten H; Hoogman, Martine; Kan, Cornelis C; Arias Vasquez, Alejandro; Buitelaar, Jan; Franke, Barbara; Norris, David G

    2016-06-01

    Attention-deficit/hyperactivity disorder (ADHD) is a common childhood psychiatric disorder that often persists into adulthood. While several studies have identified altered functional connectivity in brain networks during rest in children with ADHD, few studies have been performed on adults with ADHD. Existing studies have generally investigated small samples. We therefore investigated aberrant functional connectivity in a large sample of adult patients with childhood-onset ADHD, using a data-driven, whole-brain approach. Adults with a clinical ADHD diagnosis (N=99) and healthy, adult comparison subjects (N=113) underwent a 9-minute resting-state fMRI session in a 1.5T MRI scanner. After elaborate preprocessing including a thorough head-motion correction procedure, group independent component analysis (ICA) was applied from which we identified six networks of interest: cerebellum, executive control, left and right frontoparietal and two default-mode networks. Participant-level network maps were obtained using dual-regression and tested for differences between patients with ADHD and controls using permutation testing. Patients showed significantly stronger connectivity in the anterior cingulate gyrus of the executive control network. Trends were also observed for stronger connectivity in the cerebellum network in ADHD patients compared to controls. However, there was considerable overlap in connectivity values between patients and controls, leading to relatively low effect sizes despite the large sample size. These effect sizes were slightly larger when testing for correlations between hyperactivity/impulsivity symptoms and connectivity strength in the executive control and cerebellum networks. This study provides important insights for studies on the neurobiology of adult ADHD; it shows that resting-state functional connectivity differences between adult patients and controls exist, but have smaller effect sizes than existing literature suggested.

  2. Distribution of polybrominated diphenyl ethers in Japanese autopsy tissue and body fluid samples.

    PubMed

    Hirai, Tetsuya; Fujimine, Yoshinori; Watanabe, Shaw; Nakano, Takeshi

    2012-09-01

    Brominated flame retardants are components of many plastics and are used in products such as cars, textiles, televisions, and personal computers. Human exposure to polybrominated diphenyl ether (PBDE) flame retardants has increased exponentially during the last three decades. Our objective was to measure the body burden and distribution of PBDEs and to determine the concentrations of the predominant PBDE congeners in samples of liver, bile, adipose tissue, and blood obtained from Japanese autopsy cases. Tissues and body fluids obtained from 20 autopsy cases were analyzed. The levels of 25 PBDE congeners, ranging from tri- to hexa-BDEs, were assessed. The geometric means of the sum of the concentrations of PBDE congeners having detection frequencies >50 % (ΣPBDE) in the blood, liver, bile, and adipose tissue were 2.4, 2.6, 1.4, and 4.3 ng/g lipid, respectively. The most abundant congeners were BDE-47 and BDE-153, followed by BDE-100, BDE-99, and BDE-28+33. These concentrations of PBDE congeners were similar to other reports of human exposure in Japan but were notably lower than concentrations than those reported in the USA. Significant positive correlations were observed between the concentrations of predominant congeners and ΣPBDE among the samples analyzed. The ΣPBDE concentration was highest in the adipose tissue, but PBDEs were distributed widely among the tissues and body fluids analyzed. The PBDE levels observed in the present study are similar to those reported in previous studies in Japan and significantly lower than those reported in the USA. PMID:22544599

  3. The drug-crime nexus in a community sample of adults.

    PubMed

    Newcomb, M D; Galaif, E R; Carmona, V

    2001-09-01

    The authors tested several theories using prospective data to explain the associations between drug abuse and crime in a community sample of 470 adults. Polydrug problems in early adulthood predicted both criminal behavior and polydrug problems in adulthood. Consequences of drug problems as a young adult included arrests and convictions for drug-related offenses, property damage, and driving under the influence of other drugs. Predictors of later polydrug problems included thefts, driving under the influence of alcohol and other drugs, arrests and convictions for drug-related offenses, and a lack of support for drug problems. Theoretical implications of these findings are discussed.

  4. High-efficiency immunomagnetic isolation of solid tissue-originated integrin-expressing adult stem cells.

    PubMed

    Palmon, Aaron; David, Ran; Neumann, Yoav; Stiubea-Cohen, Raluca; Krief, Guy; Aframian, Doron J

    2012-02-01

    Isolation of highly pure specific cell types is crucial for successful adult stem cell-based therapy. As the number of such cells in adult tissue is low, an extremely efficient method is needed for their isolation. Here, we describe cell-separation methodologies based on magnetic-affinity cell sorting (MACS) MicroBeads with monoclonal antibodies against specific membrane proteins conjugated to superparamagnetic particles. Cells labeled with MACS MicroBeads are retained in a magnetic field within a MACS column placed in a MACS separator, allowing fast and efficient separation. Both positively labeled and non-labeled fractions can be used directly for downstream applications as the separated cell fractions remain viable with no functional impairment. As immunomagnetic separation depends on the interaction between a cell's membrane and the magnetically labeled antibody, separation of specific cells originating from solid tissues is more complex and demands a cell-dissociating pretreatment. In this paper, we detail the use of immunomagnetic separation for the purpose of regenerating damaged salivary gland (SG) function in animal and human models of irradiated head and neck cancer. Each year 500,000 new cases of head and neck cancer occur worldwide. Most of these patients lose SG function following irradiation therapy. SGs contain integrin α6β1-expressing epithelial stem cells. We hypothesized that these cells can be isolated, multiplied in culture and auto-implanted into the irradiated SGs to regenerate damaged SG function.

  5. Optimal transport time and conditions for cartilage tissue samples and expanded chondrocyte suspensions.

    PubMed

    Yilmaz, Banu Coskun; Yilmaz, Cengiz; Yilmaz, Necat S; Balli, Ebru; Tasdelen, Bahar

    2010-01-01

    For autologous chondrocyte implantation, the chondral tissue obtained is transferred from the operating room to the laboratory using specialized carrier systems within 24 hours. Similar expenses are used for the transport of cultured chondrocytes. The purpose of this study was to find the optimal temperature, size of tissue, and time that the chondrocytes can stand without losing viability and proliferative capacity. Fresh calf cartilage was harvested and divided into 24 groups. Half of the samples were diced into 1- to 2-mm(3) pieces. All 12 groups were kept at either 4 degrees C, 25 degrees C, or 37 degrees C for 1, 3, 5, or 7 days and were seeded for cell culture. Times to reach confluence values were compared. Produced cell suspensions were grouped similarly and tested similarly. Neither the temperature nor the waiting days caused any difference in the proliferative capacity of the cells. Diced tissues yielded a shorter time to reach confluence values. Chondral tissue obtained from the patient can be transferred to the laboratory at temperatures ranging from 4 degrees C to 37 degrees C in up to 7 days. These conditions did not affect the proliferative capacity or the viability of the chondrocytes. Dicing the tissue prior to transport will shorten total culturing time. The expanded cell suspensions should be transferred at temperatures from 4 degrees C to 25 degrees C within 3 days. Specialized carrier systems to get the chondral tissue from the operating room to the laboratory and to take the expanded chondrocytes back to the operating room within hours may not be necessary.

  6. Threshold-dependent sample sizes for selenium assessment with stream fish tissue

    USGS Publications Warehouse

    Hitt, Nathaniel P.; Smith, David

    2013-01-01

    Natural resource managers are developing assessments of selenium (Se) contamination in freshwater ecosystems based on fish tissue concentrations. We evaluated the effects of sample size (i.e., number of fish per site) on the probability of correctly detecting mean whole-body Se values above a range of potential management thresholds. We modeled Se concentrations as gamma distributions with shape and scale parameters fitting an empirical mean-to-variance relationship in data from southwestern West Virginia, USA (63 collections, 382 individuals). We used parametric bootstrapping techniques to calculate statistical power as the probability of detecting true mean concentrations up to 3 mg Se/kg above management thresholds ranging from 4-8 mg Se/kg. Sample sizes required to achieve 80% power varied as a function of management thresholds and type-I error tolerance (α). Higher thresholds required more samples than lower thresholds because populations were more heterogeneous at higher mean Se levels. For instance, to assess a management threshold of 4 mg Se/kg, a sample of 8 fish could detect an increase of ∼ 1 mg Se/kg with 80% power (given α = 0.05), but this sample size would be unable to detect such an increase from a management threshold of 8 mg Se/kg with more than a coin-flip probability. Increasing α decreased sample size requirements to detect above-threshold mean Se concentrations with 80% power. For instance, at an α-level of 0.05, an 8-fish sample could detect an increase of ∼ 2 units above a threshold of 8 mg Se/kg with 80% power, but when α was relaxed to 0.2 this sample size was more sensitive to increasing mean Se concentrations, allowing detection of an increase of ∼ 1.2 units with equivalent power. Combining individuals into 2- and 4-fish composite samples for laboratory analysis did not decrease power because the reduced number of laboratory samples was compensated by increased precision of composites for estimating mean

  7. Threshold-dependent sample sizes for selenium assessment with stream fish tissue.

    PubMed

    Hitt, Nathaniel P; Smith, David R

    2015-01-01

    Natural resource managers are developing assessments of selenium (Se) contamination in freshwater ecosystems based on fish tissue concentrations. We evaluated the effects of sample size (i.e., number of fish per site) on the probability of correctly detecting mean whole-body Se values above a range of potential management thresholds. We modeled Se concentrations as gamma distributions with shape and scale parameters fitting an empirical mean-to-variance relationship in data from southwestern West Virginia, USA (63 collections, 382 individuals). We used parametric bootstrapping techniques to calculate statistical power as the probability of detecting true mean concentrations up to 3 mg Se/kg above management thresholds ranging from 4 to 8 mg Se/kg. Sample sizes required to achieve 80% power varied as a function of management thresholds and Type I error tolerance (α). Higher thresholds required more samples than lower thresholds because populations were more heterogeneous at higher mean Se levels. For instance, to assess a management threshold of 4 mg Se/kg, a sample of eight fish could detect an increase of approximately 1 mg Se/kg with 80% power (given α=0.05), but this sample size would be unable to detect such an increase from a management threshold of 8 mg Se/kg with more than a coin-flip probability. Increasing α decreased sample size requirements to detect above-threshold mean Se concentrations with 80% power. For instance, at an α-level of 0.05, an 8-fish sample could detect an increase of approximately 2 units above a threshold of 8 mg Se/kg with 80% power, but when α was relaxed to 0.2, this sample size was more sensitive to increasing mean Se concentrations, allowing detection of an increase of approximately 1.2 units with equivalent power. Combining individuals into 2- and 4-fish composite samples for laboratory analysis did not decrease power because the reduced number of laboratory samples was compensated for by increased precision of composites

  8. Determination of benzophenones in human placental tissue samples by liquid chromatography-tandem mass spectrometry.

    PubMed

    Vela-Soria, F; Jiménez-Díaz, I; Rodríguez-Gómez, R; Zafra-Gómez, A; Ballesteros, O; Navalón, A; Vílchez, J L; Fernández, M F; Olea, N

    2011-09-30

    Benzophenones (BPs) are a family of compounds widely used to protect the skin and hair from UV irradiation. Despite human exposure to BPs through dermal application of products containing sunscreen agents and the increasing evidence that BPs are able to interfere with endocrine systems, few studies have examined the occurrence of BPs in humans. In this work, we propose a new liquid chromatography-tandem mass spectrometry (LC-MS/MS) method to determine six BPs, namely, benzophenone-1 (BP-1), benzophenone-2 (BP-2), benzophenone-3 (BP-3), benzophenone-6 (BP-6), benzophenone-8 (BP-8) and 4-hydroxybenzophenone (4-OH-BP) in human placental tissue samples. The method involves an extraction step of the analytes from the samples using ethyl acetate, followed by a clean-up step using centrifugation prior to their quantification by LC-MS/MS using an atmospheric pressure chemical ionization (APCI) interface in the positive mode. Benzophenone-d(10) (BP-d(10)) was used as surrogate. Found detection limits (LOD) ranged from 0.07 to 0.3 ng g(-1) and quantification limits (LOQ) from 0.3 to 1.0 ng g(-1), while inter- and intra-day variability was under 5%. The method was validated using standard addition calibration and a recovery assay. Recovery rates for spiked samples ranged from 98 to 104%. This method was satisfactorily applied for the determination of BPs in 16 placental tissue samples collected from women who live in Granada (Spain).

  9. The relationship between decorrelation time and sample thickness in acute rat brain tissue slices (Conference Presentation)

    NASA Astrophysics Data System (ADS)

    Brake, Joshua; Jang, Mooseok; Yang, Changhuei

    2016-03-01

    The optical opacity of biological tissue has long been a challenge in biomedical optics due to the strong scattering nature of tissue in the optical regime. While most conventional optical techniques attempt to gate out multiply scattered light and use only unscattered light, new approaches in the field of wavefront shaping exploit the time reversible symmetry of optical scattering in order to focus light inside or through scattering media. While these approaches have been demonstrated effectively on static samples, it has proven difficult to apply them to dynamic biological samples since even small changes in the relative positions of the scatterers within will cause the time symmetry that wavefront shaping relies upon to decorrelate. In this paper we investigate the decorrelation curves of acute rat brain slices for thicknesses in the range 1-3 mm (1/e decorrelation time on the order of seconds) using multi-speckle diffusing wave spectroscopy (MSDWS) and compare the results with theoretical predictions. The results of this study demonstrate that the 1/L^2 relationship between decorrelation time and thickness predicted by diffusing wave spectroscopy provides a good rule of thumb for estimating how the decorrelation of a sample will change with increasing thickness. Understanding this relationship will provide insight to guide the future development of biophotonic wavefront shaping tools by giving an estimate of how fast wavefront shaping systems need to operate to overcome the dynamic nature of biological samples.

  10. Quantitative RT-PCR comparison of the urea and nitric oxide cycle gene transcripts in adult human tissues.

    PubMed

    Neill, Meaghan Anne; Aschner, Judy; Barr, Frederick; Summar, Marshall L

    2009-06-01

    The urea cycle and nitric oxide cycle play significant roles in complex biochemical and physiologic reactions. These cycles have distinct biochemical goals including the clearance of waste nitrogen; the production of the intermediates ornithine, citrulline, and arginine for the urea cycle; and the production of nitric oxide for the nitric oxide pathway. Despite their disparate functions, the two pathways share two enzymes, argininosuccinic acid synthase and argininosuccinic acid lyase, and a transporter, citrin. Studying the gene expression of these enzymes is paramount in understanding these complex biochemical pathways. Here, we examine the expression of genes involved in the urea cycle and the nitric oxide cycle in a panel of eleven different tissue samples obtained from individual adults without known inborn errors of metabolism. In this study, the pattern of co-expressed enzymes provides a global view of the metabolic activity of the urea and nitric oxide cycles in human tissues. Our results show that these transcripts are differentially expressed in different tissues. Using the co-expression profiles, we discovered that the combination of expression of enzyme transcripts as detected in our study, might serve to fulfill specific physiologic function(s) including urea production/nitrogen removal, arginine/citrulline production, nitric oxide production, and ornithine production. Our study reveals the importance of studying not only the expression profile of an enzyme of interest, but also studying the expression profiles of the other enzymes involved in a particular pathway so as to better understand the context of expression. The tissue patterns we observed highlight the variety of important functions of these enzymes and provides insight into the many clinical observations that result from their disruption. These results have implications for the management of urea cycle patients and raise considerations for the care of those patients receiving liver

  11. [An oral-ruminal probe for rumen sampling in the adult sheep].

    PubMed

    Geishauser, T; Gitzel, A

    1995-12-01

    Ten adult (height: 63-85 cm) rumen-fistulated sheep were used to test the usefulness of an ororuminal probe and a suction pump for the acquisition of ruminal fluid. The use of these instruments in 50 samplings rendered 200 ml of ruminal fluid each time. The introduction of the probe took 18 +/- 6.5 s, whereby the probe was inserted to a length of 91.3 +/- 3.5 cm. The collection of 200 ml of ruminal fluid took 7.4 +/- 2.2 s. The sampling location was the ventral ruminal sack. By comparing the fluid taken by the ororuminal probe with samples taken via ruminal fistula by a tube-like probe no significant differences in regard to pH, total acidity, reduction potential, ammonia, acetate, L-lactate, sodium and chloride concentrations were found. However, significant differences between ororuminal probe samples and fistula samples were observed in regards to the concentrations of total volatile fatty acids, propionate, n-butyrate, i-butyrate, n-valeriate, i-valeriate, calcium, phosphate, magnesium and potassium. The differences were independent of the sampling order (ororuminal probe prior to fistula and vice versa). These results indicate that samples taken by the ororuminal probe do not contain sampling-dependent saliva residues. Those samples collected via fistula had consistently higher concentrations of total volatile fatty acids, propionate, n-butyrate, i-butyrate, n-valeriate, and i-valeriate, as well as higher concentrations of calcium, phosphate, magnesium and potassium than samples taken with the ororuminal probe. These results indicate that samples taken via fistula originated from more dorsal regions than the samples taken by ororuminal probe from the ventral ruminal sack. The ororuminal probe and the suction pump used proved to be useful for the collection of ruminal fluid from the ventral ruminal sack in adult sheep (height: 63-85 cm).

  12. A single lysis solution for the analysis of tissue samples by different proteomic technologies.

    PubMed

    Gromov, Pavel; Celis, Julio E; Gromova, Irina; Rank, Fritz; Timmermans-Wielenga, Vera; Moreira, José M A

    2008-12-01

    Cancer, being a major healthcare concern worldwide, is one of the main targets for the application of emerging proteomic technologies and these tools promise to revolutionize the way cancer will be diagnosed and treated in the near future. Today, as a result of the unprecedented advances that have taken place in molecular biology, cell biology and genomics there is a pressing need to accelerate the translation of basic discoveries into clinical applications. This need, compounded by mounting evidence that cellular model systems are unable to fully recapitulate all biological aspects of human dissease, is driving scientists to increasingly use clinically relevant samples for biomarker and target discovery. Tissues are heterogeneous and as a result optimization of sample preparation is critical for generating accurate, representative, and highly reproducible quantitative data. Although a large number of protocols for preparation of tissue lysates has been published, so far no single recipe is able to provide a "one-size fits all" solubilization procedure that can be used to analyse the same lysate using different proteomics technologies. Here we present evidence showing that cell lysis buffer 1 (CLB1), a lysis solution commercialized by Zeptosens [a division of Bayer (Schweiz) AG], provides excellent sample solubilization and very high 2D PAGE protein resolution both when using carrier ampholytes and immobilized pH gradient strips. Moreover, this buffer can also be used for array-based proteomics (reverse-phase lysate arrays or direct antibody arrays), allowing the direct comparison of qualitative and quantitative data yielded by these technologies when applied to the same samples. The usefulness of the CLB1 solution for gel-based proteomics was further established by 2D PAGE analysis of a number of technically demanding specimens such as breast carcinoma core needle biopsies and problematic tissues such as brain cortex, cerebellum, skeletal muscle, kidney cortex and

  13. Fetal and adult fibroblasts display intrinsic differences in tendon tissue engineering and regeneration

    PubMed Central

    Tang, Qiao-Mei; Chen, Jia Lin; Shen, Wei Liang; Yin, Zi; Liu, Huan Huan; Fang, Zhi; Heng, Boon Chin; Ouyang, Hong Wei; Chen, Xiao

    2014-01-01

    Injured adult tendons do not exhibit optimal healing through a regenerative process, whereas fetal tendons can heal in a regenerative fashion without scar formation. Hence, we compared FFs (mouse fetal fibroblasts) and AFs (mouse adult fibroblasts) as seed cells for the fabrication of scaffold-free engineered tendons. Our results demonstrated that FFs had more potential for tendon tissue engineering, as shown by higher levels of tendon-related gene expression. In the in situ AT injury model, the FFs group also demonstrated much better structural and functional properties after healing, with higher levels of collagen deposition and better microstructure repair. Moreover, fetal fibroblasts could increase the recruitment of fibroblast-like cells and reduce the infiltration of inflammatory cells to the injury site during the regeneration process. Our results suggest that the underlying mechanisms of better regeneration with FFs should be elucidated and be used to enhance adult tendon healing. This may assist in the development of future strategies to treat tendon injuries. PMID:24992450

  14. Fetal and adult fibroblasts display intrinsic differences in tendon tissue engineering and regeneration.

    PubMed

    Tang, Qiao-Mei; Chen, Jia Lin; Shen, Wei Liang; Yin, Zi; Liu, Huan Huan; Fang, Zhi; Heng, Boon Chin; Ouyang, Hong Wei; Chen, Xiao

    2014-07-03

    Injured adult tendons do not exhibit optimal healing through a regenerative process, whereas fetal tendons can heal in a regenerative fashion without scar formation. Hence, we compared FFs (mouse fetal fibroblasts) and AFs (mouse adult fibroblasts) as seed cells for the fabrication of scaffold-free engineered tendons. Our results demonstrated that FFs had more potential for tendon tissue engineering, as shown by higher levels of tendon-related gene expression. In the in situ AT injury model, the FFs group also demonstrated much better structural and functional properties after healing, with higher levels of collagen deposition and better microstructure repair. Moreover, fetal fibroblasts could increase the recruitment of fibroblast-like cells and reduce the infiltration of inflammatory cells to the injury site during the regeneration process. Our results suggest that the underlying mechanisms of better regeneration with FFs should be elucidated and be used to enhance adult tendon healing. This may assist in the development of future strategies to treat tendon injuries.

  15. Determination of optical properties of oxidative bleaching human dental tissue samples using optical coherence tomography

    NASA Astrophysics Data System (ADS)

    Ni, Y. R.; Guo, Z. Y.; Shu, S. Y.; Zeng, C. C.; Zhong, H. Q.; Chen, B. L.; Liu, Z. M.; Bao, Y.

    2011-10-01

    Oxidative bleaching changes of human teeth induced changes in the optical properties of dental tissue. We introduced 1310 nm wavelengths of optical coherence tomography (OCT) attenuation coefficient method which is a relatively novel and rarely reported methodology to measure the correlation coefficient during the teeth oxidative bleaching procedure. And the quantitative parameters of enamel optical thickness and disruption of the entrance signal (DES) were extracted from the OCT images. The attenuation coefficient of the bleached tissue is 6.2 mm-1 which is significant (p < 0.001) higher than that unbleached sample is 1.4 mm-1. But attenuation coefficient varied significantly (p < 0.001) between 5.9 and 1.5 mm-1 in dentine which is downtrend. Furthermore, the persistence of bleaching oxidation in 35% hydrogen peroxide-induced optical thickness of enamel is similar with unbleached tissue which may indicate the refractive index of enamel is unchanged. Moreover, disruption of the entrance signal (DES) analysis showed that remarkable difference was appeared at enamel surface. The results indicate that optical properties of oxidative bleaching human dental tissue can be determined by attenuation coefficient using OCT system.

  16. Molecular strain identification of the Mycobacterium tuberculosis complex in archival tissue samples

    PubMed Central

    Zink, A R; Nerlich, A G

    2004-01-01

    Aims: To investigate the use of different molecular analyses that can identify distinct strains of human pathogenic mycobacteria in formalin fixed and paraffin wax embedded archival tissue samples to see whether it is possible to differentiate between the members of the Mycobacterium tuberculosis complex (M tuberculosis, M bovis, M africanum, M microti, or M canettii) and/or substrains in a high number of samples. This would be of interest for identifying individual infection traits and superinfection by different mycobacterial strains. Methods: Forty nine archival tissue samples with clinically and/or histologically suspected tuberculosis infection were subjected to molecular DNA analysis. Results: The molecular analysis revealed the presence of M tuberculosis complex DNA in 20 samples, whereas acid fast bacilli could be detected by Ziehl-Neelsen staining in only eight samples. All IS6110 positive samples were further characterised by spoligotyping and seven cases provided M tuberculosis specific signatures, whereas M bovis specific signatures were obtained in four cases. The analysis of mtp40, oxyR, and pncA partial gene sequences confirmed the presence of M tuberculosis in six cases and M bovis in one case. The amplification and sequencing of four further genetic regions (katG, gyrA, TbD1, RD9) characterised six “modern” M tuberculosis strains belonging to genetic groups 2 or 3. Conclusion: This study provides clear evidence that archival paraffin wax embedded material can be used for further studies on the strain identification of M tuberculosis complex strains and can therefore unequivocally be used for the study of the epidemiology and evolution of tuberculosis pathogens. PMID:15509681

  17. Use of buccal swabs for sampling DNA from nestling and adult birds

    USGS Publications Warehouse

    Handel, Colleen M.; Pajot, Lisa; Talbot, Sandra L.; Sage, George K.

    2006-01-01

    We evaluated the feasibility and efficiency of using swabs to collect buccal epithelial cells fromsmall (2‐ to 13‐g) birds as a source of DNA for genetic studies. We used commercially available buccal swab kits to collect samples from 42 adult and 39 nestling (4‐ to 8‐day‐old) black‐capped chickadees (Poecile atricapillus) and from6 4‐day‐old nestling boreal chickadees (P. hudsonica). We compared DNA from buccal epithelial samples to that fromblood samples from the same individuals. We extracted sufficient quantities of DNA for analysis from all buccalsamples, and samples remained viable even after being stored in original plastic sampling tubes at room temperature for up to 18 months. Yields were equivalent whether extracted using the proprietary quick‐extraction solution provided with buccal swab kits or using a salt‐extraction process with inexpensive reagents. Yields of DNA from buccal samples were consistently lower than those from blood samples, but quantities were sufficient for all analyses. Assignment of sex, based on DNA extracted from paired buccal and blood samples, was identical for all 87 birds. We found no difference in the genotypes obtained from buccal and blood samples for 12 individuals tested using 5 microsatellite loci and found perfect concordance in sequencing of an 823‐base‐pair segment within the control region of mitochondrial DNA for 7 individuals tested. Use of buccal swabs is highly recommended as a rapid, noninvasive technique for sampling avian genomic DNA, especially for extremely young altricial nestlings or small‐bodied adults, or for any birds for which blood sampling may be impossible or stressful.

  18. A workflow to preserve genome-quality tissue samples from plants in botanical gardens and arboreta1

    PubMed Central

    Gostel, Morgan R.; Kelloff, Carol; Wallick, Kyle; Funk, Vicki A.

    2016-01-01

    Premise of the study: Internationally, gardens hold diverse living collections that can be preserved for genomic research. Workflows have been developed for genomic tissue sampling in other taxa (e.g., vertebrates), but are inadequate for plants. We outline a workflow for tissue sampling intended for two audiences: botanists interested in genomics research and garden staff who plan to voucher living collections. Methods and Results: Standard herbarium methods are used to collect vouchers, label information and images are entered into a publicly accessible database, and leaf tissue is preserved in silica and liquid nitrogen. A five-step approach for genomic tissue sampling is presented for sampling from living collections according to current best practices. Conclusions: Collecting genome-quality samples from gardens is an economical and rapid way to make available for scientific research tissue from the diversity of plants on Earth. The Global Genome Initiative will facilitate and lead this endeavor through international partnerships. PMID:27672517

  19. A workflow to preserve genome-quality tissue samples from plants in botanical gardens and arboreta1

    PubMed Central

    Gostel, Morgan R.; Kelloff, Carol; Wallick, Kyle; Funk, Vicki A.

    2016-01-01

    Premise of the study: Internationally, gardens hold diverse living collections that can be preserved for genomic research. Workflows have been developed for genomic tissue sampling in other taxa (e.g., vertebrates), but are inadequate for plants. We outline a workflow for tissue sampling intended for two audiences: botanists interested in genomics research and garden staff who plan to voucher living collections. Methods and Results: Standard herbarium methods are used to collect vouchers, label information and images are entered into a publicly accessible database, and leaf tissue is preserved in silica and liquid nitrogen. A five-step approach for genomic tissue sampling is presented for sampling from living collections according to current best practices. Conclusions: Collecting genome-quality samples from gardens is an economical and rapid way to make available for scientific research tissue from the diversity of plants on Earth. The Global Genome Initiative will facilitate and lead this endeavor through international partnerships.

  20. Liquid microjunction surface sampling of acetaminophen, terfenadine and their metabolites in thin tissue sections

    SciTech Connect

    Kertesz, Vilmos; Paranthaman, Nithya; Moench, Paul; Catoire, Alexandre; Flarakos, Jimmy; Van Berkel, Gary J.

    2014-10-01

    The aim of this paper was to evaluate the analytical performance of a fully automated droplet-based surface-sampling system for determining the distribution of the drugs acetaminophen and terfenadine, and their metabolites, in rat thin tissue sections. The following are the results: The rank order of acetaminophen concentration observed in tissues was stomach > small intestine > liver, while the concentrations of its glucuronide and sulfate metabolites were greatest in the liver and small intestine. Terfenadine was most concentrated in the liver and kidney, while its major metabolite, fexofenadine, was found in the liver and small intestine. In conclusion, the spatial distributions of both drugs and their respective metabolites observed in this work were consistent with previous studies using radiolabeled drugs.

  1. Liquid microjunction surface sampling of acetaminophen, terfenadine and their metabolites in thin tissue sections

    DOE PAGES

    Kertesz, Vilmos; Paranthaman, Nithya; Moench, Paul; Catoire, Alexandre; Flarakos, Jimmy; Van Berkel, Gary J.

    2014-10-01

    The aim of this paper was to evaluate the analytical performance of a fully automated droplet-based surface-sampling system for determining the distribution of the drugs acetaminophen and terfenadine, and their metabolites, in rat thin tissue sections. The following are the results: The rank order of acetaminophen concentration observed in tissues was stomach > small intestine > liver, while the concentrations of its glucuronide and sulfate metabolites were greatest in the liver and small intestine. Terfenadine was most concentrated in the liver and kidney, while its major metabolite, fexofenadine, was found in the liver and small intestine. In conclusion, the spatialmore » distributions of both drugs and their respective metabolites observed in this work were consistent with previous studies using radiolabeled drugs.« less

  2. Determination of /sup 35/S-aminoacyl-transfer ribonucleic acid specific radioactivity in small tissue samples

    SciTech Connect

    Samarel, A.M.; Ogunro, E.A.; Ferguson, A.G.; Lesch, M.

    1981-11-15

    Rate determination of protein synthesis utilizing tracer amino acid incorporation requires accurate assessment of the specific radioactivity of the labeled precursor aminoacyl-tRNA pool. Previously published methods presumably useful for the measurement of any aminoacyl-tRNA were unsuccessful when applied to (/sup 35/S)methionine, due to the unique chemical properties of this amino acid. Herein we describe modifications of these methods necessary for the measurement of /sup 35/S-aminoacyl-tRNA specific radioactivity from small tissue samples incubated in the presence of (/sup 35/S)methionine. The use of (/sup 35/S)methionine of high specific radioactivity enables analysis of the methionyl-tRNA from less than 100 mg of tissue. Conditions for optimal recovery of /sup 35/S-labeled dansyl-amino acid derivatives are presented and possible applications of this method are discussed.

  3. Cranial irradiation induces bone marrow-derived microglia in adult mouse brain tissue.

    PubMed

    Okonogi, Noriyuki; Nakamura, Kazuhiro; Suzuki, Yoshiyuki; Suto, Nana; Suzue, Kazutomo; Kaminuma, Takuya; Nakano, Takashi; Hirai, Hirokazu

    2014-07-01

    Postnatal hematopoietic progenitor cells do not contribute to microglial homeostasis in adult mice under normal conditions. However, previous studies using whole-body irradiation and bone marrow (BM) transplantation models have shown that adult BM cells migrate into the brain tissue and differentiate into microglia (BM-derived microglia; BMDM). Here, we investigated whether cranial irradiation alone was sufficient to induce the generation of BMDM in the adult mouse brain. Transgenic mice that express green fluorescent protein (GFP) under the control of a murine stem cell virus (MSCV) promoter (MSCV-GFP mice) were used. MSCV-GFP mice express GFP in BM cells but not in the resident microglia in the brain. Therefore, these mice allowed us to detect BM-derived cells in the brain without BM reconstitution. MSCV-GFP mice, aged 8-12 weeks, received 13.0 Gy irradiation only to the cranium, and BM-derived cells in the brain were quantified at 3 and 8 weeks after irradiation. No BM-derived cells were detected in control non-irradiated MSCV-GFP mouse brains, but numerous GFP-labeled BM-derived cells were present in the brain stem, basal ganglia and cerebral cortex of the irradiated MSCV-GFP mice. These BM-derived cells were positive for Iba1, a marker for microglia, indicating that GFP-positive BM-derived cells were microglial in nature. The population of BMDM was significantly greater at 8 weeks post-irradiation than at 3 weeks post-irradiation in all brain regions examined. Our results clearly show that cranial irradiation alone is sufficient to induce the generation of BMDM in the adult mouse.

  4. Child maltreatment and ADHD symptoms in a sample of young adults

    PubMed Central

    Sanderud, Karoline; Murphy, Siobhan; Elklit, Ask

    2016-01-01

    Objective The present study investigated the relationship between different types of childhood maltreatment (emotional, sexual, overall abuse, and no abuse) and the symptoms of attention deficit hyperactivity disorder (ADHD) in young adulthood. Method Data were collected from a Danish national study conducted by The Danish National Centre for Social Research in 2008 and 2009. A sample of 4,718 young adults (24 years of age) were randomly selected using the total birth cohort of children born in 1984. Structured interviews were conducted with a response rate of 63%, equating to a total sample size of 2,980 participants. Results Chi-square analyses revealed significant relationships between child maltreatment groups and a probable diagnosis of ADHD using the Adult ADHD Self-Report Scale (ASRS). Binary logistic regression analysis showed that the overall abuse class was more strongly associated with probable ADHD (OR=5.08), followed by emotional abuse (OR=3.09) and sexual abuse (OR=2.07). Conclusions The results showed that childhood maltreatment was associated with increased risk of ADHD symptoms in young adulthood. The findings of this study are discussed within the existing literature and suggestions for future research are outlined in order to replicate these findings in other adult populations. Highlights of the article Child maltreatment is associated with higher levels of ADHD symptoms in a nationally representative sample of young Danish adults. Co-occurring (multiple) types of maltreatment display stronger associations with ADHD symptoms with attenuated effects for sexual abuse. Males confer stronger associations with ADHD symptoms than females. Findings underscore the importance of exploring the role of childhood trauma in young adults with ADHD symptoms. PMID:27306866

  5. Extension of the Contingency Naming Test to adult assessment: psychometric analysis in a college student sample.

    PubMed

    Riddle, Tara; Suhr, Julie

    2012-01-01

    The Contingency Naming Test (CNT; Taylor, Albo, Phebus, Sachs, & Bierl, 1987) was initially designed to assess aspects of executive functioning, such as processing speed and response inhibition, in children. The measure has shown initial utility in identifying differences in executive function among child clinical groups; however, there is an absence of adequate psychometric data for use with adults. The current study expanded psychometric data upward for use with a college student sample and explored the measure's test-retest reliability and factor structure. Performance in the adult sample showed continued improvement above child norms, consistent with theories of executive function development. Exploratory factor analysis showed that the CNT is most closely related to measures of processing speed, as well as elements of response inhibition within the latter trials. Overall, results from the current study provide added support for the utility of the CNT as a measure of executive functioning in young adults. However, more research is needed to determine patterns of performance among adult clinical groups, as well as to better understand how performance patterns may change in a broader age range, including middle and older adulthood.

  6. Localization and osteoblastic differentiation potential of neural crest-derived cells in oral tissues of adult mice.

    PubMed

    Ono, Miki; Suzawa, Tetsuo; Takami, Masamichi; Yamamoto, Gou; Hosono, Tomohiko; Yamada, Atsushi; Suzuki, Dai; Yoshimura, Kentaro; Watahiki, Junichi; Hayashi, Ryuhei; Arata, Satoru; Mishima, Kenji; Nishida, Kohji; Osumi, Noriko; Maki, Koutaro; Kamijo, Ryutaro

    2015-09-01

    In embryos, neural crest cells emerge from the dorsal region of the fusing neural tube and migrate throughout tissues to differentiate into various types of cells including osteoblasts. In adults, subsets of neural crest-derived cells (NCDCs) reside as stem cells and are considered to be useful cell sources for regenerative medicine strategies. Numerous studies have suggested that stem cells with a neural crest origin persist into adulthood, especially those within the mammalian craniofacial compartment. However, their distribution as well as capacity to differentiate into osteoblasts in adults is not fully understood. To analyze the precise distribution and characteristics of NCDCs in adult oral tissues, we utilized an established line of double transgenic (P0-Cre/CAG-CAT-EGFP) mice in which NCDCs express green fluorescent protein (GFP) throughout their life. GFP-positive cells were scattered like islands throughout tissues of the palate, gingiva, tongue, and buccal mucosa in adult mice, with those isolated from the latter shown to form spheres, typical cell clusters composed of stem cells, under low-adherent conditions. Furthermore, GFP-positive cells had markedly increased alkaline phosphatase (a marker enzyme of osteoblast differentiation) activity and mineralization as shown by alizarin red staining, in the presence of bone morphogenetic protein (BMP)-2. These results suggest that NCDCs reside in various adult oral tissues and possess potential to differentiate into osteoblastic cells. NCDCs in adults may be a useful cell source for bone regeneration strategies.

  7. Evaluation of sample holders designed for long-lasting X-ray micro-tomographic scans of ex-vivo soft tissue samples

    NASA Astrophysics Data System (ADS)

    Dudak, J.; Zemlicka, J.; Krejci, F.; Karch, J.; Patzelt, M.; Zach, P.; Sykora, V.; Mrzilkova, J.

    2016-03-01

    X-ray microradiography and microtomography are imaging techniques with increasing applicability in the field of biomedical and preclinical research. Application of hybrid pixel detector Timepix enables to obtain very high contrast of low attenuating materials such as soft biological tissue. However X-ray imaging of ex-vivo soft tissue samples is a difficult task due to its structural instability. Ex-vivo biological tissue is prone to fast drying-out which is connected with undesired changes of sample size and shape producing later on artefacts within the tomographic reconstruction. In this work we present the optimization of our Timepix equipped micro-CT system aiming to maintain soft tissue sample in stable condition. Thanks to the suggested approach higher contrast of tomographic reconstructions can be achieved while also large samples that require detector scanning can be easily measured.

  8. Three-dimensional hard tissue palatal size and shape: a 10-year longitudinal evaluation in healthy adults.

    PubMed

    Ferrario, Virgilio F; Sforza, Chiarella; Dellavia, Claudia; Colombo, Anna; Ferrari, Raffaella P

    2002-01-01

    A 10-year longitudinal evaluation of the morphology (size and shape) of hard tissue palate was performed in 6 female and 6 male healthy adults (mean age at the second evaluation was 33 years, SD = 2.2). All subjects had a complete permanent dentition, including the second molars, and were free from respiratory problems. Palatal landmarks were digitized with a computerized 3D instrument, and their coordinates were used to derive a mathematical model of palatal form. Palatal shape (size-independent) was assessed by a fourth-grade polynomial in the sagittal and frontal plane projections. Palatal dimensions in the frontal and sagittal planes were computed and compared between the 2 evaluations by paired Student t tests. A great variability was observed, and no significant modifications in size were found (P > .05 for all variables). No variations in shape were observed. Sex had no significant effect for any variable (Student t for independent samples, P > .05). This study showed that in healthy subjects, hard tissue palatal morphology does not seem to change between the third and the fourth decades of life.

  9. Tissue specific uptake and elimination of perfluoroalkyl acids (PFAAs) in adult rainbow trout (Oncorhynchus mykiss) after dietary exposure.

    PubMed

    Falk, Sandy; Failing, Klaus; Georgii, Sebastian; Brunn, Hubertus; Stahl, Thorsten

    2015-06-01

    Tissue specific uptake and elimination of perfluoroalkyl acids (PFAAs) were studied in rainbow trout (Oncorhynchus mykiss). Adult trout were exposed to perfluorobutane sulfonic acid (PFBS), perfluorohexane sulfonic acid (PFHxS), perfluorooctane sulfonic acid (PFOS), perfluorooctanoic acid (PFOA) and perfluorononanoic acid (PFNA) via food over a time period of 28d. In the following 28-d depuration period the fish were fed PFAA-free food. At defined sampling times four animals were removed from the experimental tank, euthanized and dissected. Muscle, liver, kidneys, gills, blood, skin and carcass were examined individually. At the end of the accumulation phase between 0.63% (PFOA) and 15.5% (PFOS) of the absolute, applied quantity of PFAAs was recovered in the whole fish. The main target organ was the liver with recovery rates between 0.11% (PFBS) and 4.01% (PFOS) of the total amount of ingested PFAAs. Perfluoroalkyl sulfonic acids were taken up more readily and had longer estimated elimination half-lives than perfluoroalkyl carboxylic acids of the same chain length. The longest estimated elimination half-lives were found to be for PFOS between 8.4d in muscle tissue and 20.4d in the liver and for PFNA between 8.2d in the blood and 11.6d in the liver.

  10. Microscopy and elemental analysis in tissue samples using computed microtomography with synchrotron x-rays

    SciTech Connect

    Spanne, P.; Rivers, M.L.

    1988-01-01

    The initial development shows that CMT using synchrotron x-rays can be developed to ..mu..m spatial resolution and perhaps even better. This creates a new microscopy technique which is of special interest in morphological studies of tissues, since no chemical preparation or slicing of the sample is necessary. The combination of CMT with spatial resolution in the ..mu..m range and elemental mapping with sensitivity in the ppM range results in a new tool for elemental mapping at the cellular level. 7 refs., 1 fig.

  11. Assessing skin blood flow dynamics in older adults using a modified sample entropy approach.

    PubMed

    Liao, Fuyuan; Jan, Yih-Kuen

    2014-01-01

    The aging process may result in attenuated microvascular reactivity in response to environmental stimuli, which can be evaluated by analyzing skin blood flow (SBF) signals. Among various methods for analyzing physiological signals, sample entropy (SE) is commonly used to quantify the degree of regularity of time series. However, we found that for temporally correlated data, SE value depends on the sampling rate. When data are oversampled, SE may give misleading results. To address this problem, we propose to modify the definition of SE by using time-lagged vectors in the calculation of the conditional probability that any two vectors of successive data points are within a tolerance r for m points remain within the tolerance at the next point. The lag could be chosen as the first minimum of the auto mutual information function. We tested the performance of modified SE using simulated signals and SBF data. The results showed that modified SE is able to quantify the degree of regularity of the signals regardless of sampling rate. Using this approach, we observed a more regular behavior of blood flow oscillations (BFO) during local heating-induced maximal vasodilation period compared to the baseline in young and older adults and a more regular behavior of BFO in older adults compared to young adults. These results suggest that modified SE may be useful in the study of SBF dynamics.

  12. Assessment of bioburden on human and animal tissues: part 2--results of testing of human tissue and qualification of a composite sample for routine bioburden determination.

    PubMed

    Kowalski, John B; Merritt, Karen; Gocke, David; Osborne, Joel

    2012-08-01

    A quantitative method was developed and validated to assess bioburden on tissue from human donors and to compare bioburden determination results to swab culture results from the same donor. An initial study with allograft tissue from 101 donors showed a wide range of bioburden levels; values from no colony-forming units (CFU) detected to >28,000 CFU were observed. Tissues from donors that had swab cultures negative for objectionable microorganisms generally had lower bioburden than tissues from donors where objectionable microorganisms were recovered by swab culturing. In a follow-up study with 1,445 donors, a wide range of bioburden levels was again observed on tissues from donors that were swab culture negative for objectionable microorganisms. Tissues from 885 (61%) of these donors had no recoverable bioburden (<2 CFU). Importantly, tissues from 560 (39%) of the donors had recoverable bioburden which ranged from 1 to >24,000 CFU. Identification of bioburden isolates showed a diversity of genera and species. In compliance with the recent revision of the American Association of Tissue Banks K2.210 Standard, the quantitative bioburden determination method was validated with a composite tissue sample that contains bone and soft tissue sections tested together in one extraction vessel. A recovery efficiency of 68% was validated and the composite sample was shown to be representative of all of the tissues recovered from a donor. The use of the composite sample in conjunction with the quantitative bioburden determination method will facilitate an accurate assessment of the numbers and types of contaminating microorganisms on allografts prior to disinfection/sterilization. This information will ensure that disinfection/sterilization processes are properly validated and the capability of the overall allograft process is understood on a donor by donor basis.

  13. METHODS FOR USING 3-D ULTRASOUND SPECKLE TRACKING IN BIAXIAL MECHANICAL TESTING OF BIOLOGICAL TISSUE SAMPLES

    PubMed Central

    Yap, Choon Hwai; Park, Dae Woo; Dutta, Debaditya; Simon, Marc; Kim, Kang

    2014-01-01

    Being multilayered and anisotropic, biological tissues such as cardiac and arterial walls are structurally complex, making full assessment and understanding of their mechanical behavior challenging. Current standard mechanical testing uses surface markers to track tissue deformations and does not provide deformation data below the surface. In the study described here, we found that combining mechanical testing with 3-D ultrasound speckle tracking could overcome this limitation. Rat myocardium was tested with a biaxial tester and was concurrently scanned with high-frequency ultrasound in three dimensions. The strain energy function was computed from stresses and strains using an iterative non-linear curve-fitting algorithm. Because the strain energy function consists of terms for the base matrix and for embedded fibers, spatially varying fiber orientation was also computed by curve fitting. Using finite-element simulations, we first validated the accuracy of the non-linear curve-fitting algorithm. Next, we compared experimentally measured rat myocardium strain energy function values with those in the literature and found a matching order of magnitude. Finally, we retained samples after the experiments for fiber orientation quantification using histology and found that the results satisfactorily matched those computed in the experiments. We conclude that 3-D ultrasound speckle tracking can be a useful addition to traditional mechanical testing of biological tissues and may provide the benefit of enabling fiber orientation computation. PMID:25616585

  14. MicroRNA Profiling from RSV-Infected Biofluids, Whole Blood, and Tissue Samples.

    PubMed

    Anderson, Lydia; Jorquera, Patricia A; Tripp, Ralph A

    2016-01-01

    Several studies have shown that respiratory syncytial virus (RSV) can modulate the host innate immune response by dysregulation of host microRNAs (miRNAs) related to the antiviral response, a feature that also affects the memory immune response to RSV (Thornburg et al. MBio 3(6), 2012). miRNAs are small, endogenous, noncoding RNAs that function in posttranscriptional gene regulation. Here, we explain a compilation of methods for the purification, quantification, and characterization of miRNA expression profiles in biofluids, whole blood samples, and tissue samples obtained from in vivo studies. In addition, this chapter describes methods for the isolation of exosomal miRNA populations. Understanding alterations in miRNA expression profiles and identifying miRNA targets genes, and their contribution to the pathogenesis of RSV, may help elucidate novel mechanism of host-virus interaction (Rossi et al., Pediatr Pulmonol, 2015). PMID:27464696

  15. Variable ploidy of ovarian clear cell carcinomas. Implications for adequacy of tissue sampling.

    PubMed

    Listinsky, C M; Bonfiglio, T A; Leary, J

    1988-02-01

    Ten cases of clear cell (mesonephroid) adenocarcinoma of the ovary were examined for (1) variations of morphology within each tumor and its metastases, (2) ploidy of each morphologic region and (3) clinical behavior. Correlations were sought among these factors. Analysis of the ploidy in up to six morphologic regions per tumor showed variations in the ploidy in seven of the ten cases, with all seven having both diploid and nondiploid regions. The presence or absence of abnormal ploidy was not predictable based on the histomorphologic appearance of a given section. These results suggest that (1) the evaluation of a single random tissue sample may not discover aneuploidy that is present and (2) future ploidy studies on malignant tumors may require extensive tumor sampling in order to definitively exclude the presence of aneuploid populations.

  16. Rapid microarray-based genotyping of Chlamydia spp. strains from clinical tissue samples.

    PubMed

    Sachse, Konrad; Ruettger, Anke

    2015-01-01

    Pathogenic Chlamydia (C.) psittaci and C. trachomatis strains can be genotyped based on variations in the ompA genomic locus. In the present chapter, we describe rapid genotyping assays for both chlamydial agents using the ArrayStrip™ (AS) microarray platform. The test is targeting multiple discriminatory sites in the variable domains of the ompA gene by using 35 (C. psittaci) and 61 (C. trachomatis) oligonucleotide probes representing genotype-specific polymorphisms. In addition to discrimination among the established genotypes, this approach allows identification of atypical strains that were not accessible to typing using previously established techniques, such as PCR-RFLP or serotyping. The present DNA microarray assay can be conducted directly on clinical tissue samples and is suitable for tracing epidemiological chains and exploring the dissemination of particular genotypes. The procedure is easy to handle and economically affordable, and it allows genotyping of up to 32 clinical samples per day, thus lending itself for routine diagnosis as well.

  17. From the Cover: Cell-replacement therapy for diabetes: Generating functional insulin-producing tissue from adult human liver cells

    NASA Astrophysics Data System (ADS)

    Sapir, Tamar; Shternhall, Keren; Meivar-Levy, Irit; Blumenfeld, Tamar; Cohen, Hamutal; Skutelsky, Ehud; Eventov-Friedman, Smadar; Barshack, Iris; Goldberg, Iris; Pri-Chen, Sarah; Ben-Dor, Lya; Polak-Charcon, Sylvie; Karasik, Avraham; Shimon, Ilan; Mor, Eytan; Ferber, Sarah

    2005-05-01

    Shortage in tissue availability from cadaver donors and the need for life-long immunosuppression severely restrict the large-scale application of cell-replacement therapy for diabetic patients. This study suggests the potential use of adult human liver as alternate tissue for autologous beta-cell-replacement therapy. By using pancreatic and duodenal homeobox gene 1 (PDX-1) and soluble factors, we induced a comprehensive developmental shift of adult human liver cells into functional insulin-producing cells. PDX-1-treated human liver cells express insulin, store it in defined granules, and secrete the hormone in a glucose-regulated manner. When transplanted under the renal capsule of diabetic, immunodeficient mice, the cells ameliorated hyperglycemia for prolonged periods of time. Inducing developmental redirection of adult liver offers the potential of a cell-replacement therapy for diabetics by allowing the patient to be the donor of his own insulin-producing tissue. pancreas | transdifferentiation

  18. The sequential tissue distribution of duck Tembusu virus in adult ducks.

    PubMed

    Wu, Li; Liu, Jinxiong; Chen, Pucheng; Jiang, Yongping; Ding, Leilei; Lin, Yuan; Li, Qimeng; He, Xijun; Chen, Qiusheng; Chen, Hualan

    2014-01-01

    In 2010, a novel Tembusu virus (TMUV) that caused a severe decrease in the egg production of ducks was isolated in southeast China. Given the novelty of this duck pathogen, little information is available regarding its pathogenesis. Here, we systematically investigated the replication kinetics of TMUV PTD2010 in adult male and female ducks. We found that PTD2010 was detectable in most of the parenchymatous organs as well as the oviduct and intestinal tract from days 1 to 7 after inoculation. Viral titers were maintained at high levels for at least 9 days in the spleen, kidney, bursa of Fabricius, brain, and ovary. No virus was detected in any of these organs or tissues at 18 days after inoculation. PTD2010, thus, causes systemic infections in male and female ducks; its replication kinetics show similar patterns in most organs, with the exception of the ovaries and testes.

  19. Frequency of word occurrence in communication samples produced by adult communication aid users.

    PubMed

    Beukelman, D R; Yorkston, K M; Poblete, M; Naranjo, C

    1984-11-01

    Communication samples generated by five nonspeaking adults using Canon Communicators were collected for 14 consecutive days. Samples were analyzed to determine frequency of word occurrence. A core vocabulary of the 500 most frequently occurring words was analyzed further to determine spelling level and proportion of complete communication samples represented by subsets of the core vocabulary list. The 500 core vocabulary words represented 80% of the total words in the combined communication samples for the 5 subjects. Of all messages generated by the subjects, 33% could be communicated in their entirety using words from the core vocabulary list. The communication of the remaining messages required one or more words in addition to the core vocabulary. The spelling grade level of the words in the core vocabulary list did not exceed the seventh grade. The implications of the results for designing and customizing communication aids and for potential user training are discussed.

  20. Feasibility of direct digital sampling for diffuse optical frequency domain spectroscopy in tissue

    NASA Astrophysics Data System (ADS)

    Roblyer, Darren; O'Sullivan, Thomas D.; Warren, Robert V.; Tromberg, Bruce J.

    2013-04-01

    Frequency domain optical spectroscopy in the diffusive regime is currently being investigated for biomedical applications including tumor detection, therapy monitoring, exercise metabolism and others. Analog homodyne or heterodyne detection of sinusoidally modulated signals has been the predominant method for measuring phase and amplitude of photon density waves that have traversed through tissue. Here we demonstrate the feasibility of utilizing direct digital sampling of modulated signals using a 3.6 gigasample/second 12 bit analog to digital converter. Digitally synthesized modulated signals between 50 MHz and 400 MHz were measured on tissue-simulating phantoms at six near-infrared wavelengths. An amplitude and phase precision of 1% and 0.6° were achieved during drift tests. Amplitude, phase, scattering and absorption values were compared with a well-characterized network analyzer-based diffuse optical device. Optical properties measured with both systems were within 3.6% for absorption and 2.8% for scattering over a range of biologically relevant values. Direct digital sampling represents a viable method for frequency domain diffuse optical spectroscopy and has the potential to reduce system complexity, size and cost.

  1. Evaluation of convenient pretreatment protocols for RNA virus metagenomics in serum and tissue samples.

    PubMed

    Rosseel, Toon; Ozhelvaci, Orkun; Freimanis, Graham; Van Borm, Steven

    2015-09-15

    Viral metagenomic approaches are increasingly being used for viral discovery. Various strategies are applied to enrich viral sequences, but there is often a lack of knowledge about their effective influence on the viral discovery sensitivity. We evaluate some convenient and widely used approaches for RNA virus discovery in clinical samples in order to reveal their sensitivity and potential bias introduced by the enrichment or amplifications steps. An RNA virus was artificially spiked at a fixed titer in serum and lung tissue, respectively, low and high nucleic acid content matrices. For serum, a simple DNase treatment on the RNA extract gave the maximum gain in proportion of viral sequences (83×), and a subsequent ribosomal RNA removal nearly doubled once more the proportion of viral sequences. For lung tissue, a ribosomal RNA depletion step on the RNA extract had the biggest gain in proportion of viral sequences (32×). We show also that direct sequencing of cDNA is recommended above an extra random PCR amplification step, and a that the virion enrichment strategy (filtration and nuclease treatment) has a beneficial effect for sequencing-based virus discovery. Our findings provide sample-dependent guidelines for targeted virus discovery strategies.

  2. Protocol: High-throughput and quantitative assays of auxin and auxin precursors from minute tissue samples

    PubMed Central

    2012-01-01

    Background The plant hormone auxin, indole-3-acetic acid (IAA), plays important roles in plant growth and development. The signaling response to IAA is largely dependent on the local concentration of IAA, and this concentration is regulated by multiple mechanisms in plants. Therefore, the precise quantification of local IAA concentration provides insights into the regulation of IAA and its biological roles. Meanwhile, pathways and genes involved in IAA biosynthesis are not fully understood, so it is necessary to analyze the production of IAA at the metabolite level for unbiased studies of IAA biosynthesis. Results We have developed high-throughput methods to quantify plant endogenous IAA and its biosynthetic precursors including indole, tryptophan, indole-3-pyruvic acid (IPyA), and indole-3-butyric acid (IBA). The protocol starts with homogenizing plant tissues with stable-labeled internal standards added, followed by analyte purification using solid phase extraction (SPE) tips and analyte derivatization. The derivatized analytes are finally analyzed by selected reaction monitoring on a gas chromatograph-mass spectrometer (GC-MS/MS) to determine the precise abundance of analytes. The amount of plant tissue required for the assay is small (typically 2–10 mg fresh weight), and the use of SPE tips is simple and convenient, which allows preparation of large sets of samples within reasonable time periods. Conclusions The SPE tips and GC-MS/MS based method enables high-throughput and accurate quantification of IAA and its biosynthetic precursors from minute plant tissue samples. The protocol can be used for measurement of these endogenous compounds using isotope dilution, and it can also be applied to analyze IAA biosynthesis and biosynthetic pathways using stable isotope labeling. The method will potentially advance knowledge of the role and regulation of IAA. PMID:22883136

  3. Use of Adult Stem Cells for Cartilage Tissue Engineering: Current Status and Future Developments

    PubMed Central

    Baugé, Catherine; Boumédiene, Karim

    2015-01-01

    Due to their low self-repair ability, cartilage defects that result from joint injury, aging, or osteoarthritis, are the most often irreversible and are a major cause of joint pain and chronic disability. So, in recent years, researchers and surgeons have been working hard to elaborate cartilage repair interventions for patients who suffer from cartilage damage. However, current methods do not perfectly restore hyaline cartilage and may lead to the apparition of fibro- or hypertrophic cartilage. In the next years, the development of new strategies using adult stem cells, in scaffolds, with supplementation of culture medium and/or culture in low oxygen tension should improve the quality of neoformed cartilage. Through these solutions, some of the latest technologies start to bring very promising results in repairing cartilage from traumatic injury or chondropathies. This review discusses the current knowledge about the use of adult stem cells in the context of cartilage tissue engineering and presents clinical trials in progress, as well as in the future, especially in the field of bioprinting stem cells. PMID:26246809

  4. Maternal undernutrition programs tissue-specific epigenetic changes in the glucocorticoid receptor in adult offspring.

    PubMed

    Begum, Ghazala; Davies, Alison; Stevens, Adam; Oliver, Mark; Jaquiery, Anne; Challis, John; Harding, Jane; Bloomfield, Frank; White, Anne

    2013-12-01

    Epidemiological data indicate that an adverse maternal environment during pregnancy predisposes offspring to metabolic syndrome with increased obesity, and type 2 diabetes. The mechanisms are still unclear although epigenetic modifications are implicated and the hypothalamus is a likely target. We hypothesized that maternal undernutrition (UN) around conception in sheep would lead to epigenetic changes in hypothalamic neurons regulating energy balance in the offspring, up to 5 years after the maternal insult. We found striking evidence of decreased glucocorticoid receptor (GR) promoter methylation, decreased histone lysine 27 trimethylation, and increased histone H3 lysine 9 acetylation in hypothalami from male and female adult offspring of UN mothers. These findings are entirely compatible with the increased GR mRNA and protein observed in the hypothalami. The increased GR predicted the decreased hypothalamic proopiomelanocortin expression and increased obesity that we observed in the 5-year-old adult males. The epigenetic and expression changes in GR were specific to the hypothalamus. Hippocampal GR mRNA and protein were decreased in UN offspring, whereas pituitary GR was altered in a sex-specific manner. In peripheral polymorphonuclear leukocytes there were no changes in GR methylation or protein, indicating that this epigenetic analysis did not predict changes in the brain. Overall, these results suggest that moderate changes in maternal nutrition, around the time of conception, signal life-long and tissue-specific epigenetic alterations in a key gene regulating energy balance in the hypothalamus.

  5. Predicting visceral adipose tissue by MRI using DXA and anthropometry in adolescents and young adults

    PubMed Central

    Laddu, Deepika R.; Lee, Vinson R.; Blew, Robert M.; Sato, Tetsuya; Lohman, Timothy G.; Going, Scott B.

    2015-01-01

    Objective Accumulation of intra-abdominal (visceral) adipose tissue, independent of total adiposity, is associated with development of metabolic abnormalities such as insulin resistance and type-2 diabetes in children and adults. The objective of this study was to develop prediction equations for estimating visceral adiposity (VAT) measured by magnetic resonance imaging (MRI) using anthropometric variables and measures of abdominal fat mass from DXA in adolescents and young adults. Methods Cross-sectional data was collected from a multiethnic population of seventy males and females, aged 12–25 years, with BMI ranging from 14.5–38.1 kg/m2. Android (AFM; android region as defined by manufacturers instruction) and lumbar L1-L4 regional fat masses were assessed using DXA (GE Lunar Prodigy; GE Lunar Corp, Madison, WI, USA). Criterion measures of intra-abdominal visceral fat were obtained using single-slice MRI (General Electric Signa Model 5x 1.5T) and VAT area was analyzed at the level OF L4–L5. Image analysis was carried out using ZedView 3.1. Results DXA measures of AFM (r=0.76) and L1-L4 (r=0.71) were significantly (P<0.0001) correlated with MRI-measured VAT. DXA AFM, together with gender and weight, explained 62% of the variance in VAT (SEE=10.06 cm2). DXA L1-L4 fat mass with gender explained 54% of the variance in VAT (SEE=11.08 cm2). Addition of the significant interaction, gender × DXA fat mass, improved prediction of VAT from AFM (Radj2=0.61, SEE=10.10cm2) and L1-L4 (Radj2=0.59, SEE=10.39cm2). Conclusion These results demonstrate that VAT is accurately estimated from regional fat masses measured by DXA in adolescents and young adults. PMID:26097436

  6. Identification of fungal pathogens in Formalin-fixed, Paraffin-embedded tissue samples by molecular methods.

    PubMed

    Rickerts, Volker

    2016-02-01

    The etiology of invasive fungal infections (IFI) is incompletely understood due to diagnostic limitations including insensitivity of cultures and failure of histopathology to discriminate between different species. This diagnostic gap precludes the optimal use of antifungals, leading to adverse patient outcomes. The identification of fungal pathogens from Formalin-fixed, Paraffin-embedded tissue (FFPE) blocks by molecular methods is emerging as an alternative approach to study the etiology of IFI. PCR assays, including species specific- and broadrange fungal tests are used with FFPE samples from patients with proven IFI. Fungal species identification is achieved in 15-90% of the samples. This heterogeneity may be explained by the samples studied. However, comparison of different studies is impaired, as controls ruling out false positive-, false negative test results or PCR inhibition are frequently not reported. Studies using in situ hybridization also vary in the clinical samples included and the targeted fungi. In addition, target sequences, the probe chemistry and the detection of hybridization signals also account for the differences in diagnostic sensitivity. Using both approaches in parallel yields additive insights, potentially leading to a superior identification of fungal etiology and awareness of the limitations of both molecular diagnostic approaches.

  7. Liquid Microjunction Surface Sampling Probe Electrospray Mass Spectrometry for Detection of Drugs and Metabolites in Thin Tissue Sections

    SciTech Connect

    Van Berkel, Gary J; Kertesz, Vilmos; Koeplinger, Kenneth A.; Vavek, Marissa; Kong, Ah-Ng Tony

    2008-01-01

    A self-aspirating, liquid micro-junction surface sampling probe/electrospray emitter mass spectrometry system was demonstrated for use in the direct analysis of spotted and dosed drugs and their metabolites in thin tissue sections. Proof-of-principle sampling and analysis directly from tissue without the need for sample preparation was demonstrated first by raster scanning a region on a section of rat liver onto which reserpine was spotted. The mass spectral signal from selected reaction monitoring was used to develop a chemical image of the spotted drug on the tissue. The probe was also used to selectively spot sample areas of sagittal whole mouse body tissue sections that had been dosed orally (90 mg/kg) with R,S-sulforaphane 3 hrs prior to sacrifice. Sulforaphane and its glutathione and N-acetyl cysteine conjugates were monitored with selected reaction monitoring and detected in the stomach and various other tissues from the dosed mouse. No signal for these species was observed in the tissue from a control mouse. The same dosed tissue section was used to illustrate the possibility of obtaining a line scan across the whole body section. In total these results illustrate the potential for rapid screening of the distribution of drugs and metabolites in tissue sections with the micro-liquid junction surface sampling probe/electrospray mass spectrometry approach.

  8. Identification and characterization of the pumilio-2 expressed in zebrafish embryos and adult tissues.

    PubMed

    Wang, Huan Nan; Xu, Yan; Tao, Ling Jie; Zhou, Jian; Qiu, Meng Xi; Teng, Yu Hang; Deng, Feng Jiao

    2012-03-01

    Pumilio proteins regulate the translation of specific proteins required for germ cell development and morphogenesis. In the present study, we have identified the pumilio-2 in zebrafish and analyze its expression in adult tissues and early embryos. Pumilio-2 codes for the full-length Pumilio-2 protein and contains a PUF-domain. When compared to the mammalian and avian Pumilio-2 proteins, zebrafish Pumilio-2 protein was found to contain an additional sequence of 24 amino acid residues within the PUF-domain. Zebrafish pumilio-2 mRNA is expressed in the ovary, testis, liver, kidney and brain but is absent in the heart and muscle as detected by RT-PCR. The results of in situ hybridization indicate that transcripts of pumilio-2 are distributed in all blastomeres from the 1-cell stage to the sphere stage and accumulate in the head and tail during the 60%-epiboly and 3-somite stages. Transcripts were also detected in the brain and neural tube of the 24 h post-fertilization (hpf) embryos. Western blot analyses indicate that the Pumilio-2 protein is strongly expressed in the ovary, testis and brain but not in other tissues. These data suggest that pumilio-2 plays an important role in the development of the zebrafish germ cells and nervous system.

  9. Osteosarcomas and adult soft tissue sarcomas: is there a place for high LET radiation therapy?

    PubMed

    Chauvel, P

    1992-04-01

    The treatment policy for non-operable or unresectable osteosarcoma and adult soft tissue sarcoma remains unclear or controversial, despite the progress achieved in multimodality treatments. The poor results obtained by radiotherapy alone led to consider these tumours as radioresistant and to use high Linear Energy Transfer (LET) particles, such as neutrons. These particles benefit from a higher Relative Biological Efficiency (RBE) and other biological properties tending to decrease radioresistance phenomenas. From the non randomized studies previously published, neutron-therapy seems to give better local control rates, compared to photons and/or electrons. But these results are not strongly convincing, due to the large heterogeneities in patient recruitment, histological types, sizes, sites and moreover to the high complication rates encountered in some studies, even if they are mainly imputable to the use of low energy machines. The use of high-energy hospital-based accelerators combined to the possibilities of accurate dose distribution offered by conformal therapy, the potential value of light ion beam therapy combining the dose distribution advantages of protons to the biological properties of high LET particles, represent the directions in which progresses might be made for further improvement of non-operable or unresectable osteosarcoma and soft tissue sarcomas treatment results. PMID:1622850

  10. Assessment of the interstitial fluid in the subcutaneous tissue of healthy adults using ultrasonography

    PubMed Central

    Ueda-Iuchi, Terumi; Ohno, Naoki; Miyati, Tosiaki; Dai, Misako; Okuwa, Mayumi; Nakatani, Toshio; Sanada, Hiromi; Sugama, Junko

    2015-01-01

    Objective: Lymphoedema involves swelling, especially in the subcutaneous tissues. For lymphoedema management to be successful, it is necessary to remove the interstitial fluid. Subcutaneous echogenicity may be associated with interstitial fluid, but echogenicity is not an indicator for the evaluation of management because we do not directly compare echogenicity with the interstitial fluid. We aimed to identify an outcome indicator for the evaluation of interstitial fluid using ultrasonography. We assessed the correlation between echogenicity and transverse relaxation rate (R2) on magnetic resonance imaging. Methods: This was an observational study. Healthy adults with leg swelling after activity for >8 h were recruited. The legs of 13 women were evaluated using ultrasonography, magnetic resonance imaging and measurements of the limb circumference before and after an intervention to reduce the swelling. Results: Echogenicity in the oedema group was greater than that of the controls. Echogenicity decreased with reductions in oedema. The range of the strongest correlations with the changes in R2 occurred at echogenicity values of 48–144 (Pearson’s correlation coefficient: r = −0.63 and p < 0.01). Thus, it was possible to evaluate the interstitial fluid using echogenicity. Conclusion: The outcome indicators for the evaluation of interstitial fluid using ultrasonography were echogenicities in the range of 48–144, and these values were valid for assessing the interstitial fluid in the subcutaneous tissue. PMID:27092255

  11. Typology of club drug use among young adults recruited using time-space sampling.

    PubMed

    Ramo, Danielle E; Grov, Christian; Delucchi, Kevin; Kelly, Brian C; Parsons, Jeffrey T

    2010-03-01

    The present study examined patterns of recent club drug use among 400 young adults (18-29) recruited using time-space sampling in NYC. Subjects had used at least one of six club drugs (methylenedioxymethamphetamine (MDMA), ketamine, gamma-hydroxybutyrate (GHB), cocaine, methamphetamine, and D-lysergic acid diethylamide (LSD)) within the prior 3 months. We used latent class analysis (LCA) to estimate latent groups based on patterns of recent club drug use and examined differences in demographic and psychological variables by class. A 3-class model fit the data best. Patterns were: Primary cocaine users (42% of sample), Mainstream users (44% of sample), and Wide-range users (14% of sample). Those most likely to be Primary cocaine users were significantly less likely to be heterosexual males and had higher educational attainment than the other two classes. Those most likely to be Wide-range users were less likely to be heterosexual females, more likely to be gay/bisexual males, dependent on club drugs, had significantly greater drug and sexual sensation seeking, and were more likely to use when experiencing physical discomfort or pleasant times with others compared to the other two groups. Findings highlight the utility of using person-centered approaches to understand patterns of substance use, as well as highlight several patterns of club drug use among young adults. PMID:19939585

  12. In vivo facial tissue depth for Canadian Mi'kmaq adults: a case study from Nova Scotia, Canada.

    PubMed

    Peckmann, Tanya R; Harris, Mikkel; Huculak, Meaghan; Pringle, Ashleigh; Fournier, Michel

    2015-01-01

    This study examines facial tissue depth in Canadian Mi'kmaq adults. Using ultrasound, measurements were taken at 19 landmarks on the faces of 152 individuals aged 18-75 years old. The relationships between tissue thickness, age, and sex were investigated. A positive linear trend exists between tissue thickness and age for Mi'kmaq males and females at multiple landmarks. Seven landmarks show significant differences in facial tissue depth between males and females aged 18-34 years old; no landmarks show significant differences in facial tissue depth between males and females aged 35-45 years old and 46-55 years old. Significant differences were shown in facial tissue depth between Mi'kmaq and White Americans and Mi'kmaq and African Americans. These data can assist in 3-D facial reconstructions and aid in establishing the identity of unknown Mi'kmaq individuals.

  13. Mapping international practice patterns in EUS-guided tissue sampling: outcome of a global survey

    PubMed Central

    van Riet, Priscilla A.; Cahen, Djuna L.; Poley, Jan-Werner; Bruno, Marco J.

    2016-01-01

    Background and study aims: Although Endoscopic Ultrasound (EUS)-guided tissue sampling is widely used, the optimal sampling strategy remains subject of debate. We evaluated practice patterns within the international endosonographic community. Patients and methods: An online questionnaire was sent to 400 endosonographers from the United States, Europe, and Asia. Results: A total of 186 (47 %) endosonographers participated: United States 54 (29 %), Europe 85 (46 %), and Asia 47 (25 %). European (75 %) and Asian (84 %) respondents routinely check coagulation status, whereas US respondents only check on indication (64 %, P = 0.007). While propofol sedation is standard in the United States (83 %), conscious sedation is still widely used in Europe (52 %) and Asia (84 %, P < 0.001). Overall, the 22-gauge needle is most commonly used (52 %). For fine-needle aspiration (FNA) of solid pancreatic lesions, 22-gauge (45 %) and 25-gauge (49 %) needles are used equally. For fine-needle biopsy (FNB) of solid masses, the 25-gauge device is less favored than the 22-gauge FNA device (49 % versus 21 %). The 19-gauge needle is generally used for FNB of submucosal masses (62 %). Rapid on-site pathological evaluation (ROSE) is utilized more often by US (98 %) than by European and Asian respondents (51 %, P < 0.001). Cytolyt (52 %), formalin (15 %) and alcohol (15 %) are used for FNA specimen preservation in the United States and Europe, while saline (27 %) and alcohol (38 %) are widely used in Asia (P < 0.001). Conclusions: EUS-guided tissue sampling practices vary substantially within the international endosonographic community and differ considerably from recommendations expressed in guidelines. Because the clinical relevance of these variations is largely unknown, the outcome of this survey suggests a need for further studies. PMID:27227103

  14. Associations between Exposure to Alienating Behaviors, Anxiety, and Depression in an Italian Sample of Adults.

    PubMed

    Verrocchio, Maria Cristina; Baker, Amy J L; Bernet, William

    2016-05-01

    The aim of this study was to examine associations between exposure to alienating behaviors (ABs) and anxiety and depression as mediated through psychological maltreatment and parental bonding in a sample of Italian adults in the community. Five hundred and nine adults were given a measure of exposure to ABs, the Baker Strategy Questionnaire; the Psychological Maltreatment Measure; the Parental Bonding Instrument; the State-Trait Anxiety Inventory-Y; and the Beck Depression Inventory-II. Exposure to ABs was associated with psychological maltreatment, which was associated with parental bonding, which was associated with each of the three mental health outcomes: depression, state anxiety, and trait anxiety. The authors conclude that exposure to ABs in childhood represents a risk factor for subsequent poor mental health.

  15. Exploring the effect of depressive symptoms and ageing on metamemory in an Italian adult sample.

    PubMed

    Fastame, Maria Chiara

    2014-01-01

    The current study aimed to investigate the effect of depression and age-related factors on metamemory measures in an Italian adult sample. Fifty-eight healthy participants were recruited in Northern Italy and were, respectively, assigned to the following groups: Young (20-30 years old), old (60-70 years old), and Very Old (71-84 years old). Participants were administered a battery of tests, including a word recall task, self-referent mnestic efficiency scales, general beliefs about memory, and depression measures. General beliefs about memory, self-efficacy, and beliefs about the control of personal memory were predicted by age, education, depression, and mnestic and cognitive efficiency. Finally, age-related differences were found in metamemory measures: the accuracy of mnestic control processes is thought to be lower by very old adults than by old and young individuals. PMID:23731341

  16. Symptom Checklist-90-Revised scores in adult children exposed to alienating behaviors: an Italian sample.

    PubMed

    Bernet, William; Baker, Amy J L; Verrocchio, Maria C

    2015-03-01

    This study addresses a particular form of child psychological maltreatment, exposing a child to alienating behaviors in the context of a high degree of conflict between the parents. The objective of this research was to identify retrospectively the alienating behaviors that occurred in an Italian sample of children and the reported associated psychosocial symptoms. Seven hundred and thirty-nine adults in Chieti, Italy, completed an anonymous and confidential survey regarding their childhood exposure to parental alienating behaviors and measures of current symptomatology. About 75% of the sample reported some exposure to parental alienating behaviors; 15% of the sample endorsed the item, "tried to turn me against the other parent." The results revealed strong and statistically significant associations between reported exposure to parental alienating behaviors and reports of current symptomatology.

  17. Multimodal Raman-fluorescence spectroscopy of formalin fixed samples is able to discriminate brain tumors from dysplastic tissue

    NASA Astrophysics Data System (ADS)

    Anand, Suresh; Cicchi, Riccardo; Giordano, Flavio; Buccoliero, Anna Maria; Pavone, Francesco Saverio

    2014-05-01

    In the recent years, there has been a considerable surge in the application of spectroscopy for disease diagnosis. Raman and fluorescence spectra provide characteristic spectral profile related to biochemical and morphological changes when tissues progress from normal state towards malignancy. Spectroscopic techniques offer the advantage of being minimally invasive compared to traditional histopathology, real time and quantitative. In biomedical optical diagnostics, freshly excised specimens are preferred for making ex-vivo spectroscopic measurements. With regard to fresh tissues, if the lab is located far away from the clinic it could pose a problem as spectral measurements have to be performed immediately after dissection. Tissue samples are usually placed in a fixative agent such as 4% formaldehyde to preserve the samples before processing them for routine histopathological studies. Fixation prevents the tissues from decomposition by arresting autolysis. In the present study, we intend to investigate the possibility of using formalin fixed samples for discrimination of brain tumours from dysplastic tissue using Raman and fluorescence spectroscopy. Formalin fixed samples were washed with phosphate buffered saline for about 5 minutes in order to remove the effects of formalin during spectroscopic measurements. In case of fluorescence spectroscopy, changes in spectral profile have been observed in the region between 550-670 nm between dysplastic and tumor samples. For Raman measurements, we found significant differences in the spectral profiles between dysplasia and tumor. In conclusion, formalin fixed samples can be potentially used for the spectroscopic discrimination of tumor against dysplastic tissue in brain samples.

  18. One-step labeling of degenerative neurons in unfixed brain tissue samples using Fluoro-Jade C.

    PubMed

    Gu, Qiang; Schmued, Larry C; Sarkar, Sumit; Paule, Merle G; Raymick, Bryan

    2012-06-30

    Neurodegeneration is the underlying cause of a vast majority of neurological disorders and often a result of brain trauma, stroke, or neurotoxic insult. Here we describe a simple method for labeling degenerating neurons in unfixed brain tissue samples. This method could provide a new avenue for identifying and harvesting degenerative neurons from unfixed brain tissues for subsequent molecular analyses.

  19. Sources of Technical Variability in Quantitative LC-MS Proteomics: Human Brain Tissue Sample Analysis.

    SciTech Connect

    Piehowski, Paul D.; Petyuk, Vladislav A.; Orton, Daniel J.; Xie, Fang; Moore, Ronald J.; Ramirez Restrepo, Manuel; Engel, Anzhelika; Lieberman, Andrew P.; Albin, Roger L.; Camp, David G.; Smith, Richard D.; Myers, Amanda J.

    2013-05-03

    To design a robust quantitative proteomics study, an understanding of both the inherent heterogeneity of the biological samples being studied as well as the technical variability of the proteomics methods and platform is needed. Additionally, accurately identifying the technical steps associated with the largest variability would provide valuable information for the improvement and design of future processing pipelines. We present an experimental strategy that allows for a detailed examination of the variability of the quantitative LC-MS proteomics measurements. By replicating analyses at different stages of processing, various technical components can be estimated and their individual contribution to technical variability can be dissected. This design can be easily adapted to other quantitative proteomics pipelines. Herein, we applied this methodology to our label-free workflow for the processing of human brain tissue. For this application, the pipeline was divided into four critical components: Tissue dissection and homogenization (extraction), protein denaturation followed by trypsin digestion and SPE clean-up (digestion), short-term run-to-run instrumental response fluctuation (instrumental variance), and long-term drift of the quantitative response of the LC-MS/MS platform over the 2 week period of continuous analysis (instrumental stability). From this analysis, we found the following contributions to variability: extraction (72%) >> instrumental variance (16%) > instrumental stability (8.4%) > digestion (3.1%). Furthermore, the stability of the platform and its’ suitability for discovery proteomics studies is demonstrated.

  20. Cellulase activity screening using pure carboxymethylcellulose: application to soluble cellulolytic samples and to plant tissue prints.

    PubMed

    Johnsen, Hanne R; Krause, Kirsten

    2014-01-01

    Reliable, rapid and inexpensive detection of cellulolytic enzymes that can be used for a wide variety of biological and environmental samples are currently in high demand. Here, a new cellulase detection protocol is described that circumvents problems observed with popular agar-based methods by exploiting the ability of carboxymethylcellulose (CMC) to form gel-like surfaces on its own. These pure CMC-layers are sensitive to cellulolytic degradation and stainable by Gram's iodine without showing unwelcome reactions with other enzymes. The staining intensity negatively correlates with the enzyme activity and can be used for quantification. Cellulase activities are not obstructed by high sugar contents (e.g., in plant material) which limit the applicability of other quantification methods, making our new method particularly attractive for screening of plant extracts. A useful variant of this new method is its applicability to plant tissue prints for spatial mapping of the cellulolytic activity in a zymogram-like fashion. PMID:24413752

  1. Tissue response of defined collagen-elastin scaffolds in young and adult rats with special attention to calcification.

    PubMed

    Daamen, W F; Nillesen, S T M; Hafmans, T; Veerkamp, J H; van Luyn, M J A; van Kuppevelt, T H

    2005-01-01

    Collagen-elastin scaffolds may be valuable biomaterials for tissue engineering because they combine tensile strength with elasticity. In this study, the tissue response to and the calcification of these scaffolds were evaluated. In particular, the hypothesis was tested that calcification, a common phenomenon in biomaterials, may be due to microfibrils within the elastic fibre, and that these microfibrils might generate a tissue response. Four scaffolds were subcutaneously implanted, viz. collagen, collagen + pure elastin, collagen+microfibril-containing, and collagen + pulverised elastic ligament (the source for elastin). Explants were evaluated at day 3, 7 and 21. In young Sprague Dawley rats, collagen + ligament calcified substantially, whereas collagen + elastin (with and without microfibrils) calcified less, and collagen did not. Calcification started at elastic fibres. In both Sprague Dawley and Wistar adult rats, however, none of the scaffolds calcified. Mononuclear cell infiltration was prominent in young and adult Sprague Dawley rats. In adult Wistar rats, this infiltration was associated with the presence of microfibrils. Degradation of scaffolds and new matrix formation were related with cellular influx and degree of vascularisation. In conclusion, absence of microfibrils from the elastic fibre does not prevent calcification in young Sprague Dawley rats, but does reduce the tissue response in adult Wistar rats. Cellular response and calcification differs with age and strain and therefore the choice of animal model is of key importance in biomaterial evaluation.

  2. In vivo optical detection of pH in microscopic tissue samples of Arabidopsis thaliana.

    PubMed

    Kašík, Ivan; Podrazký, Ondřej; Mrázek, Jan; Martan, Tomáš; Matějec, Vlastimil; Hoyerová, Klára; Kamínek, Miroslav

    2013-12-01

    Minimally invasive in vivo measurement of pH in microscopic biological samples of μm or μl size, e.g. plant cells, tissues and saps, may help to explain complex biological processes. Consequently, techniques to achieve such measurements are a focus of interest for botanists. This paper describes a technique for the in vivo measurement of pH in the range pH5.0 to pH7.8 in microscopic plant tissue samples of Arabidopsis thaliana based on a ratiometric fluorescence method using low-loss robust tapered fiber probes. For this purpose tapered fiber probes were prepared and coated with a detection layer containing ion-paired fluorescent pH-transducer 8-hydroxypyrene-1,3,6-trisulfonic acid trisodium salt (c-HPTS). A fluorescence ratiometric approach was employed based on excitation at 415 nm and 450 nm and on the comparison of the fluorescence response at 515 nm. The suitability of tapered fiber probes for local detection of pH between 5.0 and 7.8 was demonstrated. A pH sensitivity of 0.15 pH units was achieved within the pH ranges 5.0-5.9 and 7.1-7.8, and this was improved to 0.04 pH units within the pH range 5.9-7.1. Spatial resolution of the probes was better than 20 μm and a time response within 15-20s was achieved. Despite the minute dimensions of the tapered fiber probes the setup developed was relatively robust and compact in construction and performed reliably. It has been successfully employed for the in vivo local determination of pH of mechanically resistant plant tissues of A. thaliana of microscopic scale. The detection of momentary pH gradients across the intact plant seems to be a good tool for the determination of changes in pH in response to experimental treatments affecting for example enzyme activities, availability of mineral nutrients, hormonal control of plant development and plant responses to environmental cues.

  3. Life-space mobility, perceived health, and depression symptoms in a sample of Mexican older adults.

    PubMed

    González, Bertha Cecilia Salazar; Delgado, Leticia Hernández; Quevedo, Juana Edith Cruz; Gallegos Cabriales, Esther C

    2013-01-01

    Mobility in older adults is essential to preserving their physical independence and health. Changes in mobility are related to cognitive, physical, and emotional factors, among others. We explored symptoms of depression as a mediator variable between chronic diseases and comorbidities and the outcomes of perceived health and life-space mobility in a convenience sample of 135 older Mexican adults. A cross-sectional design was used. Simple and multiple linear regression models were adjusted to verify the assumptions of mediation using Baron and Kenny's model. Chronic diseases and comorbidities served as independent variables in two separate models, perceived health and life-space mobility served as dependent variables, and depressive symptoms as the mediator variable. Results showed that perceived health and life-space mobility are affected by chronic diseases and comorbidities. However, when symptoms of depression enter the equation, the β coefficients decreased suggesting partial mediation. It is important to assess and treat depression symptoms in older adults rather than assuming that, at their age, depression is normal.

  4. The prevalence of child sexual abuse victimization in an adult sample of Texas residents.

    PubMed

    Kercher, G A; McShane, M

    1984-01-01

    Attempts to identify characteristics of children who have been sexually abused have generally been based on cases reported to clinics, hospitals, law enforcement and social service agencies. In this study a questionnaire was mailed to a representative sample of the adult population in Texas. Child sexual abuse was defined as sexual interaction between a child and an adult or between two minors when the perpetrator is significantly older than the victim or is in a position of power over the victim. A return rate of 53% was obtained. The surveys returned varied from previous studies of the same population in that the respondents were slightly older, more often married and from a slightly higher socioeconomic level. They were, however, closely matched on the variables of sex, race and educational level. Childhood sexual victimization was reported by 7.4% of the respondents. Female victims comprised 82% of this group. The racial/ethnic distribution of victims paralleled that of the Texas census with 77% white, 5% black, and 16% Hispanic. Victimization rates were highest for Hispanic females followed by black and white females in that order. Hispanic and black males reported the lowest victimization rates. Based on these findings, it was estimated that 821,359 adult Texas residents were sexually abused as children. These results were compared with those of other studies along with their implications for policy-making. PMID:6335060

  5. Empathy across the adult lifespan: Longitudinal and experience-sampling findings.

    PubMed

    Grühn, Daniel; Rebucal, Kristine; Diehl, Manfred; Lumley, Mark; Labouvie-Vief, Gisela

    2008-12-01

    This study examined change in self-reported empathy in a four-wave longitudinal study spanning 12 years (1992-2004) and the association between empathy and other measures, including daily reports of relationship experiences. Participants initially ranged in age from 10 years to 87 years. Cross-sectional and longitudinal associations of age with empathy revealed divergent patterns. Whereas cross-sectional analyses suggested that older adults scored lower in empathy than younger adults, longitudinal analyses showed no age-related decline in empathy. This combined pattern suggests that the cross-sectional age-differences reflect a cohort rather than an age effect, with older cohorts reporting lower levels of empathy than younger ones. Independent of age, empathy was associated with a positive well-being (e.g., life satisfaction) and interaction profile (e.g., positive relations with others). In addition, a subsample of participants (n = 114) conducted experience-sampling about social interactions for a week. People with high self-reported empathy perceived their interactions as more meaningful, felt more positive in these interactions, and thought that their interaction partner felt also more positive. Thus, self-reported empathy was meaningfully associated with adults' actual social interactions.

  6. Elastic moduli of normal and pathological human breast tissues: an inversion-technique-based investigation of 169 samples

    NASA Astrophysics Data System (ADS)

    Samani, Abbas; Zubovits, Judit; Plewes, Donald

    2007-03-01

    Understanding and quantifying the mechanical properties of breast tissues has been a subject of interest for the past two decades. This has been motivated in part by interest in modelling soft tissue response for surgery planning and virtual-reality-based surgical training. Interpreting elastography images for diagnostic purposes also requires a sound understanding of normal and pathological tissue mechanical properties. Reliable data on tissue elastic properties are very limited and those which are available tend to be inconsistent, in part as a result of measurement methodology. We have developed specialized techniques to measure tissue elasticity of breast normal tissues and tumour specimens and applied them to 169 fresh ex vivo breast tissue samples including fat and fibroglandular tissue as well as a range of benign and malignant breast tumour types. Results show that, under small deformation conditions, the elastic modulus of normal breast fat and fibroglandular tissues are similar while fibroadenomas were approximately twice the stiffness. Fibrocystic disease and malignant tumours exhibited a 3-6-fold increased stiffness with high-grade invasive ductal carcinoma exhibiting up to a 13-fold increase in stiffness compared to fibrogalndular tissue. A statistical analysis showed that differences between the elastic modulus of the majority of those tissues were statistically significant. Implications for the specificity advantages of elastography are reviewed.

  7. Subcutaneous adipose tissue topography (SAT-Top) development in children and young adults.

    PubMed

    Tafeit, Erwin; Möller, Reinhard; Jurimae, Toivo; Sudi, Karl; Wallner, Sandra Johanna

    2007-06-01

    The importance of body composition measurements to elucidate the dynamics of related diseases in pediatrics is gaining recognition. The methods used should not expose subjects to high doses of radiation and require substantial cooperation. The Lipometer is a new optical device that enables the non-invasive, quick and safe determination of the thickness of subcutaneous adipose tissue (SAT) layers (in mm) at any site of the human body. The topographic specification of 15 evenly distributed body sites, which makes it possible to precisely measure subcutaneous body fat distribution, is called subcutaneous adipose tissue topography (SAT-Top). SAT-Top was determined in more than 1000 children and young adults between the ages of 7 and 21. In this paper we describe the SAT-Top development of these subjects through different age groups and the differences between male and female SAT-Top development in each age group. SAT layer profiles (medians of the 15 body sites) for boys and girls in age group 1 (7-9 yrs) show a very similar pattern for both sexes, followed by slightly decreasing SAT layer thicknesses in boys and increasing values in girls in the subsequent age groups. Between age group 3 (11-13 yrs) and age group 7 (19-21 yrs) male and female SAT-Top is significantly different. The discriminating power between male and female SAT-Top was investigated by stepwise discriminant analysis, which provided no significant results for age group 1 (7-9 yrs), about 73% correct classification for age group 2 (9-11 yrs) and 3 (11-13 yrs), 83% for age group 4 (13-15 yrs), and about 91-93% for the following age groups (15-21 yrs). It is known that SAT development is the same in both sexes until puberty, when girls gain relatively more fat mass than boys to reach a higher body-fat percentage as adults. This paper presents a precise description of SAT development in boys and girls from childhood to adolescence, which provides a basis for further investigations. PMID:17847915

  8. Adjuvant Radiotherapy for Pediatric and Young Adult Nonrhabdomyosarcoma Soft-Tissue Sarcoma

    SciTech Connect

    Smith, Kristy B.; Indelicato, Daniel J.; Knapik, Jacquelyn A.; Lagmay, Joanne P.; Morris, Christopher; Kirwan, Jessica M.; Zlotecki, Robert A.; Scarborough, Mark T.; Gibbs, C. Parker; Marcus, Robert B.

    2011-09-01

    Purpose: To evaluate the prognostic factors, outcomes, and complications in patients aged {<=}30 years with resectable nonrhabdomyosarcoma soft-tissue sarcoma treated at the University of Florida with radiotherapy (RT) during a 34-year period. Methods and Materials: A total of 95 pediatric or young adult patients with nonrhabdomyosarcoma soft-tissue sarcoma were treated with curative intent with surgery and RT at the University of Florida between 1973 and 2007. The most common histologic tumor subtypes were synovial sarcoma in 22 patients, malignant fibrous histiocytoma in 19, and malignant peripheral nerve sheath tumor in 11 patients. The mean age at RT was 22 years (range, 6-30). Of the 95 patients, 73 had high-grade tumors; 45 had undergone preoperative RT and 50 postoperative RT. The prognostic factors for survival, local recurrence, and distant recurrence were analyzed. Results: The median follow-up was 7.2 years (range, 0.4-30.5). The actuarial 5-year local control rate was 88%. A microscopically negative margin was associated with superior local control. Although 83% of local recurrence cases initially developed in the absence of metastases, all patients with local failure ultimately died of their disease. The actuarial estimate of 5-year overall survival and disease-free survival was 65% and 63%, respectively. Of all the deaths, 92% were disease related. An early American Joint Committee on Cancer stage, tumor <8 cm, and the absence of neurovascular invasion were associated with superior disease-free survival. The National Cancer Institute Common Toxicity Criteria, version 3, Grade 3-4 treatment complication rate was 9%. No secondary malignancies were observed. Conclusion: In the present large single-institution study, we found positive margins and locally advanced features to be poor prognostic factors for both local progression and survival. The results from the present study have helped to characterize the therapeutic ratio of RT in pediatric and young

  9. Attitudes Towards the Sexuality of Adults with an Intellectual Disability: Parents, Support Staff, and a Community Sample

    ERIC Educational Resources Information Center

    Cuskelly, Monica; Bryde, Rachel

    2004-01-01

    Attitudes toward the sexuality of adults with intellectual disability were assessed in parents and carers of adults with intellectual disability and in a community sample. An instrument that contained items relating to eight aspects of sexuality (sexual feelings, sex education, masturbation, personal relationships, sexual intercourse,…

  10. Mental Disorders and Problem Behavior in a Community Sample of Adults with Intellectual Disability: Three-Month Prevalence and Comorbidity

    ERIC Educational Resources Information Center

    Hove, Oddbjorn; Havik, Odd E.

    2008-01-01

    The aim of this study was to investigate the prevalence of mental disorders in a Norwegian sample of adults with intellectual disability (ID) using the "Psychopathology Checklists for Adults With Intellectual Disability" (P-AID; Hove & Havik, 2008), a screening instrument adopting diagnostic criteria from the "Diagnostic Criteria for Psychiatric…

  11. Comparison of teen and adult driver crash scenarios in a nationally representative sample of serious crashes.

    PubMed

    McDonald, Catherine C; Curry, Allison E; Kandadai, Venk; Sommers, Marilyn S; Winston, Flaura K

    2014-11-01

    Motor vehicle crashes are the leading cause of death and acquired disability during the first four decades of life. While teen drivers have the highest crash risk, few studies examine the similarities and differences in teen and adult driver crashes. We aimed to: (1) identify and compare the most frequent crash scenarios-integrated information on a vehicle's movement prior to crash, immediate pre-crash event, and crash configuration-for teen and adult drivers involved in serious crashes, and (2) for the most frequent scenarios, explore whether the distribution of driver critical errors differed for teens and adult drivers. We analyzed data from the National Motor Vehicle Crash Causation Survey, a nationally representative study of serious crashes conducted by the U.S. National Highway Traffic Safety Administration from 2005 to 2007. Our sample included 642 16- to 19-year-old and 1167 35- to 54-year-old crash-involved drivers (weighted n=296,482 and 439,356, respectively) who made a critical error that led to their crash's critical pre-crash event (i.e., event that made the crash inevitable). We estimated prevalence ratios (PR) and 95% confidence intervals (CI) to compare the relative frequency of crash scenarios and driver critical errors. The top five crash scenarios among teen drivers, accounting for 37.3% of their crashes, included: (1) going straight, other vehicle stopped, rear end; (2) stopped in traffic lane, turning left at intersection, turn into path of other vehicle; (3) negotiating curve, off right edge of road, right roadside departure; (4) going straight, off right edge of road, right roadside departure; and (5) stopped in lane, turning left at intersection, turn across path of other vehicle. The top five crash scenarios among adult drivers, accounting for 33.9% of their crashes, included the same scenarios as the teen drivers with the exception of scenario (3) and the addition of going straight, crossing over an intersection, and continuing on a

  12. Temporally and spatially controlled expression of transgenes in embryonic and adult tissues

    PubMed Central

    Zhang, Qian; Triplett, Aleata A.; Harms, Don W.; Lin, Wan-chi; Creamer, Bradley A.; Rizzino, Angie; Wagner, Kay-Uwe

    2009-01-01

    Using ES cell-mediated transgenesis, we generated a novel mouse strain that permits a temporally and spatially controlled expression of responder genes in embryonic and multiple adult tissues. The transgene was constructed in a way that a CMV enhancer linked to the chicken β-actin promoter (CAG) drives the expression of the tetracycline-controlled transactivator (tTA) in particular tissues upon Cre-mediated excision of a floxed βgeo marker located between the promoter and the tTA. Based on the enzymatic activity of lacZ, the CAG-βgeo-tTA construct exhibits a widespread expression and appears to be very strong in the brain, heart, muscle, pancreas, and skin. Like the embryonic stem cell line that was used to generate this strain, the CAG-βgeo-tTA transgene is already highly active in preimplantation embryos. Using in vivo bioluminescence imaging on MMTV-Cre, CAG-βgeo-tTA, TetO-Luciferase triple transgenic mice and their controls, we demonstrated that the expression of the tTA, which is strictly dependent on the presence of Cre recombinase, induces the activation of the reporter transgene in the absence of any ligands. The tTA-mediated transactivation can be completely ablated through administration of doxycycline, and its subsequent withdrawal lifts the transcriptional block. Based on these characteristics, this novel strain may be useful in experiments that require a sustained expression of transgenes in particular cell types over a prolonged period followed by a rapid downregulation, for example in studies that examine the therapeutic value of cancer-initiating oncogenes during disease progression. PMID:19821046

  13. New aspects in fenestrated capillary and tissue dynamics in the sensory circumventricular organs of adult brains

    PubMed Central

    Miyata, Seiji

    2015-01-01

    The blood–brain barrier (BBB) generally consists of endothelial tight junction barriers that prevent the free entry of blood-derived substances, thereby maintaining the extracellular environment of the brain. However, the circumventricular organs (CVOs), which are located along the midlines of the brain ventricles, lack these endothelial barriers and have fenestrated capillaries; therefore, they have a number of essential functions, including the transduction of information between the blood circulation and brain. Previous studies have demonstrated the extensive contribution of the CVOs to body fluid and thermal homeostasis, energy balance, the chemoreception of blood-derived substances, and neuroinflammation. In this review, recent advances have been discussed in fenestrated capillary characterization and dynamic tissue reconstruction accompanied by angiogenesis and neurogliogenesis in the sensory CVOs of adult brains. The sensory CVOs, including the organum vasculosum of the lamina terminalis (OVLT), subfornical organ (SFO), and area postrema (AP), have size-selective and heterogeneous vascular permeabilities. Astrocyte-/tanycyte-like neural stem cells (NSCs) sense blood- and cerebrospinal fluid-derived information through the transient receptor potential vanilloid 1, a mechanical/osmotic receptor, Toll-like receptor 4, a lipopolysaccharide receptor, and Nax, a Na-sensing Na channel. They also express tight junction proteins and densely and tightly surround mature neurons to protect them from blood-derived neurotoxic substances, indicating that the NSCs of the CVOs perform BBB functions while maintaining the capacity to differentiate into new neurons and glial cells. In addition to neurogliogenesis, the density of fenestrated capillaries is regulated by angiogenesis, which is accompanied by the active proliferation and sprouting of endothelial cells. Vascular endothelial growth factor (VEGF) signaling may be involved in angiogenesis and neurogliogenesis, both

  14. Population patterns of Mexican corn rootworm (Coleoptera: Chrysomelidae) adults indicated by different sampling methods.

    PubMed

    Spurgeon, D W; Esquivel, J F; Suh, C P C

    2004-04-01

    The Mexican corn rootworm, Diabrotica virgifera zeae Krysan & Smith, is a serious pest of corn, Zea mays L., in several areas of Texas. Recent demonstrations of areawide adult control suggest this tactic has promise for rootworm management, but additional information regarding treatment thresholds and sampling methodology is needed. In 2000 and 2001 we examined the influence of distance into the field on rootworm captures by CRW and Pherocon AM traps, the fidelity of trap captures to population estimates from visual counts of beetles on plants (whole plant samples), and the seasonal population patterns indicated by each sampling method. Only the CRW trap consistently indicated reduced trap captures at the field margin compared with other distances. However, trends for the AM trap and whole plant samples suggested sampling on the field margin should be avoided. Population estimates at other distances into the field (2-30 m) were usually statistically similar. Thus, monitoring does not require trap placement far into the field. Both trap types indicated population peaks after flowering in corn, whereas plant samples indicated peak populations during tasseling and flowering. Both the CRW trap and plant samples showed the proportion of female beetles increased as the season progressed, but the CRW trap underestimated the proportion of females until after flowering. Regressions relating captures by traps to counts from plant samples indicated efficiency of both traps increased with increasing plant development. Our findings should increase acceptance of the CRW trap by producers and consultants and provide a rationale for development of improved, plant growth stage-specific treatment thresholds.

  15. Alaska Marine Mammal Tissue Archival Project: Sample inventory and results of analyses of selected samples for organic compounds and trace elements

    SciTech Connect

    Becker, P.R.; Wise, S.A.; Schantz, M.M.; Koster, B.J.; Zeisler, R.

    1992-02-01

    In 1987, the Alaska Marine Mammal Tissue Archival Project (AMMTAP) was established as part of the National Biomonitoring Specimen Bank (NBSB) program at the National Institute of Standards and Technology (NIST).The purpose of the AMMTAP was to establish a representative collection of Alaska marine mammal tissues for future contaminant analyses and documentation of long-term trends in environmental quality. Since 1987, specimens have been collected from 65 animals (seven species) from six different sites. The report contains the current sample inventory and the results of the analysis of selected samples for the measurement of inorganic and organic compounds.

  16. Respondent-Driven Sampling with Hard-to-Reach Emerging Adults: An Introduction and Case Study with Rural African Americans

    ERIC Educational Resources Information Center

    Kogan, Steven M.; Wejnert, Cyprian; Chen, Yi-fu; Brody, Gene H.; Slater, LaTrina M.

    2011-01-01

    Obtaining representative samples from populations of emerging adults who do not attend college is challenging for researchers. This article introduces respondent-driven sampling (RDS), a method for obtaining representative samples of hard-to-reach but socially interconnected populations. RDS combines a prescribed method for chain referral with a…

  17. Analysis of protein biomarkers in human clinical tumor samples: critical aspects to success from tissue acquisition to analysis.

    PubMed

    Warren, Madhuri V; Chan, W Y Iris; Ridley, John M

    2011-04-01

    There has been increased interest in the analysis of protein biomarkers in clinical tumor tissues in recent years. Tissue-based biomarker assays can add value and aid decision-making at all stages of drug development, as well as being developed for use as predictive biomarkers and for patient stratification and prognostication in the clinic. However, there must be an awareness of the legal and ethical issues related to the sourcing of human tissue samples. This article also discusses the limits of scope and critical aspects on the successful use of the following tissue-based methods: immunohistochemistry, tissue microarrays and automated image analysis. Future advances in standardization of tissue biobanking methods, immunohistochemistry and quantitative image analysis techniques are also discussed. PMID:21473728

  18. Walking, body mass index, and self-rated health in a representative sample of Spanish adults.

    PubMed

    Romo-Perez, Vicente; Souto, Dilia; Mota, Jorge

    2016-01-01

    Obesity and physical inactivity (PI) are risk factors for chronic diseases and are associated with lifestyle and environmental factors. The study tested the association between PI, body mass index (BMI), and self-rated health in a representative sample of the Spanish adult population (N = 21,486). The sample included 41.5% men, with mean age 52.3 years (± 18.03), and age range 20-82 years. Prevalence of overweight/obesity was 34.2%/12.7% in women and 52.1%/12.7% in men (p < 0.001 for obesity in both sexes). 53% of women and 57.5% of men met recommended levels of physical activity by walking (≥ 150 minutes/week). According to logistic regression analysis, individuals that walked less had higher risk of overweight or obesity. Data from the population-based surveillance study support suggestions that regular walking by adults is associated with positive self-rated health and better BMI profile. Obesity and low/very low self-rated health have low prevalence rates to meet the recommendations. PMID:26886367

  19. Prevalence and Correlates of Sex Exchange Among a Nationally Representative Sample of Adolescents and Young Adults.

    PubMed

    Ulloa, Emilio; Salazar, Marissa; Monjaras, Lidia

    2016-07-01

    The present study examines prevalence and correlates of exchanging sex for drugs, money, food shelter, or other favors (sex exchange) among a nationally representative sample of youth and young adults. Adolescents and young adults (n = 11,620, 53% female, 47% male) from the National Longitudinal Study of Adolescent Health were used for the current sample. Participants completed in-home interviews at both waves. Results revealed that sex exchange was reported by 4.9% (n = 569) of the population in wave 2 or wave 3, and 4.6% (n = 26) of those who exchanged sex did so at both waves. More males reported exchanging sex than females (n = 332 versus n = 237). Respondents who reported child sexual abuse were more likely to exchange sex (95% CI 2.51-4.28, p < .05) than respondents who reported any other form of child abuse. Both males and females who engaged in sex exchange were at greater risk for sexually transmitted infections; however, the odds of ever exchanging sex were highest among males who ever had gonorrhea (OR = 6.2; 95% CI 3.75-10.3). Although sex exchange has been studied extensively among homeless and runaway youth, the current study reveals sex exchange also occurs in the general population. PMID:27266400

  20. Prevalence and Correlates of Sex Exchange Among a Nationally Representative Sample of Adolescents and Young Adults.

    PubMed

    Ulloa, Emilio; Salazar, Marissa; Monjaras, Lidia

    2016-07-01

    The present study examines prevalence and correlates of exchanging sex for drugs, money, food shelter, or other favors (sex exchange) among a nationally representative sample of youth and young adults. Adolescents and young adults (n = 11,620, 53% female, 47% male) from the National Longitudinal Study of Adolescent Health were used for the current sample. Participants completed in-home interviews at both waves. Results revealed that sex exchange was reported by 4.9% (n = 569) of the population in wave 2 or wave 3, and 4.6% (n = 26) of those who exchanged sex did so at both waves. More males reported exchanging sex than females (n = 332 versus n = 237). Respondents who reported child sexual abuse were more likely to exchange sex (95% CI 2.51-4.28, p < .05) than respondents who reported any other form of child abuse. Both males and females who engaged in sex exchange were at greater risk for sexually transmitted infections; however, the odds of ever exchanging sex were highest among males who ever had gonorrhea (OR = 6.2; 95% CI 3.75-10.3). Although sex exchange has been studied extensively among homeless and runaway youth, the current study reveals sex exchange also occurs in the general population.

  1. Loneliness and mental health in a representative sample of community-dwelling Spanish older adults.

    PubMed

    Losada, Andrés; Márquez-González, María; García-Ortiz, Luis; Gómez-Marcos, Manuel Angel; Fernández-Fernández, Virginia; Rodríguez-Sánchez, Emiliano

    2012-01-01

    Research seems to support loneliness as a risk factor for mental health problems in the elderly. Most studies analyzing the effects of loneliness on older adults' mental health have relied on convenience samples. In this study, the prevalence and predictors of feelings of loneliness were studied in a representative sample of 272 community-dwelling Spanish older adults. The potential of feelings of loneliness to significantly contribute to the explanation of mental health of the elderly was also explored. The percentage of people reporting feelings of loneliness was 23.1%. Being a woman, being older, living alone, having fewer economic resources, having lower perceived health, and being dissatisfied with the frequency of contact with relatives and friends were found to be significant predictors of feelings of loneliness. Loneliness contributed significantly to the explanation of mental health, even when other significant variables were statistically controlled. The results of this study suggest that loneliness is a relevant factor for the analysis and understanding of mental health in the elderly.

  2. HEAD LICE IN HAIR SAMPLES FROM YOUTHS, ADULTS AND THE ELDERLY IN MANAUS, AMAZONAS STATE, BRAZIL

    PubMed Central

    NUNES, Suellen Cristina Barbosa; MORONI, Raquel Borges; MENDES, Júlio; JUSTINIANO, Sílvia Cássia Brandão; MORONI, Fábio Tonissi

    2015-01-01

    A study of head lice infestations among young people, adults and elderly individuals was conducted from August 2010 to July 2013 in Manaus, AM, Northern Brazil. Hair samples collected from 1,860 individuals in 18 barber shops and beauty parlors were examined for the ectoparasite. The occurrence of pediculosis and its association with factors, such as sex, age, ethnicity, hair characteristics and the socioeconomic profile of salon customers, salon location and seasonal variation were determined. The overall occurrence rate was 2.84%. Occurrence was higher in hair samples from non-blacks and the elderly. Higher occurrence was also observed during kindergarten, elementary and junior education school holidays. The results indicate that the occurrence of head lice among young people, adults and the elderly in Manaus is relatively low compared to that determined in children and in other regions of the country. After children, the elderly were the most affected. The study also indicated the need to adopt additional procedures to improve surveys among the population with low or no purchasing power, which is usually the most affected by this ectoparasitic disease. PMID:26200965

  3. HEAD LICE IN HAIR SAMPLES FROM YOUTHS, ADULTS AND THE ELDERLY IN MANAUS, AMAZONAS STATE, BRAZIL.

    PubMed

    Nunes, Suellen Cristina Barbosa; Moroni, Raquel Borges; Mendes, Júlio; Justiniano, Sílvia Cássia Brandão; Moroni, Fábio Tonissi

    2015-01-01

    A study of head lice infestations among young people, adults and elderly individuals was conducted from August 2010 to July 2013 in Manaus, AM, Northern Brazil. Hair samples collected from 1,860 individuals in 18 barber shops and beauty parlors were examined for the ectoparasite. The occurrence of pediculosis and its association with factors, such as sex, age, ethnicity, hair characteristics and the socioeconomic profile of salon customers, salon location and seasonal variation were determined. The overall occurrence rate was 2.84%. Occurrence was higher in hair samples from non-blacks and the elderly. Higher occurrence was also observed during kindergarten, elementary and junior education school holidays. The results indicate that the occurrence of head lice among young people, adults and the elderly in Manaus is relatively low compared to that determined in children and in other regions of the country. After children, the elderly were the most affected. The study also indicated the need to adopt additional procedures to improve surveys among the population with low or no purchasing power, which is usually the most affected by this ectoparasitic disease.

  4. Balancing Sodium and Potassium: Estimates of Intake in a New Zealand Adult Population Sample

    PubMed Central

    McLean, Rachael; Edmonds, Julia; Williams, Sheila; Mann, Jim; Skeaff, Sheila

    2015-01-01

    Dietary intakes of sodium and potassium are important determinants of blood pressure. We assessed sodium and potassium intake in a cross-sectional survey which included a random sample of New Zealand Adults aged 18 to 64 years from two New Zealand cities: Dunedin and Wellington. Participants completed a short questionnaire, had height, weight and blood pressure measured, and collected a 24 h urine sample. Mean 24 h sodium excretion was 3386 mg/day (95% CI 3221, 3551): 3865 mg/day for men and for 2934 mg/day women. Mean 24 h potassium excretion was 2738 mg/day (95% CI 2623, 2855): 3031 mg/day for men and 2436 mg/day for women. Mean sodium:potassium ratio was 1.32 (95% CI 1.26, 1.39); 1.39 for men and 1.26 for women. Sodium intake was higher among younger people, men, those with a higher BMI and higher potassium excretion. Potassium excretion was higher among older people, men and those with a higher sodium excretion. New Zealand adults have high sodium intakes and low potassium intakes compared to recommended levels. This is likely to adversely affect population blood pressure levels as well as incidence of cardiovascular disease. A comprehensive public health programme to reduce dietary sodium intake and increase intake of fruit and vegetables is warranted. PMID:26516912

  5. Balancing Sodium and Potassium: Estimates of Intake in a New Zealand Adult Population Sample.

    PubMed

    McLean, Rachael; Edmonds, Julia; Williams, Sheila; Mann, Jim; Skeaff, Sheila

    2015-10-28

    Dietary intakes of sodium and potassium are important determinants of blood pressure. We assessed sodium and potassium intake in a cross-sectional survey which included a random sample of New Zealand Adults aged 18 to 64 years from two New Zealand cities: Dunedin and Wellington. Participants completed a short questionnaire, had height, weight and blood pressure measured, and collected a 24 h urine sample. Mean 24 h sodium excretion was 3386 mg/day (95% CI 3221, 3551): 3865 mg/day for men and for 2934 mg/day women. Mean 24 h potassium excretion was 2738 mg/day (95% CI 2623, 2855): 3031 mg/day for men and 2436 mg/day for women. Mean sodium:potassium ratio was 1.32 (95% CI 1.26, 1.39); 1.39 for men and 1.26 for women. Sodium intake was higher among younger people, men, those with a higher BMI and higher potassium excretion. Potassium excretion was higher among older people, men and those with a higher sodium excretion. New Zealand adults have high sodium intakes and low potassium intakes compared to recommended levels. This is likely to adversely affect population blood pressure levels as well as incidence of cardiovascular disease. A comprehensive public health programme to reduce dietary sodium intake and increase intake of fruit and vegetables is warranted.

  6. Season of birth and population schizotypy: Results from a large sample of the adult general population.

    PubMed

    Konrath, Lisa; Beckius, Danièle; Tran, Ulrich S

    2016-08-30

    Although the last years have seen an increasing interest in schizotypy and its pathogenesis, there exist only a handful of studies examining the possible interaction between season of birth (SOB) and schizotypic personality structure. Available research used differing screening instruments, rendering comparisons between studies difficult, and sample sizes in adult populations may have been too small to detect a mild effect. The current study examined the association between SOB and psychometric schizotypy in the so far single-largest sample from the adult general population (N=8114), balanced for men and women, and utilizing a valid and reliable instrument for the assessment of schizotypy. Using the 12 most informative items of the Schizotypal Personality Questionnaire Brief, we obtained evidence of a small, but significant, effect of late winter and early spring births (February/March) on psychometric schizotypy. The effect was not constrained to women, but affected men and women alike. The observed association between SOB and schizotypy appears compatible with seasonal variations of temperature and influenza prevalence, and with recent evidence on seasonal variability in the activity of the human immune system. Our findings lend support to the continuum hypothesis of schizotypy and schizophrenia, for which SOB effects have been previously established. PMID:27310922

  7. Balancing Sodium and Potassium: Estimates of Intake in a New Zealand Adult Population Sample.

    PubMed

    McLean, Rachael; Edmonds, Julia; Williams, Sheila; Mann, Jim; Skeaff, Sheila

    2015-11-01

    Dietary intakes of sodium and potassium are important determinants of blood pressure. We assessed sodium and potassium intake in a cross-sectional survey which included a random sample of New Zealand Adults aged 18 to 64 years from two New Zealand cities: Dunedin and Wellington. Participants completed a short questionnaire, had height, weight and blood pressure measured, and collected a 24 h urine sample. Mean 24 h sodium excretion was 3386 mg/day (95% CI 3221, 3551): 3865 mg/day for men and for 2934 mg/day women. Mean 24 h potassium excretion was 2738 mg/day (95% CI 2623, 2855): 3031 mg/day for men and 2436 mg/day for women. Mean sodium:potassium ratio was 1.32 (95% CI 1.26, 1.39); 1.39 for men and 1.26 for women. Sodium intake was higher among younger people, men, those with a higher BMI and higher potassium excretion. Potassium excretion was higher among older people, men and those with a higher sodium excretion. New Zealand adults have high sodium intakes and low potassium intakes compared to recommended levels. This is likely to adversely affect population blood pressure levels as well as incidence of cardiovascular disease. A comprehensive public health programme to reduce dietary sodium intake and increase intake of fruit and vegetables is warranted. PMID:26516912

  8. Low invasive in vivo tissue sampling for monitoring biomarkers and drugs during surgery.

    PubMed

    Bojko, Barbara; Gorynski, Krzysztof; Gomez-Rios, German A; Knaak, Jan M; Machuca, Tiago; Cudjoe, Erasmus; Spetzler, Vinzent N; Hsin, Michael; Cypel, Marcelo; Selzner, Markus; Liu, Mingyao; Keshjavee, Shaf; Pawliszyn, Janusz

    2014-05-01

    The techniques currently used for drug, metabolite, and biomarker determination are based on sample collection, and therefore they are not suitable for repeated analysis because of the high invasiveness. Here, we present a novel method of biochemical analysis directly in organ during operation without need of a separate sample collection step: solid-phase microextraction (SPME). The approach is based on flexible microprobe coated with biocompatible extraction phase that is inserted to the tissue with no damage or disturbance of the organ. The method was evaluated during lung and liver transplantations using normothermic ex vivo liver perfusion (NEVLP) and ex vivo lung perfusion (EVLP). The study demonstrated feasibility of the method to extract wide range of endogenous compounds and drugs. Statistical analysis allowed observing metabolic changes of lung during cold ischemic time, perfusion, and reperfusion. It was also demonstrated that the level of drugs and their metabolites can be monitored over time. Based on the methylprednisolone as a selected example, the impairment of enzymatic properties of liver was detected in the injured organs but not in healthy control. This finding was supported by changes in pathways of endogenous metabolites. The SPME probe was also used for analysis of perfusion fluid using stopcock connection. The evaluation of biochemical profile of perfusates demonstrated potential of the approach for monitoring organ function during ex vivo perfusion. The simplicity of the device makes it convenient to use by medical personnel. With the microprobe, different areas of the organ or various organs can be sampled simultaneously. The technology allows assessment of organ function by biochemical profiling, determination of potential biomarkers, and drug monitoring. The use of this method for preintervention analysis could enhance the decision-making process for the best possible personalized approach, whereas post-transplantation monitoring would be

  9. Effects of MRTI sampling characteristics on estimation of HIFU SAR and tissue thermal diffusivity

    NASA Astrophysics Data System (ADS)

    Dillon, C. R.; Todd, N.; Payne, A.; Parker, D. L.; Christensen, D. A.; Roemer, R. B.

    2013-10-01

    While the non-invasive and three-dimensional nature of magnetic-resonance temperature imaging (MRTI) makes it a valuable tool for high-intensity focused ultrasound (HIFU) treatments, random and systematic errors in MRTI measurements may propagate into temperature-based parameter estimates used for pretreatment planning. This study assesses the MRTI effects of zero-mean Gaussian noise (SD = 0.0-2.0 °C), temporal sampling (tacq = 1.0-8.0 s), and spatial averaging (Res = 0.5-2.0 mm isotropic) on HIFU temperature measurements and temperature-based estimates of the amplitude and full width half maximum (FWHM) of the HIFU specific absorption rate and of tissue thermal diffusivity. The ultrasound beam used in simulations and ex vivo pork loin experiments has lateral and axial FWHM dimensions of 1.4 mm and 7.9 mm respectively. For spatial averaging simulations, beams with lateral FWHM varying from 1.2-2.2 mm are also assessed. Under noisy conditions, parameter estimates are improved by fitting to data from larger voxel regions. Varying the temporal sampling results in minimal changes in measured temperatures (<2% change) and parameter estimates (<5% change). For the HIFU beams studied, a spatial resolution of 1 × 1 × 3 mm3 or smaller is required to keep errors in temperature and all estimated parameters less than 10%. By quantifying the errors associated with these sampling characteristics, this work provides researchers with appropriate MRTI conditions for obtaining estimates of parameters essential to pretreatment modeling of HIFU thermal therapies.

  10. A mystery unraveled: nontumorigenic pluripotent stem cells in human adult tissues

    PubMed Central

    Simerman, Ariel A; Perone, Marcelo J; Gimeno, María L; Dumesic, Daniel A; Chazenbalk, Gregorio D

    2014-01-01

    Introduction: Embryonic stem cells and induced pluripotent stem cells have emerged as the gold standard of pluripotent stem cells and the class of stem cell with the highest potential for contribution to regenerative and therapeutic application; however, their translational use is often impeded by teratoma formation, commonly associated with pluripotency. We discuss a population of nontumorigenic pluripotent stem cells, termed Multilineage Differentiating Stress Enduring (Muse) cells, which offer an innovative and exciting avenue of exploration for the potential treatment of various human diseases. Areas covered: This review discusses the origin of Muse cells, describes in detail their various unique characteristics, and considers future avenues of their application and investigation with respect to what is currently known of adult pluripotent stem cells in scientific literature. We begin by defining cell potency, then discuss both mesenchymal and various reported populations of pluripotent stem cells, and finally delve into Muse cells and the characteristics that set them apart from their contemporaries. Expert opinion: Muse cells derived from adipose tissue (Muse-AT) are efficiently, routinely and painlessly isolated from human lipoaspirate material, exhibit tripoblastic differentiation both spontaneously and under media-specific induction, and do not form teratomas. We describe qualities specific to Muse-AT cells and their potential impact on the field of regenerative medicine and cell therapy. PMID:24745973

  11. Organotypical tissue cultures from adult murine colon as an in vitro model of intestinal mucosa

    PubMed Central

    Bareiss, Petra M.; Metzger, Marco; Sohn, Kai; Rupp, Steffen; Frick, Julia S.; Autenrieth, Ingo B.; Lang, Florian; Schwarz, Heinz; Skutella, Thomas

    2008-01-01

    Together with animal experiments, organotypical cell cultures are important models for analyzing cellular interactions of the mucosal epithelium and pathogenic mechanisms in the gastrointestinal tract. Here, we introduce a three-dimensional culture model from the adult mouse colon for cell biological investigations in an in vivo-like environment. These explant cultures were cultured for up to 2 weeks and maintained typical characteristics of the intestinal mucosa, including a high-prismatic epithelium with specific epithelial cell-to-cell connections, a basal lamina and various connective tissue cell types, as analyzed with immunohistological and electron microscopic methods. The function of the epithelium was tested by treating the cultures with dexamethasone, which resulted in a strong upregulation of the serum- and glucocorticoid-inducible kinase 1 similar to that found in vivo. The culture system was investigated in infection experiments with the fungal pathogen Candida albicans. Wildtype but not Δcph1/Δefg1-knockout Candida adhered to, penetrated and infiltrated the epithelial barrier. The results demonstrate the potential usefulness of this intestinal in vitro model for studying epithelial cell-cell interactions, cellular signaling and microbiological infections in a three-dimensional cell arrangement. PMID:18320204

  12. Freezing skeletal muscle tissue does not affect its decomposition in soil: evidence from temporal changes in tissue mass, microbial activity and soil chemistry based on excised samples.

    PubMed

    Stokes, Kathryn L; Forbes, Shari L; Tibbett, Mark

    2009-01-10

    The study of decaying organisms and death assemblages is referred to as forensic taphonomy, or more simply the study of graves. This field is dominated by the fields of entomology, anthropology and archaeology. Forensic taphonomy also includes the study of the ecology and chemistry of the burial environment. Studies in forensic taphonomy often require the use of analogues for human cadavers or their component parts. These might include animal cadavers or skeletal muscle tissue. However, sufficient supplies of cadavers or analogues may require periodic freezing of test material prior to experimental inhumation in the soil. This study was carried out to ascertain the effect of freezing on skeletal muscle tissue prior to inhumation and decomposition in a soil environment under controlled laboratory conditions. Changes in soil chemistry were also measured. In order to test the impact of freezing, skeletal muscle tissue (Sus scrofa) was frozen (-20 degrees C) or refrigerated (4 degrees C). Portions of skeletal muscle tissue (approximately 1.5 g) were interred in microcosms (72 mm diameter x 120 mm height) containing sieved (2mm) soil (sand) adjusted to 50% water holding capacity. The experiment had three treatments: control with no skeletal muscle tissue, microcosms containing frozen skeletal muscle tissue and those containing refrigerated tissue. The microcosms were destructively harvested at sequential periods of 2, 4, 6, 8, 12, 16, 23, 30 and 37 days after interment of skeletal muscle tissue. These harvests were replicated 6 times for each treatment. Microbial activity (carbon dioxide respiration) was monitored throughout the experiment. At harvest the skeletal muscle tissue was removed and the detritosphere soil was sampled for chemical analysis. Freezing was found to have no significant impact on decomposition or soil chemistry compared to unfrozen samples in the current study using skeletal muscle tissue. However, the interment of skeletal muscle tissue had a

  13. Detection of Cryptococcus neoformans DNA in Tissue Samples by Nested and Real-Time PCR Assays

    PubMed Central

    Bialek, Ralf; Weiss, Michael; Bekure-Nemariam, Kubrom; Najvar, Laura K.; Alberdi, Maria B.; Graybill, John R.; Reischl, Udo

    2002-01-01

    Two PCR protocols targeting the 18S rRNA gene of Cryptococcus neoformans were established, compared, and evaluated in murine cryptococcal meningitis. One protocol was designed as a nested PCR to be performed in conventional block thermal cyclers. The other protocol was designed as a quantitative single-round PCR adapted to LightCycler technology. One hundred brain homogenates and dilutions originating from 20 ICR mice treated with different azoles were examined. A fungal burden of 3 × 101 to 2.9 × 104 CFU per mg of brain tissue was determined by quantitative culture. Specific PCR products were amplified by the conventional and the LightCycler methods in 86 and 87 samples, respectively, with products identified by DNA sequencing and real-time fluorescence detection. An analytical sensitivity of 1 CFU of C. neoformans per mg of brain tissue and less than 10 CFU per volume used for extraction was observed for both PCR protocols, while homogenates of 70 organs from mice infected with other fungi were PCR negative. Specificity testing was performed with genomic DNA from 31 hymenomycetous fungal species and from the ustilaginomycetous yeast Malassezia furfur, which are phylogenetically related to C. neoformans. Twenty-four strains, including species of human skin flora like M. furfur and Trichosporon spp., were PCR negative. Amplification was observed with Cryptococcus amylolentus, Filobasidiella depauperata, Cryptococcus laurentii, and five species unrelated to clinical specimens. LightCycler PCR products from F. depauperata and Trichosporon faecale could be clearly discriminated by melting curve analysis. The sensitive and specific nested PCR assay as well as the rapid and quantitative LightCycler PCR assay might be useful for the diagnosis and monitoring of human cryptococcal infections. PMID:11874894

  14. Comparative potential of juvenile and adult human articular chondrocytes for cartilage tissue formation in three-dimensional biomimetic hydrogels.

    PubMed

    Smeriglio, Piera; Lai, Janice H; Dhulipala, Lakshmi; Behn, Anthony W; Goodman, Stuart B; Smith, Robert L; Maloney, William J; Yang, Fan; Bhutani, Nidhi

    2015-01-01

    Regeneration of human articular cartilage is inherently limited and extensive efforts have focused on engineering the cartilage tissue. Various cellular sources have been studied for cartilage tissue engineering including adult chondrocytes, and embryonic or adult stem cells. Juvenile chondrocytes (from donors below 13 years of age) have recently been reported to be a promising cell source for cartilage regeneration. Previous studies have compared the potential of adult and juvenile chondrocytes or adult and osteoarthritic (OA) chondrocytes. To comprehensively characterize the comparative potential of young, old, and diseased chondrocytes, here we examined cartilage formation by juvenile, adult, and OA chondrocytes in three-dimensional (3D) biomimetic hydrogels composed of poly(ethylene glycol) and chondroitin sulfate. All three human articular chondrocytes were encapsulated in the 3D biomimetic hydrogels and cultured for 3 or 6 weeks to allow maturation and extracellular matrix formation. Outcomes were analyzed using quantitative gene expression, immunofluorescence staining, biochemical assays, and mechanical testing. After 3 and 6 weeks, juvenile chondrocytes showed a greater upregulation of chondrogenic gene expression than adult chondrocytes, while OA chondrocytes showed a downregulation. Aggrecan and type II collagen deposition and glycosaminoglycan accumulation were high for juvenile and adult chondrocytes but not for OA chondrocytes. Similar trend was observed in the compressive moduli of the cartilage constructs generated by the three different chondrocytes. In conclusion, the juvenile, adult and OA chondrocytes showed differential responses in the 3D biomimetic hydrogels. The 3D culture model described here may also provide a useful tool to further study the molecular differences among chondrocytes from different stages, which can help elucidate the mechanisms for age-related decline in the intrinsic capacity for cartilage repair. PMID:25054343

  15. Preservation of nucleic acids and tissue morphology in paraffin-embedded clinical samples: comparison of five molecular fixatives.

    PubMed

    Staff, Synnöve; Kujala, Paula; Karhu, Ritva; Rökman, Annika; Ilvesaro, Joanna; Kares, Saara; Isola, Jorma

    2013-09-01

    Formalin fixation preserves tissue morphology at the expense of macromolecule integrity. Freshly frozen samples are the golden standard for DNA and RNA analyses but require laborious deep-freezing and frozen sectioning for morphological studies. Alternative tissue stabilisation methods are therefore needed. We analysed the preservation of nucleic acids, immunohistochemical staining properties and tissue morphology in paraffin-embedded clinical tissue samples fixed with Z7, RCL2, PAXgene, Allprotect and RNAlater. Formalin-fixed and deep-frozen samples were used as controls. Immunohistochemical analyses showed good preservation of antigenicity in all except Allprotect and RNAlater-fixed samples. RNA quality, based on RNA integrity number value by Bioanalyzer, was comparable with freshly frozen samples only in PAXgene-fixed samples. According to quantitative reverse transcription-polymerase chain reaction (qRT-PCR) analyses, RNA from PAXgene samples yielded results similar to freshly frozen samples. No difference between fixatives was seen in DNA analyses (PCR and real-time PCR). In conclusion, PAXgene seems to be superior to other molecular fixatives and formaldehyde.

  16. Design and implementation of coded aperture coherent scatter spectral imaging of cancerous and healthy breast tissue samples.

    PubMed

    Lakshmanan, Manu N; Greenberg, Joel A; Samei, Ehsan; Kapadia, Anuj J

    2016-01-01

    A scatter imaging technique for the differentiation of cancerous and healthy breast tissue in a heterogeneous sample is introduced in this work. Such a technique has potential utility in intraoperative margin assessment during lumpectomy procedures. In this work, we investigate the feasibility of the imaging method for tumor classification using Monte Carlo simulations and physical experiments. The coded aperture coherent scatter spectral imaging technique was used to reconstruct three-dimensional (3-D) images of breast tissue samples acquired through a single-position snapshot acquisition, without rotation as is required in coherent scatter computed tomography. We perform a quantitative assessment of the accuracy of the cancerous voxel classification using Monte Carlo simulations of the imaging system; describe our experimental implementation of coded aperture scatter imaging; show the reconstructed images of the breast tissue samples; and present segmentations of the 3-D images in order to identify the cancerous and healthy tissue in the samples. From the Monte Carlo simulations, we find that coded aperture scatter imaging is able to reconstruct images of the samples and identify the distribution of cancerous and healthy tissues (i.e., fibroglandular, adipose, or a mix of the two) inside them with a cancerous voxel identification sensitivity, specificity, and accuracy of 92.4%, 91.9%, and 92.0%, respectively. From the experimental results, we find that the technique is able to identify cancerous and healthy tissue samples and reconstruct differential coherent scatter cross sections that are highly correlated with those measured by other groups using x-ray diffraction. Coded aperture scatter imaging has the potential to provide scatter images that automatically differentiate cancerous and healthy tissue inside samples within a time on the order of a minute per slice. PMID:26962543

  17. Premature death of adult adoptees: analyses of a case-cohort sample.

    PubMed

    Petersen, Liselotte; Andersen, Per Kragh; Sørensen, Thorkild I A

    2005-05-01

    Genetic and environmental influence on risk of premature death in adulthood was investigated by estimating the associations in total and cause-specific mortality of adult Danish adoptees and their biological and adoptive parents. Among all 14,425 non-familial adoptions formally granted in Denmark during the period 1924 through 1947, we selected the study population according to a case-cohort sampling design. As the case-control design, the case-cohort design has the advantage of economic data collection and little loss in statistical efficiency, but the case-cohort sample has the additional advantages that rate ratio estimates may be obtained, and re-use of the cohort sample in future studies of other outcomes is possible. Analyses were performed using Kalbfleisch and Lawless's estimator for hazard ratio, and robust estimation for variances. In the main analyses the sample was restricted to birth years of the adoptees 1924 and after, and age of transfer to the adoptive parents before 7 years, and age at death was restricted to 16 to 70 years. The results showed a higher mortality among adoptees, whose biological parents died in the age range of 16 to 70 years; this was significant for deaths from natural causes, vascular causes and all causes. No influence was seen from early death of adoptive parents, regardless of cause of death.

  18. Moderators of implicit and explicit drinking identity in a large US adult sample.

    PubMed

    Lindgren, Kristen P; Gasser, Melissa L; Werntz, Alexandra; Namaky, Nauder; Baldwin, Scott A; Teachman, Bethany A

    2016-09-01

    Drinking identity (viewing oneself as a drinker) is a potential risk factor for problematic drinking in US undergraduate samples. Whether that risk extends to a broader, more general US sample is unknown. Additionally, there are critical, unanswered questions with respect to moderators of the drinking identity-problematic drinking relationship; an important issue for designing prevention efforts. Study aims were to assess the unique associations and interactive effects of implicit and explicit measures of drinking identity on problematic drinking, and to evaluate age and sex as potential moderators of the drinking identity-problematic drinking relationship. A sample of 11,320 adults aged 18-98 completed measures of implicit and explicit drinking identity and problematic drinking (the Alcohol Use Disorder Identification Test; AUDIT). Implicit and explicit drinking identity had positive, significant associations with AUDIT scores, as expected. Moderation analyses indicated small, but significant, interactions. There was an implicit by explicit identity interaction consistent with a synergistic effect: lower implicit and explicit identity was linked to a greater probability of being a non-drinker. Age moderated explicit but not implicit identity: lower drinking identity appeared to be more protective for younger individuals. Sex moderated implicit but not explicit identity: a weaker positive association with implicit identity and AUDIT scores was observed among men, potentially reflecting stigma against women's drinking. Findings suggest that drinking identity's potential as a risk factor for problematic drinking extends to a more general US sample and that both implicit and explicit identity should be targeted in prevention efforts. PMID:27156218

  19. Moderators of implicit and explicit drinking identity in a large US adult sample.

    PubMed

    Lindgren, Kristen P; Gasser, Melissa L; Werntz, Alexandra; Namaky, Nauder; Baldwin, Scott A; Teachman, Bethany A

    2016-09-01

    Drinking identity (viewing oneself as a drinker) is a potential risk factor for problematic drinking in US undergraduate samples. Whether that risk extends to a broader, more general US sample is unknown. Additionally, there are critical, unanswered questions with respect to moderators of the drinking identity-problematic drinking relationship; an important issue for designing prevention efforts. Study aims were to assess the unique associations and interactive effects of implicit and explicit measures of drinking identity on problematic drinking, and to evaluate age and sex as potential moderators of the drinking identity-problematic drinking relationship. A sample of 11,320 adults aged 18-98 completed measures of implicit and explicit drinking identity and problematic drinking (the Alcohol Use Disorder Identification Test; AUDIT). Implicit and explicit drinking identity had positive, significant associations with AUDIT scores, as expected. Moderation analyses indicated small, but significant, interactions. There was an implicit by explicit identity interaction consistent with a synergistic effect: lower implicit and explicit identity was linked to a greater probability of being a non-drinker. Age moderated explicit but not implicit identity: lower drinking identity appeared to be more protective for younger individuals. Sex moderated implicit but not explicit identity: a weaker positive association with implicit identity and AUDIT scores was observed among men, potentially reflecting stigma against women's drinking. Findings suggest that drinking identity's potential as a risk factor for problematic drinking extends to a more general US sample and that both implicit and explicit identity should be targeted in prevention efforts.

  20. Association between dietary patterns and metabolic syndrome in a sample of portuguese adults

    PubMed Central

    2012-01-01

    Background There is scarce evidence regarding the association between diet and metabolic syndrome (MetS) in Portuguese population. We aim to evaluate the association between a posteriori dietary patterns (DPs) and MetS and its features. Methods Using random digit dialing, a sample of 2167 adults was selected between 1999 and 2003, in Porto. During a face-to-face interview, a questionnaire was applied, anthropometric measures were taken, blood pressure measured and a fasting blood sample collected. Diet was assessed using a validated food frequency questionnaire, and four DPs were identified in each sex by multivariate finite mixture models. Results After adjustment for age and daily energy intake, comparing to the “healthy” DP, women with the “low fruit and vegetables” DP had a higher odds of high waist circumference (OR = 1.88 95% CI 1.17-3.01) and low HDL-cholesterol (OR = 1.78 95% IC 1.12-2.82) and women in the “red meat and alcohol” DP had higher odds of high waist circumference (OR = 1.45 95% CI 1.01-2.07) and of MetS (OR = 1.57 95% CI 1.07-2.29); men with the “fish” DP had a higher odds of high triglycerides (OR = 1.57 95% CI 1.05-2.35). After further adjustments (education, physical activity, smoking, alcohol drinking, BMI, and menopausal status) no significant associations remained. Conclusions Four distinct DPs were identified in a community sample of Portuguese adults and there was no association with the prevalence of MetS. PMID:22943133

  1. Critical comparison of sample preparation strategies for shotgun proteomic analysis of formalin-fixed, paraffin-embedded samples: insights from liver tissue

    PubMed Central

    2014-01-01

    Background The growing field of formalin-fixed paraffin-embedded (FFPE) tissue proteomics holds promise for improving translational research. Direct tissue trypsinization (DT) and protein extraction followed by in solution digestion (ISD) or filter-aided sample preparation (FASP) are the most common workflows for shotgun analysis of FFPE samples, but a critical comparison of the different methods is currently lacking. Experimental design DT, FASP and ISD workflows were compared by subjecting to the same label-free quantitative approach three independent technical replicates of each method applied to FFPE liver tissue. Data were evaluated in terms of method reproducibility and protein/peptide distribution according to localization, MW, pI and hydrophobicity. Results DT showed lower reproducibility, good preservation of high-MW proteins, a general bias towards hydrophilic and acidic proteins, much lower keratin contamination, as well as higher abundance of non-tryptic peptides. Conversely, FASP and ISD proteomes were depleted in high-MW proteins and enriched in hydrophobic and membrane proteins; FASP provided higher identification yields, while ISD exhibited higher reproducibility. Conclusions These results highlight that diverse sample preparation strategies provide significantly different proteomic information, and present typical biases that should be taken into account when dealing with FFPE samples. When a sufficient amount of tissue is available, the complementary use of different methods is suggested to increase proteome coverage and depth. PMID:25097466

  2. Determination of malachite green residues in the eggs, fry, and adult muscle-tissue of rainbow-trout (Oncorhynchus-mykiss)

    USGS Publications Warehouse

    Allen, John L.; Gofus, J.E.; Meinertz, Jeffery R.

    1994-01-01

    Malachite green, an effective antifungal therapeutant used in fish culture, is a known teratogen. We developed a method to simultaneously detect both the chromatic and leuco forms of malachite green residues in the eggs, fry, and adult muscle tissue of rainbow trout (oncorhynchus mykiss). Homogenates of these tissues were fortified with [c-14] malachite green chloride and extracted with 1% (v/v) acetic acid in acetonitrile or in methanol. The extracts were partitioned with chloroform, dried, redissolved in mobile phase, and analyzed by liquid chromatography (lc) with postcolumn oxidation of leuco malachite green to the chromatic form. Lc fractions were collected every 30 s for quantitation by scintillation counting. Recoveries of total [c-14] malachite green chloride residue were 85 and 98% in eggs fortified with labeled malachite green at concentrations of 0.5 And 1.00 Mug/g, respectively; 68% in fry similarly fortified at a concentration of 0.65 Mug/g; and 66% in muscle homogenate similarly fortified at a level of 1.00 Mug/g. The method was tested under operational conditions by exposing adult rainbow trout to 1.00 Mg/l [c-14] malachite green chloride bath for 1 h. Muscle samples analyzed by sample oxidation and scintillation counting contained 1.3 And 0.5 Mug/g total malachite green chloride residues immediately after exposure and after a 5-day withdrawal period, respectively.

  3. Integrated genome-wide analysis of genomic changes and gene regulation in human adrenocortical tissue samples

    PubMed Central

    Gara, Sudheer Kumar; Wang, Yonghong; Patel, Dhaval; Liu-Chittenden, Yi; Jain, Meenu; Boufraqech, Myriem; Zhang, Lisa; Meltzer, Paul S.; Kebebew, Electron

    2015-01-01

    To gain insight into the pathogenesis of adrenocortical carcinoma (ACC) and whether there is progression from normal-to-adenoma-to-carcinoma, we performed genome-wide gene expression, gene methylation, microRNA expression and comparative genomic hybridization (CGH) analysis in human adrenocortical tissue (normal, adrenocortical adenomas and ACC) samples. A pairwise comparison of normal, adrenocortical adenomas and ACC gene expression profiles with more than four-fold expression differences and an adjusted P-value < 0.05 revealed no major differences in normal versus adrenocortical adenoma whereas there are 808 and 1085, respectively, dysregulated genes between ACC versus adrenocortical adenoma and ACC versus normal. The majority of the dysregulated genes in ACC were downregulated. By integrating the CGH, gene methylation and expression profiles of potential miRNAs with the gene expression of dysregulated genes, we found that there are higher alterations in ACC versus normal compared to ACC versus adrenocortical adenoma. Importantly, we identified several novel molecular pathways that are associated with dysregulated genes and further experimentally validated that oncostatin m signaling induces caspase 3 dependent apoptosis and suppresses cell proliferation. Finally, we propose that there is higher number of genomic changes from normal-to-adenoma-to-carcinoma and identified oncostatin m signaling as a plausible druggable pathway for therapeutics. PMID:26446994

  4. Integrated genome-wide analysis of genomic changes and gene regulation in human adrenocortical tissue samples.

    PubMed

    Gara, Sudheer Kumar; Wang, Yonghong; Patel, Dhaval; Liu-Chittenden, Yi; Jain, Meenu; Boufraqech, Myriem; Zhang, Lisa; Meltzer, Paul S; Kebebew, Electron

    2015-10-30

    To gain insight into the pathogenesis of adrenocortical carcinoma (ACC) and whether there is progression from normal-to-adenoma-to-carcinoma, we performed genome-wide gene expression, gene methylation, microRNA expression and comparative genomic hybridization (CGH) analysis in human adrenocortical tissue (normal, adrenocortical adenomas and ACC) samples. A pairwise comparison of normal, adrenocortical adenomas and ACC gene expression profiles with more than four-fold expression differences and an adjusted P-value < 0.05 revealed no major differences in normal versus adrenocortical adenoma whereas there are 808 and 1085, respectively, dysregulated genes between ACC versus adrenocortical adenoma and ACC versus normal. The majority of the dysregulated genes in ACC were downregulated. By integrating the CGH, gene methylation and expression profiles of potential miRNAs with the gene expression of dysregulated genes, we found that there are higher alterations in ACC versus normal compared to ACC versus adrenocortical adenoma. Importantly, we identified several novel molecular pathways that are associated with dysregulated genes and further experimentally validated that oncostatin m signaling induces caspase 3 dependent apoptosis and suppresses cell proliferation. Finally, we propose that there is higher number of genomic changes from normal-to-adenoma-to-carcinoma and identified oncostatin m signaling as a plausible druggable pathway for therapeutics.

  5. Stable isotope analysis of 1987-1991 zooplankton samples and bowhead whale tissues. Final report

    SciTech Connect

    Schell, D.M.

    1992-06-01

    Stable isotope analyses of bowhead whale tissue samples and bowhead whale prey organisms collected over the years 1987 to 1991 were used to provide detail on the isotope ratio gradients evident in the arctic Alaskan zooplankton and to verify previous findings regarding the growth rates and age determination techniques developed for bowhead whales. Zooplankton of the Bering and Chukchi seas are enriched in (13)C relative to the eastern Beaufort Sea. The analysis of baleen from bowhead whales taken between 1987 to 1990 indicate that the whales are slow-growing and the young animals between year one and about six to seven years of age, undergo a period of little or no linear growth. The authors estimate that bowheads require 16-18 years to reach the length of sexual maturity, i.e., 13-14 m. From baleen Delta(13C) cycles, a 20 year record of the isotope ratios in the phytoplankton of the northern Bering and Chukchi seas was constructed. The long-term record has been compared with the temperature anomalies in surface waters of the Bering Sea. The Delta(13C) of the zooplankton is inversely correlated with temperature and refutes current models attempting to relate ocean temperature, and atmospheric carbon dioxide levels with the Delta(13C) of ocean sediment organic matter.

  6. Gene expression profiling of human breast tissue samples using SAGE-Seq.

    PubMed

    Wu, Zhenhua Jeremy; Meyer, Clifford A; Choudhury, Sibgat; Shipitsin, Michail; Maruyama, Reo; Bessarabova, Marina; Nikolskaya, Tatiana; Sukumar, Saraswati; Schwartzman, Armin; Liu, Jun S; Polyak, Kornelia; Liu, X Shirley

    2010-12-01

    We present a powerful application of ultra high-throughput sequencing, SAGE-Seq, for the accurate quantification of normal and neoplastic mammary epithelial cell transcriptomes. We develop data analysis pipelines that allow the mapping of sense and antisense strands of mitochondrial and RefSeq genes, the normalization between libraries, and the identification of differentially expressed genes. We find that the diversity of cancer transcriptomes is significantly higher than that of normal cells. Our analysis indicates that transcript discovery plateaus at 10 million reads/sample, and suggests a minimum desired sequencing depth around five million reads. Comparison of SAGE-Seq and traditional SAGE on normal and cancerous breast tissues reveals higher sensitivity of SAGE-Seq to detect less-abundant genes, including those encoding for known breast cancer-related transcription factors and G protein-coupled receptors (GPCRs). SAGE-Seq is able to identify genes and pathways abnormally activated in breast cancer that traditional SAGE failed to call. SAGE-Seq is a powerful method for the identification of biomarkers and therapeutic targets in human disease.

  7. Apicomplexa primers amplify Proteromonas (Stramenopiles, Slopalinida, Proteromonadidae) in tissue and blood samples from lizards.

    PubMed

    Maia, João P M C; Gómez-Díaz, Elena; Harris, D James

    2012-12-01

    Microscopy has traditionally been the most common method in parasitological studies, but in recent years molecular screening has become increasingly frequent to detect protozoan parasites in a wide range of vertebrate hosts and vectors. During routine molecular screening of apicomplexan parasites in reptiles using the 18S rRNA gene, we have amplified and sequenced Proteromonas parasites from three lizard hosts (less than 1% prevalence). We conducted phylogenetic analysis to confirm the taxonomic position and infer their relationships with other stramenopiles. Although our phylogeny is limited due to scarcity of molecular data on these protists, our results confirm they are closely related to Proteromonas lacertae. Our findings show that unexpected parasites can be amplified from host samples (blood and tissue) using general procedures to detect hemoparasites, and stress that positive PCR amplifications alone should not be considered as definitive proof of infection by particular parasites. Further validation by sequence confirmation and thorough phylogenetic assessment will not only avoid false positives and biased prevalence estimates but also provide valuable information on the biodiversity and phylogenetic relationships of other parasitic organisms. More generally, our results illustrate the perils of general diagnosis protocols in parasitological studies and the need of cross-validation procedures.

  8. Weight loss expectations and goals in a population sample of overweight and obese US adults.

    PubMed

    Fabricatore, Anthony N; Wadden, Thomas A; Rohay, Jeffrey M; Pillitteri, Janine L; Shiffman, Saul; Harkins, Andrea M; Burton, Steven L

    2008-11-01

    The purpose of this study was to investigate weight loss expectations and goals in a population sample of US adults who planned to make a weight loss attempt, and to examine predictors of those expectations and goals. Participants were 658 overweight and obese adults (55% women, mean age = 47.9 years, BMI = 31.8 kg/m(2)) who responded to a telephone survey about weight loss. Respondents reported weight loss expectations (i.e., reductions they realistically expected) and goals (i.e., reductions they ideally desired) for an upcoming "serious and deliberate" weight loss attempt. They also reported the expectations they had, and the reductions they actually achieved, in a previous attempt. Respondents' weight loss expectations for their upcoming attempt (8.0% reduction in initial weight) were significantly more modest than their goals for that attempt (16.8%), and smaller than the losses that they expected (12.0%), and achieved (8.9%) in their most recent past attempt (Ps

  9. Violence victimization after HIV infection in a US probability sample of adult patients in primary care.

    PubMed Central

    Zierler, S; Cunningham, W E; Andersen, R; Shapiro, M F; Nakazono, T; Morton, S; Crystal, S; Stein, M; Turner, B; St Clair, P; Bozzette, S A

    2000-01-01

    OBJECTIVES: This study estimated the proportion of HIV-infected adults who have been assaulted by a partner or someone important to them since their HIV diagnosis and the extent to which they reported HIV-seropositive status as a cause of the violence. METHODS: Study participants were from a nationally representative probability sample of 2864 HIV-infected adults who were receiving medical care and were enrolled in the HIV Costs and Service Utilization Study. All interviews (91% in person, 9% by telephone) were conducted with computer-assisted personal interviewing instruments. Interviews began in January 1996 and ended 15 months later. RESULTS: Overall, 20.5% of the women, 11.5% of the men who reported having sex with men, and 7.5% of the heterosexual men reported physical harm since diagnosis, of whom nearly half reported HIV-seropositive status as a cause of violent episodes. CONCLUSIONS: HIV-related care is an appropriate setting for routine assessment of violence. Programs to cross-train staff in antiviolence agencies and HIV care facilities need to be developed for men and women with HIV infection. PMID:10667181

  10. Prevalence and Cognitive Bases of Subjective Memory Complaints in Older Adults: Evidence from a Community Sample.

    PubMed

    Fritsch, Thomas; McClendon, McKee J; Wallendal, Maggie S; Hyde, Trevor F; Larsen, Janet D

    2014-01-01

    Objectives. To estimate the prevalence of subjective memory complaints (SMCs) in a sample of community-dwelling, older adults and to examine cognitive bases of these complaints. Participants. 499 community-dwelling adults, 65 and older. Measurements. A telephone survey consisting of cognitive tests and clinical and sociodemographic variables. SMCs were based on subjects' evaluations and subjects' perceptions of others' evaluations. Analysis. Logistic regression was used to model the risk for SMCs as a function of the cognitive, clinical, and sociodemographic variables. We tested for interactions of the cognitive variables with age, education, and gender. Results. 27.1% reported memory complaints. Among the younger age, better objective memory performance predicted lower risk for SMCs, while among the older age, better memory had no effect on risk. Among the better-educated people, better global cognitive functioning predicted lower risk for SMCs, while among the less-educated people, better global cognitive functioning had no effect on SMC risk. When predicting others' perceptions, better objective memory was associated with lower risk for SMCs. Conclusion. Objective memory performance and global cognitive functioning are associated with lower risk for SMCs, but these relationships are the strongest for the younger age and those with more education, respectively. Age and education may affect the ability to accurately appraise cognitive functioning. PMID:26317004

  11. Empathy Across the Adult Lifespan: Longitudinal and Experience-Sampling Findings

    PubMed Central

    Grühn, Daniel; Rebucal, Kristine; Diehl, Manfred; Lumley, Mark; Labouvie-Vief, Gisela

    2009-01-01

    This study examined change in self-reported empathy in a four-wave longitudinal study spanning 12 years (1992–2004) and the association between empathy and other measures, including daily reports of relationship experiences. Participants initially ranged in age from 10 years to 87 years. Cross-sectional and longitudinal associations of age with empathy revealed divergent patterns. Whereas cross-sectional analyses suggested that older adults scored lower in empathy than younger adults, longitudinal analyses showed no age-related decline in empathy. This combined pattern suggests that the cross-sectional age-differences reflect a cohort rather than an age effect, with older cohorts reporting lower levels of empathy than younger ones. Independent of age, empathy was associated with a positive well-being (e.g., life satisfaction) and interaction profile (e.g., positive relations with others). In addition, a subsample of participants (n= 114) conducted experience-sampling about social interactions for a week. People with high self-reported empathy perceived their interactions as more meaningful, felt more positive in these interactions, and thought that their interaction partner felt also more positive. Thus, self-reported empathy was meaningfully associated with adults’ actual social interactions. PMID:19102586

  12. Grinding and polishing instead of sectioning for the tissue samples with a graft: Implications for light and electron microscopy.

    PubMed

    Mukhamadiyarov, Rinat A; Sevostyanova, Victoria V; Shishkova, Daria K; Nokhrin, Andrey V; Sidorova, Olga D; Kutikhin, Anton G

    2016-06-01

    A broad use of the graft replacement requires a detailed investigation of the host-graft interaction, including both histological examination and electron microscopy. A high quality sectioning of the host tissue with a graft seems to be complicated; in addition, it is difficult to examine the same tissue area by both of the mentioned microscopy techniques. To solve these problems, we developed a new technique of epoxy resin embedding with the further grinding, polishing, and staining. Graft-containing tissues prepared by grinding and polishing preserved their structure; however, sectioning frequently required the explantation of the graft and led to tissue disintegration. Moreover, stained samples prepared by grinding and polishing may then be assessed by both light microscopy and backscattered scanning electron microscopy. Therefore, grinding and polishing outperform sectioning when applied to the tissues with a graft. PMID:27023831

  13. The adult brain tissue response to hollow fiber membranes of varying surface architecture with or without cotransplanted cells

    NASA Astrophysics Data System (ADS)

    Zhang, Ning

    A variety of biomaterials have been chronically implanted into the central nervous system (CNS) for repair or therapeutic purposes. Regardless of the application, chronic implantation of materials into the CNS induces injury and elicits a wound healing response, eventually leading to the formation of a dense extracellular matrix (ECM)-rich scar tissue that is associated with the segregation of implanted materials from the surrounding normal tissue. Often this reaction results in impaired performance of indwelling CNS devices. In order to enhance the performance of biomaterial-based implantable devices in the CNS, this thesis investigated whether adult brain tissue response to implanted biomaterials could be manipulated by changing biomaterial surface properties or further by utilizing the biology of co-transplanted cells. Specifically, the adult rat brain tissue response to chronically implanted poly(acrylonitrile-vinylchloride) (PAN-PVC) hollow fiber membranes (HFMs) of varying surface architecture were examined temporally at 2, 4, and 12 weeks postimplantation. Significant differences were discovered in the brain tissue response to the PAN-PVC HFMs of varying surface architecture at 4 and 12 weeks. To extend this work, whether the soluble factors derived from a co-transplanted cellular component further affect the brain tissue response to an implanted HFM in a significant way was critically exploited. The cells used were astrocytes, whose ability to influence scar formation process following CNS injury by physical contact with the host tissue had been documented in the literature. Data indicated for the first time that astrocyte-derived soluble factors ameliorate the adult brain tissue reactivity toward HFM implants in an age-dependent manner. While immature astrocytes secreted soluble factors that suppressed the brain tissue reactivity around the implants, mature astrocytes secreted factors that enhanced the gliotic response. These findings prove the feasibility

  14. Feasibility and Utility of Experience Sampling to Assess Alcohol Consumption Among Older Adults

    PubMed Central

    Sacco, Paul; Smith, Cristan A.; Harrington, Donna; Svoboda, Deborah V.; Resnick, Barbara

    2015-01-01

    In the literature on alcohol use and aging, drinking has often been conceptualized as a means of coping with negative feelings, such as stress, yet much of the literature on older adults and drinking has utilized cross-sectional or other data ill-suited for exploring dynamic processes. Experience sampling methods have the ability to measure and analyze dynamic processes in real time, such as relations between alcohol use and mood states. Nonetheless, these approaches are intensive and may burden respondents. Therefore, this study evaluated the feasibility, acceptability, and validity of a modified daily diary to measure alcohol use and explored alternate methods of collecting diary data. Findings suggest that a modified diary was acceptable and not burdensome. Respondents were reluctant to consider technology (e.g., cellphone)-based means of data collection. Measures of alcohol use showed little within-person variation suggesting that for those who drink at all, drinking is a daily habit. PMID:24652928

  15. Incremental Validity of Mindfulness Skills in relation to Emotional Dysregulation among a Young Adult Community Sample

    PubMed Central

    Vujanovic, Anka A.; Bonn-Miller, Marcel O.; Bernstein, Amit; McKee, Laura G.; Zvolensky, Michael J.

    2009-01-01

    The present investigation examined the incremental predictive validity of mindfulness skills, as measured by the Kentucky Inventory of Mindfulness Skills (KIMS), in relation to multiple facets of emotional dysregulation, as indexed by the Difficulties in Emotion Regulation Scale (DERS), above and beyond variance explained by negative affectivity, anxiety sensitivity, and distress tolerance. Participants were a non-clinical community sample of 193 young adults (106 women; Mage = 23.91). The KIMS - Accepting without Judgment subscale was incrementally negatively predictive of all facets of emotional dysregulation, as measured by the DERS. Furthermore, KIMS - Acting with Awareness was incrementally negatively related to Difficulties Engaging in Goal-Directed Behavior. Additionally, both Observing and Describing mindfulness skills were incrementally negatively related to Lack of Emotional Awareness, and Describing skills also were incrementally negatively related to a Lack of Emotional Clarity. Findings are discussed in relation to advancing scientific understanding of emotional dysregulation, from a mindfulness skills-based framework. PMID:20182933

  16. Linking typologies of childhood adversity to adult incarceration: Findings from a nationally representative sample.

    PubMed

    Roos, Leslie E; Afifi, Tracie O; Martin, Christina Gamache; Pietrzak, Robert H; Tsai, Jack; Sareen, Jitender

    2016-01-01

    Ecologically valid typologies of adverse child experiences (ACEs) were identified to investigate the link between ACEs and adult incarceration. In a nationally representative sample (N = 34,653, age 20+), latent class analysis (LCA) was conducted with childhood maltreatment (physical, sexual, and emotional abuse, interpersonal violence [IPV] exposure, physical neglect) and caregiver maladjustment (substance use, incarceration, mental illness, and suicidal behavior) indicators. LCA identified a 5-typology model (1. ; 2. ; 3. ; 4. ; and 5. ). Controlling for sociodemographics and substance use problems, logistic regression analyses determined that, compared with the typology, all typologies (except ) had elevated incarceration risk (adjusted odds ratios: 1.76-4.18). Maltreatment experiences were more predictive of incarceration for women versus men. Childhood maltreatment confers risk for incarceration beyond established risk factors, but caregiver maladjustment, alone, does not. Preventative efforts should focus on understanding and targeting pathways to delinquency for individuals with childhood maltreatment. (PsycINFO Database Record PMID:27078049

  17. Polytraumatization in an adult national sample and its association with psychological distress and self-esteem

    PubMed Central

    Nilsson, Doris; Dahlstöm, Örjan; Priebe, Gisela; Svedin, Carl Göran

    2015-01-01

    Objective The objective of this study was to examine the prevalence of self-reported experiences of potential childhood traumas and polytraumatization, and to find cut-off values for different kinds of potential traumatic events in a national representative sample of adults in Sweden. In addition, to analyse the association between polytraumatization and both psychological distress and global self-esteem. Method A web-based survey - containing SCL-25 and Rosenberg Self-Esteem Scale, and Linköping Difficult Life Events Scale - Adult - was sent out to a nationally reprative sample and 5062 people chose to participate in the study. Results Results showed that almost everyone (97%) has experienced at least one potential traumatic event and that polytraumatization (the 10% of the participants with most reported traumas) was significantly (Z = 12.57, P < 0.001, r = 0.18) associated with psychological distress and global self-esteem. Gender differences were significant (Z = 8.44, P < 0.001, r = 0.12), in that men experience more noninterpersonal traumas but women report more symptoms. The effect sizes regarding the impact of potential trauma on self-esteem were largest for women with experience of polytraumatization in the age group 18–25 (r = 0.48). There was almost linear increase in psychological distress and linear decrease in self-esteem with increasing number of traumatic events experienced. Conclusion Experience of polytrauma can be considered an important factor to take into account in psychiatric settings as well. PMID:25722950

  18. Substance Use and Treatment of Substance Use Disorders in a Community Sample of Transgender Adults

    PubMed Central

    Keuroghlian, Alex S.; Reisner, Sari L.; White, Jaclyn M.; Weiss, Roger D.

    2015-01-01

    Background Transgender people have elevated substance use prevalence compared with the U.S. general population, however no studies have comprehensively examined the relationship of psychosocial risk factors to substance use and substance use disorder (SUD) treatment among both male-to-female (MTF) and female-to-male (FTM) transgender adults. Methods Secondary data analysis of a 2013 community-based survey of transgender adults in Massachusetts (N=452) was conducted. Adjusted multivariable logistic regression models were fit to examine the relationship of four risk factor domains with SUD treatment history and recent substance use: (1) demographics; (2) gender-related characteristics; (3) mental health; (4) socio-structural factors. Adjusted Odds Ratios (aOR) and 95% Confidence Intervals (95% CI) were estimated. Results Ten percent of the sample reported lifetime SUD treatment. Factors associated with significant increase in odds of lifetime SUD treatment alongside recent substance use (all p<0.05) were: (1) older age (aOR=1.02; 95% CI=1.01–1.04), higher educational attainment (aOR=3.59; 95% CI=2.35–5.50), low income (aOR=0.58; 95% CI=0.39–0.86); (2) MTF identity (aOR=3.03; 95% CI=1.95–4.67), gender-affirming medical care (aOR=1.99; 95% CI=1.32–3.00); (3) intimate partner violence (aOR=1.68; 95% CI=1.13–2.49), posttraumatic stress disorder (aOR = 2.56; 95% CI=1.69–3.88), depression (aOR=2.30; 95% CI=1.58–3.35), mental health treatment (aOR=1.65; 95% CI=1.11–2.45); (4) discrimination (aOR=1.90; 95% CI=1.22–2.95), unstable housing (aOR=1.80; 95% CI=1.21–2.67), and sex work (aOR=2.48; 95% CI=1.24–4.95). Conclusions Substance use and SUD treatment among transgender adults are associated with demographic, gender-related, mental health, and socio-structural risk factors. Studies are warranted that identify SUD treatment barriers, and integrate SUD treatment with psychosocial and structural interventions for a diverse spectrum of transgender adults

  19. Comparison of the relationship between bone marrow adipose tissue and volumetric bone mineral density in children and adults.

    PubMed

    Shen, Wei; Velasquez, Gilbert; Chen, Jun; Jin, Ye; Heymsfield, Steven B; Gallagher, Dympna; Pi-Sunyer, F Xavier

    2014-01-01

    Several large-scale studies have reported the presence of an inverse relationship between bone mineral density (BMD) and bone marrow adipose tissue (BMAT) in adults. We aim to determine if there is an inverse relationship between pelvic volumetric BMD (vBMD) and pelvic BMAT in children and to compare this relationship in children and adults. Pelvic BMAT and bone volume (BV) was evaluated in 181 healthy children (5-17yr) and 495 healthy adults (≥18yr) with whole-body magnetic resonance imaging (MRI). Pelvic vBMD was calculated using whole-body dual-energy X-ray absorptiometry to measure pelvic bone mineral content and MRI-measured BV. An inverse correlation was found between pelvic BMAT and pelvic vBMD in both children (r=-0.374, p<0.001) and adults (r=-0.650, p<0.001). In regression analysis with pelvic vBMD as the dependent variable and BMAT as the independent variable, being a child or adult neither significantly contribute to the pelvic BMD (p=0.995) nor did its interaction with pelvic BMAT (p=0.415). The inverse relationship observed between pelvic vBMD and pelvic BMAT in children extends previous findings that found the inverse relationship to exist in adults and provides further support for a reciprocal relationship between adipocytes and osteoblasts.

  20. [Bacteria isolated from urine and renal tissue samples and their relation to renal histology].

    PubMed

    Gökalp, A; Gültekin, E Y; Bakici, M Z; Ozdeşlik, B

    1988-01-01

    The bacteria from the urine and renal biopsy specimens of 40 patients undergoing renal surgery were isolated and their relations with renal histology investigated. The urine cultures were positive in 14 patients, the same organisms being isolated from the renal tissue in 7 cases. In 6 patients with negative urine cultures, bacteria were isolated from renal tissues. Of the 28 cases pathologically diagnosed as chronic pyelonephritis, bacteria were isolated from the renal tissue in 13 cases, the urine cultures being positive in only 11 cases. E. coli was the most commonly encountered bacteria in both the urine and renal tissues.

  1. Comparison between whole mount tissue preparations and virtual tissue microarray samples for measuring Ki-67 and apoptosis indices in human bladder cancer: A cross-sectional study.

    PubMed

    Oshiro, Hisashi; Czerniak, Bogdan A; Sakamaki, Kentaro; Tsuta, Koji; Bondaruk, Jolanta; Keyhani, Afsaneh; Dinney, Colin P; Nagai, Takeshi; Kamat, Ashish M

    2016-08-01

    Recent tissue microarray (TMA)-based studies have shown that cell proliferation- and apoptosis-related biomarkers are associated with clinical outcomes in patients with bladder urothelial carcinoma. However, little is known about the differences in these biomarker measurements between whole mount tissue preparations and TMAs. This study aimed to elucidate the discrepancy in the measurements of Ki-67 indices (KIs) and apoptosis indices (AIs) between whole mount tissue preparations and TMAs of bladder urothelial carcinoma samples.Whole mount tissue preparations for Ki-67 immunohistochemistry and terminal deoxynucleotidyl transferase dUTP nick end labeling were made from 30 patients who underwent transurethral resection of bladder urothelial carcinoma. Digital microscopy-assisted virtual TMAs, consisting of 3 small round areas (1 or 0.6 mm in diameter), were generated from the same whole mount tissue preparations. The measurement results in highly reactive areas of biomarkers were compared between the whole mount tissue preparation- and the TMA-based methods. Bland-Altman plot analysis, regression analysis, and Kendall τ were performed to investigate differences in the measurement results, systematic biases, and correlations between biomarkers.Although the Bland-Altman plot analysis demonstrated that almost all the plots were within the limits of agreement, fixed biases were detected in the 1- and 0.6-mm TMAs for the KI (0.181 and 0.222, respectively) and the AI (0.055 and 0.063, respectively). Proportional biases were also detected in the 1- and 0.6-mm TMAs for the AI (P < 0.001 and P < 0.001, respectively). Furthermore, positive correlations between KIs and AIs were observed in whole mount tissue preparations (r = 0.260, P = 0.044) and in the 1 mm TMAs (r = 0.375, P = 0.004); however, no such correlation was observed in the 0.6 mm TMAs.Our study suggests that the measurement results for certain biomarkers of bladder urothelial

  2. Comparison between whole mount tissue preparations and virtual tissue microarray samples for measuring Ki-67 and apoptosis indices in human bladder cancer

    PubMed Central

    Oshiro, Hisashi; Czerniak, Bogdan A.; Sakamaki, Kentaro; Tsuta, Koji; Bondaruk, Jolanta; Keyhani, Afsaneh; Dinney, Colin P.; Nagai, Takeshi; Kamat, Ashish M.

    2016-01-01

    Abstract Recent tissue microarray (TMA)-based studies have shown that cell proliferation- and apoptosis-related biomarkers are associated with clinical outcomes in patients with bladder urothelial carcinoma. However, little is known about the differences in these biomarker measurements between whole mount tissue preparations and TMAs. This study aimed to elucidate the discrepancy in the measurements of Ki-67 indices (KIs) and apoptosis indices (AIs) between whole mount tissue preparations and TMAs of bladder urothelial carcinoma samples. Whole mount tissue preparations for Ki-67 immunohistochemistry and terminal deoxynucleotidyl transferase dUTP nick end labeling were made from 30 patients who underwent transurethral resection of bladder urothelial carcinoma. Digital microscopy-assisted virtual TMAs, consisting of 3 small round areas (1 or 0.6 mm in diameter), were generated from the same whole mount tissue preparations. The measurement results in highly reactive areas of biomarkers were compared between the whole mount tissue preparation- and the TMA-based methods. Bland–Altman plot analysis, regression analysis, and Kendall τ were performed to investigate differences in the measurement results, systematic biases, and correlations between biomarkers. Although the Bland–Altman plot analysis demonstrated that almost all the plots were within the limits of agreement, fixed biases were detected in the 1- and 0.6-mm TMAs for the KI (0.181 and 0.222, respectively) and the AI (0.055 and 0.063, respectively). Proportional biases were also detected in the 1- and 0.6-mm TMAs for the AI (P < 0.001 and P < 0.001, respectively). Furthermore, positive correlations between KIs and AIs were observed in whole mount tissue preparations (r = 0.260, P = 0.044) and in the 1 mm TMAs (r = 0.375, P = 0.004); however, no such correlation was observed in the 0.6 mm TMAs. Our study suggests that the measurement results for certain biomarkers of bladder

  3. An Unsupervised MVA Method to Compare Specific Regions in Human Breast Tumor Tissue Samples Using ToF-SIMS

    PubMed Central

    Bluestein, Blake M.; Morrish, Fionnuala; Graham, Daniel J.; Guenthoer, Jamie; Hockenbery, David; Porter, Peggy; Gamble, Lara J.

    2016-01-01

    Imaging time-of-flight secondary ion mass spectrometry (ToF-SIMS) and principal component analysis (PCA) were used to investigate two sets of pre- and post-chemotherapy human breast tumor tissue sections to characterize lipids associated with tumor metabolic flexibility and response to treatment. The micron spatial resolution imaging capability of ToF-SIMS provides a powerful approach to attain spatially-resolved molecular and cellular data from cancerous tissues not available with conventional imaging techniques. Three ca. 1 mm2 areas per tissue section were analyzed by stitching together 200 μm × 200 μm raster area scans. A method to isolate and analyze specific tissue regions of interest by utilizing PCA of ToF-SIMS images is presented, which allowed separation of cellularized areas from stromal areas. These PCA-generated regions of interest were then used as masks to reconstruct representative spectra from specifically stromal or cellular regions. The advantage of this unsupervised selection method is a reduction in scatter in the spectral PCA results when compared to analyzing all tissue areas or analyzing areas highlighted by a pathologist. Utilizing this method, stromal and cellular regions of breast tissue biopsies taken pre- versus post-chemotherapy demonstrate chemical separation using negatively-charged ion species. In this sample set, the cellular regions were predominantly all cancer cells. Fatty acids (i.e. palmitic, oleic, and stearic), monoacylglycerols, diacylglycerols and vitamin E profiles were distinctively different between the pre- and post-therapy tissues. These results validate a new unsupervised method to isolate and interpret biochemically distinct regions in cancer tissues using imaging ToF-SIMS data. In addition, the method developed here can provide a framework to compare a variety of tissue samples using imaging ToF-SIMS, especially where there is section-to-section variability that makes it difficult to use a serial hematoxylin and

  4. Optical coherence tomography detection of shear wave propagation in inhomogeneous tissue equivalent phantoms and ex-vivo carotid artery samples

    PubMed Central

    Razani, Marjan; Luk, Timothy W.H.; Mariampillai, Adrian; Siegler, Peter; Kiehl, Tim-Rasmus; Kolios, Michael C.; Yang, Victor X.D.

    2014-01-01

    In this work, we explored the potential of measuring shear wave propagation using optical coherence elastography (OCE) in an inhomogeneous phantom and carotid artery samples based on a swept-source optical coherence tomography (OCT) system. Shear waves were generated using a piezoelectric transducer transmitting sine-wave bursts of 400 μs duration, applying acoustic radiation force (ARF) to inhomogeneous phantoms and carotid artery samples, synchronized with a swept-source OCT (SS-OCT) imaging system. The phantoms were composed of gelatin and titanium dioxide whereas the carotid artery samples were embedded in gel. Differential OCT phase maps, measured with and without the ARF, detected the microscopic displacement generated by shear wave propagation in these phantoms and samples of different stiffness. We present the technique for calculating tissue mechanical properties by propagating shear waves in inhomogeneous tissue equivalent phantoms and carotid artery samples using the ARF of an ultrasound transducer, and measuring the shear wave speed and its associated properties in the different layers with OCT phase maps. This method lays the foundation for future in-vitro and in-vivo studies of mechanical property measurements of biological tissues such as vascular tissues, where normal and pathological structures may exhibit significant contrast in the shear modulus. PMID:24688822

  5. Association between subcutaneous white adipose tissue and serum 25-hydroxyvitamin D in overweight and obese adults

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Background: Cholecalciferol is known to be deposited in human adipose tissue, but the distribution of 25-hydroxyvitamin D (25(OH)D) in adipose tissue is not known. Objectives: To determine whether 25(OH)D is detectable in subcutaneous white adipose tissue (SWAT) in overweight and obese persons an...

  6. Effects of sampling design on the estimation of adult mosquito abundance.

    PubMed

    Reisen, W K; Lothrop, H D

    1999-06-01

    During 1994-5, Culex tarsalis comprised 75% of the 902,643 adult female mosquitoes collected by 63 dry-ice-baited Centers for Disease Control (CDC)-style traps operated biweekly in a uniform sampling grid that covered the southern Coachella Valley, Riverside County, California. The ln(y + 1) transformation successfully controlled the variance and normalized the distribution of catch size among trap nights. When tested by analysis of variance, abundance varied significantly among months, years, and trap sites. Although the trap by months interaction was not significant, female distribution changed seasonally as larval habitats shifted from wetlands along the Salton Sea to agriculture to managed duck marshes. Conditional simulations utilized subsets of trap sites to compare sampling designs that required no (uniform, random, and transect designs) or prior (best-estimate and stratified random designs) knowledge of mosquito spatial distribution. All designs provided similar information on population seasonal trends, but a stratified random design provided the most accurate and precise simulation. A uniform trap grid that employed every 2nd trap site subsequently was adopted by the Coachella Valley Mosquito and Vector Control District to provide information on focal changes in abundance indicative of missed or newly created larval habitats or control failures. PMID:10412106

  7. Circadian typology, age, and the alternative five-factor personality model in an adult women sample.

    PubMed

    Muro, Anna; Gomà-i-Freixanet, Montserrat; Adan, Ana; Cladellas, Ramon

    2011-10-01

    Research on personality and circadian typology indicates evening-type women are more impulsive and novelty seeking, neither types are more anxious, and morning types tend to be more active, conscientious, and persistent. The purpose of this study is to examine the differences between circadian typologies in the light of the Zuckerman's Alternative Five-Factor Model (AFFM) of personality, which has a strong biological basis, in an adult sample of 412 women 18 to 55 yrs of age. The authors found morning-type women had significant higher scores than evening-type and neither-type women on Activity, and its subscales General Activity and Work Activity. In contrast, evening-type women scored significantly higher than morning-type women on Aggression-Hostility, Impulsive Sensation Seeking, and its subscale Sensation Seeking. In all groups, results were independent of age. These findings are in accordance with those previously obtained in female student samples and add new data on the AFFM. The need of using personality models that are biologically based in the study of circadian rhythms is discussed.

  8. Direct sampling of resting codling moth (Lepidoptera: Tortricidae) adults in apple tree canopies and surrounding habitats.

    PubMed

    Epstein, David L; Miller, James R; Grieshop, Matthew J; Stelinski, Lukasz L; Gut, Larry J

    2011-06-01

    Field investigations were conducted to determine the resting locations of codling moth (Cydia pomonella [L.]) (Lepidoptera: Tortricidae) males and females in mating disrupted and nondisrupted apple (Malus domestica Borkh.) orchard plots. A custom-made sampling device, consisting of a leaf blower converted into a powerful vacuum, yielded 20-24% success in recovering marked moths, released in the tree canopy in orchards. Four collections each were made between 0900 and 1800 hours and 1800 and 2200 hours in 2005. Ninety-four moths were collected during the 1800-2200 hours samples. In mating disruption plots, 42% of females and 22% of males were found in the top third of the tree canopy (3.0-4.5m), 46% females and 43% males in the middle third (1.5-3.0m), and 12% female and 35% male in the lower third (0-1.5m). In nondisrupted plots 36.4% of females and 40% of males were in the top third of the canopy, 36.4% females and 52% males in the middle third, and 27.2% females and 8% males in the lower third of the tree canopy. Daylight vacuum sampling recovered only one female and two male moths from the top, four males from the middle and one male from the lower third of the tree canopy. Release-recapture studies of marked adult codling moths were conducted in 2006-2007 in screened tents to determine within orchard habitats for adult moths during 0900-1800 hours. Of moths recaptured, 14.6% of females and 13.5% of males were from the ground (herbicide strip and drive-row grass) and 32.9% of females and 24.6% of males were captured in the tree canopy 16-h post release, 17.4% of females and 3.4% of males from the ground and 26.5% of females and 38.2% of males in the tree 40-h post release, and 15.1% of females and 18.6% of males from the ground and 15.7 of females and 25.5% of males in the tree 64-h post release. Application of pyrethrum + PBO by using an orchard blast sprayer in 2007 resulted in the recapture of 28% and 37% of laboratory reared male and female moths

  9. Pancreatic-derived pathfinder cells enable regeneration of critically damaged adult pancreatic tissue and completely reverse streptozotocin-induced diabetes.

    PubMed

    Stevenson, Karen; Chen, Daxin; MacIntyre, Alan; McGlynn, Liane M; Montague, Paul; Charif, Rawiya; Subramaniam, Murali; George, W D; Payne, Anthony P; Davies, R Wayne; Dorling, Anthony; Shiels, Paul G

    2011-04-01

    We demonstrate that intravenous delivery of human, or rat, pancreas-derived pathfinder (PDP) cells can totally regenerate critically damaged adult tissue and restore normal function across a species barrier. We have used a mouse model of streptozotocin (STZ)-induced diabetes to demonstrate this. Normoglycemia was restored and maintained for up to 89 days following the induction of diabetes and subsequent intravenous delivery of PDP cells. Normal pancreatic histology also appeared to be restored, and treated diabetic animals gained body weight. Regenerated tissue was primarily of host origin, with few rat or human cells detectable by fluorescent in situ hybridization (FISH). Crucially, the insulin produced by these animals was overwhelmingly murine in origin and was both types I and II, indicative of a process of developmental recapitulation. These results demonstrate the feasibility of using intravenous administration of adult cells to regenerate damaged tissue. Critically, they enhance our understanding of the mechanisms relating to such repair and suggest a means for novel therapeutic intervention in loss of tissue and organ function with age.

  10. C-Myb(+) erythro-myeloid progenitor-derived fetal monocytes give rise to adult tissue-resident macrophages.

    PubMed

    Hoeffel, Guillaume; Chen, Jinmiao; Lavin, Yonit; Low, Donovan; Almeida, Francisca F; See, Peter; Beaudin, Anna E; Lum, Josephine; Low, Ivy; Forsberg, E Camilla; Poidinger, Michael; Zolezzi, Francesca; Larbi, Anis; Ng, Lai Guan; Chan, Jerry K Y; Greter, Melanie; Becher, Burkhard; Samokhvalov, Igor M; Merad, Miriam; Ginhoux, Florent

    2015-04-21

    Although classified as hematopoietic cells, tissue-resident macrophages (MFs) arise from embryonic precursors that seed the tissues prior to birth to generate a self-renewing population, which is maintained independently of adult hematopoiesis. Here we reveal the identity of these embryonic precursors using an in utero MF-depletion strategy and fate-mapping of yolk sac (YS) and fetal liver (FL) hematopoiesis. We show that YS MFs are the main precursors of microglia, while most other MFs derive from fetal monocytes (MOs). Both YS MFs and fetal MOs arise from erythro-myeloid progenitors (EMPs) generated in the YS. In the YS, EMPs gave rise to MFs without monocytic intermediates, while EMP seeding the FL upon the establishment of blood circulation acquired c-Myb expression and gave rise to fetal MOs that then seeded embryonic tissues and differentiated into MFs. Thus, adult tissue-resident MFs established from hematopoietic stem cell-independent embryonic precursors arise from two distinct developmental programs.

  11. Temperament and Character Profiles of Sasang Typology in an Adult Clinical Sample

    PubMed Central

    Park, Soo Hyun; Kim, Myoung-geun; Lee, Soo Jin; Kim, Jong Yeol; Chae, Han

    2011-01-01

    The purpose of this study was to examine the biopsychological personality profiles of traditional Korean Sasang typology based on the Temperament and Character Inventory (TCI) in a Korean adult clinical sample. A total of 97 adults completed the Korean version of the TCI. The participants were classified as one of three traditional Korean Sasang types (31 So-Yang, 41 Tae-Eum, 25 So-Eum) by three specialists in Sasang typology. The seven dimensions of TCI were compared between the different Sasang types using analysis of variance (ANOVA) and profile analysis. There were no significant differences in age, gender and education across the Sasang types. The TCI profile for each of the Sasang types was significantly different (profile analysis, df = 5.038, F = 3.546, P = .004). There were significant differences in the temperament dimensions of Novelty Seeking (F = 3.43, P = .036) and Harm Avoidance (F = 5.43, P = .006) among the Sasang types. The Novelty Seeking score of the So-Yang type (31.90 ± 9.87) was higher than that of the So-Eum type (25.24 ± 9.21; P = .019) while the So-Eum type (44.64 ± 8.47) scored higher on the Harm Avoidance score compared to the So-Yang type (35.16 ± 11.50; P = .003). There were no significant differences in the temperament dimension of Reward Dependence and Persistence, and the three character dimensions of Self-Directedness, Cooperativeness and Self-Transcendence. Results demonstrated distinct temperament traits associated with traditional Korean Sasang types using an objective biopsychological personality inventory. With further study, the Sasang typology may lead to enhanced clinical safety and efficacy as part of personalized medicine with traditional medicine. PMID:19380352

  12. Cadmium levels in a representative sample of the Spanish adult population: The BIOAMBIENT.ES project.

    PubMed

    López-Herranz, Ana; Cutanda, Francisco; Esteban, Marta; Pollán, Marina; Calvo, Eva; Pérez-Gómez, Beatriz; Victoria Cortes, Maria; Castaño, Argelia

    2016-09-01

    Urinary cadmium levels (U-Cd) were measured in 1770 adults (aged 18-65 years) as a representative sample of the Spanish workforce. The geometric mean (GM) was 0.28 μg/l with 95% CI: 0.27-0.32 μg/l (GM: 0.20 μg/g 95% CI: 0.18-022 μg/g creatinine). The 95% percentile was 1.03 μg/l. U-Cd increased with age, with women showing higher U-Cd than men (p<0.001; 0.24 μg/g vs 0.17 μg/g). A multivariate analysis confirmed that sex, age and smoking habit significantly influence U-Cd. Smoking habit increases U-Cd by ∼90% per 10 years of age, almost twice the increase observed for non-smoking. Female smokers had 85% higher U-Cd than non-smokers, whereas the corresponding value for male smokers and non-smokers was 45%. No regional differences were observed with respect to the national reference level. The Spanish population studied here exhibits similar urinary cadmium levels to its European counterparts in Germany and slightly lower levels than in France, the Czech Republic, Italy and the United Kingdom. This paper provides the first baseline information concerning cadmium exposure in the Spanish adult population on a national scale. As such, these findings will help us to establish reference levels, follow temporal trends and identify high-exposure groups, thereby enabling comparisons with other countries and contributing to the improvement of public health and environmental quality.

  13. Executive Cognitive Functioning and Cardiovascular Autonomic Regulation in a Population-Based Sample of Working Adults

    PubMed Central

    Stenfors, Cecilia U. D.; Hanson, Linda M.; Theorell, Töres; Osika, Walter S.

    2016-01-01

    Objective: Executive cognitive functioning is essential in private and working life and is sensitive to stress and aging. Cardiovascular (CV) health factors are related to cognitive decline and dementia, but there is relatively few studies of the role of CV autonomic regulation, a key component in stress responses and risk factor for cardiovascular disease (CVD), and executive processes. An emerging pattern of results from previous studies suggest that different executive processes may be differentially associated with CV autonomic regulation. The aim was thus to study the associations between multiple measures of CV autonomic regulation and measures of different executive cognitive processes. Method: Participants were 119 healthy working adults (79% women), from the Swedish Longitudinal Occupational Survey of Health. Electrocardiogram was sampled for analysis of heart rate variability (HRV) measures, including the Standard Deviation of NN, here heart beats (SDNN), root of the mean squares of successive differences (RMSSD), high frequency (HF) power band from spectral analyses, and QT variability index (QTVI), a measure of myocardial repolarization patterns. Executive cognitive functioning was measured by seven neuropsychological tests. The relationships between CV autonomic regulation measures and executive cognitive measures were tested with bivariate and partial correlational analyses, controlling for demographic variables, and mental health symptoms. Results: Higher SDNN and RMSSD and lower QTVI were significantly associated with better performance on cognitive tests tapping inhibition, updating, shifting, and psychomotor speed. After adjustments for demographic factors however (age being the greatest confounder), only QTVI was clearly associated with these executive tests. No such associations were seen for working memory capacity. Conclusion: Poorer CV autonomic regulation in terms of lower SDNN and RMSSD and higher QTVI was associated with poorer executive

  14. Localization of 5'-ribonucleotide phosphohydrolase in regenerating (and normal) limb tissues of the adult newt Notophthalmus viridescens.

    PubMed

    Schmidt, A J; Woerthwein, K F

    1975-08-11

    The regenerating forelimb of the adult newt, Notophthalmus viridescens was investigated for 5'-nucleotidase (5' ribonucleotide phosphohydrolase, 3.1.3.5) acitivity. The newt's humeri were surgically removed, and after a twenty-one-day recovery period, the forelimbs amputated above the elbows. Regenerates were sampled at predetermined times for specific phases in the progress of regeneration, frozen, sectioned in a cryostat, and the sections fixed in 10% cold formol calcium. The Wachstein and Meisel [25] lead procedure at neutral pH was used predominately in these experiments, although tests were also conducted with Gomori's [14] calcium, Allen's [21] highly alkaline procedures. The substrates used to obtain specific enzyme reactions were adenine, cytosine, guanine, uracil and inosine 5'-monophosphate nucleotides. Sodium beta-glycerophosphate served as a non-specific phosphomonoesterase substrate, distilled water replaced substrate, and inhibitors such as zinc and cyanide ions were used as control measures to assist in increasing the precision in interpreting the results obtained. The most reactive 5'-nucleotidase (5'-Nase) loci were in the walls of the blood vascular system, mysial and neural sheaths, dermis, and periosteum: the principal cells involved were macrophages, endothelium of blood vessels, and fibrocytes of connective tissues. A moderate enzyme response was elicited from secretory cells of some of the subcutaneous glands, hypertrophied chondrocytes and osteogenic centers, chondrocytes in the articular regions and within red blood cells and leucocytes. Normal, injured and degenerating, or regenerating striated muscle and nerve fibers were judged unreactive for 5'-Nase. The epidermis and wound epithelium displayed negative responses for 5'-Nase. Cells forming the regeneration blastema were 5'-Nase reactive during the early formative phase, but with growth and development of the blastema into bulb and conic forms, these cells did not respond for this enzyme

  15. Single-pixel hyperspectral imaging for real-time cancer detection: detecting damage in ex vivo porcine tissue samples

    NASA Astrophysics Data System (ADS)

    Peller, Joseph; Farahi, Faramarz; Trammell, Susan R.

    2016-03-01

    We are developing a single-pixel hyperspectral imaging system based on compressive sensing that acquires spatial and spectral information simultaneously. Our spectral imaging system uses autofluorescencent emission from collagen (400 nm) and NAD(P)H (475 nm), as well as, differences in the optical reflectance spectra as diagnostics for differentiating between healthy and diseased tissue. In this study, we demonstrate the ability of our imaging system to discriminate between healthy and damaged porcine epidermal tissue. Healthy porcine epidermal tissue samples (n=11) were imaged ex vivo using our hyperspectral system. The amount of NAD(P)H emission and the reflectance properties were approximately constant across the surface of healthy tissue samples. The tissue samples were then thermally damaged using an 1850 nm thulium fiber laser and re-imaged after laser irradiation. The damaged regions were clearly visible in the hyperspectral images as the thermal damage altered the fluorescent emission of NAD(P)H and changed the scattering properties of the tissue. The extent of the damaged regions was determined based on the hyperspectral images and these estimates were compared to damage extents measured in white light images acquired with a traditional camera. The extent of damage determined via hyperspectral imaging was in good agreement with estimates based on white light imaging indicating that our system is capable of differentiating between healthy and damaged tissue. Possible applications of our single pixel hyperspectral imaging system range from real-time determination of tumor margins during surgery to the use of this technique in the pathology lab to aid with cancer diagnosis and staging.

  16. Characterization and antimicrobial resistance of Salmonella isolated from internal tissues, ceca and rinse samples from commercial broiler chickens

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The presence, serotype, and antimicrobial resistance profile of Salmonella from internal tissues (spleen, liver/gall bladder, thymus, Meckel’s diverticulum, and free floating yolk), ceca and carcass rinse samples were determined from six-week-old (n=30) and eight-week-old (n=40) commercial broilers ...

  17. Comparative evaluation of specific methods for labeling of Encephalitozoon cuniculi in paraffin wax-embedded tissue samples.

    PubMed

    Habenbacher, Bettina; Klang, Andrea; Fragner, Karin; Dinhopl, Nora; Künzel, Frank; Weissenböck, Herbert

    2012-03-01

    Detection of the microsporidian Encephalitozoon cuniculi in tissue samples is considered difficult. The aim of the current study was to determine whether immunohistochemistry (IHC) and in situ hybridization (ISH) represent reliable methods for the detection of E. cuniculi in postmortem tissue samples of rabbits. Paraffin-embedded tissue sections of brain and kidneys of 48 naturally infected pet rabbits, 10 negative controls, and the eyes of 3 further rabbits were used for all investigations. By IHC in 19 animals (37.3%), spores could be clearly detected and were all equally stained. By ISH using a digoxigenin-labeled oligonucleotide probe, only 6 animals (11.8%) proved undoubtedly positive. In these cases, many parasite-like objects revealed strong typical purple-black positive signals. However, several of the examined samples showed only partial staining of the pathogen or unclear results. Thus, in order to find an explanation for these inconsistent ISH results and to take a more detailed look at the different developmental stages of the organism, electron microscopy was applied. Empty spores, which had already discharged their polar filaments, prevailed in total number. Taken together, both techniques are rather insensitive, but under the condition that sufficient numbers of microsporidia are present, IHC can be recommended for specific identification of E. cuniculi in tissue samples. In contrast, ISH failed to detect some developmental stages of the organism, and, as such, ISH is therefore considered an inappropriate diagnostic method.

  18. Development of a sensitive method to extract and detect low numbers of Cryptosporidium oocysts from adult cattle faecal samples.

    PubMed

    Wells, B; Thomson, S; Ensor, H; Innes, E A; Katzer, F

    2016-08-30

    Cryptosporidium transmission studies to date have concluded that adult cattle are not a significant source of oocysts contributing to clinical cryptosporidiosis in calves on farm. However current methods of sample processing have been optimised for calf faecal samples and may be less sensitive when used on adult samples due to lower numbers of oocysts and larger size of samples. A modified and novel method of oocyst extraction and concentration was developed and applied in an experiment involving spiking adult cattle faecal samples with known concentrations of Cryptosporidium oocysts. The results showed an increased sensitivity of detection from 100oocysts/g of faecal sample using conventional protocols to 5oocysts/g using the newly developed method. As it is important to be able to accurately assess the contribution of adult ruminants to the transmission of Cryptosporidium, both on farm and in the environment, the development of the techniques described here is likely to make an important contribution to Cryptosporidium transmission studies in future and in subsequent control strategies aimed at the reduction of Cryptosporidium infection in calves on farm. PMID:27523933

  19. Development of a sensitive method to extract and detect low numbers of Cryptosporidium oocysts from adult cattle faecal samples.

    PubMed

    Wells, B; Thomson, S; Ensor, H; Innes, E A; Katzer, F

    2016-08-30

    Cryptosporidium transmission studies to date have concluded that adult cattle are not a significant source of oocysts contributing to clinical cryptosporidiosis in calves on farm. However current methods of sample processing have been optimised for calf faecal samples and may be less sensitive when used on adult samples due to lower numbers of oocysts and larger size of samples. A modified and novel method of oocyst extraction and concentration was developed and applied in an experiment involving spiking adult cattle faecal samples with known concentrations of Cryptosporidium oocysts. The results showed an increased sensitivity of detection from 100oocysts/g of faecal sample using conventional protocols to 5oocysts/g using the newly developed method. As it is important to be able to accurately assess the contribution of adult ruminants to the transmission of Cryptosporidium, both on farm and in the environment, the development of the techniques described here is likely to make an important contribution to Cryptosporidium transmission studies in future and in subsequent control strategies aimed at the reduction of Cryptosporidium infection in calves on farm.

  20. Development and application of specific cytokine assays in tissue samples from a bottlenose dolphin with hyperinsulinemia

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Chronic inflammation has been associated with insulin resistance and type 2 diabetes in humans. Postmortem hepatic and splenic tissue from a 46-year old geriatric male bottlenose dolphin (Tursiops truncatus) with insulin resistance (chronic hyperinsulinemia with hyperglycemia) , chronic = inflamma...

  1. Characterizing active and inactive brown adipose tissue in adult humans using PET-CT and MR imaging.

    PubMed

    Gifford, Aliya; Towse, Theodore F; Walker, Ronald C; Avison, Malcolm J; Welch, E Brian

    2016-07-01

    Activated brown adipose tissue (BAT) plays an important role in thermogenesis and whole body metabolism in mammals. Positron emission tomography (PET)-computed tomography (CT) imaging has identified depots of BAT in adult humans, igniting scientific interest. The purpose of this study is to characterize both active and inactive supraclavicular BAT in adults and compare the values to those of subcutaneous white adipose tissue (WAT). We obtained [(18)F]fluorodeoxyglucose ([(18)F]FDG) PET-CT and magnetic resonance imaging (MRI) scans of 25 healthy adults. Unlike [(18)F]FDG PET, which can detect only active BAT, MRI is capable of detecting both active and inactive BAT. The MRI-derived fat signal fraction (FSF) of active BAT was significantly lower than that of inactive BAT (means ± SD; 60.2 ± 7.6 vs. 62.4 ± 6.8%, respectively). This change in tissue morphology was also reflected as a significant increase in Hounsfield units (HU; -69.4 ± 11.5 vs. -74.5 ± 9.7 HU, respectively). Additionally, the CT HU, MRI FSF, and MRI R2* values are significantly different between BAT and WAT, regardless of the activation status of BAT. To the best of our knowledge, this is the first study to quantify PET-CT and MRI FSF measurements and utilize a semiautomated algorithm to identify inactive and active BAT in the same adult subjects. Our findings support the use of these metrics to characterize and distinguish between BAT and WAT and lay the foundation for future MRI analysis with the hope that some day MRI-based delineation of BAT can stand on its own. PMID:27166284

  2. An exploratory survey of professionals on the use of stored tissue samples from minors for genetic research.

    PubMed

    Hens, K; Snoeck, J; Nys, H; Cassiman, J-J; Dierickx, K

    2010-01-01

    he ethical aspects of the use of stored tissue samples collected from minors are of topical interest. However, the views of professionals working in the field of genetics have not been investigated in depth anywhere. We conducted a survey among 194 such professionals in Belgium. This list was composed of the members of the High Council for Anthropogenetics, supplemented with all professionals working in the field of genetics that we found on the websites of the eight Belgian centers of human genetics and of the associated university registries. We achieved a response rate of 35.5%. The vast majority (92%) think that research on stored tissue samples is useful. Most respondents stated that parental consent is valid (82.5%), and 76.5% thought that children should also be given the right to assent when they are able to comprehend the implications of the storage of biological samples and of genetic research. Slightly more than half put the age at which young people can understand storage or research rather high: 16-18 years (51 and 53.1%, respectively). Although there is some consensus in the literature that donors should be allowed to give broad consent for future research on their biological samples, only 47.6% in our survey thought that parents should be allowed to consent to any future research on their children's samples. The aim of our study was to give some basis for future ethical reflections and policies on the subject of stored tissue samples from minors for genetic research. We concluded that a large majority of Belgian researchers and clinicians in the field of genetic research think research on stored tissue samples from minors is useful. They also think that parental consent for such research is valid, but that children should be allowed to assent as they grow older.

  3. Simultaneous determination of perfluorinated compounds and their potential precursors in mussel tissue and fish muscle tissue and liver samples by liquid chromatography-electrospray-tandem mass spectrometry.

    PubMed

    Zabaleta, I; Bizkarguenaga, E; Prieto, A; Ortiz-Zarragoitia, M; Fernández, L A; Zuloaga, O

    2015-03-27

    An analytical method for the simultaneous determination in fish liver and muscle tissue and mussel samples of 14 perfluorinated compounds (PFCs), including three perfluoroalkylsulfonates (PFSAs), seven perfluorocarboxylic acids (PFCAs), three perfluorophosphonic acids (PFPAs) and perfluorooctanesulfonamide (PFOSA), and 10 potential precursors, including four polyfluoroalkyl phosphates (PAPs), four fluorotelomer saturated acids (FTCAs) and two fluorotelomer unsaturated acids (FTUCAs), was developed in the present work. Different clean-up strategies by means of solid-phase extraction (SPE) using a mix-mode weak anion exchanger (WAX), reverse phase Envi-Carb or a combination of them was optimized and evaluated for the clean-up of focused ultrasonic solid-liquid (FUSLE) extracts before the analysis by liquid chromatography-triple quadrupole tandem mass spectrometry (LC-MS/MS). Mix-mode WAX coupled in-line to Envi-Carb was finally selected since it rendered the cleanest extracts and minimum matrix effect. The FUSLE-SPE-LC-MS/MS methodology was validated in terms of recovery, precision and method detection limits (MDLs). Apparent recovery values in the 65-116%, 59-119% and 67-126% range and MDLs in the 0.1-2.7 ng/g, 0.1-3.8 ng/g and 0.2-3.1ng/g range were obtained for liver, mussel and fish muscle tissue samples, respectively. The method developed was applied to the analysis of grey mullet liver (Chelon labrosus) and mussel (Mytilus galloprovincialis) samples from the Basque Coast (North of Spain) and Yellowfin tuna muscle tissue (Thunnus albacares) samples from the Indian Ocean. To the best of our knowledge this is the first method that describes the simultaneous determination of 14 PFCs and 10 potential precursors in fish liver, fish muscle tissue and mussel samples. Besides, this is the first time that 8:2 monosubstituted polyfluorodecyl phosphate (8:2 monoPAP) and 8:2 disubstituted polyfluorodecyl phosphate (8:2 diPAP) were detected in mussel and tuna samples

  4. Measurement of characteristic prompt gamma rays emitted from oxygen and carbon in tissue-equivalent samples during proton beam irradiation

    NASA Astrophysics Data System (ADS)

    Polf, Jerimy C.; Panthi, Rajesh; Mackin, Dennis S.; McCleskey, Matt; Saastamoinen, Antti; Roeder, Brian T.; Beddar, Sam

    2013-09-01

    The purpose of this work was to characterize how prompt gamma (PG) emission from tissue changes as a function of carbon and oxygen concentration, and to assess the feasibility of determining elemental concentration in tissues irradiated with proton beams. For this study, four tissue-equivalent water-sucrose samples with differing densities and concentrations of carbon, hydrogen, and oxygen were irradiated with a 48 MeV proton pencil beam. The PG spectrum emitted from each sample was measured using a high-purity germanium detector, and the absolute detection efficiency of the detector, average beam current, and delivered dose distribution were also measured. Changes to the total PG emission from 12C (4.44 MeV) and 16O (6.13 MeV) per incident proton and per Gray of absorbed dose were characterized as a function of carbon and oxygen concentration in the sample. The intensity of the 4.44 MeV PG emission per incident proton was found to be nearly constant for all samples regardless of their carbon concentration. However, we found that the 6.13 MeV PG emission increased linearly with the total amount (in grams) of oxygen irradiated in the sample. From the measured PG data, we determined that 1.64 × 107 oxygen PGs were emitted per gram of oxygen irradiated per Gray of absorbed dose delivered with a 48 MeV proton beam. These results indicate that the 6.13 MeV PG emission from 16O is proportional to the concentration of oxygen in tissue irradiated with proton beams, showing that it is possible to determine the concentration of oxygen within tissues irradiated with proton beams by measuring 16O PG emission.

  5. Measurement of characteristic prompt gamma rays emitted from oxygen and carbon in tissue-equivalent samples during proton beam irradiation

    PubMed Central

    Polf, Jerimy C; Panthi, Rajesh; Mackin, Dennis S; McCleskey, Matt; Saastamoinen, Antti; Roeder, Brian T; Beddar, Sam

    2013-01-01

    The purpose of this work was to characterize how prompt gamma (PG) emission from tissue changes as a function of carbon and oxygen concentration, and to assess the feasibility of determining elemental concentration in tissues irradiated with proton beams. For this study, four tissue-equivalent water-sucrose samples with differing densities and concentrations of carbon, hydrogen, and oxygen were irradiated with a 48 MeV proton pencil beam. The PG spectrum emitted from each sample was measured using a high-purity germanium detector, and the absolute detection efficiency of the detector, average beam current, and delivered dose distribution were also measured. Changes to the total PG emission from 12C (4.44 MeV) and 16O (6.13 MeV) per incident proton and per Gray of absorbed dose were characterized as a function of carbon and oxygen concentration in the sample. The intensity of the 4.44 MeV PG emission per incident proton was found to be nearly constant for all samples regardless of their carbon concentration. However, we found that the 6.13 MeV PG emission increased linearly with the total amount (in grams) of oxygen irradiated in the sample. From the measured PG data, we determined that 1.64 × 107 oxygen PGs were emitted per gram of oxygen irradiated per Gray of absorbed dose delivered with a 48 MeV proton beam. These results indicate that the 6.13 MeV PG emission from 16O is proportional to the concentration of oxygen in tissue irradiated with proton beams, showing that it is possible to determine the concentration of oxygen within tissues irradiated with proton beams by measuring 16O PG emission. PMID:23920051

  6. Pathology Tissue-quantitative Mass Spectrometry Analysis to Profile Histone Post-translational Modification Patterns in Patient Samples*

    PubMed Central

    Noberini, Roberta; Uggetti, Andrea; Pruneri, Giancarlo; Minucci, Saverio

    2016-01-01

    Histone post-translational modifications (hPTMs) generate a complex combinatorial code that has been implicated with various pathologies, including cancer. Dissecting such a code in physiological and diseased states may be exploited for epigenetic biomarker discovery, but hPTM analysis in clinical samples has been hindered by technical limitations. Here, we developed a method (PAThology tissue analysis of Histones by Mass Spectrometry - PAT-H-MS) that allows to perform a comprehensive, unbiased and quantitative MS-analysis of hPTM patterns on formalin-fixed paraffin-embedded (FFPE) samples. In pairwise comparisons, histone extracted from formalin-fixed paraffin-embedded tissues showed patterns similar to fresh frozen samples for 24 differentially modified peptides from histone H3. In addition, when coupled with a histone-focused version of the super-SILAC approach, this method allows the accurate quantification of modification changes among breast cancer patient samples. As an initial application of the PAThology tissue analysis of Histones by Mass Spectrometry method, we analyzed breast cancer samples, revealing significant changes in histone H3 methylation patterns among Luminal A-like and Triple Negative disease subtypes. These results pave the way for retrospective epigenetic studies that combine the power of MS-based hPTM analysis with the extensive clinical information associated with formalin-fixed paraffin-embedded archives. PMID:26463340

  7. Psychometric Properties of the Theory of Mind Assessment Scale in a Sample of Adolescents and Adults.

    PubMed

    Bosco, Francesca M; Gabbatore, Ilaria; Tirassa, Maurizio; Testa, Silvia

    2016-01-01

    This research aimed at the evaluation of the psychometric properties of the Theory of Mind Assessment Scale (Th.o.m.a.s.). Th.o.m.a.s. is a semi-structured interview meant to evaluate a person's Theory of Mind (ToM). It is composed of several questions organized in four scales, each focusing on one of the areas of knowledge in which such faculty may manifest itself: Scale A (I-Me) investigates first-order first-person ToM; Scale B (Other-Self) investigates third-person ToM from an allocentric perspective; Scale C (I-Other) again investigates third-person ToM, but from an egocentric perspective; and Scale D (Other-Me) investigates second-order ToM. The psychometric proprieties of Th.o.m.a.s. were evaluated in a sample of 156 healthy persons: 80 preadolescent and adolescent (aged 11-17 years, 42 females) and 76 adults (aged from 20 to 67 years, 35 females). Th.o.m.a.s. scores show good inter-rater agreement and internal consistency; the scores increase with age. Evidence of criterion validity was found as Scale B scores were correlated with those of an independent instrument for the evaluation of ToM, the Strange Stories task. Confirmatory factor analysis (CFA) showed good fit of the four-factors theoretical model to the data, although the four factors were highly correlated. For each of the four scales, Rasch analyses showed that, with few exceptions, items fitted the Partial credit model and their functioning was invariant for gender and age. The results of this study, along with those of previous researches with clinical samples, show that Th.o.m.a.s. is a promising instrument to assess ToM in different populations. PMID:27242563

  8. Association of telomere length and mitochondrial DNA copy number in a community sample of healthy adults.

    PubMed

    Tyrka, Audrey R; Carpenter, Linda L; Kao, Hung-Teh; Porton, Barbara; Philip, Noah S; Ridout, Samuel J; Ridout, Kathryn K; Price, Lawrence H

    2015-06-01

    Cellular aging plays a role in longevity and senescence, and has been implicated in medical and psychiatric conditions, including heart disease, cancer, major depression and posttraumatic stress disorder. Telomere shortening and mitochondrial dysfunction are thought to be central to the cellular aging process. The present study examined the association between mitochondrial DNA (mtDNA) copy number and telomere length in a sample of medically healthy adults. Participants (total n=392) were divided into 4 groups based on the presence or absence of early life adversity and lifetime psychopathology: No Adversity/No Disorder, n=136; Adversity/No Disorder, n=91; No Adversity/Disorder, n=46; Adversity/Disorder, n=119. Telomere length and mtDNA copy number were measured using quantitative polymerase chain reaction. There was a positive correlation between mtDNA and telomere length in the entire sample (r=0.120, p<0.001) and in each of the four groups of participants (No Adversity/No Disorder, r=0.291, p=0.001; Adversity/No Disorder r=0.279, p=0.007; No Adversity/Disorder r=0.449, p=0.002; Adversity/Disorder, r=0.558, p<0.001). These correlations remained significant when controlling for age, smoking, and body mass index and establish an association between mtDNA and telomere length in a large group of women and men both with and without early adversity and psychopathology, suggesting co-regulation of telomeres and mitochondrial function. The mechanisms underlying this association may be important in the pathophysiology of age-related medical conditions, such as heart disease and cancer, as well as for stress-associated psychiatric disorders.

  9. Psychometric Properties of the Theory of Mind Assessment Scale in a Sample of Adolescents and Adults

    PubMed Central

    Bosco, Francesca M.; Gabbatore, Ilaria; Tirassa, Maurizio; Testa, Silvia

    2016-01-01

    This research aimed at the evaluation of the psychometric properties of the Theory of Mind Assessment Scale (Th.o.m.a.s.). Th.o.m.a.s. is a semi-structured interview meant to evaluate a person's Theory of Mind (ToM). It is composed of several questions organized in four scales, each focusing on one of the areas of knowledge in which such faculty may manifest itself: Scale A (I-Me) investigates first-order first-person ToM; Scale B (Other-Self) investigates third-person ToM from an allocentric perspective; Scale C (I-Other) again investigates third-person ToM, but from an egocentric perspective; and Scale D (Other-Me) investigates second-order ToM. The psychometric proprieties of Th.o.m.a.s. were evaluated in a sample of 156 healthy persons: 80 preadolescent and adolescent (aged 11–17 years, 42 females) and 76 adults (aged from 20 to 67 years, 35 females). Th.o.m.a.s. scores show good inter-rater agreement and internal consistency; the scores increase with age. Evidence of criterion validity was found as Scale B scores were correlated with those of an independent instrument for the evaluation of ToM, the Strange Stories task. Confirmatory factor analysis (CFA) showed good fit of the four-factors theoretical model to the data, although the four factors were highly correlated. For each of the four scales, Rasch analyses showed that, with few exceptions, items fitted the Partial credit model and their functioning was invariant for gender and age. The results of this study, along with those of previous researches with clinical samples, show that Th.o.m.a.s. is a promising instrument to assess ToM in different populations. PMID:27242563

  10. Probing focal cortical dysplasia in formalin fixed samples using tissue optical spectroscopy

    NASA Astrophysics Data System (ADS)

    Anand, Suresh; Cicchi, Riccardo; Giordano, Flavio; Buccoliero, Anna Maria; Conti, Valerio; Guerrini, Renzo; Pavone, Francesco Saverio

    2016-03-01

    Focal cortical dysplasia (FCD) is one of most common causes of intractable epilepsy in pediatric population and these are often insensitive to anti-epileptic drugs. FCD is characterized by a disarray in localized regions of the cerebral cortex and abnormal neurons which results them to misfire with incorrect signals. Resective neurosurgery to remove or disconnect the affected parts from the rest of the brain seems to be a viable option to treat FCD. Before neurosurgery the subject could undergo imaging studies including magnetic resonance imaging (MRI) or computed tomography (CT) scans. On the downside FCD could be elusive in MRI images and may be practically invisible in CT scans. Furthermore, unnecessary removal of normal tissues is to be taken into consideration as this could lead to neurological defects. In this context, optical spectroscopy have been widely investigated as an alternative technique for the detection of abnormal tissues in different organ sites. Disease progression is accompanied by a number of architectural, biochemical and morphological changes. These variations are reflected in the spectral intensity and line shape. Here, in this proof of concept study we propose to investigate the application of tissue optical spectroscopy based on fluorescence excitation at two wavelength 378 and 445 nm coupled along with Raman spectroscopy for the detection of FCD on formalin fixed tissue specimens from pediatric subjects. For fluorescence at both the excitation wavelengths FCD showed a decreased intensity at longer wavelength when compared to normal tissues. Also, differences exist in the Raman spectral profiles of normal and FCD.

  11. Comparison of treated and untreated major depressive disorder in a nationwide sample of Korean adults.

    PubMed

    Park, Subin; Cho, Maeng Je; Bae, Jae Nam; Chang, Sung Man; Jeon, Hong Jin; Hahm, Bong-Jin; Son, Jung-Woo; Kim, Shin Gyeom; Bae, Ahn; Hong, Jin Pyo

    2012-06-01

    We examined factors associated with lifetime treatment of major depressive disorder (MDD) in a nationwide sample of Korean adults. Of the 6,510 subjects aged 18-64 years who participated in the Korean Epidemiologic Catchment Area study, 362 (5.6%) with a lifetime diagnosis of MDD were analyzed. Diagnostic assessments were based on the Korean version of the Composite International Diagnostic Interview administered by lay interviewers. Of the 362 respondents with a lifetime diagnosis of MDD, 117 (32.3%) had been treated for psychiatric problems. Treated individuals with MDD were more likely to have chronic episode(s), more symptoms of depression, insomnia, and suicidal ideation, and were less likely to have feelings of guilt. In addition, treated individuals were more likely to have comorbid anxiety disorders, especially obsessive-compulsive disorder, post-traumatic stress disorder, and generalized anxiety disorder. Treatment-seeking by individuals with MDD is affected by socio-cultural factors such as misconception and stigma of mental illness, as well as severity of depression and comorbid conditions.

  12. Detecting Superior Face Recognition Skills in a Large Sample of Young British Adults

    PubMed Central

    Bobak, Anna K.; Pampoulov, Philip; Bate, Sarah

    2016-01-01

    The Cambridge Face Memory Test Long Form (CFMT+) and Cambridge Face Perception Test (CFPT) are typically used to assess the face processing ability of individuals who believe they have superior face recognition skills. Previous large-scale studies have presented norms for the CFPT but not the CFMT+. However, previous research has also highlighted the necessity for establishing country-specific norms for these tests, indicating that norming data is required for both tests using young British adults. The current study addressed this issue in 254 British participants. In addition to providing the first norm for performance on the CFMT+ in any large sample, we also report the first UK specific cut-off for superior face recognition on the CFPT. Further analyses identified a small advantage for females on both tests, and only small associations between objective face recognition skills and self-report measures. A secondary aim of the study was to examine the relationship between trait or social anxiety and face processing ability, and no associations were noted. The implications of these findings for the classification of super-recognizers are discussed. PMID:27713706

  13. Post traumatic stress disorder and coping in a sample of adult survivors of the Italian earthquake.

    PubMed

    Cofini, V; Carbonelli, A; Cecilia, M R; Binkin, N; di Orio, F

    2015-09-30

    The aim was to investigate the prevalence of post traumatic stress disorder (PTSD) in people who had left their damaged homes and were still living in temporary housing more than a year after the April 2009 L'Aquila (Italy) earthquake. In addition, we evaluated the differences in coping strategies implemented by persons who had and who did not have PTSD. A cross-sectional prevalence study was carried out on a sample of 281 people aged >18 years and living in temporary housing after the earthquake. The questionnaires used include the Davidson Trauma Scale and the Brief Cope. The prevalence of PTSD was 43%. Women and the non-employed were more vulnerable to PTSD, while, age and level of education were not associated with PTSD. Those with PTSD symptoms often employed maladaptive coping strategies for dealing with earthquake and had the highest scores in the domains of denial, venting, behavioral disengagement, self-blame. By contrast, those without PTSD generally had more adaptive coping mechanisms. Adults who were living in temporary housing after the earthquake experienced high rates of PTSD. The difference in coping mechanisms between those who have PTSD and those who do not also suggests that they influence the likeliness of developing PTSD. PMID:26162658

  14. Epidemiology of undiagnosed trichomoniasis in a probability sample of urban young adults.

    PubMed

    Rogers, Susan M; Turner, Charles F; Hobbs, Marcia; Miller, William C; Tan, Sylvia; Roman, Anthony M; Eggleston, Elizabeth; Villarroel, Maria A; Ganapathi, Laxminarayana; Chromy, James R; Erbelding, Emily

    2014-01-01

    T. vaginalis infection (trichomoniasis) is the most common curable sexually transmitted infection (STI) in the U.S. It is associated with increased HIV risk and adverse pregnancy outcomes. Trichomoniasis surveillance data do not exist for either national or local populations. The Monitoring STIs Survey Program (MSSP) collected survey data and specimens which were tested using nucleic acid amplification tests to monitor trichomoniasis and other STIs in 2006-09 among a probability sample of young adults (N = 2,936) in Baltimore, Maryland--an urban area with high rates of reported STIs. The estimated prevalence of trichomoniasis was 7.5% (95% CI 6.3, 9.1) in the overall population and 16.1% (95% CI 13.0, 19.8) among Black women. The overwhelming majority of infected men (98.5%) and women (73.3%) were asymptomatic. Infections were more common in both women (OR = 3.6, 95% CI 1.6, 8.2) and men (OR = 9.0, 95% CI 1.8, 44.3) with concurrent chlamydial infection. Trichomoniasis did not vary significantly by age for either men or women. Women with two or more partners in the past year and women with a history of personal or partner incarceration were more likely to have an infection. Overall, these results suggest that routine T vaginalis screening in populations at elevated risk of infection should be considered.

  15. Post traumatic stress disorder and coping in a sample of adult survivors of the Italian earthquake.

    PubMed

    Cofini, V; Carbonelli, A; Cecilia, M R; Binkin, N; di Orio, F

    2015-09-30

    The aim was to investigate the prevalence of post traumatic stress disorder (PTSD) in people who had left their damaged homes and were still living in temporary housing more than a year after the April 2009 L'Aquila (Italy) earthquake. In addition, we evaluated the differences in coping strategies implemented by persons who had and who did not have PTSD. A cross-sectional prevalence study was carried out on a sample of 281 people aged >18 years and living in temporary housing after the earthquake. The questionnaires used include the Davidson Trauma Scale and the Brief Cope. The prevalence of PTSD was 43%. Women and the non-employed were more vulnerable to PTSD, while, age and level of education were not associated with PTSD. Those with PTSD symptoms often employed maladaptive coping strategies for dealing with earthquake and had the highest scores in the domains of denial, venting, behavioral disengagement, self-blame. By contrast, those without PTSD generally had more adaptive coping mechanisms. Adults who were living in temporary housing after the earthquake experienced high rates of PTSD. The difference in coping mechanisms between those who have PTSD and those who do not also suggests that they influence the likeliness of developing PTSD.

  16. The reporting of maltreatment experiences during the Adult Attachment Interview in a sample of pregnant adolescents.

    PubMed

    Madigan, Sheri; Vaillancourt, Kyla; McKibbon, Amanda; Benoit, Diane

    2012-01-01

    This present student examines maltreatment experiences reported by 55 high-risk pregnant adolescents in response to a slightly adapted version of the Adult Attachment Interview (AAI; George, Kaplan, & Main, 1996 ). Previous research has suggested that the rates of unresolved states of mind regarding trauma in response to the AAI may be underestimated due to the lack of direct questions and associated probes regarding physical, sexual, and emotional abuse. We address this concern by including behaviorally phrased questions and probes regarding maltreatment experiences into the original format of the AAI and examine the concordance between reports of maltreatment experiences in response to the AAI and the Childhood Trauma Questionnaire (CTQ). Maltreatment experiences in response to the AAI were evaluated using the Maltreatment Classification Scale developed by Barnett, Manly, and Cicchetti (1993). We also examine the association between unresolved states of mind and dissociation using the Adolescent Dissociative Experience Scale. Results revealed a significant concordance between reports of maltreatment in response to the AAI and CTQ measures. Reports of maltreatment were prevalent in this sample: across the AAI and CTQ measures, 96% of pregnant adolescents reported some form of emotional abuse, 84% physical abuse, 59% sexual abuse, and 88% reported neglect. Sexual abuse history uniquely predicted unresolved status in response to the AAI. Self-reports of dissociation were significantly associated with unresolved states of mind. Results suggest that the inclusion of behaviorally focused questions and probes regarding maltreatment in the AAI protocol can further contribute to the clinical and theoretical value of this tool. PMID:22385310

  17. How You Can Help Medical Research: Donating Your Blood, Tissue, and Other Samples

    MedlinePlus

    ... sample to make sure the proposed research is ethical, useful, and based on good science. After the ... discuss further? • Am I comfortable with experts making decisions about how my samples will be used in ...

  18. Optical parameter measurement of highly diffusive tissue body phantoms with specifically designed sample holder for photo diagnostic and PDT applications

    NASA Astrophysics Data System (ADS)

    Rehman, A.; Rehman, K.; Anwar, S.; Firdous, S.; Nawaz, M.

    2015-12-01

    Knowledge of optical properties (absorption coefficients, scattering Coefficients, and anisotropy) is necessary for understanding light tissue interactions. Optical parameters define the behavior of light in the tissues. Intralipid and Indian ink are well-established tissue body phantoms. Quantitative characterization of biological tissues in terms of optical properties is achieved with integrating sphere. However, samples having significantly higher scattering and absorption coefficients such as malignant tissues potentially reduce the signal to noise ratio (SNR) and accuracy of integrating sphere. We have measured the diffuse reflection and transmission of these phantoms by placing them in integrating sphere at 632.8 nm and then applied IAD method to determine the optical properties tissue phantoms composed of Indian ink (1.0%) and Intralipid (20%). We have fabricated a special sample holder with thin microscopic cover slips which can be used to measure signal from highly concentrated intralipid and Indian ink solutions. Experiments conducted with various phantoms reveal significant improvement of SNR for a wide range of optical properties. This approach opens up a field for potential applications in measurement of optical properties of highly diffusive biological tissues. For 20% intralipid μa =0.112+/-0.046 cm-1 and μs =392.299+/-10.090 cm-1 at 632.8 nm and for 1.0% Indian ink μa =9.808+/-0.490 cm-1 and μs =1.258+/-0.063 cm-1 at same wavelength. System shows good repeatability and reproducibility within 4.9% error. Work may have important biomedical applications in photo-diagnosis and Photodynamic therapy.

  19. Towards a minimally invasive sampling tool for high resolution tissue analytical mapping

    NASA Astrophysics Data System (ADS)

    Gottardi, R.

    2015-09-01

    Multiple spatial mapping techniques of biological tissues have been proposed over the years, but all present limitations either in terms of resolution, analytical capacity or invasiveness. Ren et al (2015 Nanotechnology 26 284001) propose in their most recent work the use of a picosecond infrared laser (PIRL) under conditions of ultrafast desorption by impulsive vibrational excitation (DIVE) to extract small amounts of cellular and molecular components, conserving their viability, structure and activity. The PIRL DIVE technique would then work as a nanobiopsy with minimal damage to the surrounding tissues, which could potentially be applied for high resolution local structural characterization of tissues in health and disease with the spatial limit determined by the laser focus.

  20. Physical activity and sedentary behavior among adults 60 years and older: New York City residents compared with a national sample.

    PubMed

    Evenson, Kelly R; Morland, Kimberly B; Wen, Fang; Scanlin, Kathleen

    2014-10-01

    This study describes moderate to vigorous physical activity (MVPA) and sedentary behavior among New York City (NYC) residents 60 years and older and compared with national United States' estimates. Adults aged 60 or older living in NYC (n = 760) were compared with similar aged adults from the National Health and Nutrition Examination Survey (NHANES; n = 2,451 adults). Both groups wore an ActiGraph accelerometer for one week. The NYC sample recorded 13.2, 23.8, and 37.8 mean min/day of MVPA and the NHANES sample recorded 10.6, 21.1, and 39.3, depending on the definition. Sedentary behavior averaged 9.6 hr/day for the NYC sample and 9.3 hr/day for the NHANES sample. The NYC sample spent a longer proportion of time in sedentary behavior and light activities, but more time in MVPA than the NHANES sample. Urbanicity may explain some of the differences between the two samples.

  1. Measurement of intravenously administered γ-Fe2O3 particle amount in mice tissues using vibrating sample magnetometer.

    PubMed

    Kishimoto, Mikio; Miyamoto, Ryoichi; Oda, Tatsuya; Ohara, Yusuke; Yanagihara, Hideto; Ohkohchi, Nobuhiro; Kita, Eiji

    2014-12-01

    Dispersions of platelet γ-Fe2O3 particles 30-50nm in size were intravenously administered to mice and the amount of particles accumulated in each tissue was obtained by magnetization measurement using a vibrating sample magnetometer. Background noise was greatly reduced by measuring dried tissues under a magnetic field of 500 Oe so that the effect of diamagnetism was slight. Remarkable particle accumulation was observed in the liver and spleen. Considerable particle accumulation was observed in the lung when a large quantity of γ-Fe2 O3 particles was administered. There was no significant particle accumulation in the kidney and heart.

  2. Immunobiotic Lactobacillus strains augment NLRP3 expression in newborn and adult porcine gut-associated lymphoid tissues.

    PubMed

    Tohno, Masanori; Shimosato, Takeshi; Aso, Hisashi; Kitazawa, Haruki

    2011-12-15

    We isolated cDNA encoding porcine nucleotide-binding domain-like receptor family, pryin domain containing 3 (NLRP3) from Peyer's patches. The complete nucleotide open reading frame of porcine NLRP3 contains 3108-bp encoding a deduced polypeptide of 1036-amino acid residues. The porcine NLRP3 amino acid sequence is more similar to the longest isoform of human than the mouse counterpart. The predicted amino acid sequence of porcine NLRP3 presented nine C-terminal leucine-rich repeat domains. In newborn swine, the expression of NLRP3 was detected at higher levels in spleen and mesenteric lymph nodes, while lower levels were observed in intestinal tissues. In adult swine, NLRP3 was strongly expressed in Peyer's patches and the mesenteric lymph nodes, and the expression level in the lower intestinal tissues was comparable to that in spleen. Toll-like receptor and nucleotide-binding domain ligands, as well as Lactobacillus delbrueckii subsp. bulgaricus and Lactobacillus gasseri, enhanced NLRP3 expression in gut-associated lymphoid tissues (GALT) of newborn and adult swine. Our results should aid in understanding the intestinal immunoregulatory mechanisms underlying NLRP3 activation and the priming ability of immunobiotic lactic acid bacteria in porcine GALT.

  3. Current Understanding of the Pathways Involved in Adult Stem and Progenitor Cell Migration for Tissue Homeostasis and Repair.

    PubMed

    Goichberg, Polina

    2016-08-01

    With the advancements in the field of adult stem and progenitor cells grows the recognition that the motility of primitive cells is a pivotal aspect of their functionality. There is accumulating evidence that the recruitment of tissue-resident and circulating cells is critical for organ homeostasis and effective injury responses, whereas the pathobiology of degenerative diseases, neoplasm and aging, might be rooted in the altered ability of immature cells to migrate. Furthermore, understanding the biological machinery determining the translocation patterns of tissue progenitors is of great relevance for the emerging methodologies for cell-based therapies and regenerative medicine. The present article provides an overview of studies addressing the physiological significance and diverse modes of stem and progenitor cell trafficking in adult mammalian organs, discusses the major microenvironmental cues regulating cell migration, and describes the implementation of live imaging approaches for the exploration of stem cell movement in tissues and the factors dictating the motility of endogenous and transplanted cells with regenerative potential. PMID:27209167

  4. Comparison of Accuracy of Whole-Exome Sequencing with Formalin-Fixed Paraffin-Embedded and Fresh Frozen Tissue Samples

    PubMed Central

    Kwon, Mi Jeong; Kim, Ryong Nam; Kim, Yu Jin; Song, Ji-Young; Jung, Kyung Soo; Shin, Young Kee

    2015-01-01

    Formalin fixing with paraffin embedding (FFPE) has been a standard sample preparation method for decades, and archival FFPE samples are still very useful resources. Nonetheless, the use of FFPE samples in cancer genome analysis using next-generation sequencing, which is a powerful technique for the identification of genomic alterations at the nucleotide level, has been challenging due to poor DNA quality and artificial sequence alterations. In this study, we performed whole-exome sequencing of matched frozen samples and FFPE samples of tissues from 4 cancer patients and compared the next-generation sequencing data obtained from these samples. The major differences between data obtained from the 2 types of sample were the shorter insert size and artificial base alterations in the FFPE samples. A high proportion of short inserts in the FFPE samples resulted in overlapping paired reads, which could lead to overestimation of certain variants; >20% of the inserts in the FFPE samples were double sequenced. A large number of soft clipped reads was found in the sequencing data of the FFPE samples, and about 30% of total bases were soft clipped. The artificial base alterations, C>T and G>A, were observed in FFPE samples only, and the alteration rate ranged from 200 to 1,200 per 1M bases when sequencing errors were removed. Although high-confidence mutation calls in the FFPE samples were compatible to that in the frozen samples, caution should be exercised in terms of the artifacts, especially for low-confidence calls. Despite the clearly observed artifacts, archival FFPE samples can be a good resource for discovery or validation of biomarkers in cancer research based on whole-exome sequencing. PMID:26641479

  5. Comparison of Accuracy of Whole-Exome Sequencing with Formalin-Fixed Paraffin-Embedded and Fresh Frozen Tissue Samples.

    PubMed

    Oh, Ensel; Choi, Yoon-La; Kwon, Mi Jeong; Kim, Ryong Nam; Kim, Yu Jin; Song, Ji-Young; Jung, Kyung Soo; Shin, Young Kee

    2015-01-01

    Formalin fixing with paraffin embedding (FFPE) has been a standard sample preparation method for decades, and archival FFPE samples are still very useful resources. Nonetheless, the use of FFPE samples in cancer genome analysis using next-generation sequencing, which is a powerful technique for the identification of genomic alterations at the nucleotide level, has been challenging due to poor DNA quality and artificial sequence alterations. In this study, we performed whole-exome sequencing of matched frozen samples and FFPE samples of tissues from 4 cancer patients and compared the next-generation sequencing data obtained from these samples. The major differences between data obtained from the 2 types of sample were the shorter insert size and artificial base alterations in the FFPE samples. A high proportion of short inserts in the FFPE samples resulted in overlapping paired reads, which could lead to overestimation of certain variants; >20% of the inserts in the FFPE samples were double sequenced. A large number of soft clipped reads was found in the sequencing data of the FFPE samples, and about 30% of total bases were soft clipped. The artificial base alterations, C>T and G>A, were observed in FFPE samples only, and the alteration rate ranged from 200 to 1,200 per 1M bases when sequencing errors were removed. Although high-confidence mutation calls in the FFPE samples were compatible to that in the frozen samples, caution should be exercised in terms of the artifacts, especially for low-confidence calls. Despite the clearly observed artifacts, archival FFPE samples can be a good resource for discovery or validation of biomarkers in cancer research based on whole-exome sequencing.

  6. Comparison of Accuracy of Whole-Exome Sequencing with Formalin-Fixed Paraffin-Embedded and Fresh Frozen Tissue Samples.

    PubMed

    Oh, Ensel; Choi, Yoon-La; Kwon, Mi Jeong; Kim, Ryong Nam; Kim, Yu Jin; Song, Ji-Young; Jung, Kyung Soo; Shin, Young Kee

    2015-01-01

    Formalin fixing with paraffin embedding (FFPE) has been a standard sample preparation method for decades, and archival FFPE samples are still very useful resources. Nonetheless, the use of FFPE samples in cancer genome analysis using next-generation sequencing, which is a powerful technique for the identification of genomic alterations at the nucleotide level, has been challenging due to poor DNA quality and artificial sequence alterations. In this study, we performed whole-exome sequencing of matched frozen samples and FFPE samples of tissues from 4 cancer patients and compared the next-generation sequencing data obtained from these samples. The major differences between data obtained from the 2 types of sample were the shorter insert size and artificial base alterations in the FFPE samples. A high proportion of short inserts in the FFPE samples resulted in overlapping paired reads, which could lead to overestimation of certain variants; >20% of the inserts in the FFPE samples were double sequenced. A large number of soft clipped reads was found in the sequencing data of the FFPE samples, and about 30% of total bases were soft clipped. The artificial base alterations, C>T and G>A, were observed in FFPE samples only, and the alteration rate ranged from 200 to 1,200 per 1M bases when sequencing errors were removed. Although high-confidence mutation calls in the FFPE samples were compatible to that in the frozen samples, caution should be exercised in terms of the artifacts, especially for low-confidence calls. Despite the clearly observed artifacts, archival FFPE samples can be a good resource for discovery or validation of biomarkers in cancer research based on whole-exome sequencing. PMID:26641479

  7. Evaluation of a novel tagging and tissue preservation system for potential use in forensic sample collection.

    PubMed

    Grassberger, Martin; Stein, Christina; Hanslik, Stefan; Hochmeister, Manfred

    2005-07-16

    The authors describe a new, easy-to-use barcode-based tissue collection, preservation and body tracking system, which might prove instrumental in the containment of mass fatalities such as aircraft accidents, war related accidents, environmental disasters (e.g. earthquakes, hurricanes, and floods) terrorist bombings or mass murders.

  8. A novel approach in personal identification from tissue samples undergone different processes through STR typing.

    PubMed

    Staiti, N; Di Martino, D; Saravo, L

    2004-12-01

    Short tandem repeats (STRs) or microsatellites have been recognized worldwide as a powerful tool for human identification. They have become widely used in human identification especially in criminal cases and mass disasters. Police departments are often interested in cases where tissues are already decomposed and only do bones remain to let them perform laboratory analyses. Bone is the most resistant tissue in animal body to time depending degradation and putrefaction, but it is often hard to extract DNA from it because of its highly mineralized structure, which makes DNA extraction and/or amplification hard to carry out. We have performed human nuclear DNA extraction and STR typing in three different cases, on bones and bone fragments from long time dead persons found buried, in the sea, almost completely burnt and whose tissues were already decomposed. We report these caseworks as we would like to show how forensic scientists are improving their skill in identifying people whose corps have undergone several kinds of processes, even independently on the time passed and the level of putrefaction of their tissues.

  9. Evaluation of a novel tagging and tissue preservation system for potential use in forensic sample collection.

    PubMed

    Grassberger, Martin; Stein, Christina; Hanslik, Stefan; Hochmeister, Manfred

    2005-07-16

    The authors describe a new, easy-to-use barcode-based tissue collection, preservation and body tracking system, which might prove instrumental in the containment of mass fatalities such as aircraft accidents, war related accidents, environmental disasters (e.g. earthquakes, hurricanes, and floods) terrorist bombings or mass murders. PMID:15939157

  10. The focus on sample quality: Influence of colon tissue collection on reliability of qPCR data

    PubMed Central

    Korenkova, Vlasta; Slyskova, Jana; Novosadova, Vendula; Pizzamiglio, Sara; Langerova, Lucie; Bjorkman, Jens; Vycital, Ondrej; Liska, Vaclav; Levy, Miroslav; Veskrna, Karel; Vodicka, Pavel; Vodickova, Ludmila; Kubista, Mikael; Verderio, Paolo

    2016-01-01

    Successful molecular analyses of human solid tissues require intact biological material with well-preserved nucleic acids, proteins, and other cell structures. Pre-analytical handling, comprising of the collection of material at the operating theatre, is among the first critical steps that influence sample quality. The aim of this study was to compare the experimental outcomes obtained from samples collected and stored by the conventional means of snap freezing and by PAXgene Tissue System (Qiagen). These approaches were evaluated by measuring rRNA and mRNA integrity of the samples (RNA Quality Indicator and Differential Amplification Method) and by gene expression profiling. The collection procedures of the biological material were implemented in two hospitals during colon cancer surgery in order to identify the impact of the collection method on the experimental outcome. Our study shows that the pre-analytical sample handling has a significant effect on the quality of RNA and on the variability of qPCR data. PAXgene collection mode proved to be more easily implemented in the operating room and moreover the quality of RNA obtained from human colon tissues by this method is superior to the one obtained by snap freezing. PMID:27383461

  11. Identification of immune cell infiltration in hematoxylin-eosin stained breast cancer samples: texture-based classification of tissue morphologies

    NASA Astrophysics Data System (ADS)

    Turkki, Riku; Linder, Nina; Kovanen, Panu E.; Pellinen, Teijo; Lundin, Johan

    2016-03-01

    The characteristics of immune cells in the tumor microenvironment of breast cancer capture clinically important information. Despite the heterogeneity of tumor-infiltrating immune cells, it has been shown that the degree of infiltration assessed by visual evaluation of hematoxylin-eosin (H and E) stained samples has prognostic and possibly predictive value. However, quantification of the infiltration in H and E-stained tissue samples is currently dependent on visual scoring by an expert. Computer vision enables automated characterization of the components of the tumor microenvironment, and texture-based methods have successfully been used to discriminate between different tissue morphologies and cell phenotypes. In this study, we evaluate whether local binary pattern texture features with superpixel segmentation and classification with support vector machine can be utilized to identify immune cell infiltration in H and E-stained breast cancer samples. Guided with the pan-leukocyte CD45 marker, we annotated training and test sets from 20 primary breast cancer samples. In the training set of arbitrary sized image regions (n=1,116) a 3-fold cross-validation resulted in 98% accuracy and an area under the receiver-operating characteristic curve (AUC) of 0.98 to discriminate between immune cell -rich and - poor areas. In the test set (n=204), we achieved an accuracy of 96% and AUC of 0.99 to label cropped tissue regions correctly into immune cell -rich and -poor categories. The obtained results demonstrate strong discrimination between immune cell -rich and -poor tissue morphologies. The proposed method can provide a quantitative measurement of the degree of immune cell infiltration and applied to digitally scanned H and E-stained breast cancer samples for diagnostic purposes.

  12. Bisulfite-Based DNA Methylation Analysis from Recent and Archived Formalin-Fixed, Paraffin Embedded Colorectal Tissue Samples.

    PubMed

    Kalmár, Alexandra; Péterfia, Bálint; Hollósi, Péter; Wichmann, Barnabás; Bodor, András; Patai, Árpád V; Schöller, Andrea; Krenács, Tibor; Tulassay, Zsolt; Molnár, Béla

    2015-09-01

    We aimed to test the applicability of formalin-fixed and paraffin-embedded (FFPE) tissue samples for gene specific DNA methylation analysis after using two commercially available DNA isolation kits. Genomic DNA was isolated from 5 colorectal adenocarcinomas and 5 normal adjacent tissues from "recent", collected within 6 months, and "archived", collected more than 5 years ago, FFPE tissues using either High Pure FFPET DNA Isolation kit or QIAamp DNA FFPE Tissue kit. DNA methylation analysis of MAL, SFRP1 and SFRP2 genes, known to be hypermethylated in CRC, was performed using methylation-sensitive high resolution melting (MS-HRM) analysis and sequencing. QIAamp (Q) method resulted in slightly higher recovery in archived (HP: 1.22 ± 3.18 μg DNA; Q: 3.00 ± 4.04 μg DNA) and significantly (p < 0.05) higher recovery in recent samples compared to High Pure method (HP) (HP: 4.10 ± 2.91 μg DNA; Q: 11.51 ± 7.50 μg DNA). Both OD260/280 and OD260/230 ratios were lower, but still high in the High Pure isolated archived and recent samples compared to those isolated with QIAamp. Identical DNA methylation patterns were detected for all 3 genes tested by MS-HRM with both isolation kits in the recent group. However, despite of higher DNA recovery in QIAamp slightly more reproducible methylation results were obtained from High Pure isolated archived samples. Sequencing confirmed DNA hypermethylation in CRCs. In conclusion, reproducible DNA methylation patterns were obtained from recent samples using both isolation kits. However, long term storage may affect the reliability of the results leading to moderate differences between the efficiency of isolation kits.

  13. Bisulfite-Based DNA Methylation Analysis from Recent and Archived Formalin-Fixed, Paraffin Embedded Colorectal Tissue Samples.

    PubMed

    Kalmár, Alexandra; Péterfia, Bálint; Hollósi, Péter; Wichmann, Barnabás; Bodor, András; Patai, Árpád V; Schöller, Andrea; Krenács, Tibor; Tulassay, Zsolt; Molnár, Béla

    2015-09-01

    We aimed to test the applicability of formalin-fixed and paraffin-embedded (FFPE) tissue samples for gene specific DNA methylation analysis after using two commercially available DNA isolation kits. Genomic DNA was isolated from 5 colorectal adenocarcinomas and 5 normal adjacent tissues from "recent", collected within 6 months, and "archived", collected more than 5 years ago, FFPE tissues using either High Pure FFPET DNA Isolation kit or QIAamp DNA FFPE Tissue kit. DNA methylation analysis of MAL, SFRP1 and SFRP2 genes, known to be hypermethylated in CRC, was performed using methylation-sensitive high resolution melting (MS-HRM) analysis and sequencing. QIAamp (Q) method resulted in slightly higher recovery in archived (HP: 1.22 ± 3.18 μg DNA; Q: 3.00 ± 4.04 μg DNA) and significantly (p < 0.05) higher recovery in recent samples compared to High Pure method (HP) (HP: 4.10 ± 2.91 μg DNA; Q: 11.51 ± 7.50 μg DNA). Both OD260/280 and OD260/230 ratios were lower, but still high in the High Pure isolated archived and recent samples compared to those isolated with QIAamp. Identical DNA methylation patterns were detected for all 3 genes tested by MS-HRM with both isolation kits in the recent group. However, despite of higher DNA recovery in QIAamp slightly more reproducible methylation results were obtained from High Pure isolated archived samples. Sequencing confirmed DNA hypermethylation in CRCs. In conclusion, reproducible DNA methylation patterns were obtained from recent samples using both isolation kits. However, long term storage may affect the reliability of the results leading to moderate differences between the efficiency of isolation kits. PMID:25991403

  14. Super's Stages and the Four-Factor Structure of the Adult Career Concerns Inventory in an Australian Sample.

    ERIC Educational Resources Information Center

    Smart, Roslyn M.; Peterson, Candida C.

    1994-01-01

    Examined four-factor structure of Adult Career Concerns Inventory (ACCI) in sample of 457 employed Australians. Findings support factorial validity of ACCI by showing its congruence with Super et al.'s (1957) hypothesized 4 stages and 12 substages of career development. Results also demonstrated substantial cross-cultural support for ACCI factor…

  15. Reliability and Structural Validity of Cross Racial Identity Scale Scores in a Sample of African American Adults

    ERIC Educational Resources Information Center

    Worrell, Frank C.; Vandiver, Beverly J.; Cross, William E., Jr.; Fhagen-Smith, Peony E.

    2004-01-01

    In this article, the authors examine the internal consistency and structural validity of scores on the Cross Racial Identity Scale (CRIS; Vandiver et al., 2000; Worrell, Vandiver, & Cross, 2000) in a sample of 105 adults. Exploratory factor analyses provided support for the six-factor structure of the CRIS. Reliability estimates for the scores…

  16. Effect of sampling method on the species composition and abundance of adult mosquitoes in a Florida swamp.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Samples of the adult mosquito populations in a Florida swamp (Sumter Co.) were obtained using suction traps and portable CDC light traps (augmented with CO2) and the results compared with mosquitoes captured by mechanical aspirator when landing on a human subject. Sixteen mosquito species total wer...

  17. The Relationship between Intelligence and Multiple Domains of Religious Belief: Evidence from a Large Adult US Sample

    ERIC Educational Resources Information Center

    Lewis, Gary J.; Ritchie, Stuart J.; Bates, Timothy C.

    2011-01-01

    High levels of religiosity have been linked to lower levels of intelligence in a number of recent studies. These results have generated both controversy and theoretical interest. Here in a large sample of US adults we address several issues that restricted the generalizability of these previous results. We measured six dimensions of religiosity…

  18. The Relationship between Child Abuse, Parental Divorce, and Lifetime Mental Disorders and Suicidality in a Nationally Representative Adult Sample

    ERIC Educational Resources Information Center

    Afifi, Tracie O.; Boman, Jonathan; Fleisher, William; Sareen, Jitender

    2009-01-01

    Objectives: To determine how the experiences of child abuse and parental divorce are related to long-term mental health outcomes using a nationally representative adult sample after adjusting for sociodemographic variables and parental psychopathology. Methods: Data were drawn from the National Comorbidity Survey (NCS, n=5,877; age 15-54 years;…

  19. Concurrent Validity of Wechsler Adult Intelligence Scales-Third Edition Index Score Short Forms in the Canadian Standardization Sample

    ERIC Educational Resources Information Center

    Lange, Rael T.; Iverson, Grant L.

    2008-01-01

    This study evaluated the concurrent validity of estimated Wechsler Adult Intelligence Scales-Third Edition (WAIS-III) index scores using various one- and two-subtest combinations. Participants were the Canadian WAIS-III standardization sample. Using all possible one- and two-subtest combinations, an estimated Verbal Comprehension Index (VCI), an…

  20. The Kuder Skills Assessment-College and Adult Version: Development and Initial Validation in a College Business Sample

    ERIC Educational Resources Information Center

    Rottinghaus, Patrick J.

    2009-01-01

    This article introduces the Kuder Skills Assessment-College and Adult version (KSA-CA; Rottinghaus, 2006), a new measure incorporating advances in the measurement of self-efficacy across 16 basic occupational domains (e.g., finance, information technology) and the six Kuder Clusters. Similar to the original development sample, all scales of the…

  1. Examining the Relationship between At-Risk Gambling and Suicidality in a National Representative Sample of Young Adults

    ERIC Educational Resources Information Center

    Feigelman, William; Gorman, Bernard S.; Lesieur, Henry

    2006-01-01

    Although many clinical studies document a relationship between gambling and suicidality, evidence of this association in general population surveys has been mixed. Probing this association in a nationally representative sample of young adults with data from the National Longitudinal Survey of Adolescent Health, we made same gender comparisons of…

  2. Long-Term Neurotoxicity of Chemotherapy in Adolescents and Young Adults Treated for Bone and Soft Tissue Sarcomas

    PubMed Central

    Connolly, Sean; Latoufis, Christos; Eagle, Karen; Ash, Catherine M.; Fowler, Clare; Souhami, Robert L.

    1998-01-01

    Purpose. To study the long-term neurotoxicity of chemotherapy in adolescents and young adults treated for bone and soft tissue sarcomas. Patients and Methods. Thirty-six adolescents and young adults (median age 17 years) were examined following chemotherapy for bone and soft tissue sarcomas. Twenty-nine (29/36) had received cisplatin (median 400 mg/m2), 15/36 ifosfamide (median 20 g/m2), and 12/36 vincristine (median 16 mg). Neurotoxicity was assessed at a median of 8 months (range, 1–54 months) after completion of chemotherapy by clinical examination, nerve conduction studies, audiograms and autonomic function tests. The same nerve conduction studies were carried out in 20 normal volunteers to define normal ranges in this age group. Results. Sixteen patients (44%) had a significant reduction in deep tendon reflexes, and this clinical parameter correlated well with abnormalities detected in nerve conduction studies. Vibration perception threshold (VPT) was raised in 20/36 patients (55%) and this was the most sensitive single test in the assessment of neuropathy. There was a significant correlation between VPT and cumulative cisplatin dose received in mg/m-2 (r=0.607, p<0.01). Ten of 29 patients (35%) had abnormal nerve conduction studies with a pattern characteristic of sensory axonal neuropathy. No patient complained of auditory symptoms, but minor high tone hearing loss was detected by audiograms in 5/28 patients who had received cisplatin. No patients had symptoms of autonomic neuropathy, but autonomic function tests showed minor abnormalities in 4/22 patients tested, and all had received cisplatin. Conclusions. This study demonstrates significant, although asymptomatic, long-term neurotoxicity of cisplatin in adolescents and young adults receiving chemotherapy for bone and soft tissue sarcomas. Follow-up studies are planned to assess whether these neurological deficits improve with time. PMID:18521240

  3. Preweaning growth hormone treatment ameliorates adipose tissue insulin resistance and inflammation in adult male offspring following maternal undernutrition.

    PubMed

    Reynolds, C M; Li, M; Gray, C; Vickers, M H

    2013-08-01

    It is well established that early-life nutritional alterations lead to increased risk of obesity and metabolic disorders in adult life. Although it is clear that obesity gives rise to chronic low-grade inflammation, there is little evidence regarding the role of inflammation in the adipose tissue of undernourished (UN) offspring. GH reduces fat mass and has antiinflammatory properties. The present study examined the effect of maternal UN on adipose inflammation in adult offspring and whether GH treatment during a critical period of developmental plasticity could ameliorate metabolic dysfunction associated with a poor start to life. Sprague Dawley rats were assigned to chow (C) or UN (50% ad libitum; UN) diet throughout gestation. Male C and UN pups received saline (control saline [CS]/UN) or GH (2.5 μg/g/d; control growth hormone [CGH]/undernourished growth hormone [UNGH]) from days 3-21. Postweaning males were further randomized and fed either chow or high-fat diet until day 160. An ex vivo glucose uptake assay demonstrated adipose tissue from UN offspring displayed attenuated insulin-stimulated glucose uptake compared with CS, CGH, and UNGH. This was associated with increased insulin receptor, glucose transporter 4, and insulin receptor substrate 1 gene expression. Furthermore, UN demonstrated enhanced TNFα and IL-1β secretion from adipose explants and stromal vascular fraction cultures accompanied by increased adipose tissue gene expression of several key proinflammatory genes and markers of macrophage infiltration. Overall, UN offspring displayed a more potent immunophenotype, which correlated with decreased insulin sensitivity. Preweaning GH treatment negates these detrimental effects, indicating the potential for reversing metabolic dysfunction in UN adult offspring.

  4. Histological image data of limb skeletal tissue from larval and adult Ambystoma mexicanum.

    PubMed

    McCusker, Catherine D; Diaz-Castillo, Carlos; Sosnik, Julian; Phan, Anne; Gardiner, David M

    2016-09-01

    The data presented in this article are related to the article entitled "Cartilage and bone cells do not participate in skeletal regeneration in Ambystoma mexicanum limbs" [1]. Here we present image data of the post-embryonic development of the forelimb skeletal tissue of Ambystoma Mexicanum. Histological staining was performed on sections from the intact limbs of young (6.5 cm) and old (25 cm) animals, and on dissected skeletal tissues (cartilage, bone, and periosteum) from these animals.

  5. Dietary patterns and food choices of a population sample of adults on Guam.

    PubMed

    Pobocik, Rebecca S; Trager, Alison; Monson, Lora Morrell

    2008-01-01

    This study examined dietary patterns of adults on Guam. Four hundred subjects were selected via a multistage procedure to represent the island's population. A 24-hour dietary recall was administered via telephone in 1995 to 1996. Diets were computer analyzed. There were 4,913 food items reported, representing 1,042 individual foods. Most respondents, 51.3%, had a "more-frequent" eating pattern, 36.9% had "regular" and 11.8% had "less-frequent." Energy consumption increased (p<0.05) with eating frequency. Highest levels of all nutrients (p<0.05) were at the evening meal. The sexes did not differ in nutrient proportions by meal. Grains, meats, and beverages were eaten most frequently. Meat/fish, mixed dishes, and grains supplied 68% of the energy and 47% to 91% of the micronutrients. Twenty five percent of the carbohydrate was from sweetened drinks and desserts. Rice was the most frequently consumed food. More than half of the subjects had no fruit, a third had no vegetables, and only 38.5% had dairy foods. Calcium came from multiple sources: dairy (27.6%), mixed dishes (23.8%), fish (12.6%), desserts (8.5%) and vegetables (6.6%). Most people, 91.8%, had fiesta meals with median consumption at 6 meals per year (range: 0 to 200). Betel nut was used by 12.3% of the sample with median consumption 0 whole nuts per day (range: 0 to 25). Ethnic differences (p<0.01) were observed in both fiesta meal and betel nut consumption. The Guamanian diet includes limited use of traditional foods and dietary patterns associated with increased risk for chronic disease.

  6. Condom use errors and problems in a national sample of young Croatian adults.

    PubMed

    Baćak, Valerio; Stulhofer, Aleksandar

    2012-08-01

    In this study, we examined the correlates of condom use errors and problems in a population-based study conducted in 2010 among young Croatian adults aged 18-25 years. Out of a total sample of 1,005 participants, 679 reported condom use in the preceding year. The analyses focused on four outcomes: condom breakage, condom slippage, condom-related erection loss, and delayed condom application. Eighteen percent of participants experienced breakage, 13% reported slippage, 17% reported erection loss, and 34% applied a condom after intercourse started. Multivariate logistic regression analyses were performed to examine the correlates of these condom use errors and problems. Condom breakage was less likely to be reported by women and older participants. The odds of breakage were increased for participants who reported being under the influence of drugs during sex and who reported other condom use errors and problems in the past year. Condom slippage was more likely to occur among younger participants and those who reported condom breakage and delayed condom application. Condom-related erection loss was positively associated with a higher number of sexual partners in the preceding year, condom breakage, and a higher score on the Anti-Erotic Obstacles to Condom Use Scale. Odds of delayed condom application were increased for participants who experienced condom breakage and for those who consumed alcohol before sex in the past year. Having used a condom at first sex significantly reduced the odds of applying a condom after intercourse started. In comparison to non-habitual condom users, habitual users were found less likely to report any of the assessed condom use errors and problems. Improving condom use skills remains an important task in Croatia, which is currently hampered by the absence of evidence-based sex education in schools. PMID:21882054

  7. Psychometric properties of the Trauma and Distress Scale, TADS, in an adult community sample in Finland

    PubMed Central

    Salokangas, Raimo K. R.; Schultze-Lutter, Frauke; Patterson, Paul; von Reventlow, Heinrich Graf; Heinimaa, Markus; From, Tiina; Luutonen, Sinikka; Hankala, Juha; Kotimäki, Mika; Tuominen, Lauri

    2016-01-01

    Background There is increasing evidence that a history of childhood abuse and neglect is not uncommon among individuals who experience mental disorder and that childhood trauma experiences are associated with adult psychopathology. Although several interview and self-report instruments for retrospective trauma assessment have been developed, many focus on sexual abuse (SexAb) rather than on multiple types of trauma or adversity. Methods Within the European Prediction of Psychosis Study, the Trauma and Distress Scale (TADS) was developed as a new self-report assessment of multiple types of childhood trauma and distressing experiences. The TADS includes 43 items and, following previous measures including the Childhood Trauma Questionnaire, focuses on five core domains: emotional neglect (EmoNeg), emotional abuse (EmoAb), physical neglect (PhyNeg), physical abuse (PhyAb), and SexAb. This study explores the psychometric properties of the TADS (internal consistency and concurrent validity) in 692 participants drawn from the general population who completed a mailed questionnaire, including the TADS, a depression self-report and questions on help-seeking for mental health problems. Inter-method reliability was examined in a random sample of 100 responders who were reassessed in telephone interviews. Results After minor revisions of PhyNeg and PhyAb, internal consistencies were good for TADS totals and the domain raw score sums. Intra-class coefficients for TADS total score and the five revised core domains were all good to excellent when compared to the interviewed TADS as a gold standard. In the concurrent validity analyses, the total TADS and its all core domains were significantly associated with depression and help-seeking for mental problems as proxy measures for traumatisation. In addition, robust cutoffs for the total TADS and its domains were calculated. Conclusions Our results suggest the TADS as a valid, reliable, and clinically useful instrument for assessing

  8. Concurrent and Simultaneous Polydrug Use: Latent Class Analysis of an Australian Nationally Representative Sample of Young Adults

    PubMed Central

    Quek, Lake-Hui; Chan, Gary C. K.; White, Angela; Connor, Jason P.; Baker, Peter J.; Saunders, John B.; Kelly, Adrian B.

    2013-01-01

    Background: Alcohol use and illicit drug use peak during young adulthood (around 18–29 years of age), but comparatively little is known about polydrug use in nationally representative samples of young adults. Drawing on a nationally representative cross-sectional survey (Australian National Drug Strategy Household Survey), this study examines polydrug use patterns and associated psychosocial risk factors among young adults (n = 3,333; age 19–29). Method: The use of a broad range of licit and illicit drugs were examined, including alcohol, tobacco, cannabis, cocaine, hallucinogens, ecstasy, ketamine, GHB, inhalants, steroids, barbiturates, meth/amphetamines, heroin, methadone/buprenorphine, other opiates, painkillers, and tranquilizers/sleeping pills. Latent class analysis was employed to identify patterns of polydrug use. Results: Polydrug use in this sample was best described using a 5-class solution. The majority of young adults predominantly used alcohol only (52.3%), alcohol and tobacco (34.18%). The other classes were cannabis, ecstasy, and licit drug use (9.4%), cannabis, amphetamine derivative, and licit drug use (2.8%), and sedative and alcohol use (1.3%). Young adult males with low education and/or high income were most at risk of polydrug use. Conclusion: Almost half of young adults reported polydrug use, highlighting the importance of post-high school screening for key risk factors and polydrug use profiles, and the delivery of early intervention strategies targeting illicit drugs. PMID:24350230

  9. Limit of Detection in X-ray Diffraction Measurements of Tissue Equivalent Samples

    NASA Astrophysics Data System (ADS)

    Zheng, Y.; Vassiljev, N.; Konstantinidis, A.; Griffiths, J.; Speller, R.

    2015-09-01

    There is a suggestion of a new approach to mammography whereby following a conventional mammogram, the radiologist could interrogate suspicious regions using X-ray diffraction whilst the patient is still present and to establish the true extent of disease. A starting point for this work is to quantify the minimum detectable amount of breast cancer within a realistic thickness phantom. Perspex has a similar diffraction pattern to healthy breast tissue whilst water is similar to breast tumour, hence these two materials are used as tissue equivalent test objects for X-ray diffraction measurements. The preliminary results show linear agreement between the ratio of Perspex to water and the ratio of the diffraction peak intensities at 0.7 nm-1 and 1.5 nm-1. The minimum detectable limit for a component of the two ‘tissue’ mix was found to be 4.1%. This suggests that X-ray diffraction can be used to quantify tissue like mixtures down to the 4.1% / 95.9% mix level and hence has a strong potential for delineating the extent of infiltration disease.

  10. Tissue localization of the endosymbiotic bacterium "Candidatus Blochmannia floridanus" in adults and larvae of the carpenter ant Camponotus floridanus.

    PubMed

    Sauer, Christina; Dudaczek, Dieter; Hölldobler, Bert; Gross, Roy

    2002-09-01

    The distribution of endosymbiotic bacteria in different tissues of queens, males, and workers of the carpenter ant Camponotus floridanus was investigated by light and electron microscopy and by in situ hybridization. A large number of bacteria could be detected in bacteriocytes within the midguts of workers, young virgin queens, and males. Large amounts of bacteria were also found in the oocytes of workers and queens. In contrast, bacteria were not present in oocyte-associated cells or in the spermathecae of mature queens, although occasionally a small number of bacteria could be detected in the testis follicles of males. Interestingly, the number of bacteriocytes in mature queens was strongly reduced and the bacteriocytes contained only very few or no bacteria at all, although the endosymbionts were present in huge amounts in the ovaries of the same animals. During embryogenesis of the deposited egg, the bacteria were concentrated in a ring of endodermal tissue destined to become the midgut in later developmental stages. However, during larval development, bacteria could also be detected in other tissues although to a lesser extent. Only in the last-instar larvae were bacteria found exclusively in the midgut tissue within typical bacteriocytes. Tetracycline and rifampin efficiently cleansed C. floridanus workers of their symbionts and the bacteriocytes of these animals still remained empty several months after treatment had ceased. Despite the lack of their endosymbionts, these adult animals were able to survive without any obvious negative effect under normal cultivation conditions.

  11. In vivo adeno-associated viral vector-mediated genetic engineering of white and brown adipose tissue in adult mice.

    PubMed

    Jimenez, Veronica; Muñoz, Sergio; Casana, Estefania; Mallol, Cristina; Elias, Ivet; Jambrina, Claudia; Ribera, Albert; Ferre, Tura; Franckhauser, Sylvie; Bosch, Fatima

    2013-12-01

    Adipose tissue is pivotal in the regulation of energy homeostasis through the balance of energy storage and expenditure and as an endocrine organ. An inadequate mass and/or alterations in the metabolic and endocrine functions of adipose tissue underlie the development of obesity, insulin resistance, and type 2 diabetes. To fully understand the metabolic and molecular mechanism(s) involved in adipose dysfunction, in vivo genetic modification of adipocytes holds great potential. Here, we demonstrate that adeno-associated viral (AAV) vectors, especially serotypes 8 and 9, mediated efficient transduction of white (WAT) and brown adipose tissue (BAT) in adult lean and obese diabetic mice. The use of short versions of the adipocyte protein 2 or uncoupling protein-1 promoters or micro-RNA target sequences enabled highly specific, long-term AAV-mediated transgene expression in white or brown adipocytes. As proof of concept, delivery of AAV vectors encoding for hexokinase or vascular endothelial growth factor to WAT or BAT resulted in increased glucose uptake or increased vessel density in targeted depots. This method of gene transfer also enabled the secretion of stable high levels of the alkaline phosphatase marker protein into the bloodstream by transduced WAT. Therefore, AAV-mediated genetic engineering of adipose tissue represents a useful tool for the study of adipose pathophysiology and, likely, for the future development of new therapeutic strategies for obesity and diabetes. PMID:24043756

  12. Tissue localization of the endosymbiotic bacterium "Candidatus Blochmannia floridanus" in adults and larvae of the carpenter ant Camponotus floridanus.

    PubMed

    Sauer, Christina; Dudaczek, Dieter; Hölldobler, Bert; Gross, Roy

    2002-09-01

    The distribution of endosymbiotic bacteria in different tissues of queens, males, and workers of the carpenter ant Camponotus floridanus was investigated by light and electron microscopy and by in situ hybridization. A large number of bacteria could be detected in bacteriocytes within the midguts of workers, young virgin queens, and males. Large amounts of bacteria were also found in the oocytes of workers and queens. In contrast, bacteria were not present in oocyte-associated cells or in the spermathecae of mature queens, although occasionally a small number of bacteria could be detected in the testis follicles of males. Interestingly, the number of bacteriocytes in mature queens was strongly reduced and the bacteriocytes contained only very few or no bacteria at all, although the endosymbionts were present in huge amounts in the ovaries of the same animals. During embryogenesis of the deposited egg, the bacteria were concentrated in a ring of endodermal tissue destined to become the midgut in later developmental stages. However, during larval development, bacteria could also be detected in other tissues although to a lesser extent. Only in the last-instar larvae were bacteria found exclusively in the midgut tissue within typical bacteriocytes. Tetracycline and rifampin efficiently cleansed C. floridanus workers of their symbionts and the bacteriocytes of these animals still remained empty several months after treatment had ceased. Despite the lack of their endosymbionts, these adult animals were able to survive without any obvious negative effect under normal cultivation conditions. PMID:12200264

  13. In vivo adeno-associated viral vector-mediated genetic engineering of white and brown adipose tissue in adult mice.

    PubMed

    Jimenez, Veronica; Muñoz, Sergio; Casana, Estefania; Mallol, Cristina; Elias, Ivet; Jambrina, Claudia; Ribera, Albert; Ferre, Tura; Franckhauser, Sylvie; Bosch, Fatima

    2013-12-01

    Adipose tissue is pivotal in the regulation of energy homeostasis through the balance of energy storage and expenditure and as an endocrine organ. An inadequate mass and/or alterations in the metabolic and endocrine functions of adipose tissue underlie the development of obesity, insulin resistance, and type 2 diabetes. To fully understand the metabolic and molecular mechanism(s) involved in adipose dysfunction, in vivo genetic modification of adipocytes holds great potential. Here, we demonstrate that adeno-associated viral (AAV) vectors, especially serotypes 8 and 9, mediated efficient transduction of white (WAT) and brown adipose tissue (BAT) in adult lean and obese diabetic mice. The use of short versions of the adipocyte protein 2 or uncoupling protein-1 promoters or micro-RNA target sequences enabled highly specific, long-term AAV-mediated transgene expression in white or brown adipocytes. As proof of concept, delivery of AAV vectors encoding for hexokinase or vascular endothelial growth factor to WAT or BAT resulted in increased glucose uptake or increased vessel density in targeted depots. This method of gene transfer also enabled the secretion of stable high levels of the alkaline phosphatase marker protein into the bloodstream by transduced WAT. Therefore, AAV-mediated genetic engineering of adipose tissue represents a useful tool for the study of adipose pathophysiology and, likely, for the future development of new therapeutic strategies for obesity and diabetes.

  14. Aspergillus PCR-Based Investigation of Fresh Tissue and Effusion Samples in Patients with Suspected Invasive Aspergillosis Enhances Diagnostic Capabilities

    PubMed Central

    Reinwald, M.; Spiess, B.; Heinz, W. J.; Heussel, C. P.; Bertz, H.; Cornely, O. A.; Hahn, J.; Lehrnbecher, T.; Kiehl, M.; Laws, H. J.; Wolf, H. H.; Schwerdtfeger, R.; Schultheis, B.; Burchardt, A.; Klein, M.; Dürken, M.; Claus, B.; Schlegel, F.; Hummel, M.; Hofmann, W.-K.

    2013-01-01

    Although it is a severe complication in immunocompromised patients, diagnosing invasive fungal disease (IFD), especially invasive aspergillosis (IA), remains difficult. In certain clinical scenarios, examining tissue samples for identification of the infectious organism becomes important. As culture-based methods rarely yield results, the performance of an Aspergillus-specific nested PCR in fresh tissue or pleural effusion samples was evaluated. Fresh tissue (n = 59) and effusion (n = 47) specimens from 79 immunocompromised patients were subjected to an Aspergillus-specific PCR assay. Twenty-six patients had proven (n = 20) or probable (n = 6) IFD, according to the European Organization for Research and Treatment of Cancer/Invasive Fungal Infections Cooperative Group and National Institute of Allergy and Infectious Diseases Mycoses Study Group (EORTC/MSG) criteria, while the remaining patients were classified as having either possible IFD (n = 30) or no IFD (n = 23). IA was identified as the underlying IFD in 21/26 proven/probable cases. PCR positivity was observed for 18/21 proven/probable and 6 possible IA cases; cases classified as no IA did not show positive signals. Patients with proven IFD (n = 5) with cultures positive for non-Aspergillus molds also had negative Aspergillus PCR results. Aspergillus PCR performance analysis yielded sensitivity and specificity values of 86% (95% confidence interval [CI], 65% to 95%) and 100% (95% CI, 86% to 100%), respectively, thus leading to a diagnostic odds ratio of >200. In this analysis, good diagnostic performance of the PCR assay for detection of IA was observed for tissue samples, while effusion samples showed lower sensitivity rates. PCR testing represents a complementary tool; a positive PCR result strengthens the likelihood of IA, whereas IA seems unlikely in cases with negative results but findings could indicate non-Aspergillus IFD. Thus, PCR testing of these specimens enhances the diagnostic capabilities. PMID

  15. Storage and shipping of tissue samples for DNA analyses: A case study on earthworms☆

    PubMed Central

    Straube, Daniela; Juen, Anita

    2013-01-01

    Nowadays, molecular analyses play an important role in studies of soil dwelling animals, for example in taxonomy, phylogeography or food web analyses. The quality of the DNA, used for later molecular analyses, is an important factor and depends on collection and preservation of samples prior to DNA extraction. Ideally, DNA samples are frozen immediately upon collection, but if samples are collected in the field, suitable preservation methods might be limited due to unavailability of resources or remote field sites. Moreover, shipping samples over long distances can cause loss of DNA quality e.g. by thawing or leaking of preservation liquid. In this study we use earthworms, a key organism in soil research, to compare three different DNA preservation methods – freezing at −20 °C, storing in 75% ethanol, and freeze drying. Samples were shipped from the United States of America to Austria. The DNA of the samples was extracted using two different extraction methods, peqGOLD™ and Chelex® 100. The DNA amplification success was determined by amplifying four DNA fragments of different length. The PCR amplification success is significantly influenced by preservation method and extraction method and differed significantly depending on the length of the DNA fragment. Freeze drying samples was the best preservation method when samples were extracted using the silica based extraction method peqGOLD™. For samples that were extracted with Chelex® 100, storage in ethanol was the best preservation method. However, the overall amplification success was significantly lower for the extraction procedure based on Chelex® 100. The detection of the small DNA fragments was higher and independent from the extraction method, while the amplification success was significantly reduced for the longer DNA fragments. We recommend freeze drying of DNA samples, especially when they have to be shipped for longer distances. No special packaging or declaration is needed for freeze dried

  16. Polychlorinated dibenzo-p-dioxins, dibenzofurans, and polychlorinated biphenyls in human tissues, meat, fish, and wildlife samples from India.

    PubMed

    Kumar, K S; Kannan, K; Paramasivan, O N; Shanmuga Sundaram, V P; Nakanishi, J; Masunaga, S

    2001-09-01

    Concentrations of polychlorinated dibenzo-p-dioxins (PCDDs), dibenzofurans (PCDFs), and non- and mono-ortho-substituted polychlorinated biphenyls (dioxin-like PCBs) were measured in tissues of humans, fishes, chicken, lamb, goat, predatory birds, and Ganges River dolphins collected from various locations in India. PCDDs/DFs were found in most of the samples analyzed with the liver of spotted owlet containing the highest concentration of 3,300 pg/g, fat wt. 2,3,7,8-Substituted PCDDs and PCDFs were found in human fattissues at concentrations ranging from 170 to 1,300 pg/g, fat wt. Concentrations of PCDDs were generally greaterthan those of PCDFs in human tissues, fishes, animal fat, and dolphin. Among fishes, meat, and wildlife samples analyzed, concentrations of PCDDs/DFs were found in the following order: country chicken < goat/lamb fat < fishes < river dolphins < predatory birds. Hepta-CDDs and OCDD were the major PCDD homologues found in humans, fishes, meat products, and dolphins. 2,3,7,8-Tetrachlorodibenzo-p-dioxin equivalents of PCDDs/DFs were greater than those of PCBs in selected fish, dolphin, and human samples. To our knowledge, this is the first report of PCDDs and PCDFs in human tissues, fishes, meat, and wildlife collected from India. PMID:11563645

  17. Using population screening for recruitment of young adults engaged in illicit drug use: methodological issues and sampling outcomes.

    PubMed

    Smirnov, Andrew; Kemp, Robert; Wells, Helene; Legosz, Margot; Najman, Jake M

    2014-05-01

    Social stigma, legal sanctions and the associated lack of sampling frames create barriers to the probabilistic sampling of those engaged in a variety of behaviour, including illicit drug use. We used a novel sampling approach to recruit respondents into a longitudinal study examining amphetamine-type stimulant use. A young adult population was screened for lifetime drug use to create a sampling frame of amphetamine-type stimulant users and non-users. We posted 12,118 screening questionnaires to a random selection of young adults listed on the electoral roll for Brisbane and the Gold Coast, Australia (N=107,275). Using a small pre-paid incentive and intensive telephone and postal reminders we attained a screening response rate of 49.9%. Eligible amphetamine-type stimulant users (used ecstasy or methamphetamine⩾3 times in past 12 months) and non-users (never used ecstasy or methamphetamine) were identified by screening responses. About two-thirds of each selected group took part in the longitudinal study. Comparisons with large-scale population survey data suggest the sample was broadly representative of young adult amphetamine-type stimulant users in Australia.

  18. Mass spectral determination of phenylacetonitrile (PAN) levels in body tissues of adult desert locust, Schistocerca gregaria

    Technology Transfer Automated Retrieval System (TEKTRAN)

    : Wings and legs of the gregarious desert locust, Schistocerca gregaria have been shown to be release sites of phenylacetonitrile (PAN), the major adult male-produced pheromone. However, there is limited information on the distribution of PAN within the locust. Here we show, using gas chromatograph...

  19. Hemodialysis outcomes in a global sample of children and young adult hemodialysis patients: the PICCOLO MONDO cohort

    PubMed Central

    Ferris, Maria; Gibson, Keisha; Plattner, Brett; Gipson, Debbie S.; Kotanko, Peter; Marcelli, Daniele; Marelli, Cristina; Etter, Michael; Carioni, Paola; von Gersdorff, Gero; Xu, Xiaoqi; Kooman, Jeroen P.; Xiao, Qingqing; van der Sande, Frank M.; Power, Albert; Picoits-Filho, Roberto; Sylvestre, Lucimary; Westreich, Katherine; Usvyat, Len

    2016-01-01

    Background The aim of this study was to describe the experience of pediatric and young adult hemodialysis (HD) patients from a global cohort. Methods The Pediatric Investigation and Close Collaborative Consortium for Ongoing Life Outcomes for MONitoring Dialysis Outcomes (PICCOLO MONDO) study provided de-identified electronic information of 3244 patients, ages 0–30 years from 2000 to 2012 in four regions: Asia, Europe, North America and South America. The study sample was categorized into pediatric (≤18 years old) and young adult (19–30 years old) groups based on the age at dialysis initiation. Results For those with known end-stage renal disease etiology, glomerular disease was the most common diagnosis in children and young adults. Using Europe as a reference group, North America [odds ratio (OR) 2.69; CI 1.29, 5.63] and South America (OR 4.21; CI 2.32, 7.63) had the greatest mortality among young adults. North America also had higher rates of overweight, obesity, hypertension, cardiovascular disease, hospitalizations and secondary diabetes compared with all other regions. Initial catheter use was greater for North American (86.4% in pediatric patients and 75.2% in young adults) and South America (80.6% in pediatric patients and 75.9% in young adults). Catheter use at 1-year follow-up was most common in North American children (77.3%) and young adults (62.9%). Asia had the lowest rate of catheter use. For both age groups, dialysis adequacy (equilibrated Kt/V) ranged between 1.4 and 1.5. In Asia, patients in both age groups had significantly longer treatment times than in any other region. Conclusions The PICCOLO MONDO study has provided unique baseline and 1-year follow-up information on children and young adults receiving HD around the globe. This cohort has brought to light aspects of care in these age groups that warrant further investigation. PMID:26985383

  20. Incidence of mycobacterial infections in cats in Great Britain: estimate from feline tissue samples submitted to diagnostic laboratories.

    PubMed

    Gunn-Moore, D A; Gaunt, C; Shaw, D J

    2013-08-01

    The aim of this study was to estimate the incidence of mycobacterial infections in cats in Great Britain (GB). This was performed using the proxy measure of feline tissue samples submitted to diagnostic laboratories in GB that were found to have histopathological changes typical of mycobacterial infection ('MYC'). Sixteen primary diagnostic laboratories were asked for information on the number of feline samples submitted in 2009, the number with MYC, the number undergoing Ziehl-Neelsen (ZN) staining and, for comparison, the number diagnosed with lymphoma. Eight laboratories provided full data for the whole year: 11,782 samples; lymphoma 3.2% (mean, 95% CI: 2.89, 3.5), MYC 1.16% (0.98; 1.37) and ZN-positive 0.31% (0.22; 0.43). Data on 1569 samples from seven laboratories that provided partial data on samples for the whole year revealed similar results, although all changes were more frequent: lymphoma 5.42% (4.35; 6.66), MYC 2.36% (1.66; 3.23) and ZN-positive 0.77% (0.40; 1.33). One laboratory only provided data for part of the year (4.5 months), reporting all three types of histopathology less frequently: 18,232 samples; lymphoma 0.2% (0.18; 0.32), MYC 0.07% (0.04; 0.12) and ZN-positive 0.05% (0.02; 0.09). The reasons for low reporting rates in this high-throughput laboratory are unclear. In total, 187 samples were reported as having MYC. Five Reference laboratories were also contacted, reporting 174 feline tissue submissions in 2009, with mycobacteria being cultured from 90. The study shows that MYC are frequently reported in tissue samples from cats in GB, being reported in ~1% of samples, with confirmation as ZN-positive in ~0.3%. Lymphoma is recognized as a common disease in cats, being seen in ~3% of samples in this study. When compared against MYC, lymphoma was reported only twice as frequently. This confirms that far from being rare, clinically significant mycobacterial infections occur commonly in cats in GB.

  1. Lifetime Prevalence of Suicide Attempts Among Sexual Minority Adults by Study Sampling Strategies: A Systematic Review and Meta-Analysis

    PubMed Central

    Bogaert, Laura; Rhodes, Anne E.; Brennan, David J.; Gesink, Dionne

    2016-01-01

    Background. Previous reviews have demonstrated a higher risk of suicide attempts for lesbian, gay, and bisexual (LGB) persons (sexual minorities), compared with heterosexual groups, but these were restricted to general population studies, thereby excluding individuals sampled through LGB community venues. Each sampling strategy, however, has particular methodological strengths and limitations. For instance, general population probability studies have defined sampling frames but are prone to information bias associated with underreporting of LGB identities. By contrast, LGB community surveys may support disclosure of sexuality but overrepresent individuals with strong LGB community attachment. Objectives. To reassess the burden of suicide-related behavior among LGB adults, directly comparing estimates derived from population- versus LGB community–based samples. Search methods. In 2014, we searched MEDLINE, EMBASE, PsycInfo, CINAHL, and Scopus databases for articles addressing suicide-related behavior (ideation, attempts) among sexual minorities. Selection criteria. We selected quantitative studies of sexual minority adults conducted in nonclinical settings in the United States, Canada, Europe, Australia, and New Zealand. Data collection and analysis. Random effects meta-analysis and meta-regression assessed for a difference in prevalence of suicide-related behavior by sample type, adjusted for study or sample-level variables, including context (year, country), methods (medium, response rate), and subgroup characteristics (age, gender, sexual minority construct). We examined residual heterogeneity by using τ2. Main results. We pooled 30 cross-sectional studies, including 21 201 sexual minority adults, generating the following lifetime prevalence estimates of suicide attempts: 4% (95% confidence interval [CI] = 3%, 5%) for heterosexual respondents to population surveys, 11% (95% CI = 8%, 15%) for LGB respondents to population surveys, and 20% (95% CI

  2. A low dimensional entropy-based descriptor of several tissues in skin cancer histopathology samples

    NASA Astrophysics Data System (ADS)

    Álvarez, Pablo; Corredor, Germán.; García-Arteaga, Juan D.; Romero, Eduardo

    2015-12-01

    The use of low-level visual features to assign high level labels in datasets of histopathology images is a possible solution to the problems derived from manual labeling by experts. However, in many cases, the visual cues are not enough. In this article we propose the use of features derived exclusively from the spatial distribution of the cell nuclei. These features are calculated using the weight of k-nn graphs constructed from the distances between cells. Results show that there are k values with enhanced discriminatory power, especially when comparing cancerous and non-cancerous tissue.

  3. Comparing paraffined and deparaffinized breast cancer tissue samples and an analysis of Raman spectroscopy and infrared methods

    NASA Astrophysics Data System (ADS)

    Depciuch, J.; Kaznowska, E.; Szmuc, K.; Zawlik, I.; Cholewa, M.; Heraud, P.; Cebulski, J.

    2016-05-01

    Breast cancer makes up a quarter of all cancer in women, which is why research into new diagnostic methods and sample preparations need to be developed at an accelerated pace. Researchers are looking for diagnostic tools to detect when an individual has cancer cells and use that information to see what measurements and approaches can be used to take further diagnostic steps. The most common method of sample preparation is the imbibing of tumor tissue in paraffin, which can produce a background for spectroscopic measurements in the range of 500-3500 cm-1. In this study we demonstrated that proper preparation of paraffin-embedded specimens and the measurement methodology can eliminate paraffin vibration, as was done in the work Depciuch et al. 2015. Thanks to this spectroscopic technique there may become a reliable and accurate method of diagnosing breast cancer based on the evidence found from the prepared samples. The study compared the results obtained through Raman spectroscopy and FTIR (Fourier Transform Infrared) measurements of healthy and cancerous breast tissues that were either embedded in paraffin or deparaffinized. The resulting spectrum and accurate analysis led to the conclusion that the appropriate measurement of the background and the elimination of peaks from the paraffin had the greatest impact on the reliability of results. Furthermore, after the accurate, detailed studies FTIR and Raman spectroscopy on samples of breast tissue that were deparaffinized or embedded in paraffin, including a complete analysis of the peak after transformation Kramers-Kröning (KK), it was found that sample preparation did not affect the result obtained by measuring the reflectance in the mid-infrared range, and that this only had a minimal effect relating to the intensity obtained by the measurement of the Raman peak. Only in special cases, when Raman spectroscopic methods are used for research to find the peculiarities of the spectra, are deparaffinization recommended

  4. Pathologic evaluation of a new endoscopic ultrasound needle designed to obtain core tissue samples: A pilot study

    PubMed Central

    Adler, Douglas G.; Witt, Benjamin; Chadwick, Barbara; Wells, Jason; Taylor, Linda Jo; Dimaio, Christopher; Schmidt, Robert

    2016-01-01

    Background and Objectives: Standard endoscopic ultrasound-fine-needle aspiration (EUS-FNA) needles are in widespread use. Meaningful differences between the available needles have been difficult to identify. Recently, a new EUS needle (Shark Core®, Covidien, Dublin, Leinster, Ireland), has been introduced in an attempt to improve diagnostic accuracy, tissue yield, and to potentially obtain a core tissue sample. We performed a pilot study prospectively to evaluate this new needle when compared to a standard EUS-FNA needle. Materials and Methods: Analysis of the first 15 patients undergoing EUS-FNA with the Shark Core needle was performed and it was compared to EUS-FNA in 15 patients who underwent EUS-FNA with a standard needle. Results: The Shark Core needle required fewer needle passes to obtain diagnostic adequacy than the standard needle [(χ2(1) = 11.3, P < 0.001]. The Shark Core needle required 1.5 passes to reach adequacy, whereas the standard needle required three passes. For cases with cell blocks, the Shark Core needle produced diagnostic material in 85% of cases [95% confidence interval (CI): 54–98], whereas the standard needle produced diagnostic material in 38% of the cases (95% CI: 9-76). The Shark Core needle produced actual tissue cores 82% of the time (95% CI: 48–98) and the standard needle produced no tissue cores (95% CI: 0-71) (P = 0.03). Conclusion: This pilot study found that the Shark Core needle had a high rate of producing adequate cytologic material for the diagnosis of pancreatic and peri-pancreatic lesions sampled by EUS with fewer passes required to obtain a definitive diagnosis and with a high rate of tissue cores being obtained when compared to a standard FNA needle. PMID:27386475

  5. Histological image data of limb skeletal tissue from larval and adult Ambystoma mexicanum.

    PubMed

    McCusker, Catherine D; Diaz-Castillo, Carlos; Sosnik, Julian; Phan, Anne; Gardiner, David M

    2016-09-01

    The data presented in this article are related to the article entitled "Cartilage and bone cells do not participate in skeletal regeneration in Ambystoma mexicanum limbs" [1]. Here we present image data of the post-embryonic development of the forelimb skeletal tissue of Ambystoma Mexicanum. Histological staining was performed on sections from the intact limbs of young (6.5 cm) and old (25 cm) animals, and on dissected skeletal tissues (cartilage, bone, and periosteum) from these animals. PMID:27547798

  6. Evidence of presence of Mycobacterium tuberculosis in bovine tissue samples by multiplex PCR: possible relevance to reverse zoonosis.

    PubMed

    Mittal, M; Chakravarti, S; Sharma, V; Sanjeeth, B S; Churamani, C P; Kanwar, N S

    2014-04-01

    Bovine tuberculosis, caused by Mycobacterium bovis, remains one of the most important zoonotic health concerns worldwide. The transmission of Mycobacterium tuberculosis from humans to animals also occurs especially in countries where there is close interaction of humans with the animals. In the present study, thirty bovine lung tissue autopsy samples from an organized dairy farm located in North India were screened for the presence of Mycobacterium tuberculosis complex by smear microscopy, histopathological findings and PCR. Differential diagnosis of M. tuberculosis and M. bovis was made based on the deletion of mce-3 operon in M. bovis. The present study found eight of these samples positive for M. tuberculosis by multiplex PCR. Sequencing was performed on two PCR-positive representative samples and on annotation, and BLAST analysis confirmed the presence of gene fragment specific to Mycobacterium tuberculosis. The presence of M. tuberculosis in all the positive samples raises the possibility of human-to-cattle transmission and possible adaptation of this organism in bovine tissues. This study accentuates the importance of screening and differential diagnosis of Mycobacterium tuberculosis complex in humans and livestock for adopting effective TB control and eradication programmes.

  7. Simulating an investigative study of clinical cancer samples: use of tissue slides and PCR-based promoter-hypermethylation analysis.

    PubMed

    Chew, Yuan Yuan; Chin, Cheen Fei; Yeong, Foong May

    2015-01-01

    Topics on the molecular basis underlying cancer are quite popular among students. Also, excellent textbooks abound that provide interesting materials for discussion during lectures and tutorials about major events leading to cancer formation and progression. However, much less is available for students to conduct experiments for the analysis of cancer samples in undergraduate modules where there is a limited time-frame. Given the difficulty of working with cancer samples and the scarcity of good samples even in the clinical laboratories, it is impossible to run large-class practicals using patients' samples. Here, we describe the use of tissue slides in combination with polymerase-chain reaction (PCR) as a means of simulating an investigative approach to supplement students' learning of clinical research. By using tissue slides for histo-pathological examinations and specific budding yeast genomic DNA and primers adapted to demonstrate methylation-specific PCR, we designed an inquiry-based lab session to simulate the clinical investigation of a cohort of biopsies that students could analyze in a one-session practical.

  8. Determination of protein carbonyls in plasma, cell extracts, tissue homogenates, isolated proteins: Focus on sample preparation and derivatization conditions

    PubMed Central

    Weber, Daniela; Davies, Michael J.; Grune, Tilman

    2015-01-01

    Protein oxidation is involved in regulatory physiological events as well as in damage to tissues and is thought to play a key role in the pathophysiology of diseases and in the aging process. Protein-bound carbonyls represent a marker of global protein oxidation, as they are generated by multiple different reactive oxygen species in blood, tissues and cells. Sample preparation and stabilization are key steps in the accurate quantification of oxidation-related products and examination of physiological/pathological processes. This review therefore focuses on the sample preparation processes used in the most relevant methods to detect protein carbonyls after derivatization with 2,4-dinitrophenylhydrazine with an emphasis on measurement in plasma, cells, organ homogenates, isolated proteins and organelles. Sample preparation, derivatization conditions and protein handling are presented for the spectrophotometric and HPLC method as well as for immunoblotting and ELISA. An extensive overview covering these methods in previously published articles is given for researchers who plan to measure protein carbonyls in different samples. PMID:26141921

  9. Anterior eye tissue morphology: Scleral and conjunctival thickness in children and young adults

    PubMed Central

    Read, Scott A.; Alonso-Caneiro, David; Vincent, Stephen J.; Bremner, Alexander; Fothergill, Annabel; Ismail, Brittney; McGraw, Rebecca; Quirk, Charlotte J.; Wrigley, Elspeth

    2016-01-01

    The sclera and conjunctiva form part of the eye’s tough, protective outer coat, and play important roles in the eye’s mechanical protection and immune defence, as well as in determining the size and shape of the eye globe. Advances in ocular imaging technology now allow these tissues in the anterior eye to be imaged non-invasively and with high resolution, however there is a paucity of data examining the dimensions of these tissues in paediatric populations. In this study, we have used optical coherence tomography (OCT) imaging to examine the normal in vivo thickness profile of the anterior sclera and overlying conjunctiva in 111 healthy young participants, including a large proportion of paediatric subjects. We demonstrate that the thickness of the anterior sclera varies significantly with measurement location and meridian. Tissue thickness also varied significantly with age, with younger subjects exhibiting significantly thinner scleras and significantly greater conjunctival thickness. Males were also found to exhibit significantly greater scleral thickness. Refractive error however was not significantly associated with either scleral or conjunctival thickness in this population. These findings provide new data describing the normative dimensions of anterior eye tissues in children and the factors that can influence these dimensions in young populations. PMID:27646956

  10. Anterior eye tissue morphology: Scleral and conjunctival thickness in children and young adults.

    PubMed

    Read, Scott A; Alonso-Caneiro, David; Vincent, Stephen J; Bremner, Alexander; Fothergill, Annabel; Ismail, Brittney; McGraw, Rebecca; Quirk, Charlotte J; Wrigley, Elspeth

    2016-01-01

    The sclera and conjunctiva form part of the eye's tough, protective outer coat, and play important roles in the eye's mechanical protection and immune defence, as well as in determining the size and shape of the eye globe. Advances in ocular imaging technology now allow these tissues in the anterior eye to be imaged non-invasively and with high resolution, however there is a paucity of data examining the dimensions of these tissues in paediatric populations. In this study, we have used optical coherence tomography (OCT) imaging to examine the normal in vivo thickness profile of the anterior sclera and overlying conjunctiva in 111 healthy young participants, including a large proportion of paediatric subjects. We demonstrate that the thickness of the anterior sclera varies significantly with measurement location and meridian. Tissue thickness also varied significantly with age, with younger subjects exhibiting significantly thinner scleras and significantly greater conjunctival thickness. Males were also found to exhibit significantly greater scleral thickness. Refractive error however was not significantly associated with either scleral or conjunctival thickness in this population. These findings provide new data describing the normative dimensions of anterior eye tissues in children and the factors that can influence these dimensions in young populations. PMID:27646956

  11. MicroRNA Stability in Postmortem FFPE Tissues: Quantitative Analysis Using Autoptic Samples from Acute Myocardial Infarction Patients.

    PubMed

    Kakimoto, Yu; Kamiguchi, Hiroshi; Ochiai, Eriko; Satoh, Fumiko; Osawa, Motoki

    2015-01-01

    MicroRNAs (miRNAs) are very short (18-24 nucleotides) nucleic acids that are expressed in a number of biological tissues and have been shown to be more resistant to extreme temperatures and pH compared to longer RNA molecules, like mRNAs. As miRNAs contribute to diverse biological process and respond to various kinds of cellular stress, their utility as diagnostic biomarkers and/or therapeutic targets has recently been explored. Here, we have evaluated the usefulness of miRNA quantification during postmortem examination of cardiac tissue from acute myocardial infarction (AMI) patients. Cardiac tissue was collected within one week of the patient's death and either frozen (19 samples) or fixed in formalin for up to three years (36 samples). RNA integrity was evaluated with an electropherogram, and it appears that longer RNAs are fragmented after death in the long-term fixed samples. Quantitative PCR was also performed for seven miRNAs and three other small RNAs in order to determine the appropriate controls for our postmortem analysis. Our data indicate that miR-191 and miR-26b are more suitable than the other types of small RNA molecules as they are stably detected after death and long-term fixation. Further, we also applied our quantitation method, using these endogenous controls, to evaluate the expression of three previously identified miRNA biomarkers, miR-1, miR-208b, and miR-499a, in formalin-fixed tissues from AMI patients. Although miR-1 and miR-208b decreased (1.4-fold) and increased (1.2-fold), respectively, in the AMI samples compared to the controls, the significance of these changes was limited by our sample size. In contrast, the relative level of miR-499a was significantly decreased in the AMI samples (2.1-fold). This study highlights the stability of miRNAs after death and long-term fixation, validating their use as reliable biomarkers for AMI during postmortem examination. PMID:26046358

  12. Epidemiology and survivorship of soft tissue sarcomas in adults: a national cancer database reporta

    PubMed Central

    Corey, Robert M; Swett, Katrina; Ward, William G

    2014-01-01

    The National Cancer Data Base (NCDB) of the American College of Surgeons gather demographic and survival data on ∼70% of cancers in the USA. We wanted to investigate the demographic and survivorship data on this potentially more representative cohort of patients with soft tissue sarcomas. We selected 34 of the most commonly encountered soft tissue sarcomas reported to the NCDB, provided that each entity contained a minimum of 50 cases. This report summarizes the demographic and survivorship data on 63,714 patients with these 34 histologically distinct soft tissue sarcomas reported to the NCDB from 1998 to 2010. The overall survivorships of these sarcomas were near the lower limits of many prior reports due to the all-inclusive, minimally biased inclusion criteria. The overall best prognosis was Dermatofibrosarcoma NOS (not otherwise specified). (5-year survivorship 92%). The worst prognosis was Dedifferentiated Chondrosarcoma (5-year survivorship 19%). New observations included Biphasic Synovial Sarcoma demonstrating a better 5-year survivorship (65%) compared to spindle-cell synovial sarcoma (56%, P < 0.031) and Synovial Sarcoma, NOS (52%, P < 0.001). The demographic and 2- and 5-year survivorship data for all 34 soft tissue sarcomas are presented herein. This extent of demographic and survival data in soft tissue sarcomas is unprecedented. Because of the large number of cases and the inclusive nature of the NCDB, without restriction to certain stages, categories, or treatments, it is less subject to selection bias. Therefore, these data are thought to be more reflective of the true overall prognosis given the current management of sarcoma across the NCDB contributing sites. PMID:25044961

  13. Determination of five nitroimidazole residues in artificial porcine muscle tissue samples by capillary electrophoresis.

    PubMed

    Lin, Yingyun; Su, Yan; Liao, Xiulin; Yang, Na; Yang, Xiupei; Choi, Martin M F

    2012-01-15

    A capillary electrophoresis (CE) method with ultraviolet detection has been developed for simultaneous detection and quantification of five nitroimidazoles including benzoylmetronidazole, dimetridazole, metronidazole, ronidazole, and secnidazole in porcine muscles. Nitroimidazoles in samples were extracted by ethyl acetate with subsequent clean-up by a strong cation exchange solid phase extraction column. The clean extracts were subjected to CE separation with optimal experimental conditions: pH 3.0 running buffer containing 25mM sodium phosphate and 0.10mM tetrabutylammonium bromide, 5s hydrodynamic injection at 0.5psi and 28kV separation voltage. The nitroimidazoles could be monitored and detected at 320nm within 18min. The limits of detection were below 1.0μg/kg and limits of quantification were lower than 3.2μg/kg for all nitroimidazoles in the muscle samples. The recoveries and relative standard deviations were 85.4-96.0, 83.5-92.5, 1.3-3.9, and 1.1-4.2%, respectively for the intra-day and inter-day analyses. The proposed CE method has been successfully applied to determine nitroimidazoles in artificial porcine muscle samples with good accuracy and recovery, demonstrating that it has potential for detection and quantification of multi-nitroimidazole residue in real muscle samples. PMID:22265553

  14. Neural correlates of inhibitory control in adult ADHD: Evidence from the Milwaukee longitudinal sample

    PubMed Central

    Mulligan, Richard C.; Knopik, Valerie S.; Sweet, Lawrence H.; Fischer, Mariellen; Seidenberg, Michael; Rao, Stephen M.

    2011-01-01

    Only a few studies have investigated the neural substrate of response inhibition in adult ADHD using Stop-Signal and Go/No-Go tasks. Inconsistencies and methodological limitations in the existing literature have resulted in limited conclusions regarding underlying pathophysiology. We examined the neural basis of response inhibition in a group of adults diagnosed with ADHD in childhood and who continue to meet criteria for ADHD while addressing limitations present in earlier studies. Adults with ADHD (n=12) and controls (n=12) were recruited from an ongoing longitudinal study and were matched for age, IQ, and education. Individuals with comorbid conditions were excluded. Functional MRI was used to identify and compare the brain activation patterns during correct trials of a response inhibition task (Go/No-Go). Our results showed that the control group recruited a more extensive network of brain regions than the ADHD group during correct inhibition trials. Adults with ADHD showed reduced brain activation in the right frontal eye field, pre-supplementary motor area, left precentral gyrus, and the inferior parietal lobe bilaterally. During successful inhibition of an inappropriate response, adults with ADHD display reduced activation in fronto-parietal networks previously implicated in working memory, goal-oriented attention, and response selection. This profile of brain activation may be specifically associated with ADHD in adulthood. PMID:21937201

  15. Biodistribution Analysis of Oncolytic Adenoviruses in Patient Autopsy Samples Reveals Vascular Transduction of Noninjected Tumors and Tissues

    PubMed Central

    Koski, Anniina; Bramante, Simona; Kipar, Anja; Oksanen, Minna; Juhila, Juuso; Vassilev, Lotta; Joensuu, Timo; Kanerva, Anna; Hemminki, Akseli

    2015-01-01

    In clinical trials with oncolytic adenoviruses, there has been no mortality associated with treatment vectors. Likewise, in the Advanced Therapy Access Program (ATAP), where 290 patients were treated with 10 different viruses, no vector-related mortality was observed. However, as the patient population who received adenovirus treatments in ATAP represented heavily pretreated patients, often with very advanced disease, some patients died relatively soon after receiving their virus treatment mandating autopsy to investigate cause of death. Eleven such autopsies were performed and confirmed disease progression as the cause of death in each case. The regulatory requirement for investigating the safety of advanced therapy medical products presented a unique opportunity to study tissue samples collected as a routine part of the autopsies. Oncolytic adenoviral DNA was recovered in a wide range of tissues, including injected and noninjected tumors and various normal tissues, demonstrating the ability of the vector to disseminate through the vascular route. Furthermore, we recovered and cultured viable virus from samples of noninjected brain metastases of an intravenously treated patient, confirming that oncolytic adenovirus can reach tumors through the intravascular route. Data presented here give mechanistic insight into mode of action and biodistribution of oncolytic adenoviruses in cancer patients. PMID:26156245

  16. Child maltreatment and psychological symptoms in a Portuguese adult community sample: the harmful effects of emotional abuse.

    PubMed

    Dias, Aida; Sales, Luísa; Hessen, David J; Kleber, Rolf J

    2015-07-01

    Child maltreatment (CM) is associated with poor long-term health outcomes. However, knowledge about CM prevalence and related consequences is scarce among adults in South European countries. We examined the self-reported prevalence of five different forms of CM in a community sample of 1,200 Portuguese adults; we compared the results with similar samples from three other countries, using the same instrument. We also explored the relationship between CM and psychological symptoms. Cross-sectional data using the Childhood Trauma Questionnaire-Short Form and the Brief Symptom Inventory were analyzed. Moderate or severe CM exposure was self-reported by 14.7% of the sample, and 67% was exposed to more than one form of CM. Emotional neglect was the most endorsed experience, with women reporting greater emotional abuse and men reporting larger physical abuse. Physical and sexual abuse was less self-reported by Portuguese than by American or German subjects. CM exposure predicted 12.8% of the psychological distress. Emotional abuse was the strongest predictor for psychological symptoms, namely for paranoid ideation, depression, and interpersonal sensitivity. Emotional abuse overlapped with the exposure to all other CM forms, and interacted with physical abuse, physical neglect, and emotional neglect to predict psychological distress. Low exposure to emotional abuse was directly associated with the effects of physical abuse, physical neglect, and emotional neglect to predict adult psychological distress. Verbal abuse experiences were frequently reported and had the highest correlations with adult psychological distress. Our results underline the potential hurtful effects of child emotional abuse among Portuguese adults in the community. They also highlight the need to improve prevention and intervention actions to reduce exposure and consequences of CM, particularly emotional abuse. PMID:25270111

  17. IHC Profiler: An Open Source Plugin for the Quantitative Evaluation and Automated Scoring of Immunohistochemistry Images of Human Tissue Samples

    PubMed Central

    Malhotra, Renu; De, Abhijit

    2014-01-01

    In anatomic pathology, immunohistochemistry (IHC) serves as a diagnostic and prognostic method for identification of disease markers in tissue samples that directly influences classification and grading the disease, influencing patient management. However, till today over most of the world, pathological analysis of tissue samples remained a time-consuming and subjective procedure, wherein the intensity of antibody staining is manually judged and thus scoring decision is directly influenced by visual bias. This instigated us to design a simple method of automated digital IHC image analysis algorithm for an unbiased, quantitative assessment of antibody staining intensity in tissue sections. As a first step, we adopted the spectral deconvolution method of DAB/hematoxylin color spectra by using optimized optical density vectors of the color deconvolution plugin for proper separation of the DAB color spectra. Then the DAB stained image is displayed in a new window wherein it undergoes pixel-by-pixel analysis, and displays the full profile along with its scoring decision. Based on the mathematical formula conceptualized, the algorithm is thoroughly tested by analyzing scores assigned to thousands (n = 1703) of DAB stained IHC images including sample images taken from human protein atlas web resource. The IHC Profiler plugin developed is compatible with the open resource digital image analysis software, ImageJ, which creates a pixel-by-pixel analysis profile of a digital IHC image and further assigns a score in a four tier system. A comparison study between manual pathological analysis and IHC Profiler resolved in a match of 88.6% (P<0.0001, CI = 95%). This new tool developed for clinical histopathological sample analysis can be adopted globally for scoring most protein targets where the marker protein expression is of cytoplasmic and/or nuclear type. We foresee that this method will minimize the problem of inter-observer variations across labs and further help in

  18. Formaldehyde exposure induces autophagy in testicular tissues of adult male rats.

    PubMed

    Han, Shui-Ping; Zhou, Dang-Xia; Lin, Pu; Qin, Zhen; An, Lu; Zheng, Lie-Rui; Lei, Li

    2015-03-01

    Formaldehyde, a ubiquitous environmental pollutant, has long been suspected of causing adverse male reproductive effects. However, the molecular and cellular mechanisms underlying this phenomenon remain elusive. The overall aim of this study is to clarify the role of autophagy in male reproductive injuries induced by formaldehyde exposure, by which we can further understand the molecular mechanism of spermatogenesis and develop new targets for prevention and treatment of male infertility. In this study, electron microscopy, Western blot, and RT-PCR analysis were used to detect autophagy in testicular tissues. Moreover, testicular weights, histopathology, and morphometry were used to evaluate the reproductive injuries of formaldehyde exposure. We found that formaldehyde exposure-induced autophagy in testicular tissues was dose dependent. Increasing autophagosomes in spermatogenetic cells was observed by electron microscopy in formaldehyde exposure group. In addition, RT-PCR and Western blot analysis showed the transcription levels of the LC3-II, as well as the conversion from LC3-I to LC3-II, an indicator of autophagy, significantly increased in testicular tissue of formaldehyde exposure group in a dose dependent manner when compared with those in control group. Furthermore, the alterations of autophage were basically consistent with the changes in testicular weight and morphologic findings. In summary, formaldehyde exposure triggered autophagy, and autophagy may be a scathing factor responsible for male reproductive impairment induced by formaldehyde.

  19. A CT-scan database for the facial soft tissue thickness of Taiwan adults.

    PubMed

    Chung, Ju-Hui; Chen, Hsiao-Ting; Hsu, Wan-Yi; Huang, Guo-Shu; Shaw, Kai-Ping

    2015-08-01

    Facial reconstruction is a branch of forensic anthropology used to assist in the identification of skeletal remains. The majority of facial reconstruction techniques use facial soft tissue depth chart data to recreate facial tissue on a skull or a model of a skull through the use of modeling clay. This study relied on 193 subjects selected from the Taiwanese population on the basis of age and gender to determine the average values of 32 landmarks, include midline and bilateral measures, by means of CT scans. The mean age of the subjects was 46.9±16.4 years, with a mean age of 43.8±16.6 for males and 49.9±15.8 for females respectively. There were 16 landmarks with statistically significant differences between male and female subjects, namely S, G, N, Na, Ph, Sd and Id in the midline portion, FE, LO, ZA and Sub M2 in the bilateral-right and left portion, and IM point in the bilateral-left portion (abbreviations adapted from Karen T. Taylor's work). The mean soft tissue depth was greater in males than in females, and there was significant difference between the right and left sides of the face in Za point. This study's findings were compared with those of Bulut et al. PMID:26028278

  20. Polytobacco use and multiple-product smoking among a random community sample of African-American adults

    PubMed Central

    Corral, Irma; Landrine, Hope; Simms, Denise Adams; Bess, Jukelia J

    2013-01-01

    Objectives Little is known about polytobacco use among African-American adults. This study is the first to explore this among a random, statewide, community sample of African-American adults. Setting Community-based sampling obtained a random, household-probability sample of African-American adults from California, surveyed door to door in randomly selected census tracts statewide. Participants Participants were a statewide, random-household sample of N=2118 African-American adults from California who completed a survey on past 30-day smoking of cigarettes, blunts, bidis, kreteks, cigarillos, marijuana and cigars. Results Almost half (49.3%) of the African-American cigarette-smokers and 14.9% of the cigarette non-smokers had smoked at least one non-cigarette product in the past 30 days. Smokers had a substantial prevalence of smoking cigarillos (28.7%) and blunts (27.7%). Logistic regressions revealed that the odds of smoking most of the non-cigarette products were higher for cigarette smokers and men, inversely related to age, and unrelated to socioeconomic status. However, smoking of blunts, bidis and kreteks was not predicted by cigarette smoking. Conclusions Smoking of cigarillos (eg, Phillies, Black & Mild) and blunts may be prevalent among African-American cigarette-smokers and non-smokers alike, but such products are not examined in most population-level smoking research. Smoking of these products should be included in surveillance studies, in cancer prevention programmes and in healthcare provider-assessment of smoking, and addressed in smoking cessation programmes as well. PMID:24334154

  1. Micro-scaled high-throughput digestion of plant tissue samples for multi-elemental analysis

    PubMed Central

    Hansen, Thomas H; Laursen, Kristian H; Persson, Daniel P; Pedas, Pai; Husted, Søren; Schjoerring, Jan K

    2009-01-01

    Background Quantitative multi-elemental analysis by inductively coupled plasma (ICP) spectrometry depends on a complete digestion of solid samples. However, fast and thorough sample digestion is a challenging analytical task which constitutes a bottleneck in modern multi-elemental analysis. Additional obstacles may be that sample quantities are limited and elemental concentrations low. In such cases, digestion in small volumes with minimum dilution and contamination is required in order to obtain high accuracy data. Results We have developed a micro-scaled microwave digestion procedure and optimized it for accurate elemental profiling of plant materials (1-20 mg dry weight). A commercially available 64-position rotor with 5 ml disposable glass vials, originally designed for microwave-based parallel organic synthesis, was used as a platform for the digestion. The novel micro-scaled method was successfully validated by the use of various certified reference materials (CRM) with matrices rich in starch, lipid or protein. When the micro-scaled digestion procedure was applied on single rice grains or small batches of Arabidopsis seeds (1 mg, corresponding to approximately 50 seeds), the obtained elemental profiles closely matched those obtained by conventional analysis using digestion in large volume vessels. Accumulated elemental contents derived from separate analyses of rice grain fractions (aleurone, embryo and endosperm) closely matched the total content obtained by analysis of the whole rice grain. Conclusion A high-throughput micro-scaled method has been developed which enables digestion of small quantities of plant samples for subsequent elemental profiling by ICP-spectrometry. The method constitutes a valuable tool for screening of mutants and transformants. In addition, the method facilitates studies of the distribution of essential trace elements between and within plant organs which is relevant for, e.g., breeding programmes aiming at improvement of the

  2. Association between Subcutaneous White Adipose Tissue and Serum 25-Hydroxyvitamin D in Overweight and Obese Adults

    PubMed Central

    Piccolo, Brian D.; Dolnikowski, Gregory; Seyoum, Elias; Thomas, Anthony P.; Gertz, Erik R.; Souza, Elaine C.; Woodhouse, Leslie R.; Newman, John W.; Keim, Nancy L.; Adams, Sean H.; Van Loan, Marta D.

    2013-01-01

    Cholecalciferol is known to be deposited in human adipose tissue, but it is not known whether 25-hydroxyvitamin D (25(OH)D) is found in detectable concentrations. Therefore, our objective was to determine whether 25(OH)D is detectable in subcutaneous white adipose tissue (SWAT) in overweight and obese persons enrolled in a twelve week energy restricted diet. Baseline and post-intervention gluteal SWAT biopsies were collected from 20 subjects participating in a larger clinical weight loss intervention. LC-MS/MS was utilized to determine SWAT 25(OH)D concentrations. Serum 25(OH)D and 1,25(OH)2D were measured by RIA. Body composition was assessed by dual energy x-ray absorptiometry. SWAT 25(OH)D concentrations were 5.8 ± 2.6 nmol/kg tissue and 6.2 ± 2.7 nmol/kg tissue pre- and post-intervention SWAT, respectively. There was a significant positive association between SWAT 25(OH)D concentration and serum 25(OH)D concentration (r = 0.52, P < 0.01). Both SWAT and serum 25(OH)D concentrations did not significantly change after a twelve-week period of energy restriction with approximately 5 kg of fat loss. In conclusion, we have demonstrated our LC-MS/MS method can detect 25(OH)D3 in human subcutaneous fat tissue from overweight and obese individuals and is consistent with previously reported concentrations in swine. Additionally, our findings of no significant changes in SWAT 25(OH)D3 or serum 25(OH)D after a 6% loss of total body weight and 13% reduction in total fat provides the first human evidence that adipose 25(OH)D does not likely contribute to serum 25(OH)D with moderate weight loss; whether this is also the case with larger amounts of weight loss is unknown. Weight loss alone is not sufficient to increase serum 25(OH)D and increases in dietary or dermal biosynthesis of vitamin D appear to be the most critical contributors to in vitamin D status. PMID:24067385

  3. Use of Curcuma longa L. extract to stain various tissue samples for histological studies

    PubMed Central

    Kumar, Sachin; Singh, Narendra Nath; Singh, Arun; Singh, Neelakshi; Sinha, Raman Kant

    2014-01-01

    Background: Curcuma longa L. is a perennial herb and a member of the Zingiberaceae (ginger) family, which is used extensively in foods as well as in Ayurvedic and Chinese systems of medicine. Current researches have focused on its antioxidant, hepatoprotective, anti-inflammatory, anticarcinogenic and antimicrobial properties. Until now, very few studies suggested its role as a histological stain. Aim: To ascertain its efficacy to be used as a counterstain after hematoxylin, to compare it's staining ability with that of routinely used eosin dye and also to ascertain its role in various collagen diseases. Materials and Methods: Turmeric rhizomes were cut into small pieces and were dried. These dried turmeric rhizomes were milled to form fine powder, which was then processed to form dye for staining tissue structures. Results: It revealed that turmeric can be used as a counterstain after hematoxylin, its staining ability was also good and comparable to that of eosin dye with a special affinity for collagen and muscle fibers. Conclusion: Turmeric dye can be used as a histological stain, which stains similar to eosin dye and its specific affinity for collagen and muscle fibers authenticates its role in the treatment of collagen and muscle disorders. PMID:26195911

  4. Premilitary Adult Sexual Assault Victimization and Perpetration in a Navy Recruit Sample

    ERIC Educational Resources Information Center

    Stander, Valerie A.; Merrill, Lex L.; Thomsen, Cynthia J.; Crouch, Julie L.; Milner, Joel S.

    2008-01-01

    Female (n = 5,226) and male (n = 5,969) U.S. Navy recruits completed a survey assessing their premilitary histories of adult sexual assault (SA), defined as attempted or completed rape since the age of 14. The survey was completed under anonymous or identified conditions. Overall, 39% of women reported premilitary SA victimization and 13% of men…

  5. Relationships between Childhood and Adult Physical Activity Patterns in a Community Sample.

    ERIC Educational Resources Information Center

    Lamarine, Roland J.; Polkinghorne, Ori

    This study examined the relationship between adult physical activity levels and patterns of activity that were established during childhood. A random digit telephone survey was conducted of noninstitutionalized residents in a medium sized California city. Subjects ages 18 and over who volunteered to participate were questioned about their…

  6. Factors Associated with Paranoid Symptoms in a Community Sample of Older Adults.

    ERIC Educational Resources Information Center

    Blazer, Dan G.; And Others

    1996-01-01

    Investigates paranoia in older adults. In cross-sectional analyses, symptoms were strongly associated with black race, lower income and education, less exercise, and more depressive symptoms. In longitudinal analysis, three years following initial interview, symptoms were predicted by baseline paranoid symptoms, education and depressive symptoms…

  7. A Measure of Severe Psychological Abuse Normed on a Nationally Representative Sample of Adults

    ERIC Educational Resources Information Center

    Follingstad, Diane R.

    2011-01-01

    A psychological abuse scale representing truly egregious psychological actions that could occur between adult intimate partners was constructed. To insure that the component behaviors would be viewed as highly problematic, the likely malignant intent of the actions was included in item descriptions. Fourteen categories of psychological abuse were…

  8. Determinants of Domain Knowledge and Independent Study Learning in an Adult Sample

    ERIC Educational Resources Information Center

    Ackerman, Phillip L.; Beier, Margaret E.

    2006-01-01

    The ability (fluid and crystallized intelligence) and nonability (personality, interests, self-concept, etc.) determinants of domain knowledge before and after an independent learning opportunity were evaluated in the context of a study of 141 adults between the ages of 18 and 69. The domain knowledge under consideration included an array of…

  9. Congregational Participation of a National Sample of Adults with Intellectual and Developmental Disabilities

    ERIC Educational Resources Information Center

    Carter, Erik W.; Kleinert, Harold L.; LoBianco, Tony F.; Sheppard-­Jones, Kathleen; Butler, Laura N.; Tyree, Milton S.

    2015-01-01

    Supporting people with intellectual and developmental disabilities (IDD) to thrive requires careful consideration of multiple avenues of community involvement. Yet little attention has focused on the place of faith community participation in the lives of adults with IDD. We examined attendance at religious services using National Core Indicator…

  10. The Relationships between Cognitive Ability and Dental Status in a National Sample of USA Adults

    ERIC Educational Resources Information Center

    Sabbah, Wael; Sheiham, Aubrey

    2010-01-01

    There are very few studies on the relationship between cognitive ability and dental status in middle aged and younger adults. We postulate that lower cognitive ability is directly related to poorer dental status and that this relationship operates through the relationship between cognitive ability and health-related behaviors. The objectives of…

  11. Patterns of Learning in a Sample of Adult Returners to Higher Education

    ERIC Educational Resources Information Center

    Anderson, Anthony; Johnston, Bill; McDonald, Alexandra

    2014-01-01

    This article presents empirical research exploring adult returner students' patterns of learning via qualitative analysis of a series of semi-structured interviews. Interviewees' comments shed light on the relation between patterns of learning on the one hand, and study skills, epistemological issues and attitudes to peer interaction on…

  12. Sexuality in a Community Based Sample of Adults with Autism Spectrum Disorder

    ERIC Educational Resources Information Center

    Gilmour, Laura; Schalomon, P. Melike; Smith, Veronica

    2012-01-01

    Few studies have examined the sexual attitudes and behaviours of individuals with high functioning autism spectrum disorders (ASDs) living in community settings. A total of 82 (55 female and 17 male) adults with autism were contrasted with 282 members of the general population on their responses to an online survey of sexual knowledge and…

  13. Item Parameter Invariance of the Kaufman Adolescent and Adult Intelligence Test across Male and Female Samples

    ERIC Educational Resources Information Center

    Immekus, Jason C.; Maller, Susan J.

    2009-01-01

    The Kaufman Adolescent and Adult Intelligence Test (KAIT[TM]) is an individually administered test of intelligence for individuals ranging in age from 11 to 85+ years. The item response theory-likelihood ratio procedure, based on the two-parameter logistic model, was used to detect differential item functioning (DIF) in the KAIT across males and…

  14. Non-lethal sampling of liver tissue for toxicologic evaluation of Florida cottonmouths snakes, Agkistrodon piscivorus conanti.

    PubMed

    Quesada, Rolando J; McCleary, Ryan J R; Heard, Darryl J; Lillywhite, Harvey B

    2014-01-01

    Due to their longevity, strong site tenure, poikilothermic metabolism, and low-energy specializations, reptiles might serve as excellent environmental sentinels. Cottonmouth snakes are generalist predators and scavengers, and as such, may have higher exposure to persistent environmental contaminants as a result of bioaccumulation. Traditionally, assessment and monitoring of contaminant exposure in reptiles have involved lethal sampling techniques. In this paper, we describe a non-destructive technique for sampling liver tissue in live anesthetized Florida cottonmouths. Wild-caught snakes (n = 21) were anesthetized with propofol, and a liver wedge biopsy was obtained by clamping the edge of the organ with two small hemostatic mosquito forceps via right-sided coeliotomy incision. A minimum required tissue sample weighing >100 mg was harvested from all except one of the animals. No mortalities occurred during the procedures or recovery from anesthesia, and all snakes were released back into the field after the animal had consumed prey and defecated, usually within 2 weeks following surgery. Hemorrhage was a minor complication in most snakes, especially those with friable discolored livers. The procedure appeared to have no short-term deleterious effects, and two biopsied individuals were captured after being released into the field and appeared to be normal and healthy. However, follow-up studies and recapture of more snakes are needed to assess long-term survivability. Our non-destructive liver sampling technique might be implemented in toxicological studies of other squamates and could help to minimize the lethal sampling of threatened species. PMID:24197420

  15. Comparative analysis of paracrine factor expression in human adult mesenchymal stem cells derived from bone marrow, adipose, and dermal tissue.

    PubMed

    Hsiao, Sarah Tzu-Feng; Asgari, Azar; Lokmic, Zerina; Sinclair, Rodney; Dusting, Gregory James; Lim, Shiang Yong; Dilley, Rodney James

    2012-08-10

    Human adult mesenchymal stem cells (MSCs) support the engineering of functional tissue constructs by secreting angiogenic and cytoprotective factors, which act in a paracrine fashion to influence cell survival and vascularization. MSCs have been isolated from many different tissue sources, but little is known about how paracrine factor secretion varies between different MSC populations. We evaluated paracrine factor expression patterns in MSCs isolated from adipose tissue (ASCs), bone marrow (BMSCs), and dermal tissues [dermal sheath cells (DSCs) and dermal papilla cells (DPCs)]. Specifically, mRNA expression analysis identified insulin-like growth factor-1 (IGF-1), vascular endothelial growth factor-D (VEGF-D), and interleukin-8 (IL-8) to be expressed at higher levels in ASCs compared with other MSC populations whereas VEGF-A, angiogenin, basic fibroblast growth factor (bFGF), and nerve growth factor (NGF) were expressed at comparable levels among the MSC populations examined. Analysis of conditioned media (CM) protein confirmed the comparable level of angiogenin and VEGF-A secretion in all MSC populations and showed that DSCs and DPCs produced significantly higher concentrations of leptin. Functional assays examining in vitro angiogenic paracrine activity showed that incubation of endothelial cells in ASC(CM) resulted in increased tubulogenic efficiency compared with that observed in DPC(CM). Using neutralizing antibodies we concluded that VEGF-A and VEGF-D were 2 of the major growth factors secreted by ASCs that supported endothelial tubulogenesis. The variation in paracrine factors of different MSC populations contributes to different levels of angiogenic activity and ASCs maybe preferred over other MSC populations for augmenting therapeutic approaches dependent upon angiogenesis.

  16. Effect of alpha lipoic acid co-administration on structural and immunohistochemical changes in subcutaneous tissue of anterior abdominal wall of adult male albino rat in response to polypropylene mesh implantation

    PubMed Central

    Mazroa, Shireen A; Asker, Samar A; Asker, Waleed; Abd Ellatif, Mohamed

    2015-01-01

    Polypropylene mesh is commonly used in the treatment of abdominal hernia. Different approaches were addressed to improve their tissue integration and consequently reduce long-term complications. This study aimed to investigate the effect of alpha-lipoic acid (ALA) co-administration on structural and immunohistochemical (IHC) changes in the subcutaneous tissues of the anterior abdominal wall of the adult rat in response to polypropylene mesh implantation. Forty adult male albino rats were divided into: group I (control), group II (receiving ALA), group III (polypropylene mesh implantation) and group IV (mesh implantation + ALA co-administration). After 4 weeks, subcutaneous tissue samples were prepared for light microscopy and IHC study of CD34 as a marker for angiogenesis. In groups I and II rats, positive CD34 expression was demonstrated by IHC reaction, localized to endothelial cells lining small blood vessels. Group III showed an excess inflammatory reaction, deposition of both regular and irregularly arranged collagen fibres around mesh pores and few elastic fibres. CD34-positive was detected not only in cells lining small blood vessels but also in other cells scattered in the connective tissue indicating angiogenesis. In group IV, ALA co-administration resulted in less inflammatory reaction, regular collagen deposition, enhanced elastic fibres synthesis and a significant increase in CD34-positive cells and small blood vessels reflecting improved angiogenesis. ALA co-administration with polypropylene mesh implantation controlled the inflammatory reaction, helped regular collagen deposition, enhanced elastic fibres synthesis and improved angiogenesis in the subcutaneous tissue of anterior abdominal wall of adult albino rats, suggesting a possible role of ALA in optimizing mesh integration in subcutaneous tissue. PMID:25891652

  17. Effect of alpha lipoic acid co-administration on structural and immunohistochemical changes in subcutaneous tissue of anterior abdominal wall of adult male albino rat in response to polypropylene mesh implantation.

    PubMed

    Mazroa, Shireen A; Asker, Samar A; Asker, Waleed; Abd Ellatif, Mohamed

    2015-06-01

    Polypropylene mesh is commonly used in the treatment of abdominal hernia. Different approaches were addressed to improve their tissue integration and consequently reduce long-term complications. This study aimed to investigate the effect of alpha-lipoic acid (ALA) co-administration on structural and immunohistochemical (IHC) changes in the subcutaneous tissues of the anterior abdominal wall of the adult rat in response to polypropylene mesh implantation. Forty adult male albino rats were divided into: group I (control), group II (receiving ALA), group III (polypropylene mesh implantation) and group IV (mesh implantation + ALA co-administration). After 4 weeks, subcutaneous tissue samples were prepared for light microscopy and IHC study of CD34 as a marker for angiogenesis. In groups I and II rats, positive CD34 expression was demonstrated by IHC reaction, localized to endothelial cells lining small blood vessels. Group III showed an excess inflammatory reaction, deposition of both regular and irregularly arranged collagen fibres around mesh pores and few elastic fibres. CD34-positive was detected not only in cells lining small blood vessels but also in other cells scattered in the connective tissue indicating angiogenesis. In group IV, ALA co-administration resulted in less inflammatory reaction, regular collagen deposition, enhanced elastic fibres synthesis and a significant increase in CD34-positive cells and small blood vessels reflecting improved angiogenesis. ALA co-administration with polypropylene mesh implantation controlled the inflammatory reaction, helped regular collagen deposition, enhanced elastic fibres synthesis and improved angiogenesis in the subcutaneous tissue of anterior abdominal wall of adult albino rats, suggesting a possible role of ALA in optimizing mesh integration in subcutaneous tissue.

  18. Museums and disease: using tissue archive and museum samples to study pathogens.

    PubMed

    Tsangaras, Kyriakos; Greenwood, Alex D

    2012-01-20

    Molecular studies of archival and fossil samples have traditionally focused on the nucleic acids derived from the host species. However, there has recently been an increase in ancient DNA research on the identification and characterization of infectious agents within the hosts. The study of pathogens from the past provides great opportunities for discovering the causes of historical infection events, characterizing host-microorganism co-evolution and directly investigating the evolution of specific pathogens. Several research teams have been able to isolate and characterize a variety of different bacterial, parasite and viral microorganisms. However, this emerging field is not without obstacles. The diagenetic processes that make ancient DNA research generally difficult are also impediments to ancient pathogen research and perhaps more so given that their DNA may represent an even rarer proportion of the remaining nucleic acids in a fossil sample than host DNA. However, studies performed under controlled conditions and following stringent ancient DNA protocols can and have yielded reliable and often surprising results. This article reviews the advantages, problems, and failures of ancient microbiological research.

  19. Genome-wide detection of DNase I hypersensitive sites in single cells and FFPE tissue samples.

    PubMed

    Jin, Wenfei; Tang, Qingsong; Wan, Mimi; Cui, Kairong; Zhang, Yi; Ren, Gang; Ni, Bing; Sklar, Jeffrey; Przytycka, Teresa M; Childs, Richard; Levens, David; Zhao, Keji

    2015-12-01

    DNase I hypersensitive sites (DHSs) provide important information on the presence of transcriptional regulatory elements and the state of chromatin in mammalian cells. Conventional DNase sequencing (DNase-seq) for genome-wide DHSs profiling is limited by the requirement of millions of cells. Here we report an ultrasensitive strategy, called single-cell DNase sequencing (scDNase-seq) for detection of genome-wide DHSs in single cells. We show that DHS patterns at the single-cell level are highly reproducible among individual cells. Among different single cells, highly expressed gene promoters and enhancers associated with multiple active histone modifications display constitutive DHS whereas chromatin regions with fewer histone modifications exhibit high variation of DHS. Furthermore, the single-cell DHSs predict enhancers that regulate cell-specific gene expression programs and the cell-to-cell variations of DHS are predictive of gene expression. Finally, we apply scDNase-seq to pools of tumour cells and pools of normal cells, dissected from formalin-fixed paraffin-embedded tissue slides from patients with thyroid cancer, and detect thousands of tumour-specific DHSs. Many of these DHSs are associated with promoters and enhancers critically involved in cancer development. Analysis of the DHS sequences uncovers one mutation (chr18: 52417839G>C) in the tumour cells of a patient with follicular thyroid carcinoma, which affects the binding of the tumour suppressor protein p53 and correlates with decreased expression of its target gene TXNL1. In conclusion, scDNase-seq can reliably detect DHSs in single cells, greatly extending the range of applications of DHS analysis both for basic and for translational research, and may provide critical information for personalized medicine. PMID:26605532

  20. The circadian clock in skin: implications for adult stem cells, tissue regeneration, cancer, aging, and immunity

    PubMed Central

    Plikus, Maksim V.; Van Spyk, Elyse Noelani; Pham, Kim; Geyfman, Mikhail; Kumar, Vivek; Takahashi, Joseph S.; Andersen, Bogi

    2015-01-01

    Historically work on peripheral circadian clocks has been focused on organs and tissues that have prominent metabolic functions, such as liver, fat and muscle. In recent years, skin is emerging as a model for studying circadian clock regulation of cell proliferation, stem cell functions, tissue regeneration, aging and carcinogenesis. Morphologically skin is complex, containing multiple cell types and structures, and there is evidence for a functional circadian clock in most, if not all, of its cell types. Despite the complexity, skin stem cell populations are well defined, experimentally tractable and exhibit prominent daily cell proliferation cycles. Hair follicle stem cells also participate in recurrent, long-lasting cycles of regeneration -- the hair growth cycles. Among other advantages of skin is a broad repertoire of available genetic tools enabling the creation of cell-type specific circadian mutants. Also, due to the accessibility of the skin, in vivo imaging techniques can be readily applied to study the circadian clock and its outputs in real time, even at the single-cell level. Skin provides the first line of defense against many environmental and stress factors that exhibit dramatic diurnal variations such as solar UV radiation and temperature. Studies have already linked the circadian clock to the control of UVB-induced DNA damage and skin cancers. Due to the important role that skin plays in the defense against microorganisms, it represents a promising model system to further explore the role of the clock in the regulation of the body's immune functions. To that end, recent studies have already linked the circadian clock to psoriasis, one of the most common immune-mediated skin disorders. The skin also provides opportunities to interrogate clock regulation of tissue metabolism in the context of stem cells and regeneration. Furthermore, many animal species feature prominent seasonal hair molt cycles, offering an attractive model for investigating the

  1. The circadian clock in skin: implications for adult stem cells, tissue regeneration, cancer, aging, and immunity.

    PubMed

    Plikus, Maksim V; Van Spyk, Elyse N; Pham, Kim; Geyfman, Mikhail; Kumar, Vivek; Takahashi, Joseph S; Andersen, Bogi

    2015-06-01

    Historically, work on peripheral circadian clocks has been focused on organs and tissues that have prominent metabolic functions, such as the liver, fat, and muscle. In recent years, skin has emerged as a model for studying circadian clock regulation of cell proliferation, stem cell functions, tissue regeneration, aging, and carcinogenesis. Morphologically, skin is complex, containing multiple cell types and structures, and there is evidence for a functional circadian clock in most, if not all, of its cell types. Despite the complexity, skin stem cell populations are well defined, experimentally tractable, and exhibit prominent daily cell proliferation cycles. Hair follicle stem cells also participate in recurrent, long-lasting cycles of regeneration: the hair growth cycles. Among other advantages of skin is a broad repertoire of available genetic tools enabling the creation of cell type-specific circadian mutants. Also, due to the accessibility of skin, in vivo imaging techniques can be readily applied to study the circadian clock and its outputs in real time, even at the single-cell level. Skin provides the first line of defense against many environmental and stress factors that exhibit dramatic diurnal variations such as solar ultraviolet (UV) radiation and temperature. Studies have already linked the circadian clock to the control of UVB-induced DNA damage and skin cancers. Due to the important role that skin plays in the defense against microorganisms, it also represents a promising model system to further explore the role of the clock in the regulation of the body's immune functions. To that end, recent studies have already linked the circadian clock to psoriasis, one of the most common immune-mediated skin disorders. Skin also provides opportunities to interrogate the clock regulation of tissue metabolism in the context of stem cells and regeneration. Furthermore, many animal species feature prominent seasonal hair molt cycles, offering an attractive model

  2. Subcutaneous adipose tissue fatty acid desaturation in adults with and without rare adipose disorders

    PubMed Central

    2012-01-01

    Background Elevated stearoyl-CoA desaturase activity has been described in obese states, with an increased desaturation index (DI) suggesting enhanced lipogenesis. Differences in the DI among various phenotypes of abnormal adiposity have not been studied. Abnormal accumulation of subcutaneous adipose tissue occurs in rare adipose disorders (RADs) including Dercum's disease (DD), multiple symmetric lipomatosis (MSL), and familial multiple lipomatosis (FML). Examining the DI in subcutaneous fat of people with DD, MSL and FML may provide information on adipose tissue fatty acid metabolism in these disorders. The aims of this pilot study were: 1) to determine if differences in adipose tissue DIs are present among RADs, and 2) to determine if the DIs correlate to clinical or biochemical parameters. Methods Subcutaneous adipose tissue was obtained from human participants with DD (n = 6), MSL (n = 5), FML (n = 8) and obese Controls (n = 6). Fatty acid composition was determined by gas chromatography/mass spectrometry. The DIs (palmitoleic/palmitic, oleic/stearic, vaccenic/stearic ratios) were calculated from the gas chromatogram peak intensities. SCD1 gene expression was determined. Spearman's correlations between the DIs and available clinical or biochemical data were performed. Results In DD subjects, the vaccenic/stearic index was lower (p < 0.05) in comparison to Controls. Percent of total of the saturated fatty acid myristic acid was higher in DD compared with Controls and FML. Percent of monounsaturated vaccenic acid in DD trended lower when compared with Controls, and was decreased in comparison to FML. In MSL, total percent of the polyunsaturated fatty acids was significantly lower than in the Control group (p < 0.05). In the total cohort of subjects, the palmitoleic/palmitic and oleic/stearic DIs positively correlated with age, BMI, and percent body fat. Conclusions The positive associations between the DIs and measures of adiposity (BMI and percent body fat

  3. Comparison of the big-five factor structure across samples of Chinese and American adults.

    PubMed

    Trull, T J; Geary, D C

    1997-10-01

    We compared the factor structure of Goldberg's (1992) 50-item Bipolar Rating Scale (50-BRS) in samples of Chinese (n = 198) and American (n = 303) students. Results confirmed the hypothesized five-factor pattern for the U.S. sample, and a simultaneous multisample confirmatory factor analysis showed that the same five-factor pattern fit the item covariances in the Chinese sample. High levels of internal consistency were found within each sample, and a high degree of congruency of corresponding item factor loadings was obtained across samples. Overall, results supported the potential utility of the Five-Factor Model and the 50-BRS for assessing personality dimensions in Chinese culture.

  4. [Prolonged effects of training on adipose tissue glucose metabolism and insulin responsiveness in adult rats (author's transl)].

    PubMed

    Gommers, A; Dehez-Delhaye, M; Caucheteux, D

    1981-06-01

    Physical training (forced swimming for 6 weeks) led in resting adult rats modifications of circulating hormones, fuels and substrates, of epididymal adipose tissue composition and of its glucose metabolism. Plasma concentrations of free fatty acids, cholesterol and insulin were decreased by training. The weight of the epididymal fat pad, its triglyceride content and the mean cell size of adipocytes were significantly diminished. Twenty four hours after the end of the final exercise period, incorporation of D-U-14C-glucose into triglycerides was reduced (P less than 0.01) but the response to insulin was greatly enhanced for swimming rats both for oxidation of labelled glucose (P less than 0.001) and its incorporation into lipids (P less than 0.001) (respectively 215 and 225%). This enhancement of insulin sensitivity by training persisted and became more marked one week after the end of the exercise period (respectively 258 and 363%).

  5. Correction of Class II malocclusion and soft tissue profile in an adult patient

    PubMed Central

    Gaur, Aditi; Maheshwari, Sandhya; Verma, Sanjeev Kumar

    2016-01-01

    Treatment of Class II malocclusion in nongrowing individuals is a challenging situation for the clinician. Class II malocclusion with bialveolar protrusion often dictates premolar extractions with maximum anchorage. The present article describes the case of an adult female with skeletal Class II malocclusion, bimaxillary protrusion, increased overjet, deep bite, lip protrusion, everted lower lip, deep mentolabial sulcus, and lip incompetence. To correct the malocclusion, all four first premolars were extracted. Direct anchorage from miniscrews was used for retraction of the anterior segment. The mandibular buccal segment was protracted into the extraction space using Class II mechanics. Ideal Class I canine and molar relation were achieved in 24 months. There was a significant improvement in facial profile and smile esthetics of the patient.

  6. Correction of Class II malocclusion and soft tissue profile in an adult patient.

    PubMed

    Gaur, Aditi; Maheshwari, Sandhya; Verma, Sanjeev Kumar

    2016-01-01

    Treatment of Class II malocclusion in nongrowing individuals is a challenging situation for the clinician. Class II malocclusion with bialveolar protrusion often dictates premolar extractions with maximum anchorage. The present article describes the case of an adult female with skeletal Class II malocclusion, bimaxillary protrusion, increased overjet, deep bite, lip protrusion, everted lower lip, deep mentolabial sulcus, and lip incompetence. To correct the malocclusion, all four first premolars were extracted. Direct anchorage from miniscrews was used for retraction of the anterior segment. The mandibular buccal segment was protracted into the extraction space using Class II mechanics. Ideal Class I canine and molar relation were achieved in 24 months. There was a significant improvement in facial profile and smile esthetics of the patient. PMID:27630505

  7. Correction of Class II malocclusion and soft tissue profile in an adult patient

    PubMed Central

    Gaur, Aditi; Maheshwari, Sandhya; Verma, Sanjeev Kumar

    2016-01-01

    Treatment of Class II malocclusion in nongrowing individuals is a challenging situation for the clinician. Class II malocclusion with bialveolar protrusion often dictates premolar extractions with maximum anchorage. The present article describes the case of an adult female with skeletal Class II malocclusion, bimaxillary protrusion, increased overjet, deep bite, lip protrusion, everted lower lip, deep mentolabial sulcus, and lip incompetence. To correct the malocclusion, all four first premolars were extracted. Direct anchorage from miniscrews was used for retraction of the anterior segment. The mandibular buccal segment was protracted into the extraction space using Class II mechanics. Ideal Class I canine and molar relation were achieved in 24 months. There was a significant improvement in facial profile and smile esthetics of the patient. PMID:27630505

  8. Soft Tissue Tumors in Adults: ESSR-Approved Guidelines for Diagnostic Imaging.

    PubMed

    Noebauer-Huhmann, Iris M; Weber, Marc-André; Lalam, Radhesh K; Trattnig, Siegfried; Bohndorf, Klaus; Vanhoenacker, Filip; Tagliafico, Alberto; van Rijswijk, Carla; Vilanova, Joan C; Afonso, P Diana; Breitenseher, Martin; Beggs, Ian; Robinson, Philip; de Jonge, Milko C; Krestan, Christian; Bloem, Johan L

    2015-12-01

    Soft tissue sarcomas are rare, but early, accurate diagnosis with subsequent appropriate treatment is crucial for the clinical outcome. The ESSR guidelines are intended to help radiologists in their decision-making and support discussion among clinicians who deal with patients with suspected or proven soft tissue tumors. Potentially malignant lesions recognized by ultrasound should be referred for magnetic resonance imaging (MRI), which also serves as a preoperative local staging modality, with specific technical requirements and mandatory radiological report elements. Radiography may add information about matrix calcification and osseous involvement. Indeterminate lesions, or lesions in which therapy is dependent on histology results, should be biopsied. For biopsy, we strongly recommend referral to a specialist sarcoma center, where an interdisciplinary tumor group, with a specialized pathologist, radiologist, and the surgeon are involved. In sarcoma, a CT scan of the chest is mandatory. Additional staging modalities are entity-specific. There are no evidence-based recommendations for routine follow-up in surgically treated sarcomas. However, we would recommend regular follow-up with intervals dependent on tumor grade, for 10 years after the initial diagnosis. PMID:26696086

  9. Adjuvant epirubicin with or without Ifosfamide for adult soft-tissue sarcoma.

    PubMed

    Petrioli, Roberto; Coratti, Andrea; Correale, Pierpaolo; D'Aniello, Carlo; Grimaldi, Luca; Tanzini, Gabriello; Civitelli, Serenella; Marsili, Stefania; Messinese, Simona; Marzocca, Giuseppe; Pirtoli, Luigi; Francini, Guido

    2002-10-01

    This randomized study compared the efficacy of epirubicin-based adjuvant chemotherapy on the disease-free interval (DFI) and overall survival of patients with high-risk soft-tissue sarcomas. After curative surgery, 43 of the 88 enrolled patients were assigned to surgery with or without radiotherapy and 45 to surgery plus chemotherapy (26 epirubicin, 19 epirubicin + ifosfamide) with or without radiotherapy. The trial closed prematurely because of poor patient accrual. There was a statistical significant difference in the 5-year disease-free survival of the patients receiving adjuvant chemotherapy with or without radiotherapy (69%) and that of those treated with surgery with or without radiotherapy (44%) ( p= 0.01). The 5-year survival of the patients treated with adjuvant chemotherapy with or without radiotherapy was 72% as against 47% of those treated with surgery with or without radiotherapy ( p= 0.06). The power of the study was 0.65 for both the DFI and overall survival. The results of the study suggest a possible advantage of epirubicin-based adjuvant chemotherapy in patients with soft-tissue sarcoma at high risk of relapse. PMID:12393986

  10. Maternal hypertension programs increased cerebral tissue damage following stroke in adult offspring.

    PubMed

    Ventura, Nicole M; Jin, Albert Y; Tse, M Yat; Peterson, Nichole T; Andrew, R David; Mewburn, Jeffrey D; Pang, Stephen C

    2015-10-01

    The maternal system is challenged with many physiological changes throughout pregnancy to prepare the body to meet the metabolic needs of the fetus and for delivery. Many pregnancies, however, are faced with pathological stressors or complications that significantly impact maternal health. A shift in this paradigm is now beginning to investigate the implication of pregnancy complications on the fetus and their continued influence on offspring disease risk into adulthood. In this investigation, we sought to determine whether maternal hypertension during pregnancy alters the cerebral response of adult offspring to acute ischemic stroke. Atrial natriuretic peptide gene-disrupted (ANP(-/-)) mothers exhibit chronic hypertension that escalates during pregnancy. Through comparison of heterozygote offspring born from either normotensive (ANP(+/-WT)) or hypertensive (ANP(+/-KO)) mothers, we have demonstrated that offspring exposed to maternal hypertension exhibit larger cerebral infarct volumes following middle cerebral artery occlusion. Observation of equal baseline cardiovascular measures, cerebrovascular structure, and cerebral blood volumes between heterozygote offspring suggests no added influences on offspring that would contribute to adverse cerebral response post-stroke. Cerebral mRNA expression of endothelin and nitric oxide synthase vasoactive systems demonstrated up-regulation of Et-1 and Nos3 in ANP(+/-KO) mice and thus an enhanced acute vascular response compared to ANP(+/-WT) counterparts. Gene expression of Na(+)/K(+) ATPase channel isoforms, Atp1a1, Atp1a3, and Atp1b1, displayed no significant differences. These investigations are the first to demonstrate a fetal programming effect between maternal hypertension and adult offspring stroke outcome. Further mechanistic studies are required to complement epidemiological evidence of this phenomenon in the literature. PMID:26169981

  11. In situ assessment of mRNA accessibility in heterogeneous tissue samples using elongation factor-1 alpha (EF-1 alpha).

    PubMed

    Gruber, A D; Levine, R A

    1997-05-01

    Elongation factor-1 alpha (EF-1 alpha) is an evolutionarily highly conserved universal cofactor of protein synthesis in all living cells. In this study, its use as a positive control in situ hybridization assays for specific detection of mRNA sequences was evaluated. Northern blot analysis of various non-neoplastic and neoplastic cultured cells of different stages of confluence, cell shape, and cell cycle status revealed that EF-1 alpha had a lower and more homogeneous expression than did beta-actin. In situ hybridization assays using digoxigenin-labeled riboprobes for the detection of EF-1 alpha mRNA in routinely formalin-fixed, paraffin-embedded tissue sections showed that EF-1 alpha is a suitable positive control in all types of cells. However, variation of protease pretreatments demonstrated distinct and sometimes mutually exclusive digestion conditions for different cell types within the same tissue sample. Our results indicate that detection of EF-1 alpha mRNA is an appropriate internal standard for in situ hybridization assays and that it is useful to control artifacts such as false negatives caused by inappropriate protease pretreatments. The observed variability of optimal protease pretreatments for different cell types within the same tissue section strengthens the importance of a positive control in in situ hybridization assays.

  12. Age at Trauma Exposure and PTSD Risk in a Young Adult Female Sample

    PubMed Central

    McCutcheon, Vivia V.; Sartor, Carolyn E.; Pommer, Nicole E.; Bucholz, Kathleen K.; Nelson, Elliot C.; Madden, Pamela A.F.; Heath, Andrew C.

    2011-01-01

    The aim of the current study was to test the independent and joint contributions of 8 different types of trauma to posttraumatic stress disorder (PTSD) risk using data from a young adult female cohort. Associations of traumatic events with PTSD onset were examined using Cox proportional hazards models. Differences in risk as a function of age at trauma were tested. Childhood sexual assault, physical abuse, and neglect were stronger predictors of PTSD onset than adolescent/early adult occurrence of these events in individual models. In a model including all traumatic events, differential risk by age remained for sexual assault and physical abuse. Early sexual assault was the strongest predictor of risk but additional traumatic events increased risk even in its presence. PMID:20963847

  13. The Relationship between Dietary Patterns and Metabolic Health in a Representative Sample of Adult Australians

    PubMed Central

    Bell, Lucinda K.; Edwards, Suzanne; Grieger, Jessica A.

    2015-01-01

    Studies assessing dietary intake and its relationship to metabolic phenotype are emerging, but limited. The aims of the study are to identify dietary patterns in Australian adults, and to determine whether these dietary patterns are associated with metabolic phenotype and obesity. Cross-sectional data from the Australian Bureau of Statistics 2011 Australian Health Survey was analysed. Subjects included adults aged 45 years and over (n = 2415). Metabolic phenotype was determined according to criteria used to define metabolic syndrome (0–2 abnormalities vs. 3–7 abnormalities), and additionally categorized for obesity (body mass index (BMI) ≥30 kg/m2 vs. BMI <30 kg/m2). Dietary patterns were derived using factor analysis. Multivariable models were used to assess the relationship between dietary patterns and metabolic phenotype, with adjustment for age, sex, smoking status, socio-economic indexes for areas, physical activity and daily energy intake. Twenty percent of the population was metabolically unhealthy and obese. In the fully adjusted model, for every one standard deviation increase in the Healthy dietary pattern, the odds of having a more metabolically healthy profile increased by 16% (odds ratio (OR) 1.16; 95% confidence interval (CI): 1.04, 1.29). Poor metabolic profile and obesity are prevalent in Australian adults and a healthier dietary pattern plays a role in a metabolic and BMI phenotypes. Nutritional strategies addressing metabolic syndrome criteria and targeting obesity are recommended in order to improve metabolic phenotype and potential disease burden. PMID:26251918

  14. Synthetic Cannabinoids: Use and predictors in a Community Sample of Young Adults

    PubMed Central

    Caviness, Celeste M.; Tzilos, Golfo; Anderson, Bradley J.; Stein, Michael, D.

    2014-01-01

    Background: Synthetic cannabinoids (SCs) are a class of drugs of abuse with deleterious consequences. Despite governmental regulations related to distribution and sale, SC variants are still available online. More research is needed to determine SC use prevalence and factors associated with SC use, especially among young adults. Methods: One thousand eighty individuals, 18-25 years old, were surveyed, between January 2012 and July 2013, during recruitment for a randomized controlled trial investigating health behaviors in young adults. Advertisements were placed online and in community locations seeking individuals “who had recently used marijuana or alcohol.” Respondents were queried about their use of alcohol and drugs, including SCs, in the last month. Results: Participants averaged 21.4 years old and were 53.4% male. Nearly 59% were Non-Hispanic White, 15% were African American, 15% were Hispanic, 11% identified as Other. Approximately 9% reported SC use in the last month, a level higher than the reported use of opioids, cocaine, or hallucinogens. SC use was significantly associated with male gender, not being enrolled in school, and with use of cigarettes, binge alcohol drinking, daily and weekly marijuana use, and other drugs of abuse. There was a significant decrease in SC use after the federal ban in July, 2012. Conclusions: SC use was common in the past month and often overlaps with other drug use, particularly marijuana use, and should be asked about during clinical encounters with young adults. PMID:25222129

  15. Tissue Mercury Concentrations and Survival of Tree Swallow Embryos, Nestlings and Young Adult Females on a Contaminated Site.

    PubMed

    Taylor, Capwell E; Cristol, Daniel A

    2015-10-01

    Tree swallows nesting on mercury-contaminated sites along the South River in Virginia, USA were monitored for reproductive success. The bodies of nestlings found deceased in their nest boxes were collected, along with blood and feather samples from the adult parents and surviving siblings. We also measured hatching and fledging success of the clutches and the annual recapture rate of adults. We found that the body feathers of deceased nestlings contained significantly higher concentrations of mercury (12.89 ± 8.42 μg/g, n = 15) than those of nestlings that survived to fledge (7.41 ± 4.79 μg/g, n = 15). However, mothers of more successful clutches (>75 % hatching) did not differ in mercury concentrations from females with less successful clutches (<50 % hatching). Additionally, adult females breeding for the first time that returned to breed the following year did not differ in blood mercury from females of the same age that bred once but never returned. Our results suggest that mercury had its greatest effect on these songbirds during the nestling stage, whereas for embryos or first-time breeding females, other factors likely played larger roles in mortality.

  16. Development of a remote laser-induced breakdown spectroscopy system for investigation of calcified tissue samples

    SciTech Connect

    Hrdlicka, Ales; Prokes, Lubomir; Stankova, Alice; Novotny, Karel; Vitesnikova, Anna; Kanicky, Viktor; Otruba, Vitezslav; Kaiser, Jozef; Novotny, Jan; Malina, Radomir; Palenikova, Katerina

    2010-05-01

    The development of a remote laser-induced breakdown spectroscopy (LIBS) setup with an off-axis Newtonian collection optics, Galilean-based focusing telescope, and a 532 nm flattop laser beam source is presented. The device was tested at a 6 m distance on a slice of bone to simulate its possible use in the field, e.g., during archaeological excavations. It is shown that this setup is sufficiently sensitive to both major (P, Mg) and minor elements (Na, Zn, Sr). The measured quantities of Mg, Zn, and Sr correspond to the values obtained by reference laser ablation-inductively coupled plasma-mass spectrometry (LA-ICP-MS) measurements within an approximately 20% range of uncertainty. A single point calibration was performed by use of a bone meal standard . The radial element distribution is almost invariable by use of LA-ICP-MS, whereas the LIBS measurement showed a strong dependence on the sample porosity. Based on these results, this remote LIBS setup with a relatively large (350 mm) collecting mirror is capable of semiquantitative analysis at the level of units of mg kg{sup -1}.

  17. Assessing the feasibility and sample quality of a national random-digit dialing cellular phone survey of young adults.

    PubMed

    Gundersen, Daniel A; ZuWallack, Randal S; Dayton, James; Echeverría, Sandra E; Delnevo, Cristine D

    2014-01-01

    The majority of adults aged 18-34 years have only cellular phones, making random-digit dialing of landline telephones an obsolete methodology for surveillance of this population. However, 95% of this group has cellular phones. This article reports on the 2011 National Young Adult Health Survey (NYAHS), a pilot study conducted in the 50 US states and Washington, DC, that used random-digit dialing of cellular phones and benchmarked this methodology against that of the 2011 Behavioral Risk Factor Surveillance System (BRFSS). Comparisons of the demographic distributions of subjects in the NYAHS and BRFSS (aged 18-34 years) with US Census data revealed adequate reach for all demographic subgroups. After adjustment for design factors, the mean absolute deviations across demographic groups were 3 percentage points for the NYAHS and 2.8 percentage points for the BRFSS, nationally, and were comparable for each census region. Two-sided z tests comparing cigarette smoking prevalence revealed no significant differences between NYAHS and BRFSS participants overall or by subgroups. The design effects of the sampling weight were 2.09 for the NYAHS and 3.26 for the BRFSS. Response rates for the NYAHS and BRFSS cellular phone sampling frames were comparable. Our assessment of the NYAHS methodology found that random-digit dialing of cellular phones is a feasible methodology for surveillance of young adults.

  18. Assessing the Feasibility and Sample Quality of a National Random-digit Dialing Cellular Phone Survey of Young Adults

    PubMed Central

    Gundersen, Daniel A.; ZuWallack, Randal S.; Dayton, James; Echeverría, Sandra E.; Delnevo, Cristine D.

    2014-01-01

    The majority of adults aged 18–34 years have only cellular phones, making random-digit dialing of landline telephones an obsolete methodology for surveillance of this population. However, 95% of this group has cellular phones. This article reports on the 2011 National Young Adult Health Survey (NYAHS), a pilot study conducted in the 50 US states and Washington, DC, that used random-digit dialing of cellular phones and benchmarked this methodology against that of the 2011 Behavioral Risk Factor Surveillance System (BRFSS). Comparisons of the demographic distributions of subjects in the NYAHS and BRFSS (aged 18–34 years) with US Census data revealed adequate reach for all demographic subgroups. After adjustment for design factors, the mean absolute deviations across demographic groups were 3 percentage points for the NYAHS and 2.8 percentage points for the BRFSS, nationally, and were comparable for each census region. Two-sided z tests comparing cigarette smoking prevalence revealed no significant differences between NYAHS and BRFSS participants overall or by subgroups. The design effects of the sampling weight were 2.09 for the NYAHS and 3.26 for the BRFSS. Response rates for the NYAHS and BRFSS cellular phone sampling frames were comparable. Our assessment of the NYAHS methodology found that random-digit dialing of cellular phones is a feasible methodology for surveillance of young adults. PMID:24100957

  19. Expression of spicule matrix protein gene SM30 in embryonic and adult mineralized tissues of sea urchin Hemicentrotus pulcherrimus

    NASA Technical Reports Server (NTRS)

    Kitajima, T.; Tomita, M.; Killian, C. E.; Akasaka, K.; Wilt, F. H.

    1996-01-01

    We have isolated a cDNA clone for spicule matrix protein, SM30, from sea urchin Hemicentrotus pulcherrimus and have studied the expression of this gene in comparison with that of another spicule matrix protein gene, SM50. In cultured micromeres as well as in intact embryos transcripts of SM30 were first detectable around the onset of spicule formation and rapidly increased with the growth of spicules, which accompanied accumulation of glycosylated SM30 protein(s). When micromeres were cultured in the presence of Zn2+, spicule formation and SM30 expression were suppressed, while both events resumed concurrently after the removal of Zn2+ from the culture medium. Expression of SM50, in contrast, started before the appearance of spicules and was not sensitive to Zn2+. Differences were also observed in adult tissues; SM30 mRNA was detected in spines and tube feet but not in the test, while SM50 mRNA was apparent in all of these mineralized tissues at similar levels. These results strongly suggest that the SM30 gene is regulated by a different mechanism to that of the SM50 gene and that the products of these two genes are differently involved in sea urchin biomineralization. A possible role of SM30 protein in skeleton formation is discussed.

  20. Immunomodulatory effect of human adipose tissue-derived adult stem cells: comparison with bone marrow mesenchymal stem cells.

    PubMed

    Puissant, Bénédicte; Barreau, Corinne; Bourin, Philippe; Clavel, Cyril; Corre, Jill; Bousquet, Christine; Taureau, Christine; Cousin, Béatrice; Abbal, Michel; Laharrague, Patrick; Penicaud, Luc; Casteilla, Louis; Blancher, Antoine

    2005-04-01

    Like mesenchymal stem cells from bone marrow (BM-MSCs), adipose tissue-derived adult stem cells (ADAS cells) can differentiate into several lineages and present therapeutical potential for repairing damaged tissues. The use of allogenic stem cells can enlarge their therapeutical interest, provided that the grafted cells could be tolerated. We investigate here, for the first time, the immunosuppressive properties of ADAS cells compared with the well-characterized immunosuppressive properties of BM-MSCs. ADAS cells did not provoke in vitro alloreactivity of incompatible lymphocytes and, moreover, suppressed mixed lymphocyte reaction (MLR) and lymphocyte proliferative response to mitogens. The impairment of inhibition when ADAS cells and BM-MSCs were separated from lymphocytes by a permeable membrane suggests that cell contact is required for a full inhibitory effect. Hepatocyte growth factor is secreted by both stem cells but, similar to interleukin-10 and transforming growth factor-beta (TGF-beta), the levels of which were undetectable in supernatants of MLR inhibited by ADAS cells or BM-MSCs, it did not seem implicated in the stem cell suppressive effect. These findings support that ADAS cells share immunosuppressive properties with BM-MSCs. Therefore, ADAS cell-based reconstructive therapy could employ allogenic cells and because of their immunosuppressive properties, ADAS cells could be an alternative source to BM-MSCs to treat allogenic conflicts.

  1. A quantitative three-dimensional assessment of soft tissue facial asymmetry of cleft lip and palate adult patients.

    PubMed

    Ferrario, Virgilio F; Sforza, Chiarella; Dellavia, Claudia; Tartaglia, Gianluca M; Colombo, Anna; Carù, Armando

    2003-09-01

    The three-dimensional coordinates of 23 selected soft-tissue facial landmarks were digitized on 18 cleft lip and palate (CLP) white patients (11 male and 7 female patients aged 19-27 years) and 161 healthy controls (73 female and 89 male subjects aged 18-30 years) by an electromagnetic instrument. Facial asymmetry was quantified by detecting a plane of symmetry and the centers of gravity (CG) of the right and left hemifaces and by calculating the distance between the two CG (distance from symmetry [DFS]). Both absolute (millimeters) and percentage (of the nasion center of gravity distance) DFS was obtained. The asymmetry of single landmarks was also quantified. Overall, asymmetry in operated CLP patients appeared only moderately larger than that measured in the healthy reference population, with the largest value being only 5% larger than the maximum normal asymmetry. Female patients had a somewhat larger lateral asymmetry than male patients, and unilateral CLP patients (particularly the men) were more asymmetrical than bilateral CLP patients. Pronasale and subnasale landmarks were asymmetrical in 8 patients, whereas endocanthion, zygion, cheilion, and gonion landmarks were symmetrical in all patients. In conclusion, the facial soft-tissue structures of CLP patients operated on as adults were only moderately more asymmetrical than those measured in a reference group of the same age, sex, and ethnicity.

  2. Comparative Analysis of the Expression Profile of Wnk1 and Wnk1/Hsn2 Splice Variants in Developing and Adult Mouse Tissues

    PubMed Central

    Shekarabi, Masoud; Lafrenière, Ron G.; Gaudet, Rébecca; Laganière, Janet; Marcinkiewicz, Martin M.; Dion, Patrick A.; Rouleau, Guy A.

    2013-01-01

    The With No lysine (K) family of serine/threonine kinase (WNK) defines a small family of kinases with significant roles in ion homeostasis. WNK1 has been shown to have different isoforms due to what seems to be largely tissue specific splicing. Here, we used two distinct in situ hybridization riboprobes on developing and adult mouse tissues to make a comparative analysis of Wnk1 and its sensory associated splice isoform, Wnk1/Hsn2. The hybridization signals in developing mouse tissues, which were prepared at embryonic day e10.5 and e12.5, revealed a homogenous expression profile with both probes. At e15.5 and in the newborn mouse, the two probes revealed different expression profiles with prominent signals in nervous system tissues and also other tissues such as kidney, thymus and testis. In adult mouse tissues, the two expression profiles appeared even more restricted to the nervous tissues, kidney, thymus and testis, with no detectable signal in the other tissues. Throughout the nervous system, sensory tissues, as well as in Cornu Ammonis 1 (CA1), CA2 and CA3 areas of the hippocampus, were strongly labeled with both probes. Hybridization signals were also strongly detected in Schwann and supporting satellite cells. Our results show that the expression profiles of Wnk1 isoforms change during the development, and that the expression of the Wnk1 splice variant containing the Hsn2 exon is prominent during developing and in adult mouse tissues, suggesting its important role in the development and maintenance of the nervous system. PMID:23451271

  3. Cellular distribution of the new growth factor pleiotrophin (HB-GAM) mRNA in developing and adult rat tissues.

    PubMed

    Vanderwinden, J M; Mailleux, P; Schiffmann, S N; Vanderhaeghen, J J

    1992-09-01

    Pleiotrophin (PTN), also known as HB-GAM, belongs to an emerging cytokine family unrelated to other growth factors. We report here the first comprehensive study using in situ hybridization on the cellular distribution of this new heparin-binding growth factor mRNA in rat tissues. PTN mRNA was developmentally expressed in many--but not all--neuroectodermal and mesodermal lineages, whilst no PTN mRNA was detected in endoderm, ectoderm and trophoblast. PTN mRNA was found in the nervous system throughout development, with a post-natal peak of expression. In the adult nervous system, significant expression persisted in hippocampal CA1 pyramidal neurons and in cortical neurons, but also in different non-neuronal cells types in various locations (olfactory nerve, cerebellar astrocytes, pituicytes, Schwann cells surrounding the neurons in sensory ganglia). PTN mRNA was also found during development in the mesenchyme of lung, gut, kidney and reproductive tract, in bone and cartilage progenitors, in dental pulp, in myoblasts, and in several other sites. Expression was differently regulated in each location, but usually faded around birth. In the adult, PTN mRNA was still present in the meninges, the iris, the Leydig cells of the testis and in the uterus. PTN mRNA was also strongly expressed in the basal layers of the tongue epithelium, which is the only epithelium and ectodermal derivative to express PTN mRNA, and this only after birth. PTN is known to be a growth factor for perinatal brain neurons and a mitogen for fibroblasts in vitro. Recently, trophic effects on epithelial cells and a role as a tumour growth factor have been reported. The mechanisms of regulation and the functions of PTN are however still uncertain. Its expression pattern during development suggests important roles in growth and differentiation. Moreover, the presence of PTN mRNA in several adult tissues and the up-regulation of PTN mRNA expression in the gravid uterus indicate that PTN also has

  4. The problem with medical research on tissue and organ samples taken in connection with forensic autopsies in France.

    PubMed

    Rougé-Maillart, C; Dupont, V; Jousset, N

    2016-02-01

    Currently, in France, it is legally impossible to conduct scientific research on tissue and organ samples taken from forensic autopsies. In fact, the law schedules the destruction of such samples at the end of the judicial investigation, and the common law rules governing cadaver research cannot be applied to the forensic context. However, nothing seems in itself to stand in the way of such research since, despite their specific nature, these samples from forensic autopsies could be subject, following legislative amendments, to common law relating to medical research on samples taken from deceased persons. But an essential legislative amendment, firstly to allow the Biomedicine Agency to become authorized to issue a research permit and secondly, to change the research conditions in terms of the non-opposition of the deceased to said research. Such an amendment would be a true breakthrough because it would allow teams to continue to move forward calmly in research, and allow this research to be placed within a legal framework, which would promote international exchanges.

  5. The problem with medical research on tissue and organ samples taken in connection with forensic autopsies in France.

    PubMed

    Rougé-Maillart, C; Dupont, V; Jousset, N

    2016-02-01

    Currently, in France, it is legally impossible to conduct scientific research on tissue and organ samples taken from forensic autopsies. In fact, the law schedules the destruction of such samples at the end of the judicial investigation, and the common law rules governing cadaver research cannot be applied to the forensic context. However, nothing seems in itself to stand in the way of such research since, despite their specific nature, these samples from forensic autopsies could be subject, following legislative amendments, to common law relating to medical research on samples taken from deceased persons. But an essential legislative amendment, firstly to allow the Biomedicine Agency to become authorized to issue a research permit and secondly, to change the research conditions in terms of the non-opposition of the deceased to said research. Such an amendment would be a true breakthrough because it would allow teams to continue to move forward calmly in research, and allow this research to be placed within a legal framework, which would promote international exchanges. PMID:26694871

  6. [Attraction of Sphenophorus levis Vaurie adults (Coleoptera: Curculionidae) to vegetal tissues at different conservation levels].

    PubMed

    Girón-Pérez, Katherine; Nakano, Octávio; Silva, Amanda C; Oda-Souza, Melissa

    2009-01-01

    The occurrence of the sugarcane weevil Sphenophorus levis Vaurie is important in sugarcane in some regions in Brazil. Damage is caused by the larvae as they bore into the nodes and can reach 30 ton/ha/year. Many control alternatives have been attempted, but none were satisfactory, except for the use of toxic baits. Therefore, it is necessary to optimize their efficiency or to propose new techniques. The objective of the current study was to evaluate the attractiveness of adults of S. levis to sugarcane nodes and pineapple peelings in an 'Y' tube olfactometer. The sugarcane internodes were treated with 10% molasses, and tested after different periods of fermentation (24, 48 e 72h), at different times of the day (diurnal and nocturnal) and with both sexes. These tests were carried out in order to correlate the response of S. levis to ethyl acetate and ethanol release as a result of the fermentation process. The release of both compounds was monitored by gas chromatography-mass spectrometry (GC-MS). Our data indicated that sugarcane internodes mixed with 10% molasses fermented for 24h and 48h were the most attractive to S. levis (up to 90%). Pineapple peelings attracted 62.5% of the tested insects. The olfactory response was higher during the day, and no differences were found between the sexes. The production of ethanol in all plant substrates was higher than ethyl acetate, but we could not establish a clear correlation with the insect response to baits. PMID:20098932

  7. Evaluation of two-dimensional electrophoresis and liquid chromatography – tandem mass spectrometry for tissue-specific protein profiling of laser-microdissected plant samples

    SciTech Connect

    Schad, Martina; Lipton, Mary S.; Giavalisco, Patrick; Smith, Richard D.; Kehr, Julia

    2005-07-14

    Laser microdissection (LM) allows the collection of homogeneous tissue- and cell specific plant samples. The employment of this technique with subsequent protein analysis has thus far not been reported for plant tissues, probably due to the difficulties associated with defining a reasonable cellular morphology and, in parallel, allowing efficient protein extraction from tissue samples. The relatively large sample amount needed for successful proteome analysis is an additional issue that complicates protein profiling on a tissue- or even cell-specific level. In contrast to transcript profiling that can be performed from very small sample amounts due to efficient amplification strategies, there is as yet no amplification procedure for proteins available. In the current study, we compared different tissue preparation techniques prior to LM/laser pressure catapulting (LMPC) with respect to their suitability for protein retrieval. Cryosectioning was identified as the best compromise between tissue morphology and effective protein extraction. After collection of vascular bundles from Arabidopsis thaliana stem tissue by LMPC, proteins were extracted and subjected to protein analysis, either by classical two-dimensional gel electrophoresis (2-DE), or by high-efficiency liquid chromatography (LC) in conjunction with tandem mass spectrometry (MS/MS). Our results demonstrate that both methods can be used with LMPC collected plant material. But because of the significantly lower sample amount required for LC-MS/MS than for 2-DE, the combination of LMPC and LC-MS/MS has a higher potential to promote comprehensive proteome analysis of specific plant tissues.

  8. Persistent organochlorine pesticides detected in blood and tissue samples of vultures from different localities in South Africa.

    PubMed

    van Wyk, E; Bouwman, H; van der Bank, H; Verdoorn, G H; Hofmann, D

    2001-07-01

    Gas chromatography was used to establish the presence of quantifiable residues of 14 persistent chlorinated hydrocarbon pollutants in whole blood, clotted blood, heart, kidney, liver and muscle samples obtained from individual African whitebacked (Pseudogyps africanus), Cape griffon (Gyps coprotheres) and Lappetfaced (Torgos tracheliotos) vultures from different localities in South Africa. The levels of pesticides measured in whole blood samples of live specimens were compared between nestlings from two natural breeding colonies, adults from a wildlife area and birds held in captivity. Statistically significant (P<0.05) differences between populations were detected in geometric means calculated for gamma-BHC (lindane), alpha(cis)-chlordane and alpha-endosulfan. Five of the organochlorine contaminants displayed significant variations between concentrations detected in the clotted blood, organs and muscles excised from vulture carcasses. This includes residues of gamma-BHC, alpha-chlordane, dieldrin, beta-endosulfan and heptachlor epoxide. Values of the respective biocides measured in vulture samples were generally low in comparison to results documented for a number of avian species. Although no threat is posed by any of the organochloride pesticides, continual monitoring of especially breeding colonies is recommended. Furthermore, the suitability of African whitebacked vulture nestlings as basic bioindicators is highly advocated. PMID:11461840

  9. Isolated Limb Perfusion of Melphalan With or Without Tumor Necrosis Factor in Treating Patients With Soft Tissue Sarcoma of the Arm or Leg

    ClinicalTrials.gov

    2012-03-14

    Stage IVB Adult Soft Tissue Sarcoma; Stage IIB Adult Soft Tissue Sarcoma; Stage IIC Adult Soft Tissue Sarcoma; Recurrent Adult Soft Tissue Sarcoma; Stage IVA Adult Soft Tissue Sarcoma; Stage III Adult Soft Tissue Sarcoma

  10. Ramelteon for Insomnia Symptoms in a Community Sample of Adults with Generalized Anxiety Disorder: An Open Label Study

    PubMed Central

    Gross, Paul K.; Nourse, Rosemary; Wasser, Thomas E.

    2009-01-01

    Objective: Prior research confirms the relationship between insomnia and psychiatric disorders, particularly anxiety and depression. The effectiveness and tolerability of ramelteon was examined in adult generalized anxiety disorder (GAD) patients with insomnia symptoms. Methods: Twenty-seven adults with sleep disturbance meeting DSM-IV diagnostic criteria for GAD and partially responsive on an SSRI or SNRI by randomization visit (as signified by a Hamilton Anxiety scale [HAMA] maximum score of 15 and minimum of 8, Clinical Global Impressions Severity of Illness [CGI-S] scale of ≤ 4 and ≥ 2 [measuring anxiety symptoms], CGI-S of ≥ 4 [measuring insomnia symptoms], ≥ 5 on the Pittsburgh Sleep Quality Index [PSQI], and ≥ 10 on the Epworth Sleepiness Scale [ESS]) were treated openly for 10 weeks on ramelteon 8 mg at bedtime. Analysis was conducted using repeated measures methodology. Patient reported sleep diaries were maintained throughout the study. Results: Significant symptom reduction was observed on all scales (HAMA, ESS, CGI-I, CGI-S), with subjects falling asleep faster and sleeping longer. Headache upon stopping ramelteon, daytime tiredness, agitation, and depression were the most commonly reported side effects and were cited as transient. Conclusion: Data from this 12-week open-label study suggests ramelteon is an effective and generally well tolerated treatment for insomnia symptoms in this community sample of adults with GAD. Citation: Gross PK; Nourse R; Wasser TE. Ramelteon for insomnia symptoms in a community sample of adults with generalized anxiety disorder: an open label study. J Clin Sleep Med 2009;5(1):28–33. PMID:19317378

  11. Individual, Interpersonal, and Structural Power: Associations With Condom Use in a Sample of Young Adult Latinos.

    PubMed

    Stokes, Lynissa R; Harvey, S Marie; Warren, Jocelyn T

    2016-01-01

    Interviews were conducted with 480 sexually active Latino young adults from four rural counties in Oregon. We examined relationships between three levels of power (individual, interpersonal, and structural) and consistent condom use. Condom use self-efficacy and sexual decision-making, examples of individual and interpersonal measures of power, respectively, were associated with increased odds of consistent condom use among both men and women. Among men only, increasing relationship control, an interpersonal measure of power, was associated with lower odds of consistent condom use. Among women only, increasing medical mistrust, a structural measure of power, was associated with increased odds of consistent condom use.

  12. The shortened dental arch: prevalence and normative treatment needs in a sample of older Canadian adults.

    PubMed

    Hawkins, R J

    1998-01-01

    The concept of the shortened dental arch (SDA) was used to classify the dentition status and normative treatment needs of older adults. From 1982 to 1992, a descriptive survey was conducted in North York, Canada, of 1531 dentate adults aged 65 and over; 69% were nursing home residents. Based on the SDA concept, a "good" guadrant was defined as one which contained all premolar and anterior teeth. A "good" arch was defined as one which had two "good" quadrants. Only 6.3% of nursing home subjects and 7.5% of independently living subjects were classified as having "good" upper and lower arches; these subjects were considered to have a "functional dentition" by the criteria of the SDA concept. For subjects of both residence types, a higher percentage had a "good" lower arch (20%, 30%) as compared with a "good" upper arch (9%, 13%), and a higher proportion of non-denture wearers had "good" arches and quadrants compared with denture wearers. The most common reason for failure to meet SDA criteria was due to the loss of one or more upper premolar teeth; loss of lower canines was least frequently the reason. For subjects of both residence types, normative need for tooth extraction and prosthetic care was significantly associated with having no "good" arches. This was found for both denture wearers and non-denture wearers. Among non-denture wearers of both residence types, the need for urgent care was significantly associated with having no "good" arches.

  13. Psychometric evaluation of responses to the NEO-PI-3 in a multi-ethnic sample of adults in India.

    PubMed

    Piedmont, Ralph L; Braganza, Dinesh J

    2015-12-01

    Cross-cultural studies have demonstrated the utility of the NEO scales in organizing research on personality development (McCrae, 2004). The NEO PI-3 is the latest updated version that has modified item content more suitable to younger populations and to adults with lower literacy levels. The present study examined the utility of the English version of the NEO PI-3 in a multiethnic sample of adults from India (N = 188) by examining mean level data, Cronbach's alpha, retest reliabilities, and construct validity with particular attention given to previous findings that used the NEO Personality Inventory-Revised (NEO PI-R) in an Indian sample. Principal components analyses employing orthogonal procrustean rotations demonstrated convergence with U.S. norms and prior Indian data. The current Cronbach's alpha and retest reliability values were slightly better than data using the NEO-PI-R in 2 Indian samples (Lodhi, Deo, & Belhekar, 2002; Singh, 2009). Scores correlated significantly, and in appropriate ways, with several psychosocial measures. These findings underscore the potential utility of the NEO PI-3 in research among those for whom English is a second language.

  14. Social and psychological resources associated with health status in a representative sample of adults affected by the 2004 Florida hurricanes.

    PubMed

    Ruggiero, Kenneth J; Amstadter, Ananda B; Acierno, Ron; Kilpatrick, Dean G; Resnick, Heidi S; Tracy, Melissa; Galea, Sandro

    2009-01-01

    Overall health status after a disaster may be associated with long-term physical morbidity and mortality. Little is known about factors associated with overall health status in the aftermath of disasters. We examined self-rated health in relation to disaster characteristics, social resources, and post-disaster outcomes in a sample of adults who experienced the 2004 Florida hurricanes. We interviewed a representative sample of 1,452 adults aged 18 years and older residing in the 33 Florida counties that were in the direct path of at least one of the 2004 hurricanes (Charley, Frances, Ivan, Jeanne). Overall health status was assessed using a self-rating format known to be predictive of mortality. Poor self-rated health was endorsed by 14.6% of the sample. Final multivariable models showed that poor self-rated health was associated with older age (p < 0.001), extreme fear during the hurricane (p = 0.03), low social support (p = 0.03), and depression (p = 0.003) since the hurricane. Self-rated health following the Florida hurricanes was strongly associated with two variables (social support and depression) that potentially can be mitigated through targeted interventions after disasters. Future work should evaluate secondary prevention strategies that can address general health-related concerns in the wake of a disaster.

  15. Mediastinitis after cardiac surgery: improvement of bacteriological diagnosis by use of multiple tissue samples and strain typing.

    PubMed

    Tammelin, Ann; Hambraeus, Anna; Ståhle, Elisabeth

    2002-08-01

    The diagnosis of postsurgical mediastinitis (PSM) among patients with sternal wound complication (SWC) after cardiac surgery is sometimes difficult, as fever, elevated C-reactive protein levels, and chest pain can be caused by a general inflammatory reaction to the operative trauma and/or sternal dehiscence without infection. The definitions of PSM usually used emphasize clinical signs and symptoms easily observed by the surgeon. The aim of the study was to investigate whether the use of standardized multiple tissue sampling, optimal culturing methods, and strain typing, together with a microbiological criterion for infection, could identify more infected patients than clinical assessment alone. Patients reexplored due to SWC after cardiac artery bypass grafting (CABG) or heart valve replacement (HVR) with or without CABG performed at the Department for Cardio-Thoracic Surgery at the Uppsala University Hospital between 10 March 1998 and 9 September 2000 were investigated prospectively. Tissue samples were taken from the sternum or adjacent mediastinal tissue, preferably before the administration of antibiotics. Culturing was performed both directly (on agar plates) and using enrichment broth. Species identification was performed by standard methods, and strain typing was performed by pulsed-field gel electrophoresis. A total of 41 cases with at least five tissue samples each were included in the study group. Of these patients, 32 were infected according to the microbiological criterion (i.e., the same strain was found in >/=50% of the samples). Staphylococcus epidermidis was the primary pathogen in 38% of the cases (12/32), S. aureus was the primary pathogen in 31% (10/32), P. acnes was the primary pathogen in 25% (8/32), and S. simulans and S. haemolyticus were the primary pathogens in 3% (1/32) each. All cases of S. aureus infection and 86% (12/14) of coagulase-negative staphylococcus (CoNS) infections were identified from primary cultures. All cases fulfilling

  16. A New Sample Substrate for Imaging and Correlating Organic and Trace Metal Composition in Biological Cells and Tissues

    SciTech Connect

    Miller,L.; Wang, Q.; Smith, R.; Zhong, H.; Elliott, D.; Warren, J.

    2007-01-01

    Many disease processes involve alterations in the chemical makeup of tissue. Synchrotron-based infrared (IR) and X-ray fluorescence (XRF) microscopes are becoming increasingly popular tools for imaging the organic and trace metal compositions of biological materials, respectively, without the need for extrinsic labels or stains. Fourier transform infrared microspectroscopy (FTIRM) provides chemical information on the organic components of a material at a diffraction-limited spatial resolution of 2-10 {mu}m in the mid-infrared region. The synchrotron X-ray fluorescence (SXRF) microprobe is a complementary technique used to probe trace element content in the same systems with a similar spatial resolution. However to be most beneficial, it is important to combine the results from both imaging techniques on a single sample, which requires precise overlap of the IR and X-ray images. In this work, we have developed a sample substrate containing a gold grid pattern on its surface, which can be imaged with both the IR and X-ray microscopes. The substrate consists of a low trace element glass slide that has a gold grid patterned on its surface, where the major and minor parts of the grid contain 25 and 12 nm gold, respectively. This grid pattern can be imaged with the IR microscope because the reflectivity of gold differs as a function of thickness. The pattern can also be imaged with the SXRF microprobe because the Au fluorescence intensity changes with gold thickness. The tissue sample is placed on top of the patterned substrate. The grid pattern's IR reflectivity image and the gold SXRF image are used as fiducial markers for spatially overlapping the IR and SXRF images from the tissue. Results show that IR and X-ray images can be correlated precisely, with a spatial resolution of less than one pixel (i.e., 2-3 microns). The development of this new tool will be presented along with applications to paraffin-embedded metalloprotein crystals, Alzheimer's disease, and hair

  17. Paralytic shellfish poisoning: post-mortem analysis of tissue and body fluid samples from human victims in the Patagonia fjords.

    PubMed

    García, Carlos; del Carmen Bravo, María; Lagos, Marcelo; Lagos, Néstor

    2004-02-01

    In July 5, 2002 fishermen working in harvesting sea urchin (Loxechinus albus) in the Patagonia Chilean fjords were intoxicated by consumption of filter-feeder bivalve Aulacomya ater. After the ingestion of 7-9 ribbed mussel, two fishermen died 3-4 h after shellfish consumption. The forensic examination in both victims did not show pathological abnormalities with the exception of the lungs conditions, crackling to the touch, pulmonary congestion and edema. The toxic mussel sample showed a toxicity measured by mouse bioassay of 8575 microg of STX (saxitoxin) equivalent by 100 g of shellfish meat. Using post-column derivatization HPLC method with fluorescent on line detection was possible to measure mass amount of each paralytic shellfish poisoning (PSP) toxin yielding individual toxin concentrations. These PSP toxins were identified in the gastric content, body fluids (urine, bile and cerebrospinal fluid) and tissue samples (liver, kidney, lung, stomach, spleen, heart, brain, adrenal glands, pancreas and thyroids glands). The toxin profiles of each body fluid and tissue samples and the amount of each PSP toxin detected are reported. The PSP toxins found in the gastric content, were STX and the gonyautoxins (GTX4, GTX1, GTX5, GTX3 and GTX2) which showed to be the major amount of PSP toxins found in the victims biological samples. The PSP toxin composition in urine and bile showed as major PSP toxins neoSaxitoxin (neoSTX) and GTX4/GTX1 epimers, both STX analogues with an hydroxyl group (-OH) in the N(1) of the tetrahydropurine nucleus. The neoSTX was not present in the gastric content sample, indicating that the oxidation of N(1) in the STX tetrahydropurine nucleus resulted neoSTX, in a similar way that GTX3/GTX2 epimers were transformed in GTX4/GTX1 epimers. Beside this metabolic transformation, also the hydrolysis of carbamoyl group from STX to form its decarbomoyl analogue decarbamoylsaxitoxin was detected in liver, kidney and lung. These two findings show that PSP

  18. Acrolein and Asthma Attack Prevalence in a Representative Sample of the United States Adult Population 2000 – 2009

    PubMed Central

    deCastro, B. Rey

    2014-01-01

    Background Acrolein is an air toxic and highly potent respiratory irritant. There is little epidemiology available, but US EPA estimates that outdoor acrolein is responsible for about 75 percent of non-cancer respiratory health effects attributable to air toxics in the United States, based on the Agency's 2005 NATA (National-Scale Air Toxics Assessment) and acrolein's comparatively potent inhalation reference concentration of 0.02 µg/m3. Objectives Assess the association between estimated outdoor acrolein exposure and asthma attack reported by a representative cross-sectional sample of the adult United States population. Methods NATA 2005 chronic outdoor acrolein exposure estimates at the census tract were linked with residences oif adults (≥18 years old) in the NHIS (National Health Interview Survey) 2000 – 2009 (n = 271,348 subjects). A sample-weighted logistic regression model characterized the association between the prevalence of reporting at least one asthma attack in the 12 months prior to survey interview and quintiles of exposure to outdoor acrolein, controlling for potential confounders. Results In the highest quintile of outdoor acrolein exposure (0.05 – 0.46 µg/m3), there was a marginally significant increase in the asthma attack pOR (prevalence-odds ratio [95% CI]  = 1.08 [0.98∶1.19]) relative to the lowest quintile. The highest quintile was also associated with a marginally significant increase in prevalence-odds (1.13 [0.98∶1.29]) in a model limited to never smokers (n = 153,820). Conclusions Chronic exposure to outdoor acrolein of 0.05 – 0.46 µg/m3 appears to increase the prevalence-odds of having at least one asthma attack in the previous year by 8 percent in a representative cross-sectional sample of the adult United States population. PMID:24816802

  19. Densely calculated facial soft tissue thickness for craniofacial reconstruction in Chinese adults.

    PubMed

    Shui, Wuyang; Zhou, Mingquan; Deng, Qingqiong; Wu, Zhongke; Ji, Yuan; Li, Kang; He, Taiping; Jiang, Haiyan

    2016-09-01

    Craniofacial reconstruction (CFR) is used to recreate a likeness of original facial appearance for an unidentified skull; this technique has been applied in both forensics and archeology. Many CFR techniques rely on the average facial soft tissue thickness (FSTT) of anatomical landmarks, related to ethnicity, age, sex, body mass index (BMI), etc. Previous studies typically employed FSTT at sparsely distributed anatomical landmarks, where different landmark definitions may affect the contrasting results. In the present study, a total of 90,198 one-to-one correspondence skull vertices are established on 171 head CT-scans and the FSTT of each corresponding vertex is calculated (hereafter referred to as densely calculated FSTT) for statistical analysis and CFR. Basic descriptive statistics (i.e., mean and standard deviation) for densely calculated FSTT are reported separately according to sex and age. Results show that 76.12% of overall vertices indicate that the FSTT is greater in males than females, with the exception of vertices around the zygoma, zygomatic arch and mid-lateral orbit. These sex-related significant differences are found at 55.12% of all vertices and the statistically age-related significant differences are depicted between the three age groups at a majority of all vertices (73.31% for males and 63.43% for females). Five non-overlapping categories are given and the descriptive statistics (i.e., mean, standard deviation, local standard deviation and percentage) are reported. Multiple appearances are produced using the densely calculated FSTT of various age and sex groups, and a quantitative assessment is provided to examine how relevant the choice of FSTT is to increasing the accuracy of CFR. In conclusion, this study provides a new perspective in understanding the distribution of FSTT and the construction of a new densely calculated FSTT database for craniofacial reconstruction. PMID:27544400

  20. Is the prevalence of child sexual abuse decreasing? Evidence from a random sample of 750 young adult women.

    PubMed

    Bagley, C

    1990-06-01

    An adult recall study in Calgary, Alberta in 1988-89 of child sexual abuse used stratified, random sampling to identify 750 women ages 18 to 27 yr., divided into 10 age cohorts of approximately 75 each. Those aged 18 and 19 yr. recalled significantly less contact abuse up to age 16 than those aged 20 to 27 yr. Experiencing print or visual media on the topic of sexual abuse was associated with a decreased prevalence. It is argued that in a climate of publicity and greater understanding of help sources and harmful effects, the actual prevalence of child sexual abuse may be decreasing.

  1. Attitudes toward Bisexual Men and Women among a Nationally Representative Probability Sample of Adults in the United States

    PubMed Central

    Herbenick, Debby; Friedman, M. Reuel; Schick, Vanessa; Fu, Tsung-Chieh (Jane); Bostwick, Wendy; Bartelt, Elizabeth; Muñoz-Laboy, Miguel; Pletta, David; Reece, Michael; Sandfort, Theo G. M.

    2016-01-01

    As bisexual individuals in the United States (U.S.) face significant health disparities, researchers have posited that these differences may be fueled, at least in part, by negative attitudes, prejudice, stigma, and discrimination toward bisexual individuals from heterosexual and gay/lesbian individuals. Previous studies of individual and social attitudes toward bisexual men and women have been conducted almost exclusively with convenience samples, with limited generalizability to the broader U.S. population. Our study provides an assessment of attitudes toward bisexual men and women among a nationally representative probability sample of heterosexual, gay, lesbian, and other-identified adults in the U.S. Data were collected from the 2015 National Survey of Sexual Health and Behavior (NSSHB), via an online questionnaire with a probability sample of adults (18 years and over) from throughout the U.S. We included two modified 5-item versions of the Bisexualities: Indiana Attitudes Scale (BIAS), validated sub-scales that were developed to measure attitudes toward bisexual men and women. Data were analyzed using descriptive statistics, gamma regression, and paired t-tests. Gender, sexual identity, age, race/ethnicity, income, and educational attainment were all significantly associated with participants' attitudes toward bisexual individuals. In terms of responses to individual scale items, participants were most likely to “neither agree nor disagree” with all attitudinal statements. Across sexual identities, self-identified other participants reported the most positive attitudes, while heterosexual male participants reported the least positive attitudes. As in previous research on convenience samples, we found a wide range of demographic characteristics were related with attitudes toward bisexual individuals in our nationally-representative study of heterosexual, gay/lesbian, and other-identified adults in the U.S. In particular, gender emerged as a significant

  2. Built environment and social environment: associations with overweight and obesity in a sample of Brazilian adults.

    PubMed

    Velásquez-Meléndez, Gustavo; Mendes, Larissa Loures; Padez, Cristina Maria Proença

    2013-10-01

    The aim of this study was to assess associations between the built environment and social environment and excess weight in an urban population. Participants were selected from the Surveillance System for Risk Factors for Chronic Diseases (VIGITEL). The study used data from the city of Belo Horizonte, Minas Gerais State, Brazil. A total of 3,425 interviews from the years 2008 and 2009 were used. Georeferenced data on parks, squares, and locations for physical exercise, population density, and food stores were used to assess the built environment. Description of the social environment used income and homicide rate for the neighborhood. Environmental variables associated independently with excess weight were population density, presence of parks, squares, and locations for physical exercise, and self-reported presence of locations for physical exercise. The findings show that residential neighborhood characteristics are associated with excess weight in urban adults.

  3. Routine sample preparation and HPLC analysis for ascorbic acid (vitamin C) determination in wheat plants and Arabidopsis leaf tissues.

    PubMed

    Szalai, Gabriella; Janda, T; Pál, Magda

    2014-06-01

    Plants have developed various mechanisms to protect themselves against oxidative stress. One of the most important non-enzymatic antioxidants is ascorbic acid. There is thus a need for a rapid, sensitive method for the analysis of the reduced and oxidised forms of ascorbic acid in crop plants. In this paper a simple, economic, selective, precise and stable HPLC method is presented for the detection of ascorbate in plant tissue. The sensitivity, the short retention time and the simple isocratic elution mean that the method is suitable for the routine quantification of ascorbate in a high daily sample number. The method has been found to be better than previously reported methods, because of the use of an economical, readily available mobile phase, UV detection and the lack of complicated extraction procedures. The method has been tested on Arabidopsis plants with different ascorbate levels and on wheat plants during Cd stress.

  4. Gas-liquid chromatographic and gas-liquid-mass spectometric determination of fenvalerate and permethrin residues in grasshoppers and duck tissue samples

    USGS Publications Warehouse

    Reichel, W.L.; Kolbe, E.J.; Stafford, C.J.

    1981-01-01

    A procedure is described for determining fenvalerate and permethrin residues in grasshoppers and duck tissues. Samples are Soxhlet-extracted with hexane and cleaned up by gel permeation chromatography with an in-line alumina column. Samples are analyzed by gas-liquid chromatography with electron capture detection, and confirmed by gas-liquid chromatography-mass spectrometry. The average recovery from fortified tissues was 97%.

  5. Temporal variability of pyrethroid metabolite levels in bedtime, morning, and 24-h urine samples for 50 adults in North Carolina.

    PubMed

    Morgan, Marsha K; Sobus, Jon R; Barr, Dana Boyd; Croghan, Carry W; Chen, Fu-Lin; Walker, Richard; Alston, Lillian; Andersen, Erik; Clifton, Matthew S

    2016-01-01

    Pyrethroid insecticides are widely used to control insects in both agricultural and residential settings worldwide. Few data are available on the temporal variability of pyrethroid metabolites in the urine of non-occupationally exposed adults. In this work, we describe the study design and sampling methodology for the Pilot Study to Estimate Human Exposures to Pyrethroids using an Exposure Reconstruction Approach (Ex-R study). Two major objectives were to quantify the concentrations of several pyrethroid metabolites in bedtime, first morning void (FMV), and 24-h urine samples as concentration (wet weight), specific-gravity (SG) corrected, creatinine (CR) corrected, and excretion rate values for 50 Ex-R adults over a six-week monitoring period and to determine if these correction approaches for urine dilution reduced the variability of the biomarker levels. The Ex-R study was conducted at the United States Environmental Protection Agency's Human Studies Facility in Chapel Hill, North Carolina USA and at participants' homes within a 40-mile radius of this facility. Recruitment of participants and field activities occurred between October 2009 and May 2011. Participants, ages 19-50 years old, provided daily food, activity, and pesticide-use diaries and collected their own urine samples (bedtime, FMV, and 24-h) during weeks 1, 2, and 6 of a six-week monitoring period. A total of 2503 urine samples were collected from the study participants. These samples were analyzed for the pyrethroid metabolites 3-phenoxybenzoic acid (3-PBA), cis/trans-3-(2,2-dichlorovinyl)-2,2-dimethyl-cyclopropane carboxylic acid (cis/trans-DCCA), and 2-methyl-3-phenylbenzoic acid (MPA) using high performance liquid chromatography/tandem mass spectrometry. Only 3-PBA was frequently detected (>50%) in the adult urine samples. Median urinary 3-PBA levels were 0.88 ng/mL, 0.96 ng/mL-SG, 1.04 ng/mg, and 1.04 ng/min for concentration, SG-corrected, CR-corrected, and excretion rate values, respectively

  6. Somatomedin-C/insulin-like growth factor-I and Insulin-like growth factor-II mRNAs in rate fetal and adult tissues

    SciTech Connect

    Lund, P.K.; Moats-Staats, B.M.; Hynes, M.A.; Simmons, J.G.; Jansen, M.; D'ercole, A.J.; Van Wyk, J.J.

    1986-11-05

    Somatomedin-C or insulin-like growth factor I (Sm-C/IGF-I) and insulin-like growth factor II (IGF-II) have been implicated in the regulation of fetal growth and development. In the present study /sup 32/P-labeled complementary DNA probes encoding human and mouse Sm-C/IGF-I and human IGF-II were used in Northern blot hybridizations to analyze rat Sm-C/IGF-I and IGF-II mRNAs in poly(A/sup +/) RNAs from intestine, liver, lung, and brain of adult rats and fetal rats between day 14 and 17 of gestation. In fetal rats, all four tissues contained a major mRNA of 1.7 kilobase (kb) that hybridized with the human Sm-C/IGF-I cDNA and mRNAs of 7.5, 4.7, 1.7, and 1.2 kb that hybridized with the mouse Sm-C/IGF-I cDNA. Adult rat intestine, liver, and lung also contained these mRNAs but Sm-C/IGF-I mRNAs were not detected in adult rat brain. These findings provide direct support for prior observations that multiple tissues in the fetus synthesize immunoreactive Sm-C/IGF-I and imply a role for Sm-C/IGF-I in fetal development as well as postnatally. Multiple IGF-II mRNAs of estimated sizes 4.7, 3.9, 2.2, 1.75, and 1.2 kb were observed in fetal rat intestine, liver, lung, and brain. The 4.7- and 3.9-kb mRNAs were the major hybridizing IGF-II mRNAs in all fetal tissues. Higher abundance of IGF-II mRNAs in rat fetal tissues compared with adult tissues supports prior hypotheses, based on serum IGF-II concentrations, that IGF-II is predominantly a fetal somatomedin. IGF-II mRNAs are present, however, in some poly(A/sup +/) RNAs from adult rat tissues. The brain was the only tissue in the adult rat where the 4.7- and 3.9-kb IGF-II mRNAs were consistently detected. These findings suggest that a role for IGF-II in the adult rat, particularly in the central nervous system, cannot be excluded.

  7. Assessing the effects of model Maillard compound intake on iron, copper and zinc retention and tissue delivery in adult rats.

    PubMed

    Roncero-Ramos, Irene; Pastoriza, Silvia; Navarro, M Pilar; Delgado-Andrade, Cristina

    2016-01-01

    The behaviour of dietary Maillard reaction compounds (MRP) as metal chelating polymers can alter mineral absorption and/or retention. Our aim in this study was to analyse the long-term effects of the consumption of model MRP from glucose-lysine heated for 90 min at 150 °C (GL) on iron, copper and zinc whole-body retention and tissue delivery. For 88 days, weaning rats were fed a Control diet or one containing 3% GL, until reaching the adult stage. During the experimental period a mineral balance was conducted to investigate the mineral retention. At day 88, the animals were sacrificed, blood was drawn for haemoglobin determination and some organs were removed. Copper and zinc balances were unaffected (Cu: 450 vs. 375 μg; Zn: 6.7 vs. 6.2 mg for Control and GL groups, respectively) and no change was observed in whole-body delivery. Iron retention, too, was unaltered (11.2 mg for Control and GL groups) but due to the tendency toward decreased body weight in the GL group (248 vs. 233 g for the Control and GL groups), whole-body iron concentration was 13% higher in the GL group than in the Control group. Absorbed iron accumulated particularly in the liver (144 vs. 190 μg g(-1) for the Control and GL groups), thus reducing haemoglobin levels. The long-term intake of MRP induced iron accumulation in the body but this did not result in enhanced iron functionality, since the haemoglobin concentration declined. Taking into account the findings of our research group's studies of young and adult rats, we now corroborate the hypothesis that the negative effect of GL MRP consumption on iron functionality takes place regardless of the animals' stage of life.

  8. Lack of Association between a 3'UTR VNTR Polymorphism of Dopamine Transporter Gene (SLC6A3) and ADHD in a Brazilian Sample of Adult Patients

    ERIC Educational Resources Information Center

    Aperecida da Silva, Maria; Cordeiro, Quirino; Louza, Mario; Vallada, Homero

    2011-01-01

    Objective: To investigate a possible association between a 3'UTR VNTR polymorphism of the dopamine transporter gene (SLC6A3) and ADHD in a Brazilian sample of adult patients. Method: Study Case-control with 102 ADHD adult outpatients ("DSM-IV" criteria) and 479 healthy controls. The primers' sequence used were: 3'UTR-Forward: 5' TGT GGT GAT GGG…

  9. Intended Sensitive and Harsh Caregiving Responses to Infant Crying: The Role of Cry Pitch and Perceived Urgency in an Adult Twin Sample

    ERIC Educational Resources Information Center

    Out, Dorothee; Pieper, Suzanne; Bakermans-Kranenburg, Marian J.; Zeskind, Philip Sanford; van IJzendoorn, Marinus H.

    2010-01-01

    Objective: To examine the underlying mechanisms of adults' intended caregiving responses to cry sounds in a behavioral genetic design and to investigate the role of cry pitch and perceived urgency in sensitive and harsh caregiving responses. Methods: The sample consisted of 184 adult twin pairs (18-69 years), including males and females, parents…

  10. Early Educational Intervention, Early Cumulative Risk, and the Early Home Environment as Predictors of Young Adult Outcomes within a High-Risk Sample

    ERIC Educational Resources Information Center

    Pungello, Elizabeth P.; Kainz, Kirsten; Burchinal, Margaret; Wasik, Barbara H.; Sparling, Joseph J.; Ramey, Craig T.; Campbell, Frances A.

    2010-01-01

    The extent to which early educational intervention, early cumulative risk, and the early home environment were associated with young adult outcomes was investigated in a sample of 139 young adults (age 21) from high-risk families enrolled in randomized trials of early intervention. Positive effects of treatment were found for education attainment,…

  11. Sphingosine-1-phosphate mediates proliferation maintaining the multipotency of human adult bone marrow and adipose tissue-derived stem cells.

    PubMed

    He, Xiaoli; H'ng, Shiau-Chen; Leong, David T; Hutmacher, Dietmar W; Melendez, Alirio J

    2010-08-01

    High renewal and maintenance of multipotency of human adult stem cells (hSCs), are a prerequisite for experimental analysis as well as for potential clinical usages. The most widely used strategy for hSC culture and proliferation is using serum. However, serum is poorly defined and has a considerable degree of inter-batch variation, which makes it difficult for large-scale mesenchymal stem cells (MSCs) expansion in homogeneous culture conditions. Moreover, it is often observed that cells grown in serum-containing media spontaneously differentiate into unknown and/or undesired phenotypes. Another way of maintaining hSC development is using cytokines and/or tissue-specific growth factors; this is a very expensive approach and can lead to early unwanted differentiation. In order to circumvent these issues, we investigated the role of sphingosine-1-phosphate (S1P), in the growth and multipotency maintenance of human bone marrow and adipose tissue-derived MSCs. We show that S1P induces growth, and in combination with reduced serum, or with the growth factors FGF and platelet-derived growth factor-AB, S1P has an enhancing effect on growth. We also show that the MSCs cultured in S1P-supplemented media are able to maintain their differentiation potential for at least as long as that for cells grown in the usual serum-containing media. This is shown by the ability of cells grown in S1P-containing media to be able to undergo osteogenic as well as adipogenic differentiation. This is of interest, since S1P is a relatively inexpensive natural product, which can be obtained in homogeneous high-purity batches: this will minimize costs and potentially reduce the unwanted side effects observed with serum. Taken together, S1P is able to induce proliferation while maintaining the multipotency of different human stem cells, suggesting a potential for S1P in developing serum-free or serum-reduced defined medium for adult stem cell cultures.

  12. Recruiting and Retaining an Ethnically Diverse Sample of Older Adults in a Longitudinal Intervention Study

    ERIC Educational Resources Information Center

    Austin-Wells, Vonnette; McDougall, Graham J.; Becker, Heather

    2006-01-01

    This paper describes strategies developed to recruit and retain an ethnically diverse sample in a longitudinal intervention of 246 participants in the SeniorWISE study. The ethnic and socioeconomic differences of these participants necessitated the use of different methods of effectively communicating with this population. Recruitment benefited…

  13. Epithelial-connective tissue interactions induced by thyroid hormone receptor are essential for adult stem cell development in the Xenopus laevis intestine.

    PubMed

    Hasebe, Takashi; Buchholz, Daniel R; Shi, Yun-Bo; Ishizuya-Oka, Atsuko

    2011-01-01

    In the amphibian intestine during metamorphosis, stem cells appear and generate the adult absorptive epithelium, analogous to the mammalian one, under the control of thyroid hormone (TH). We have previously shown that the adult stem cells originate from differentiated larval epithelial cells in the Xenopus laevis intestine. To clarify whether TH signaling in the epithelium alone is sufficient for inducing the stem cells, we have now performed tissue recombinant culture experiments using transgenic X. laevis tadpoles that express a dominant-positive TH receptor (dpTR) under a control of heat shock promoter. Wild-type (Wt) or dpTR transgenic (Tg) larval epithelium (Ep) was isolated from the tadpole intestine, recombined with homologous or heterologous nonepithelial tissues (non-Ep), and then cultivated in the absence of TH with daily heat shocks to induce transgenic dpTR expression. Adult epithelial progenitor cells expressing sonic hedgehog became detectable on day 5 in both the recombinant intestine of Tg Ep and Tg non-Ep (Tg/Tg) and that of Tg Ep and Wt non-Ep (Tg/Wt). However, in Tg/Wt intestine, they did not express other stem cell markers such as Musashi-1 and never generated the adult epithelium expressing a marker for absorptive epithelial cells. Our results indicate that, while it is unclear why some larval epithelial cells dedifferentiate into adult progenitor/stem cells, TR-mediated gene expression in the surrounding tissues other than the epithelium is required for them to develop into adult stem cells, suggesting the importance of TH-inducible epithelial-connective tissue interactions in establishment of the stem cell niche in the amphibian intestine.

  14. Age-Dependent Changes in Geometry, Tissue Composition and Mechanical Properties of Fetal to Adult Cryopreserved Human Heart Valves.

    PubMed

    van Geemen, Daphne; Soares, Ana L F; Oomen, Pim J A; Driessen-Mol, Anita; Janssen-van den Broek, Marloes W J T; van den Bogaerdt, Antoon J; Bogers, Ad J J C; Goumans, Marie-José T H; Baaijens, Frank P T; Bouten, Carlijn V C

    2016-01-01

    There is limited information about age-specific structural and functional properties of human heart valves, while this information is key to the development and evaluation of living valve replacements for pediatric and adolescent patients. Here, we present an extended data set of structure-function properties of cryopreserved human pulmonary and aortic heart valves, providing age-specific information for living valve replacements. Tissue composition, morphology, mechanical properties, and maturation of leaflets from 16 pairs of structurally unaffected aortic and pulmonary valves of human donors (fetal-53 years) were analyzed. Interestingly, no major differences were observed between the aortic and pulmonary valves. Valve annulus and leaflet dimensions increase throughout life. The typical three-layered leaflet structure is present before birth, but becomes more distinct with age. After birth, cell numbers decrease rapidly, while remaining cells obtain a quiescent phenotype and reside in the ventricularis and spongiosa. With age and maturation-but more pronounced in aortic valves-the matrix shows an increasing amount of collagen and collagen cross-links and a reduction in glycosaminoglycans. These matrix changes correlate with increasing leaflet stiffness with age. Our data provide a new and comprehensive overview of the changes of structure-function properties of fetal to adult human semilunar heart valves that can be used to evaluate and optimize future therapies, such as tissue engineering of heart valves. Changing hemodynamic conditions with age can explain initial changes in matrix composition and consequent mechanical properties, but cannot explain the ongoing changes in valve dimensions and matrix composition at older age.

  15. Effects of FGF-2 on human adipose tissue derived adult stem cells morphology and chondrogenesis enhancement in Transwell culture

    SciTech Connect

    Kabiri, Azadeh; Esfandiari, Ebrahim; Hashemibeni, Batool; Kazemi, Mohammad; Mardani, Mohammad; Esmaeili, Abolghasem

    2012-07-27

    Highlights: Black-Right-Pointing-Pointer We investigated effects of FGF-2 on hADSCs. Black-Right-Pointing-Pointer We examine changes in the level of gene expressions of SOX-9, aggrecan and collagen type II and type X. Black-Right-Pointing-Pointer FGF-2 induces chondrogenesis in hADSCs, which Bullet Increasing information will decrease quality if hospital costs are very different. Black-Right-Pointing-Pointer The result of this study may be beneficial in cartilage tissue engineering. -- Abstract: Injured cartilage is difficult to repair due to its poor vascularisation. Cell based therapies may serve as tools to more effectively regenerate defective cartilage. Both adult mesenchymal stem cells (MSCs) and human adipose derived stem cells (hADSCs) are regarded as potential stem cell sources able to generate functional cartilage for cell transplantation. Growth factors, in particular the TGF-b superfamily, influence many processes during cartilage formation, including cell proliferation, extracellular matrix synthesis, maintenance of the differentiated phenotype, and induction of MSCs towards chondrogenesis. In the current study, we investigated the effects of FGF-2 on hADSC morphology and chondrogenesis in Transwell culture. hADSCs were obtained from patients undergoing elective surgery, and then cultured in expansion medium alone or in the presence of FGF-2 (10 ng/ml). mRNA expression levels of SOX-9, aggrecan and collagen type II and type X were quantified by real-time polymerase chain reaction. The morphology, doubling time, trypsinization time and chondrogenesis of hADSCs were also studied. Expression levels of SOX-9, collagen type II, and aggrecan were all significantly increased in hADSCs expanded in presence of FGF-2. Furthermore FGF-2 induced a slender morphology, whereas doubling time and trypsinization time decreased. Our results suggest that FGF-2 induces hADSCs chondrogenesis in Transwell culture, which may be beneficial in cartilage tissue engineering.

  16. Immunohistochemical evidence that gonads and gonad-associated tissues are sites for enrichment with immunoglobul