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1.
Science ; 375(6577): 172-177, 2022 Jan 14.
Article in English | MEDLINE | ID: mdl-35025630

ABSTRACT

Water-rock interactions are relevant to planetary habitability, influencing mineralogical diversity and the production of organic molecules. We examine carbonates and silicates in the martian meteorite Allan Hills 84001 (ALH 84001), using colocated nanoscale analyses, to characterize the nature of water-rock reactions on early Mars. We find complex refractory organic material associated with mineral assemblages that formed by mineral carbonation and serpentinization reactions. The organic molecules are colocated with nanophase magnetite; both formed in situ during water-rock interactions on Mars. Two potentially distinct mechanisms of abiotic organic synthesis operated on early Mars during the late Noachian period (3.9 to 4.1 billion years ago).

2.
Astrobiology ; 17(10): 971-974, 2017 10.
Article in English | MEDLINE | ID: mdl-28920443

ABSTRACT

While it is anticipated that future human missions to Mars will increase the amount of biological and organic contamination that might be distributed on that planet, robotic missions continue to grow in capability and complexity, requiring precautions to be taken now to protect Mars, and particularly areas of Mars that might be Special Regions. Such precautionary cleanliness requirements for spacecraft have evolved over the course of the space age, as we have learned more about planetary environments, and are the subject of regular deliberations and decisions sponsored by the Committee on Space Research (COSPAR). COSPAR's planetary protection policy is maintained as an international consensus standard for spacecraft cleanliness that is recognized by the United Nations Committee on the Peaceful Uses of Outer Space. In response to the paper presented in this issue by Fairén et al. (2017), we examine both their concept of evidence for possible life on Mars and their logic in recommending that spacecraft cleanliness requirements be relaxed to access Special Regions "before it is too late." We find that there are shortcomings in their plans to look for evidence of life on Mars, that they do not support their contention that appropriate levels of spacecraft cleanliness are unaffordable, that there are major risks in assuming martian life could be identified by nucleic acid sequence comparison (especially if those sequences are obtained from a Special Region contaminated with Earth life), and that the authors do not justify their contention that exploration with dirty robots, now, is preferable to the possibility that later contamination will be spread by human exploration. We also note that the potential effects of contaminating resources and environments essential to future human occupants of Mars are both significant and not addressed by Fairén et al. (2017). Key Words: Mars-Special Region-Mission-Life detection-Planetary protection. Astrobiology 17, 971-974.


Subject(s)
Mars , Space Flight , Exobiology , Extraterrestrial Environment , Humans , Spacecraft
3.
Rev Sci Instrum ; 87(11): 114503, 2016 Nov.
Article in English | MEDLINE | ID: mdl-27910445

ABSTRACT

The two interferometers of the Laser Interferometry Gravitational-wave Observatory (LIGO) recently detected gravitational waves from the mergers of binary black hole systems. Accurate calibration of the output of these detectors was crucial for the observation of these events and the extraction of parameters of the sources. The principal tools used to calibrate the responses of the second-generation (Advanced) LIGO detectors to gravitational waves are systems based on radiation pressure and referred to as photon calibrators. These systems, which were completely redesigned for Advanced LIGO, include several significant upgrades that enable them to meet the calibration requirements of second-generation gravitational wave detectors in the new era of gravitational-wave astronomy. We report on the design, implementation, and operation of these Advanced LIGO photon calibrators that are currently providing fiducial displacements on the order of 10-18m/Hz with accuracy and precision of better than 1%.

4.
Eat Weight Disord ; 16(1): e61-4, 2011 Mar.
Article in English | MEDLINE | ID: mdl-21727784

ABSTRACT

Few studies have compared low-weight individuals with eating disorder (ED) pathology with similar-weight individuals without significant pathology despite the fact that body weight is often used as a key outcome within ED research. This study compared quality of life (QoL) in one group with high levels of ED pathology to a group with low ED pathology, matched by body mass index (BMI). The high ED group reported significantly lower ED-specific quality of life (EDQoL) than the low ED group. These findings suggest that young women with high levels of ED pathology report significantly more impaired QoL than comparable young women with no ED pathology, and that being underweight alone is not a primary contributor to poorer EDQoL.


Subject(s)
Quality of Life/psychology , Thinness/psychology , Adolescent , Adult , Body Mass Index , Case-Control Studies , Feeding and Eating Disorders/psychology , Female , Humans , Psychiatric Status Rating Scales , Statistics, Nonparametric , Surveys and Questionnaires , Thinness/complications , Young Adult
5.
Adv Space Res ; 42(6): 1072-1079, 2008 Sep 15.
Article in English | MEDLINE | ID: mdl-22146801

ABSTRACT

Traveling, living and working in space is now a reality. The number of people and length of time in space is increasing. With new horizons for exploration it becomes more important to fully understand and provide countermeasures to the effects of the space environment on the human body. In addition, space provides a unique laboratory to study how life and physiologic functions adapt from the cellular level to that of the entire organism. Caenorhabditis elegans is a genetic model organism used to study physiology on Earth. Here we provide a description of the rationale, design, methods, and space culture validation of the ICE-FIRST payload, which engaged C. elegans researchers from four nations. Here we also show C. elegans growth and development proceeds essentially normally in a chemically defined liquid medium on board the International Space Station (10.9 day round trip). By setting flight constraints first and bringing together established C. elegans researchers second, we were able to use minimal stowage space to successfully return a total of 53 independent samples, each containing more than a hundred individual animals, to investigators within one year of experiment concept. We believe that in the future, bringing together individuals with knowledge of flight experiment operations, flight hardware, space biology, and genetic model organisms should yield similarly successful payloads.

6.
Adv Space Res ; 40(4): 506-512, 2007 Apr.
Article in English | MEDLINE | ID: mdl-18084631

ABSTRACT

The European Soyuz missions have been one of the main routes for conducting scientific experiments onboard the International Space Station, which is currently in the construction phase. A relatively large number of life and physical sciences experiments as well as technology demonstrations have been carried out during these missions. Included among these experiments are the Gene experiment during the Spanish "Cervantes" Soyuz mission and the ICE-1st experiment during the Dutch "Delta" mission. In both experiments, full genome microarray analyses were carried out on RNA extracted from whole animals recovered from the flight. These experiments indicated relatively large scale changes in gene expression levels in response to spaceflight for two popular model systems, Drosophila melanogaster (Gene) and Caenorabditis elegans (ICE-1st). Here we report a comparative analysis of results from these two experiments. Finding orthologous genes between the fruit fly and the nematode was far from straightforward, reducing the number of genes that we could compare to roughly 20% of the full comparative genome. Within this sub-set of the data (2286 genes), only six genes were found to display identical changes between species (decreased) while 1809 genes displayed no change in either species. Future experiments using ground simulation techniques will allow producing a better, more comprehensive picture of the putative set of genes affected in multicellular organisms by changes in gravity and getting a deeper understanding of how animals respond and adapt to spaceflight.

8.
J Abnorm Child Psychol ; 29(5): 445-63, 2001 Oct.
Article in English | MEDLINE | ID: mdl-11695545

ABSTRACT

This paper presents an initial assessment of the Children's Attributional Style Interview (CASI), a newly designed measure for assessing attributional style in young children (age 5 and up). The CASI was used to conduct prospective tests of the reformulated helplessness and the integrated hopelessness/self-esteem theories of depression in a sample of 147 5-10-year-old children. For comparison, the same tests were also conducted with the Children's Attributional Style Questionnaire-Revised, a commonly used measure for assessing attributional style in older children (age 8 and up). The CASI evidenced support of the reformulated helplessness theory and partial support of the integrated hopelessness/self-esteem theory. The CASI also demonstrated good internal consistency. Thus, our findings provide initial support for the CASI as a methodologically sound measure of attributional style for children as young as 5 years old. Although preliminary, our findings also suggest possible developmental differences in how attributional style interacts with self-esteem and negative life stress. The CASI should prove to be a useful tool in furthering the understanding of the origins and development of attributional style in childhood, as well as its contribution to the understanding of the development and prevention of depressive symptomatology in children.


Subject(s)
Cognition , Depressive Disorder, Major/diagnosis , Interview, Psychological , Psychological Theory , Social Perception , Child , Child, Preschool , Depressive Disorder, Major/psychology , Humans , Neuropsychological Tests , Prospective Studies , Psychology, Child , Self Concept , Surveys and Questionnaires
9.
Am J Physiol Cell Physiol ; 280(6): C1645-56, 2001 Jun.
Article in English | MEDLINE | ID: mdl-11350761

ABSTRACT

Evidence is accumulating to suggest that actin filament remodeling is critical for smooth muscle contraction, which implicates actin filament ends as important sites for regulation of contraction. Tropomodulin (Tmod) and smooth muscle leiomodin (SM-Lmod) have been found in many tissues containing smooth muscle by protein immunoblot and immunofluorescence microscopy. Both proteins cofractionate with tropomyosin in the Triton-insoluble cytoskeleton of rabbit stomach smooth muscle and are solubilized by high salt. SM-Lmod binds muscle tropomyosin, a biochemical activity characteristic of Tmod proteins. SM-Lmod staining is present along the length of actin filaments in rat intestinal smooth muscle, while Tmod stains in a punctate pattern distinct from that of actin filaments or the dense body marker alpha-actinin. After smooth muscle is hypercontracted by treatment with 10 mM Ca(2+), both SM-Lmod and Tmod are found near alpha-actinin at the periphery of actin-rich contraction bands. These data suggest that SM-Lmod is a novel component of the smooth muscle actin cytoskeleton and, furthermore, that the pointed ends of actin filaments in smooth muscle may be capped by Tmod in localized clusters.


Subject(s)
Carrier Proteins/analysis , Carrier Proteins/genetics , Muscle, Smooth/chemistry , Muscle, Smooth/physiology , Actins/metabolism , Animals , Carrier Proteins/metabolism , Cytoskeleton/chemistry , Cytoskeleton/metabolism , Fluorescent Antibody Technique , Gene Expression/physiology , Microfilament Proteins/chemistry , Microfilament Proteins/metabolism , Muscle Contraction/physiology , Protein Binding/physiology , RNA, Messenger/analysis , Rabbits , Rats , Stomach/chemistry , Tropomodulin
10.
Genomics ; 73(2): 127-39, 2001 Apr 15.
Article in English | MEDLINE | ID: mdl-11318603

ABSTRACT

The 64-kDa autoantigen D1 or 1D, first identified as a potential autoantigen in Graves' disease, is similar to the tropomodulin (Tmod) family of actin filament pointed end-capping proteins. A novel gene with significant similarity to the 64-kDa human autoantigen D1 has been cloned from both humans and mice, and the genomic sequences of both genes have been identified. These genes form a subfamily closely related to the Tmods and are here named the Leiomodins (Lmods). Both Lmod genes display a conserved intron-exon structure, as do three Tmod genes, but the intron-exon structure of the Lmods and the Tmods is divergent. mRNA expression analysis indicates that the gene formerly known as the 64-kDa autoantigen D1 is most highly expressed in a variety of human tissues that contain smooth muscle, earning it the name smooth muscle Leiomodin (SM-Lmod; HGMW-approved symbol LMOD1). Transcripts encoding the novel Lmod gene are present exclusively in fetal and adult heart and adult skeletal muscle, and it is here named cardiac Leiomodin (C-Lmod; HGMW-approved symbol LMOD2). Human C-Lmod is located near the hypertrophic cardiomyopathy locus CMH6 on human chromosome 7q3, potentially implicating it in this disease. Our data demonstrate that the Lmods are evolutionarily related and display tissue-specific patterns of expression distinct from, but overlapping with, the expression of Tmod isoforms.


Subject(s)
Carrier Proteins/genetics , Microfilament Proteins , Amino Acid Sequence , Animals , Autoantigens/biosynthesis , Autoantigens/chemistry , Autoantigens/genetics , Carrier Proteins/biosynthesis , Chromosome Mapping , Chromosomes, Human, Pair 7 , Evolution, Molecular , Expressed Sequence Tags , Gene Duplication , Gene Expression , Genome, Human , Graves Disease/immunology , Humans , Mice , Molecular Sequence Data , Muscle, Smooth/metabolism , Myocardium/metabolism , Sequence Homology, Amino Acid , Tissue Distribution , Tropomodulin
11.
New Hibernia Rev ; 5(3): 66-86, 2001.
Article in English | MEDLINE | ID: mdl-19195110

Subject(s)
Civil Disorders , Cultural Characteristics , Homicide , Judicial Role , Public Health , Social Behavior , Social Problems , Statistics as Topic , Stereotyping , Acculturation , Alcoholic Intoxication/economics , Alcoholic Intoxication/ethnology , Alcoholic Intoxication/history , Alcoholic Intoxication/psychology , Anthropology, Cultural/economics , Anthropology, Cultural/education , Anthropology, Cultural/history , Anthropology, Cultural/legislation & jurisprudence , Civil Disorders/economics , Civil Disorders/ethnology , Civil Disorders/history , Civil Disorders/legislation & jurisprudence , Civil Disorders/psychology , Crime/economics , Crime/ethnology , Crime/history , Crime/legislation & jurisprudence , Crime/psychology , Crime Victims/economics , Crime Victims/education , Crime Victims/history , Crime Victims/legislation & jurisprudence , Crime Victims/psychology , Cultural Diversity , History, 19th Century , Homicide/economics , Homicide/ethnology , Homicide/history , Homicide/legislation & jurisprudence , Homicide/psychology , Ireland/ethnology , Judicial Role/history , Public Health/economics , Public Health/education , Public Health/history , Public Health/legislation & jurisprudence , Punishment/history , Punishment/psychology , Scotland/ethnology , Social Identification , Social Perception , Social Problems/economics , Social Problems/ethnology , Social Problems/history , Social Problems/legislation & jurisprudence , Social Problems/psychology , Statistics as Topic/economics , Statistics as Topic/education , Statistics as Topic/history
12.
Development ; 127(18): 3947-59, 2000 Sep.
Article in English | MEDLINE | ID: mdl-10952893

ABSTRACT

The BMP-like signaling mediated by the ligands Dpp and Gbb is required to reinforce the development of most veins in the Drosophila wing. However, the formation of the cross veins is especially sensitive to reductions in BMP-like signaling. We show here that the formation of the definitive cross veins occurs after the initial specification of the longitudinal veins in a process that requires localized BMP-like activity. Since Dpp and Gbb levels are not detectably higher in the early phases of cross vein development, other factors apparently account for this localized activity. Our evidence suggests that the product of the crossveinless 2 gene is a novel member of the BMP-like signaling pathway required to potentiate Gbb of Dpp signaling in the cross veins. crossveinless 2 is expressed at higher levels in the developing cross veins and is necessary for local BMP-like activity. The Crossveinless 2 protein contains a putative signal or transmembrane sequence, and a partial Von Willebrand Factor D domain similar to those known to regulate the formation of intramolecular and intermolecular bonds. It also contains five cysteine-rich domains, similar to the cysteine-rich domains found in Chordin, Short Gastrulation and Procollagen that are known to bind BMP-like ligands. These features strongly suggest that Crossveinless 2 acts extracelluarly or in the secretory pathway to directly potentiate Dpp or Gbb signaling.


Subject(s)
Bone Morphogenetic Proteins/metabolism , Cysteine/metabolism , Drosophila Proteins , Drosophila melanogaster/embryology , Insect Proteins/chemistry , Insect Proteins/metabolism , Transcription Factors , Veins/embryology , Alleles , Amino Acid Sequence , Animals , Base Sequence , Cloning, Molecular , Cysteine/genetics , DNA-Binding Proteins/metabolism , Drosophila melanogaster/genetics , Drosophila melanogaster/metabolism , Gene Expression Regulation, Developmental , Immunohistochemistry , In Situ Hybridization , Insect Proteins/genetics , Molecular Sequence Data , Mutation/genetics , Protein Structure, Tertiary , Sequence Homology, Amino Acid , Signal Transduction/genetics , Time Factors , Transforming Growth Factor beta/genetics , Transforming Growth Factor beta/metabolism , Veins/metabolism , Wings, Animal/blood supply , Wings, Animal/embryology , Wings, Animal/metabolism
13.
Skeletal Radiol ; 28(10): 590-3, 1999 Oct.
Article in English | MEDLINE | ID: mdl-10550538

ABSTRACT

We present a case of sarcomatoid renal cell carcinoma with multiple sclerotic skeletal metastatic lesions. Renal cell carcinoma is frequently metastatic at presentation, with a high incidence of skeletal involvement, classically described as osteolytic. However, sclerotic or osteoblastic metastatic skeletal lesions from renal cell carcinoma are rare, with only two previous reports identified in the literature, neither of which involved the sarcomatoid variant of renal cell carcinoma. In our case the sclerotic metastases were characterized by bone scan, computed tomography (CT), magnetic resonance imaging (MRI), and histologic analysis.


Subject(s)
Bone Neoplasms/secondary , Carcinoma, Renal Cell/secondary , Kidney Neoplasms/pathology , Sarcoma/secondary , Bone Neoplasms/diagnosis , Bone Neoplasms/pathology , Carcinoma, Renal Cell/diagnosis , Carcinoma, Renal Cell/pathology , Female , Humans , Magnetic Resonance Imaging , Middle Aged , Sarcoma/diagnosis , Sarcoma/pathology , Sclerosis , Tomography, X-Ray Computed
14.
Curr Eye Res ; 19(4): 313-22, 1999 Oct.
Article in English | MEDLINE | ID: mdl-10520227

ABSTRACT

PURPOSE: To evaluate the tissue-specific expression pattern of the 64kD human autoantigen D1, a tropomodulin-related protein that may be involved in thyroid-associated ophthalmopathy. METHODS: Recombinant 64kD human autoantigen D1 was generated in a bacterial expression system and used to immunize rabbits. Specific antibodies were affinity-purified and used for Western blots on normal and hyperthyroid rat and rabbit tissue, and immunofluorescence localization on cryosections of rat tissue. RESULTS: Anti-64kD human autoantigen D1 antibodies recognize specifically a approximately 70kD polypeptide in western blots of extraocular muscle, sternothyroid muscle, and smooth muscle. Immunofluorescence staining demonstrates that the 64kD human autoantigen D1 localizes to myofibrils in slow fibers from rat extraocular and sternothyroid muscle. The level of this protein is not altered in extraocular muscles from hyperthyroid rabbits. CONCLUSIONS: The 64kD human autoantigen D1 is expressed in slow fibers of extraocular and sternothyroid muscles as a component of myofibrils, and is not upregulated in conditions of hyperthyroidism.


Subject(s)
Autoantigens/metabolism , Graves Disease/metabolism , Myofibrils/metabolism , Oculomotor Muscles/metabolism , Proteins/metabolism , Actins/metabolism , Animals , Autoantigens/genetics , Autoantigens/immunology , Blotting, Western , Cytoskeletal Proteins , Electrophoresis, Polyacrylamide Gel , Escherichia coli/genetics , Fluorescent Antibody Technique, Indirect , Gene Expression , Graves Disease/pathology , Humans , Immunoglobulin G/metabolism , Microscopy, Fluorescence , Myofibrils/pathology , Myosins/metabolism , Oculomotor Muscles/pathology , Proteins/genetics , Proteins/immunology , Rabbits , Rats , Recombinant Proteins/genetics , Recombinant Proteins/immunology , Recombinant Proteins/metabolism , Tropomyosin/metabolism
15.
J Biol Chem ; 274(40): 28466-75, 1999 Oct 01.
Article in English | MEDLINE | ID: mdl-10497209

ABSTRACT

Tropomodulin (E-Tmod) is an actin filament pointed end capping protein that maintains the length of the sarcomeric actin filaments in striated muscle. Here, we describe the identification and characterization of a novel tropomodulin isoform, skeletal tropomodulin (Sk-Tmod) from chickens. Sk-Tmod is 62% identical in amino acid sequence to the previously described chicken E-Tmod and is the product of a different gene. Sk-Tmod isoform sequences are highly conserved across vertebrates and constitute an independent group in the tropomodulin family. In vitro, chicken Sk-Tmod caps actin and tropomyosin-actin filament pointed ends to the same extent as does chicken E-Tmod. However, E- and Sk-Tmods differ in their tissue distribution; Sk-Tmod predominates in fast skeletal muscle fibers, lens, and erythrocytes, while E-Tmod is found in heart and slow skeletal muscle fibers. Additionally, their expression is developmentally regulated during chicken breast muscle differentiation with Sk-Tmod replacing E-Tmod after hatching. Finally, in skeletal muscle fibers that coexpress both Sk- and E-Tmod, they are recruited to different actin filament-containing cytoskeletal structures within the cell: myofibrils and costameres, respectively. All together, these observations support the hypothesis that vertebrates have acquired different tropomodulin isoforms that play distinct roles in vivo.


Subject(s)
Actins/metabolism , Carrier Proteins/metabolism , Microfilament Proteins , Muscle, Skeletal/metabolism , Protein Isoforms/metabolism , Amino Acid Sequence , Animals , Base Sequence , Carrier Proteins/genetics , Chick Embryo , Chickens , DNA, Complementary , Molecular Sequence Data , Muscle Development , Muscle, Skeletal/growth & development , Protein Isoforms/genetics , Sequence Homology, Amino Acid , Spectrin/metabolism , Tropomodulin
16.
Mol Cell ; 4(1): 129-35, 1999 Jul.
Article in English | MEDLINE | ID: mdl-10445035

ABSTRACT

We have cloned and characterized JIL-1, a novel tandem kinase in Drosophila that associates with the chromosomes throughout the cell cycle. Antibody staining and live imaging of JIL-1-GFP transgenic flies show that JIL-1 localizes to the gene-rich interband regions of larval polytene chromosomes and is upregulated almost 2-fold on the hypertranscribed male X chromosome compared to autosomes. Phylogenetic analysis suggests that JIL-1 together with human MSKs defines a separate family of tandem kinases. That JIL-1 is a functional kinase was demonstrated by autophosphorylation and phosphorylation of histone H3 in vitro. Based on these findings, we propose that JIL-1 may play a role in transcriptional control potentially by regulating chromatin structure.


Subject(s)
Chromosomal Proteins, Non-Histone/genetics , Drosophila/enzymology , Protein Kinases/genetics , Amino Acid Sequence , Animals , Animals, Genetically Modified , Cell Cycle , Chromatin/chemistry , Chromosomal Proteins, Non-Histone/chemistry , Cloning, Molecular , Drosophila/embryology , Gene Expression Regulation , Histones/metabolism , Humans , Insect Proteins/chemistry , Insect Proteins/genetics , Male , Molecular Sequence Data , Phylogeny , Protein Kinases/chemistry , Salivary Glands/enzymology , X Chromosome/genetics
17.
Leuk Lymphoma ; 33(5-6): 593-6, 1999 May.
Article in English | MEDLINE | ID: mdl-10342588

ABSTRACT

Persistent polyclonal B-cell lymphocytosis with binucleate lymphocytes is a rare lymphoproliferative syndrome of uncertain cause that is strongly associated with HLA-DR7 positivity, cigarette smoking, and female sex. As yet, there is no explanation for the strong sex predilection. We report the third case of persistent polyclonal B-cell lymphocytosis in a male. Other notable findings in this case are lack of HLA-DR7 and strong positive CD5 markers in the polyclonal B-cell population. To our knowledge, CD5 expression has not been mentioned or reported in association with this syndrome.


Subject(s)
B-Lymphocytes/pathology , Lymphocytosis , Adult , B-Lymphocytes/immunology , CD5 Antigens , Cell Differentiation , Female , Humans , Lymphoproliferative Disorders/immunology , Lymphoproliferative Disorders/pathology , Male
19.
Mov Disord ; 13(3): 576-81, 1998 May.
Article in English | MEDLINE | ID: mdl-9613758

ABSTRACT

We report a rare case of Erdheim-Chester disease (ECD) presenting as a progressive cerebellar syndrome and diabetes insipidus. On magnetic resonance imaging, a 7-mm extraaxial, enhancing mass was seen enveloping the right vertebral artery and was confirmed at autopsy to represent an adventitial xanthoma with lipid-laden, foamy histiocytes. The cerebellar syndrome most likely resulted from extensive histiocytic infiltration of the pons, particularly the basis pontis and middle cerebellar peduncles.


Subject(s)
Cerebellar Diseases/diagnosis , Cerebellum/pathology , Granuloma/diagnosis , Histiocytosis, Langerhans-Cell/diagnosis , Lipomatosis/diagnosis , Pons/pathology , Aged , Cerebellar Ataxia/diagnosis , Cerebellar Ataxia/pathology , Cerebellar Diseases/pathology , Cerebellar Nuclei/pathology , Diagnosis, Differential , Follow-Up Studies , Granuloma/pathology , Histiocytosis, Langerhans-Cell/pathology , Humans , Lipomatosis/pathology , Magnetic Resonance Imaging , Male , Medulla Oblongata/pathology
20.
Biotechniques ; 22(3): 488, 491-6, 1997 Mar.
Article in English | MEDLINE | ID: mdl-9067027

ABSTRACT

A modification of a previously described tissue print technique has been developed in which soft tissues are frozen and sectioned in a cryostat prior to direct collection on nitrocellulose or nylon membranes. The inexpensive embedding technique described here allows accurate orientation of specimens prior to mounting, and mounted material may be stored easily after initial sectioning for future reexamination. Standard hand tissue prints of soft specimens exhibit tissue distortion and uneven delivery of material to the membrane, which limits resolution and makes interpretation difficult. Cryostat sections retain tissue fragments in their original arrangement relative to each other during printing and deliver a consistent and quantitative amount of material from all parts of the specimen. The cryostat tissue print technique is applied here to immature floral buds, demonstrating the tissue-level histochemical localization of beta-glucuronidase in transgenic plants and immunolocalization of a novel protein present only in mutant plants. This modified technique is applicable for examining both plant and animal tissues.


Subject(s)
Cryoultramicrotomy/methods , Immunohistochemistry/methods , Plant Proteins/analysis , Plants/chemistry , Frozen Sections/methods , Gene Expression Regulation, Plant , Genes, Plant/genetics , Glucuronidase/analysis , Membranes, Artificial , Plants, Genetically Modified/enzymology , Tissue Embedding/methods , Transformation, Genetic
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