Rapid Differentiation of Epithelial Cell Types in Forensic Samples Using Intrinsic Fluorescence and Morphological Signatures

AbstractEstablishing the tissue source of epithelial cells within a biological sample is an important capability for forensic laboratories. In this study we used Imaging Flow Cytometry (IFC) to analyze individual cells recovered from buccal, contact epithelial, and vaginal samples that had been dried between 24 hours and more than eight weeks. Measurements capturing the size, shape, and fluorescent properties of cells were collected in an automated manner and then used to build a multivariate statistical framework for differentiating cells based on tissue type. Results showed that cell populations from the three tissue types could be differentiated using a Discriminant Function plot of IFC measurements. Epidermal cells were distinguished from vaginal and buccal cells with an average classification accuracy of ~94%. Ultimately, cellular measurements such as these, which can be obtained non-destructively, may provide probative information for many types of biological samples and complement results from standard genetic profiling techniques.

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Telomere length varies substantially between blood cell types in a reptile

Royal Society Open Science ◽

10.1098/rsos.192136 ◽

2020 ◽

Vol 7 (6) ◽

pp. 192136 ◽

Cited By ~ 3

Author(s):

Mats Olsson ◽

Nicholas J. Geraghty ◽

Erik Wapstra ◽

Mark Wilson

Keyword(s):

Blood Cell ◽

Telomere Length ◽

Whole Blood ◽

Phylogenetic Analyses ◽

Natural Populations ◽

Cell Types ◽

Tissue Type ◽

Telomeric Dna ◽

Blood Cell Type ◽

Widespread Phenomenon

Telomeres are repeat sequences of non-coding DNA-protein molecules that cap or intersperse metazoan chromosomes. Interest in telomeres has increased exponentially in recent years, to now include their ongoing dynamics and evolution within natural populations where individuals vary in telomere attributes. Phylogenetic analyses show profound differences in telomere length across non-model taxa. However, telomeres may also differ in length within individuals and between tissues. The latter becomes a potential source of error when researchers use different tissues for extracting DNA for telomere analysis and scientific inference. A commonly used tissue type for assessing telomere length is blood, a tissue that itself varies in terms of nuclear content among taxa, in particular to what degree their thrombocytes and red blood cells (RBCs) contain nuclei or not. Specifically, when RBCs lack nuclei, leucocytes become the main source of telomeric DNA. RBCs and leucocytes differ in lifespan and how long they have been exposed to ‘senescence' and erosion effects. We report on a study in which cells in whole blood from individual Australian painted dragon lizards ( Ctenophorus pictus ) were identified using flow cytometry and their telomere length simultaneously measured. Lymphocyte telomeres were on average 270% longer than RBC telomeres, and in azurophils (a reptilian monocyte), telomeres were more than 388% longer than those in RBCs. If this variation in telomere length among different blood cell types is a widespread phenomenon, and DNA for comparative telomere analyses are sourced from whole blood, evolutionary inference of telomere traits among taxa may be seriously complicated by the blood cell type comprising the main source of DNA.

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Revision of Banded Knifefishes of the Gymnotus carapo and G. tigre clades (Gymnotidae Gymnotiformes) from the Southern Neotropics

Zootaxa ◽

10.11646/zootaxa.4379.1.3 ◽

2018 ◽

Vol 4379 (1) ◽

pp. 47 ◽

Cited By ~ 7

Author(s):

JACK M. CRAIG ◽

LUIZ R. MALABARBA ◽

WILLIAM G. R. CRAMPTON ◽

JAMES S. ALBERT

Keyword(s):

Lateral Line ◽

Electric Organ ◽

Valid Species ◽

Species Boundaries ◽

Multivariate Statistical ◽

Widespread Species ◽

Statistical Framework ◽

Fin Ray ◽

Taxonomic Confusion ◽

Electric Organ Discharges

Banded Knifefishes (Gymnotus, Gymnotidae) comprise the most species-rich, ecologically tolerant (eurytopic), and geographically widespread genus of Neotropical electric fishes (Gymnotiformes), with 40 valid species occupying most habitats and regions throughout the humid Neotropics. Despite substantial alpha-taxonomic work in recent years, parts of the genus remain characterized by taxonomic confusion. Here we describe and delimit species of the G. carapo and G. tigre clades from the southern Neotropics, using body proportions (caliper-based morphometrics), fin-ray, scale and laterosensory-pore counts (meristics), quantitative shape differences (geometric morphometrics), osteology, color patterns and electric organ discharges. We report these data from 174 Gymnotus specimens collected from 100 localities throughout the southern Neotropics, and delimit species boundaries in a multivariate statistical framework. We find six species of the G. carapo clade (G. carapo australis, G. cuia n. sp., G. chimarrao, G. omarorum, G. pantanal, and G. sylvius), and two species of the G. tigre clade (G. inaequilabiatus and G. paraguensis) in the southern Neotropics. The new species G. cuia is readily distinguished from the morphologically similar and broadly sympatric G. c. australis by a shorter head and deeper head and body, and from the morphologically similar and sympatric G. omarorum by fewer lateral-line ventral rami and fewer pored lateral-line scales anterior to the first ventral ramus. We also review the geographic distributions of all eight species of the G. carapo and G. tigre clades in the southern Neotropics, showing that G. cuia is the most widespread species in the region. These results affirm the importance of understanding the structure of variation within and between species, both geographic and ontogenetic, in delimiting species boundaries.

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Immunocytochemical demonstration of tissue-type plasminogen activator in endocrine cells of the rat pituitary gland.

The Journal of Cell Biology ◽

10.1083/jcb.101.1.305 ◽

1985 ◽

Vol 101 (1) ◽

pp. 305-311 ◽

Cited By ~ 40

Author(s):

P Kristensen ◽

L S Nielsen ◽

J Grøndahl-Hansen ◽

P B Andresen ◽

L I Larsson ◽

...

Keyword(s):

Growth Hormone ◽

Pituitary Gland ◽

Endocrine Cells ◽

Large Population ◽

Cell Types ◽

Anterior Lobe ◽

Tissue Type ◽

Intermediate Lobe ◽

Posterior Lobe ◽

Rat Pituitary

We immunocytochemically stained rat pituitary glands using antibodies against plasminogen activators of the tissue type (t-PA) and the urokinase type (u-PA). A large population of endocrine cells in the anterior lobe of the gland displayed intense cytoplasmic immunoreactivity with anti-t-PA. In some areas of the intermediate lobe we found a weak staining, and we observed weakly staining granular structures in the posterior lobe. Controls included absorption of the antibodies with highly purified t-PA. In addition, SDS PAGE followed by immunoblotting of pituitary gland extracts revealed only one band with an electrophoretic mobility similar to that of t-PA when stained with anti-t-PA IgG. No u-PA immunoreactivity was detected in the rat pituitary gland. Sequential staining experiments using antibodies against growth hormone and t-PA demonstrated that the t-PA-immunoreactive cells constitute a large subpopulation of the growth hormone-containing cells. These findings represent the first direct evidence for the presence of t-PA in cell types other than endothelial cells in the intact normal organism. In this article we discuss the implications of the results for a possible role of t-PA in the posttranslational processing of prohormones.

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Mouse connective tissue mast cell proteases tryptase and carboxypeptidase A3 play protective roles in itch induced by endothelin-1

Journal of Neuroinflammation ◽

10.1186/s12974-020-01795-4 ◽

2020 ◽

Vol 17 (1) ◽

Author(s):

Elín I. Magnúsdóttir ◽

Mirjana Grujic ◽

Jessica Bergman ◽

Gunnar Pejler ◽

Malin C. Lagerström

Keyword(s):

Mast Cell ◽

Mast Cells ◽

Connective Tissue ◽

Treatment Options ◽

Cell Types ◽

Tissue Type ◽

Human Mast Cell ◽

Knock Out ◽

Endothelin 1 ◽

Deficient Mice

Abstract Background Itch is an unpleasant sensation that can be debilitating, especially if it is chronic and of non-histaminergic origin, as treatment options are limited. Endothelin-1 (ET-1) is a potent endogenous vasoconstrictor that also has the ability to induce a burning, non-histaminergic pruritus when exogenously administered, by activating the endothelin A receptor (ETAR) on primary afferents. ET-1 is released endogenously by several cell-types found in the skin, including macrophages and keratinocytes. Mast cells express ETARs and can thereby be degranulated by ET-1, and mast cell proteases chymase and carboxypeptidase A3 (CPA3) are known to either generate or degrade ET-1, respectively, suggesting a role for mast cell proteases in the regulation of ET-1-induced itch. The mouse mast cell proteases (mMCPs) mMCP4 (chymase), mMCP6 (tryptase), and CPA3 are found in connective tissue type mast cells and are the closest functional homologs to human mast cell proteases, but little is known about their role in endothelin-induced itch. Methods In this study, we evaluated the effects of mast cell protease deficiency on scratching behavior induced by ET-1. To investigate this, mMCP knock-out and transgenic mice were injected intradermally with ET-1 and their scratching behavior was recorded and analyzed. Results CPA3-deficient mice and mice lacking all three proteases demonstrated highly elevated levels of scratching behavior compared with wild-type controls. A modest increase in the number of scratching bouts was also seen in mMCP6-deficient mice, while mMCP4-deficiency did not have any effect. Conclusion Altogether, these findings identify a prominent role for the mast cell proteases, in particular CPA3, in the protection against itch induced by ET-1.

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Detection of Circulating Tumor Cells Using Membrane-Based SERS Platform: A New Diagnostic Approach for ‘Liquid Biopsy’

Nanomaterials ◽

10.3390/nano9030366 ◽

2019 ◽

Vol 9 (3) ◽

pp. 366 ◽

Cited By ~ 11

Author(s):

Agnieszka Kamińska ◽

Tomasz Szymborski ◽

Evelin Witkowska ◽

Ewa Kijeńska-Gawrońska ◽

Wojciech Świeszkowski ◽

...

Keyword(s):

Tumor Cells ◽

Circulating Tumor Cells ◽

Principal Component ◽

Cell Types ◽

Surface Enhanced Raman Spectroscopy ◽

Statistical Technique ◽

Diagnostic Efficiency ◽

Multivariate Statistical ◽

Multi Stage ◽

Filtration Enrichment

The detection and monitoring of circulating tumor cells (CTCs) in blood is an important strategy for early cancer evidence, analysis, monitoring of therapeutic response, and optimization of cancer therapy treatments. In this work, tailor-made membranes (MBSP) for surface-enhanced Raman spectroscopy (SERS)-based analysis, which permitted the separation and enrichment of CTCs from blood samples, were developed. A thin layer of SERS-active metals deposited on polymer mat enhanced the Raman signals of CTCs and provided further insight into CTCs molecular and biochemical composition. The SERS spectra of all studied cells—prostate cancer (PC3), cervical carcinoma (HeLa), and leucocytes as an example of healthy (normal) cell—revealed significant differences in both the band positions and/or their relative intensities. The multivariate statistical technique based on principal component analysis (PCA) was applied to identify the most significant differences (marker bands) in SERS data among the analyzed cells and to perform quantitative analysis of SERS data. Based on a developed PCA algorithm, the studied cell types were classified with an accuracy of 95% in 2D PCA to 98% in 3D PCA. These results clearly indicate the diagnostic efficiency for the discrimination between cancer and normal cells. In our approach, we exploited the one-step technology that exceeds most of the multi-stage CTCs analysis methods used and enables simultaneous filtration, enrichment, and identification of the tumor cells from blood specimens.

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Internationalization of SMEs: a research note

Journal of Small Business and Enterprise Development ◽

10.1108/14626000210419473 ◽

2002 ◽

Vol 9 (1) ◽

pp. 38-48 ◽

Cited By ~ 18

Author(s):

Paul Westhead ◽

Martin Binks ◽

Deniz Ucbasaran ◽

Mike Wright

Keyword(s):

Environmental Variables ◽

Firm Growth ◽

Employment Growth ◽

Telephone Survey ◽

Multivariate Statistical ◽

Activity Data ◽

Performance Variables ◽

Statistical Framework ◽

Profit Performance ◽

Survey Responses

In 1990/91, survey responses were gathered from 621 independent businesses located in Great Britain. A follow‐on telephone survey was conducted with 150 surviving firms in 1997. This survey gathered information surrounding the propensity of firms to export their goods or services abroad as well as other performance and goal outcomes. Organizational and external environmental variables collected in 1990 are used to explain within a multivariate statistical framework the propensity of a firm to be an exporter in 1997, and the intensity of internationalization activity. Data collected in 1990 is also used to explain variations in several performance variables (i.e. whether exporting was regarded as a path to firm growth; profit performance reported in 1997 relative to competition; and the propensity to report employment growth over the 1990 to 1997 period).

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Human tissue-type plasminogen activator

Thrombosis and Haemostasis ◽

10.1160/th13-06-0517 ◽

2014 ◽

Vol 112 (08) ◽

pp. 243-254 ◽

Cited By ~ 27

Author(s):

Egbert K. O. Kruithof ◽

Sylvie Dunoyer-Geindre

Keyword(s):

Plasminogen Activator ◽

Gene Transcription ◽

Current Knowledge ◽

Plasminogen Activator Inhibitor ◽

Cell Types ◽

Tissue Type ◽

Tissue Type Plasminogen Activator ◽

Type Plasminogen Activator ◽

Central Nervous System Activity ◽

Its Gene

SummaryTissue-type plasminogen activator (t-PA ) plays an important role in the removal of intravascular fibrin deposits and has several physiological roles and pathological activities in the brain. Its production by many other cell types suggests that t-PA has additional functions outside the vascular and central nervous system. Activity of t-PA is regulated at the level of its gene transcription, its mRNA stability and translation, its storage and regulated release, its interaction with cofactors that enhance its activity, its inhibition by inhibitors such as plasminogen activator inhibitor type 1 or neuroserpin, and its removal by clearance receptors. Gene transcription of t-PA is modulated by a large number of hormones, growth factors, cytokines or drugs and t-PA gene responses may be tissue-specific. The aim of this review is to summarise current knowledge on t-PA function and regulation of its pericellular activity, with an emphasis on regulation of its gene expression.

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Soft-Tissue Analogue Design and Tissue Engineering of Liver

MRS Bulletin ◽

10.1557/s0883769400031857 ◽

1996 ◽

Vol 21 (11) ◽

pp. 52-54 ◽

Cited By ~ 5

Author(s):

Prabhas V. Moghe

Keyword(s):

Tissue Engineering ◽

Health Care Cost ◽

Three Dimensional ◽

Cell Types ◽

The United States ◽

Tissue Loss ◽

Tissue Type ◽

Design Parameters ◽

Immune Rejection ◽

Engineered Tissue

Tissue engineering involves the application of physical and life sciences to develop functional substitutes for dysfunctional organs or tissue structures. From an engineering standpoint, tissues contain two basic components—the cells that are organized into proper units, and the material surrounding the cells, called the extracellular matrix (ECM). A third, frequently overlooked feature essential to the maintenance of the activity of the engineered tissue is the three-dimensional architecture of the cell-matrix composite.A comprehensive review of the scope and impact of tissue engineering has previously appeared. Tissue-engineered devices have the potential to reduce the annual health-care cost related to tissue loss and end-stage organ failure to the order of $400 billion, eight million invasive surgical procedures, and 65 million hospital days. A common approach to engineer a functional tissue is to place cells derived from a healthy organ or tissue type (identical or similar to the dysfunctional tissue/organ) on or within matrices analogous to host-tissue ECM. These systems can then be enclosed in appropriate membranes that isolate cells from immune rejection following implantation or can be transplanted directly with the administration of drugs that prevent the immune rejection. Another application of these systems is for extracorporeal (outside the patient's body) device support of a dysfunctional organ. In either instance, the success of the engineered tissue depends critically on the interactions of cells with the tissue analogues. The objective of this article is to outline the simplest matrix-design parameters to control these interactions. While organs are comprised of very different tissue types, for the sake of simplicity, this article is primarily pertinent to the tissue engineering of one major organ, the liver. The choice of this tissue type is intended to serve as a comprehensive generalization of many different cell types since in the diversity and complexity of its activities, the liver has few parallels. The development of an artificial liver is also critically awaited, as in the United States alone, 35,000 people, including the many wait listed for the exorbitant liver organ transplants ($300,000), die each year of chronic liver disorders. In many other countries, liver disease is the second leading cause of death.

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Evidence for carbohydrate-independent endocytosis of tissue-type plasminogen activator by liver cells

Biochemical Journal ◽

10.1042/bj2850799 ◽

1992 ◽

Vol 285 (3) ◽

pp. 799-804 ◽

Cited By ~ 5

Author(s):

E Stang ◽

N Roos ◽

M Schlüter ◽

T Berg ◽

J Krause

Keyword(s):

Plasminogen Activator ◽

Liver Cell ◽

Hepatic Clearance ◽

Mannose Receptor ◽

Cell Types ◽

Tissue Type ◽

Side Chains ◽

Tissue Type Plasminogen Activator ◽

Hepatic Cells ◽

Type Plasminogen Activator

In the liver, tissue-type plasminogen activator (t-PA) is endocytosed by hepatic parenchymal (PC), endothelial (EC) and Kupffer (KC) cells. Although the endocytosis is receptor-mediated, it remains a matter of discussion which receptors are involved in this catabolic process. To evaluate the role of a protein-specific receptor, as well as the possible involvement of the galactose receptor on PC and the mannose receptor on EC, we have employed different glycosylation variants of t-PA in biochemical and immunocytochemical studies. Partial or total removal of carbohydrate side-chains by endoglycosidases did not prevent clearance and hepatic endocytosis of t-PA by either of the liver cell types. Blockade of the galactose and mannose receptors by co-application of a large excess of the glycoprotein ovalbumin remained without effect on the binding and uptake of t-PA by hepatic cells. However, the contribution of different liver cell types to the hepatic clearance of t-PA was to a certain extent dependent on the type of oligosaccharide chains removed. The mannose receptor on EC is partially responsible for the clearance of t-PA by this cell type, whereas the galactose receptor does not seem to be involved in this process. The results obtained in this study further demonstrate that the major portion of the hepatic catabolism of t-PA is independent of its carbohydrate side-chains.

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Light microscopy survey of extant gymnosperm root protophloem and comparison with basal angiosperms

Botany ◽

10.1139/cjb-2013-0284 ◽

2014 ◽

Vol 92 (5) ◽

pp. 388-401 ◽

Cited By ~ 3

Author(s):

Thomas C. Pesacreta ◽

Michael A. Purpera

Keyword(s):

Root Apex ◽

Cell Types ◽

Tissue Type ◽

Root Tip ◽

Chemical Fixation ◽

Physiological Mechanisms ◽

Sieve Elements ◽

Phloem Parenchyma ◽

Extant Species ◽

Large Central Vacuole

Gymnosperm root protophloem is not well understood. There is a question as to whether root protophloem cells mature as phloem parenchyma, or as sieve elements, or if within the protophloem there is an anatomical and evolutionary gradient having characteristics of both cell types. This question is relevant to understanding anatomical and physiological mechanisms that supply nutrients to the root tip. Anatomical data from a broad range of species show that gymnosperms have one to three layers of parenchymatous protophloem cells located at the vascular cylinder periphery between the pericyle and the metaphloem. In some species, these cells are associated with secretory idioblasts. Near the root apex, protophloem cells develop a large central vacuole and, in transverse sections, their radial walls tend to be radially elongated. When mature, these cells are highly longitudinally elongated. Only these cells exhibit surging toward the root apex during chemical fixation. These data indicate that protophloem of gymnosperm roots lacks sieve elements. Because of its distinctive anatomical characteristics and the absence of sieve elements, gymnosperm root protophloem is a vegetative synapomorphy among extant species. The restriction of this tissue type to gymnosperms supports the hypothesis that it originated in a progenitor of that clade.

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