Alkaline phosphatase activity in normal and scorbutic bones.
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Alkaline Phosphatase↗
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Complexities in ETS-domain transcription factor function and regulation: lessons from the TCF (ternary complex factor) subfamily. The Colworth Medal Lecture.
The ETS-domain transcription factor family can be divided into a series of subfamilies. Elk-1 represents the founding member of the ternary complex factor (TCF) subfamily. By focusing on the TCF subfamily, we can demonstrate the complexities that exist in the function and regulation of ETS-domain transcription factors. This article focuses on Elk-1 in detail and summarizes the functions of other TCFs. The key themes covered include the domain structure of the TCFs, the mechanisms of complex formation with serum response factor, regulation of TCFs by mitogen-activated protein kinase cascades, and transcriptional regulatory properties of the TCFs. Finally, the emerging role of the TCFs in vivo is discussed. A picture is developing indicating that, while these proteins exhibit significant sequence and functional conservation, key differences in their structure and regulation are being identified which may relate to unique functions of these proteins in vivo.
Amino Acid Sequence↗
Matrix metalloproteinases (MMPs) in fresh human prostate tumour tissue and organ-cultured prostate tissue: levels of collagenolytic and gelatinolytic MMPs are low, variable and different in fresh tissue versus organ-cultured tissue.
Prostate tissue was obtained from 22 radical prostatectomies (performed for clinical management of prostate carcinoma) immediately after surgery. A small piece of tissue was fixed immediately in formalin and used for routine histology while a second piece was frozen in OCT and used for immuno-histochemistry. Another small piece was used for isolation of epithelial and stromal cells. The remainder of the tissue was cut into 2 x 2 mm pieces and incubated in organ culture for 8 days. In organ culture, non-malignant, basal epithelial cells underwent a proliferative response. This was accompanied by de-differentiation of glandular structures and by migration of epithelial cells across the surface of the tissue. Erosion of the basement membrane could also be seen in places, but was not widespread. Invasion of epithelial cells into the adjacent stroma was not evident. Production of matrix metalloproteinases (MMPs) with gelatinolytic activity or collagenolytic activity was assessed in organ culture and compared to expression patterns in fresh tissue. MMP-1 (interstitial collagenase) and MMP-9 (92-kDa gelatinase B) were undetectable or low in fresh tissue specimens. Both enzymes were detected in organ culture and both increased over time. Even after 6 days, however, there was only a low level of gelatin-hydrolytic activity and no measurable collagen-hydrolytic activity. In past studies we used organ cultures of normal skin and malignant skin tumours (basal cell carcinomas) to help elucidate the role of collagenolytic and gelatinolytic MMPs in epithelial cell invasion (Varani et al, 2000). Compared to MMP levels observed in skin, levels of these enzymes in prostate are low. The low level of collagenolytic and gelatinolytic MMPs in fresh prostate tissue and in organ-cultured prostate tissue may help explain why there is little tissue destruction in many primary prostate tumours and why the majority of such tumours remain confined to the prostate for extended periods.
Collagen↗
[Lasers in dentistry. Part B--Interaction with biological tissues and the effect on the soft tissues of the oral cavity, the hard tissues of the tooth and the dental pulp].
The interaction of laser energy with target tissue is mainly determined by two non operator-dependent factors: the specific wavelength of the laser and the optical properties of the target tissues. Power density, energy density, pulse repetition rate, pulse duration and the mode of energy transferring to the tissue are dictated by the clinician. Combination of these factors enables to control optimal response for the clinical application. Four responses are described when the laser beam hits the target tissue: reflection, absorption, transmission and scattering. Three main mechanisms of interaction between the laser and the biological tissues exist: photothermic, photoacoustic and photochemical. The effect of lasers on the soft tissues of the oral cavity is based on transformation of light energy into thermal energy which, in turn heats the target tissue to produce the desirable effect. In comparison to the scalpel used in surgical procedures, the laser beam is characterized by tissue natural sterility and by minimum bleeding during the surgical procedures due to blood vessels welding. The various effects achieved by the temperature elevation during the laser application on the soft tissue are: I. coagulation and hemostasis II. tissue sterilization III. tissue welding IV. incision and excision V. ablation and vaporization Ablation and melting are the two basic modalities by which the effect of lasers on the hard tissues of the tooth is produced. When discussing the effect of laser on dental hard tissues, the energy absorption in the hydroxyapatite plays a major role in addition to its absorption in water. When laser energy is absorbed in the water of the hard tissues, a rapid volume expansion of the evaporating water occurs as a result of a substantial temperature elevation in the interaction site. Microexplosions are produced causing hard tissue disintegration. If pulp temperatures are raised beyond 5 degrees C level, damage to the dental pulp is irreversible. Histologically, after laser ablation, presence of odontoblastic nuclei is important. Consistency and composition of the intracellular tissue is another factor influencing cell viability. If heat is intensive and exists for an extended time, the consistency of the intracellular ground substance may not be preserved. Accordingly, the application of excessive energy densities has been shown to result in significant damage to pulp tissue and in particular to odontoblasts. Studies showed that the use of Er:YAG laser to treat dental hard tissues is both safe and effective for caries removal, cavity preparation and enamel etching.
Absorption↗
Tissue polypeptide antigen and tissue polypeptide specific antigen in primary breast cancer. Evaluation in serum and tumour tissue.
Tissue polypeptide antigen, measured by both a polyclonal antibody (TPA IRMA Prolifigen) and a monoclonal antibody prototype kit (TPA-M IRMA Prolifigen), and the tissue polypeptide specific antigen were evaluated. The markers were measured in 266 serum samples and in 291 tumour cytosols from patients with primary breast cancer. The three markers were available in matched pairs of both serum and cytosol from the same patient in 144 cases. Diagnostic sensitivity of serum levels of the three markers was not significantly different when using cut-off values calculated on the basis of healthy subjects. In the cytosol, tissue polypeptide antigen (TPA IRMA), tissue polypeptide antigen (TPA-M IRMA) and tissue polypeptide specific antigen were significantly correlated with steroid receptor status, while their serum levels were not. Cytosol and serum levels of the three markers were not significantly associated. All three were significantly correlated both in serum and in cytosol. The association was closer between tissue polypeptide antigen (TPA IRMA) and tissue polypeptide antigen (TPA-M IRMA) than between each of these two markers and tissue polypeptide specific antigen. From these findings we draw the following conclusions: 1. Tissue polypeptide specific antigen (TPA IRMA) and tissue polypeptide antigen (TPA-M IRMA) probably provide superimposable information both in serum and in cytosol; 2. Tissue polypeptide specific antigen and tissue polypeptide antigen (TPA IRMA) or tissue polypeptide antigen (TPA-M IRMA), although closely associated, probably measure in part different cytokeratins. Therefore, they should not be considered interchangeable in individual patients; 3. The determination of the markers in serum and in cytosol provides different information concerning the tumour phenotype.
Antibodies, Monoclonal↗
Antigen-induced release of histamine from rat tissues in vitro: dissociation in development of serosal mast cell, lung tissue, and tracheal tissue response capacity.
We examined the temporal development and the fading in Sprague Dawley rats, actively sensitized to ovalbumin (OA), of the capacity of serosal mast cells, chopped lung tissue, and occasionally chopped tracheal tissue, to respond at antigen challenge in vitro with histamine release. Response capacity of both serosal mast cells and lung tissue developed within 2-3 weeks after injection of 1 microgram OA or more together with 100 mg of alum. Maximum response capacity was observed in cells and tissue from animals injected with 10 micrograms OA, part of the response capacity then remained until 3 months after immunization. Development of serosal mast cell reactivity was occasionally dissociated from that of lung tissue. When low amounts of alum (1 or 10 mg) were employed as adjuvant, lung tissue reactivity could be induced in the virtual absence of serosal mast cell response capacity. Silica gel was less efficient than alum as an adjuvant for induction of a primary response, but 'secondary' tissue responses could be induced when silica gel was used as an adjuvant. Pretreatment of the animals with cyclophosphamide before the booster injection enhanced and prolonged the response capacity of lung tissue. Animals injected with OA together with Freund's complete adjuvant did not provide responding serosal mast cells; response capacity of lung tissue varied with immunization dose of antigen. Antigen-induced histamine release from chopped tracheal tissue did not correlate to response capacity of lung tissue. Thus, the development in the rat of response capacity with respect to antigen-induced histamine release dissociates from serosal mast cells, lung tissue, and tracheal tissue.
Adjuvants, Immunologic↗
Comparison of the release of adipokines by adipose tissue, adipose tissue matrix, and adipocytes from visceral and subcutaneous abdominal adipose tissues of obese humans.
The purpose of this study was to examine the source of adipokines released by the visceral and sc adipose tissues of obese humans. Human adipose tissue incubated in primary culture for 48 h released more prostaglandin E(2), IL-8, and IL-6 than adiponectin, whereas the release of plasminogen activator inhibitor 1 and hepatocyte growth factor was less than that of adiponectin but greater than that of leptin. IL-10 and TNFalpha were released in amounts less than those of leptin, whereas vascular endothelial growth factor and IL1-beta were released in much lower amounts. The accumulation of adipokines was also examined in the three fractions (adipose tissue matrix, isolated stromovascular cells, and adipocytes) obtained by collagenase digestion of adipose tissue. Over 90% of the adipokine release by adipose tissue, except for adiponectin and leptin, could be attributed to nonfat cells. Visceral adipose tissue released greater amounts of vascular endothelial growth factor, IL-6, and plasminogen activator inhibitor 1 compared with abdominal sc tissue. The greatly enhanced total release of TNFalpha, IL-8, and IL-10 by adipose tissue from individuals with a body mass index of 45 compared with 32 was due to nonfat cells. Furthermore, most of the adipokine release by the nonfat cells of adipose tissue was due to cells retained in the tissue matrix after collagenase digestion.
Abdomen↗
Comprehensive expression profiling of highly homologous 39 hox genes in 26 different human adult tissues by the modified systematic multiplex RT-pCR method reveals tissue-specific expression pattern that suggests an important role of chromosomal structure in the regulation of hox gene expression in adult tissues.
Homeobox genes play a crucial role as molecular address labels in early embryogenesis by conferring cell fate and establishing regional identity in tissues. Homeobox gene expression is not restricted to the early development, but it is also observed in the differentiated cells in adult tissues. To have a better understanding of the functionality of homeobox gene expression in adult tissues in physiological and pathological phenomena, it is important to determine the expression profiles of Hox genes. We established a system to study the expression of 39 human Hox genes by the modified Systematic Multiplex RT-PCR method. Using this system, we have systematically examined their expression in 26 different adult tissues. The results showed tissue-specific differential expression. They also revealed that the posterior tissues generally express more Hox genes than the anterior tissues and that the genes located centrally in the Hox Gene Complexes are expressed in more tissues than the genes located at the 5' or 3' end of the complexes. Instead of similar expression patterns among paralogous genes, we found that several neighboring Hox genes on the same chromosomes exhibited similar tissue-specific expression pattern, which may suggest that the regulation of Hox gene expression may be more dependent on chromosomal structure in adult tissues.
Adult↗
Elevated tissue concentrations of sialyl Lex-i in cancerous tissues compared with those in noncancerous tissues of various organs.
Sialyl Lex-i (SLX) concentrations in the extracts of noncancerous and cancerous tissues of various human organs were determined by radioimmunoassay for detailed evaluation of SLX. Cancerous tissues had significantly elevated SLX concentrations compared with noncancerous tissues of various organs. Tissue SLX concentration of the cancerous part was significantly higher than that of the adjacent noncancerous part in the same tissue. There was no significant correlation between tissue SLX concentration and serum SLX level. Positive localization of SLX was clearly observed in such cancerous tissues by immunohistochemical study, although not in any noncancerous tissues. Each of the antigens: SLX, CA 19-9, carcinoembryonic antigen, and CA 125 showed a different distribution pattern in tissue concentration or localization in various organs. These results indicate that SLX may be a valuable cancer-associated antigen produced by malignant tissues, suggesting its clinical application as a tumor marker.
Aged↗
The significance of folic acid, tissue iron stores, and tissue viability in determining iron uptake from serum by thyroid tissue slices.
This paper describes an attempt to measure in vitro iron uptake from serum by human thyroid slices and to relate the uptake to tissue iron stores, folic acid status, and tissue viability. It is an extension of work previously reported (Buchanan, 1969). Thyroids were obtained from patients undergoing partial thyroidectomy for colloid goitre and serum from clinically normal healthy adults. The haemoglobin, serum iron, and folic acid levels of both thyroid and serum donors were measured and thyroids examined histologically for the presence of stainable iron. Viable and non-viable tissue slices were incubated in sera treated with radioactive iron so as to produce high and normal levels of transferrin saturation. Iron was taken up both from sera with normal and high transferrin saturation but the amount was, in almost all cases, greater from the more highly saturated. The uptake by non-viable tissue was appreciable but did not vary to any great extent from one serum to the next, and was attributed to simple diffusion of ionic iron into the tissue. There was, however, marked variation in uptake from different sera by viable tissue. It was concluded therefore that viability is a factor affecting the uptake. As the variation in uptake by viable tissue incubated in a single serum was significantly less than tissue incubated in a number of different sera it was further concluded that there was also a factor in the serum itself affecting iron uptake. The nature of the factor was not elucidated but neither folic acid nor levels of iron stores appeared to influence uptake because no correlation was found between iron uptake and iron stores or folic acid.
Adult↗
Distribution of tissue polypeptide antigen (TPA) in normal human tissues: Immunohistochemical study on unfixed, methanol-, ethanol-, and formalin-fixed tissues.
The distribution of tissue polypeptide antigen (TPA) was studied in unfixed, methanol-, 95% ethanol-1% acetic acid (EA)-, and formalin-fixed paraffin-embedded sections of all adult human tissues using an indirect immunoperoxidase method. The specific staining patterns were virtually identical in unfixed and alcohol-fixed tissues, but in formalin-fixed tissues this similarity was found only after fixation for up to 24 hr and pretreatment with protease for 15 min. Although prolongation of formalin fixation beyond 48 hr increasingly diminished the TPA reactivity, TPA could still be demonstrated in tissues fixed in formalin for up to 6 months. TPA was found to be a cytoplasmic constituent of almost all adult human duct and cavity lining, simple, and stratified epithelia. TPA was not demonstrated in epidermis, renal proximal convoluted and testicular tubules, basket-like myoepithelial cells, nor in most glandular acini, including hepatocytes and pancreatic acinar cells. The TPA staining was also negative in all non-epithelial tissues, including lymph nodes and bone marrow. The well-defined epithelial distribution and the comparable demonstrability in differently preserved tissues make TPA a useful tool for the identification of cells of epithelial character.
Ethanol↗
Tissue distribution of arsenic species in rabbits after single and multiple parenteral administration of arsenic trioxide: tissue accumulation and the reversibility after washout are tissue-selective.
Parenteral administration of arsenic trioxide has recently been recognized as an effective antineoplastic therapy, especially for the treatment of acute promyelocytic leukemia. Its efficacy and toxicity are concentration-dependent and are related to the fractions of different arsenic species and the degree of methylation. In this study, arsenic trioxide was given parenterally to rabbits as a single dose or as a daily dose (0.2, 0.6, and 1.5 mg/kg) for 30 days. The blood and organ concentrations of the arsenic species, including As(III), dimethylarsinic acid (DMA), and monomethylarsonic acid (MMA), were studied on day 1 (single-dose study), day 30 (multiple dosing study), and day 60 (reversibility study). As(III) was the major detectable arsenic species in the blood. The pharmacokinetic parameters (total clearance, area under the curve, etc.) for As(III) indicated a limit for the capacity to eliminate As(III) at the dose of 1.5 mg/kg, and were quite the same after a single dose or chronic multiple dosing. In tissues, DMA was found to be the major metabolite and the concentrations of DMA, As(III), and MMA in general increased with the dose, with the increase most significant at a dose of 1.5 mg/kg. However, normalized tissue distribution of As(III) in the kidney on day 1, but not on day 30, was nonlinear. Along with decreased levels of As(III) and increased levels of DMA, an inducible capacity for methylating As(III) to DMA after chronic dosing in kidney was suggested. The tissue concentration of DMA was highest in lung and liver, and the normalized tissue distributions in liver on day 30 were nonlinear, suggesting a limit in eliminating DMA after a chronic high load of As(III). Tissue concentrations of As(III), DMA, and MMA in bladder increased dramatically after chronic dosing. However, after washout for 30 days, As(III), DMA, and MMA were all undetectable in bladder and liver. However, As(III) in hair and low levels of DMA in lung, kidney, heart and hair were still detected. In conclusion, in rabbits we found a similar pharmacological profile after a single dose or chronic multiple dosing of parenteral arsenic trioxide, with a limiting metabolizing capacity at a dose of 1.5 mg/kg. Tissue accumulation of arsenic species, mainly DMA, and its reversibility after washout were tissue-selective. The potential for late toxicities of arsenic trioxide in organs with a significant tendency for arsenic accumulation with low reversibility should be closely monitored.
Animals↗
An Act (No. 65 of 1983) to provide for the donation or the making available of human bodies and tissue for the purposes of medical or dental training, research or therapy or the advancement of medicine or dentistry in general; for the post-mortem examination of certain human bodies; for the removal of tissue, blood and gametes from the bodies of living persons and the use thereof for medical or dental purposes; for the control of the artificial insemination of persons; and for the regulation of the import and export of human tissue, blood and gametes; and to provide for matters connected therewith. Date of commencement: 12 July 1985 (the Human Tissue Act, 1983)
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Blood Donors↗
[Filatov's tissue therapy. III. Wound hormonal effect of Filatov's refrigerated tissues, aloes and fish-liver oil on washed kohlrabi tuber; limitation of tissue-poisoning effect of unrefrigerated tissues].
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Aloe↗
Describing patients' normal tissue reactions: concerning the possibility of individualising radiotherapy dose prescriptions based on potential predictive assays of normal tissue radiosensitivity. Steering Committee of the BioMed2 European Union Concerted Action Programme on the Development of Predictive Tests of Normal Tissue Response to Radiation Therapy.
Clinical radiotherapeutic doses are limited by the tolerance of normal tissues. Patients given a standard treatment exhibit a range of normal tissue reactions, and a better understanding of this individual variation might allow for individualisation of radiotherapeutic prescriptions, with consequent improvement in the therapeutic ratio. At present, there is no simple way to describe normal tissue reactions, which hampers communication between clinic and laboratory and between groups from different centres. There is also no method for comparing the severity of reactions in different normal tissues. This arises largely because there is no definition of a "normal" reaction, an "extreme" reaction or the particular term "over-reactor" (OR). This report proposes definitions for these terms, as well as a simple terminology for describing normal tissue reactions in patients having radiotherapy. The "normal" range represents the individual variation in normal tissue reactions amongst large numbers of patients treated in the same way which is within clinically acceptable limits. The term "OR" is applied to an individual whose reaction is more severe than the normal range but also implies that this forced a major change in the radiotherapeutic prescription or that the reactions were very severe or fatal. A "severe OR" would develop serious problems with a typical radical dose, while an "extreme OR" would have such difficulties at a much lower dose. To describe the normal range, a numerical scale is suggested, from 1 to 5, resistant to sensitive. The term "highly radiosensitive" (HR) is suggested for category 5. An "informal" relative scale, as suggested here, is quick and simple. It should allow comparison between different hospitals, compensate for differences in radiotherapeutic dose and technique and allow comparison of reactions between different anatomical sites. It should be adequate for discriminating patients at the extremes of the normal range from those at the centre. It is hoped that the definitions and terminology proposed here will aid communication in the field of predictive testing of normal tissue radiosensitivity.
Follow-Up Studies↗
Prediction of in vivo tissue distribution from in vitro data. 2. Influence of albumin diffusion from tissue pieces during an in vitro incubation on estimated tissue-to-unbound plasma partition coefficients (Kpu).
PURPOSE: To determine the extent of albumin diffusion from tissue pieces into medium during in vitro incubations, to develop and assess the utility of mathematical models describing this effect on the estimation of tissue-to-unbound plasma partition coefficients (Kpu) of drug substances and to derive factors to correct for associated errors. METHODS: Twelve separate tissues were obtained from rats sacrificed by cervical dislocation, 48 h after an intravenous dose of 125I-human albumin, and tissue pieces incubated to determine the efflux of albumin into media over 2 to 4 h. A mathematical model was developed to predict and correct for the effect of albumin diffusion on the measured Kpu values of drugs. RESULTS: The model predicted that the effect of albumin diffusion from tissue pieces during in vitro incubation (ranging from 14 to 59% remaining in tissue) on Kpu values was generally minimal, except for compounds that are highly plasma bound and have a low measured Kpu. Under these circumstances, the measured Kpu substantially underestimates the true value. An equation was derived from readily available or measurable parameters to correct for this underestimation. CONCLUSIONS: Albumin diffuses from tissue pieces into protein free media during in vitro incubations until equilibrium is reached, defined by the albumin Kpu. Model predictions indicated that for the majority of compounds albumin diffusion would have a minimal effect on the measured Kpu value and that a correction factor could be calculated to account for any deviation.
Albumins↗
Muscular tissue engineering: capillary-incorporated hybrid muscular tissues in vivo tissue culture.
Requirements for a functional hybrid muscular tissue are 1) a high density of multinucleated cells, 2) a high degree of cellular orientation, and 3) the presence of a capillary network in the hybrid tissue. Rod-shaped hybrid muscular tissues composed of C2C12 cells (skeletal muscle myoblast cell line) and type I collagen, which were prepared using the centrifugal cell-packing method reported in our previous article, were implanted into nude mice. The grafts, comprised three hybrid tissues (each dimension, diameter, approximately 0.3 mm, length, approximately 1 mm, respectively), were inserted into the subcutaneous spaces on the backs of nude mice. All nude mice that survived the implantation were sacrificed at 1, 2, and 4 wk after the implantation. The grafts were easily distinguishable from the subcutaneous tissues of host mice with implantation time. The grafts increased in size with time after implantation, and capillary networks were formed in the vicinities and on the surfaces of the grafts. One week after implantation, many capillaries formed in the vicinities of the grafts. In the central portion of the graft, few capillaries and necrotic cells were observed. Mononucleated myoblasts were densely distributed and a low number of multinucleated myotubes were scattered. Two weeks after implantation, the formation of a capillary network was induced, resulting in the surfaces of the grafts being covered by capillaries. Numerous elongated multinucleated myotubes and mononucleated myoblasts were densely distributed and numerous capillaries were observed throughout the grafts. Four weeks after implantation a dense capillary network was formed in the vicinities and on the surfaces of the grafts. In the peripheral portion of the graft, multinucleated myotubes in the vicinities of the rich capillaries were observed. Thus, hybrid muscular tissues in vitro preconstructed was remodeled in vivo, which resulted in facilitating the incorporation of capillary networks into the tissues.
Animals↗
Tissue engineering of skeletal muscle. Highly dense, highly oriented hybrid muscular tissues biomimicking native tissues.
A highly dense, highly oriented hybrid muscular tissue was devised using C2C12 cells (skeletal muscle myoblast cell line) and Type I collagen. A cold mixture of C2C12 cells suspended in DMEM (Dulbecco's modified Eagle's medium; Gibco Lab Inc., Grand Island, NY) and Type I collagen solution was poured into capillary tube molds of two different sizes (inner diameters: 0.90 mm and 0.53 mm, respectively) sealed at each end. After centrifugation (1000 RPM, 5 min) and subsequent thermal gelation, a rod shaped gel was formed. The resultant gel shrank to become a highly dense tissue after incubation on an agarose gel coated dish. Small diameter rod shaped tissues were composed of numerous multi-nucleated myotubes and a few necrotic cells. On the other hand, a ring shaped tissue fabricated by centrifugation with a specially devised agarose gel mold was subjected to cyclic stretching at 60 RPM. The resultant highly dense, highly oriented hybrid muscular tissue involved both densely accumulated cells and collagen fiber bundles, which tended to be aligned in the direction of stretching. Sequential procedures of a centrifugal cell packing method and a mechanical stress loading method facilitated fabrication of hybrid muscular tissues similar to native muscular tissues in terms of cell density and orientation.
Animals↗