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M Witt

Publications and source records attributed to M Witt.

At least 55 records · Page 3Linked to original sources
PubMed1999

Immunohistochemical investigations on the differentiation marker protein E11 in rat calvaria, calvaria cell culture and the osteoblastic cell line ROS 17/2.8.

Until now, many extracellular matrix proteins, e.g. osteopontin and osteonectin, have been used to determine a cell's osteogenic maturation. The disadvantage in evaluation of these proteins is their relative wide-ranging appearance throughout the osteogenic differentiation process. Thus, the aim of this study was to establish an immunohistochemical setup using E11, a marker that binds selectively to cells of the late osteogenic cell lineage. In addition, the histochemical expression of the bone matrix proteins osteonectin, osteopontin and fibronectin was compared to that of E11 using monoclonal antibodies. For light microscopical detection of osteogenic markers in cultured cells we developed a simple paraffin technique using a fibrin glue as embedding medium. This allows the handling of cultured cells such as a tissue sample and includes the use of stored biological specimens for further immunohistochemical experiments. We used newborn rat calvariae for whole tissue preparations and for isolation and cultivation of bone cells. In addition, we included the rat osteosarcoma cell line ROS 17/2.8 in this study. For the first time, we have localised E11 in osteocytes of rat calvaria preparations at the electron microscopical level. E11 was detected at plasma membranes of osteocytes and their processes, but not at those of osteoblasts. Accompanying experiments with cultured newborn rat calvaria cells and ROS 17/2.8 cells revealed E11 reactivity on a subset of cells. The results obtained confirm the suitability of the differentiation marker E11 as a sensitive instrument for the characterisation of bone cell culture systems.

Animals↗
PubMed1999

Distribution of cytokeratin filaments and vimentin in developing human taste buds.

Taste buds in humans originate from approximately the 8th postovulatory week under the influence of ingrowing nerve fibers. Since they develop from local epithelium, it is of interest whether or not prospective taste cells maintain or develop characteristics of epithelial cells that are different from those of the adjacent epithelium during differentiation. The aim of this study was to monitor changes of the distribution of the cytokeratin filaments (CKs) 8, 18, 19 and 20 ("gastrointestinal" type), CK 7 ("ductal" type), and CK 13 (maturation "mucosa type"), as well as vimentin in developing human taste buds and adjacent squamous epithelium. With the exception of CK13, which remains negative in taste bud anlagen and adult taste buds, all cytokeratins tested were present in taste cells. With the progress of development, the distribution of CKs becomes more and more restricted to taste cells and salivatory ducts as well as Ebner gland cells. Only CK20 is exclusively specific to taste bud anlagen and sometimes to individual bipolar cells occurring in early stages (week 8-9). Vimentin was located mainly in mesodermal derivatives but also in perigemmal epithelial cells during all stages of development. The occurrence of vimentin in "borderline" epithelia that interface with underlying connective tissue, i.e., in a region of discontinuity, may be associated with particular events in development, cell migration or even dedifferentiation.

Adult↗
PubMed1999

Immunohistochemical localization of the differentiation marker E11 in dental development of rats.

E11 antigen, originally characterized in a rat osteosarcoma cell line, is known to be expressed during late stages of the osteogenic cell lineage both in vitro and in vivo. The aim of the present study was to monitor the occurrence and distribution patterns of the E11 antigen using monoclonal antibodies (mAb E11 and MEP-1) during different stages of tooth germ development of new-born rats by means of immunohistochemistry. Both antibodies strongly bound to plasma membranes of ameloblasts in presecretory and secretory stages. In addition, odontoblasts and cells of the periodontium were immunoreactive for E11 and MEP-1. During maturation, the immunoreactivity of ameloblast plasma membranes decreased significantly. Our data suggest that E11 and MEP-1 might be important as markers for cell differentiation and mineralization processes during tooth germ development.

Ameloblasts↗
PubMed1999

The influence of low doses 131I-induced maternal hypothyroidism on the development of rat embryos.

The aim of our research was to create and verify a model for studying the effects of a low dose of 131I and 131I-induced maternal hypothyroidism on the development of the embryo's thyroid gland and brain. The given dose (150 microCi) corresponds to the absorbed dose of 0.5 Gy. This dose is similar to that dose received by large numbers of the population of the C.I.S. regions polluted by radioactive isotopes of iodine as a result of the Chernobyl accident in 1985. Thirty-five female Wistar rats and their 168 newborn pups were used for observation. The females were divided into a control group and four experimental groups (each distinguished by the time of 131I injection: group I - no less than 12 days before mating; groups II, III and IV - on 5th, 10th and 16th days of gestation, respectively). In all the experimental female groups the incorporate dose of 131I led to hypothyroidism accompanied by a 43% reduction in the thyroxin level and by a nearly 8-fold increase in the TSH level. However, the influence of maternal hypothyroidism on the development of the thyroid gland and brain of embryos depends on the time when 131I took effect. There is a reduction in the weight of the newborns' brain and thyroid gland, total body mass. The hormonal status of the newborns' thyroid gland also changes. The proposed model will allow us to study many aspects of induced changes in the brain and thyroid gland of the embryos which develop under conditions of maternal hypothyroidism resulting from a low dose of 131I, administered at the critical times of development.

Animals↗
PubMed1999

Progressive multifocal leukoencephalopathy and human immunodeficiency virus-associated white matter lesions in AIDS: magnetization transfer MR imaging.

PURPOSE: To determine the magnetization transfer features of progressive multifocal leukoencephalopathy (PML) and human immunodeficiency virus (HIV)-associated white matter lesions (WML) (hereafter, HIV-WML) on magnetic resonance (MR) images obtained in patients with acquired immunodeficiency syndrome (AIDS). MATERIALS AND METHODS: Conventional MR imaging and magnetization transfer MR imaging were performed in 21 AIDS patients with 42 areas of white matter hyperintensity on MR images (13 patients had 25 PML lesions, eight patients had 17 WML). The magnetization transfer ratio was calculated for each lesion. RESULTS: Compared with normal-appearing white matter (magnetization transfer ratio = 47.9%), both PML and HIV-WML showed reduced magnetization transfer ratio. The magnetization transfer ratio was significantly lower in PML lesions (magnetization transfer ratio = 26.1%) than in HIV-WML (magnetization transfer ratio = 38.0%, P < .0001), and there was no overlap in the magnetization transfer ratio between PML lesions and HIV-WML. The separation in magnetization transfer ratio between the two lesion types was valid for lesion as small as 0.5 cm2. CONCLUSION: The larger reduction in magnetization transfer ratio for PML lesions is most likely due to demyelination, whereas the reduction in HIV-WML may be associated primarily with gliosis. PML lesions appear to cause strong reductions in magnetization transfer ratio early in the course of disease. Magnetization transfer MR imaging is a noninvasive tool that improves the differentiation between PML and HIV-WML in patients with AIDS.

AIDS Dementia Complex↗
PubMed1999

Distribution of vimentin in the developing chick taste bud during the perihatching period.

The tissue environment within which taste bud cells develop has not been wholly elaborated. Previous studies of taste bud development in vertebrates, including the avian chick, have suggested that taste bud cells could arise from one, or several tissue sources (e.g. crest-mesenchyme, local ectoderm or endoderm). Thus, molecular markers which are present in gemmal as well as interfacing (peribud epithelium; mesenchyme-epithelium) regions, and their degree of expression during stages of taste bud development, are of special interest. The intermediate filament protein, vimentin, occurs in mesenchymal and mesodermally-derived (e.g. endothelial, fibroblast) cells as well as highly proliferating epithelium (e.g. tumors). The present study in chick gustatory tissue utilized antibodies against vimentin and the avidin-biotin-peroxidase technique to evaluate vimentin immunoreactivity (IR) within a timeframe which includes: 1) early stages of the taste bud primordium [embryonic days (E)17-E18)]; 2) the beginning of an accelerated bud cell proliferation at the time of initial, taste bud pore opening [around E19]; 3) attaining the adult complement of taste buds [around posthatch (H) day 1], and 4) completed organogenesis (H 17). During this time span, vimentin-IR was characterized in a region including and sometimes bridging taste bud and subepithelial connective tissue, whereas non-gustatory surrounding epithelium and salivary glands were vimentin-immuno-negative. Intragemmally, the proportion of vimentin-IR cells as related to total taste bud cells peaked at E19. These results indicate that vimentin expression, in part, is related to the onset of taste bud cell proliferation and suggest that mesenchyme could be one source of taste bud cells. Secondly, fibronectin, an extracellular matrix component of the epithelial basement membrane interface with mesenchyme, was expressed at or near the apical surfaces of taste bud cells projecting into the bud lumen, and in the basal gemmal region suggesting the possible role of fibronectin as a chemotactic anchor for differentiating and migrating taste bud receptor cells. Lastly, neuron-specific enolase-IR indicates that axonal varicosities are already present intragemmally at E17-E18, that is, during the incipient period of identifiable taste bud primordia.

Animals↗
PubMed1999

[Frequency of mutations and genotypes of the CFTR gene in cystic fibrosis adults in Poland].

Based on the Polish Registry of Cystic Fibrosis the analysis of alive CF adults (18 y.a. and more) was performed. Out of 156 Polish CF adults, in 106 (68.0%) identification of the CFTR gene mutations was performed. Relative frequencies of 24 mutations of CFTR gene mutations is 72.2%. Frequencies of the most common mutations are: delta F508 49.06%, 3849 + 10kbCT 8.96%, G542X 1.89%; in Polish CF adults mutations other than delta F508 comprise 23.13% of CF alleles. The most common genotype delta F508/delta F508 occurs with the frequency 22.6%, significantly lower than in CF children; genotype delta F508/3849 + 10kbCT occurs with the frequency 11.3%, and this value is significantly higher than in other age groups.

Adult↗
PubMed1998

Immunohistochemical study of fibronectin and thyroglobulin in the thyroid gland of female rats after exposure to radioactive iodine.

The aim of this work was to study the effect of a dose of 150 microCi 131I on the barrier properties of the thyroid epithelium in pregnant female rats. Thirty-five female Wistar rats were divided into a control and four experimental groups (each distinguished by the time of 131I injection: group I--no less then 12 days before mating; groups II, III, and IV--on 5th, 10th, and 16th days of gestation, respectively). The thyroid glands were fixed in Bouin's fluid, embedded in paraffin, and stained immunohistochemically for thyroglobulin and fibronectin. In group IV the appearance of follicles with fibronectin-positive colloid demonstrates the penetration of blood plasma into the follicular lumen. There are more fibronectin positive follicles in group III. Regardless of the nature of the follicles' contents, numerous thyrocytes with an intensive fibronectin positive reaction begin to appear in the follicles. In group II the number of fibronectin positive follicles and thyrocytes is clearly reduced, and in group I only a few remain. In group IV there is a noticeable reduction in the quantity of colloid inside the follicles and often an absence of any thyroglobulin positive reaction. There are thyrocytes in which thyroglobulin positive granules localized in the basal zone. There is thyroglobulin positive staining in the stroma and blood vessels. In group II thyroglobulin is no longer found in the stroma. Small doses of 131I provoke a serious breakdown in the thyroid epithelium's barrier properties, although these changes are of a transient nature. The central zone of the thyroid gland reacts more actively and dynamically to exposure to radioactive iodine than the peripheral zone.

Animals↗
PubMed1998

Innervation of developing human taste buds. An immunohistochemical study.

Morphological changes in developing human gustatory papillae during the 6th to the 23rd postovulatory week have been studied. The general innervation pattern of taste papillae and taste bud primordia was revealed immunohistochemically using antibodies against protein gene product 9.5 (PGP9.5), neurofilament H (NFH), neurofilament L (NFL), neurone-specific enolase (NSE), and tubulin. The autonomic and somatosensory nerve supply has been investigated using antibodies against substance P (SP), calcitonin gene-related peptide (CGRP), tyrosine hydroxylase (TH), neuropeptide Y (NPY), the neuronal form of nitric oxide synthase (n-NOS), and, enzyme histochemically, NADPH-diaphorase. Nerve fibers approach the basal membrane of the lingual epithelium around the 7th postovulatory week and invade the epithelium of papilla-like structures at the 8th week, but some also penetrate the basal membrane of the non-papillary epithelium. They are in close contact with slender epithelial cells that are considered to be the taste bud's progenitor cells. Early human taste buds situated at the anterior part of the tongue do not necessarily require a dermal (later fungiform) papilla. The NADPH-diaphorase reaction revealed positive results in dermal nerve fibers, but the immunohistochemical reaction against n-NOS was negative. Immunohistochemical detection of neuropeptides and vasoactive substances rendered negative results for developmental stages of 7-18 postovulatory weeks. By the 18th week, only SP was detected in dermal papillae, but not in the vicinity of taste buds' primordia. Thus, autonomic and somatosensory nerves seem not to play a key role in formation and maintenance of early human taste buds.

Biomarkers↗
PubMed1998

Immunohistochemical distribution of CD44 and some of its isoforms during human taste bud development.

Taste buds are accumulations of elongated bipolar cells situated on lingual papillae. The factors that determine the sites where a taste bud may develop are largely obscure, although it is known that the early invasion of nerve fibers plays one of the key roles in taste bud development and maturation. The conditions under which taste bud primordium cells develop are influenced by the interaction between epithelial cells and extracellular matrix molecules of the mesenchyma, such as hyaluronan. Thus, we investigated immunohistochemically the distribution pattern of the receptor for hyaluronan, CD44s, and its epithelial variant isoforms CD44v6 and CD44v9, in taste buds of human embryonic, fetal, perinatal, and adult tongues. Furthermore, we wanted to determine the temporal and spatial relationships of CD44 to sensory innervation of taste bud primordia. In early gestational stages (weeks 7-9), CD44 and its isoforms are expressed on membranes of apical perigemmal (marginal) cells covering taste bud primordia. It seems that CD44 serves as a marker for marginal cells (perigemmal cells) in early developmental stages. The expression of CD44 follows rather than precedes the invasion of sensory nerve fibers and the development of taste bud primordia (weeks 7-8). In new-born and adult taste bud cells, only the standard molecule, CD44s, is expressed; the variant isoforms, CD44v6 and CD44v9, occur only in the adjacent epithelium. From these results it is likely that marginal cells are of the utmost importance for the development and maturation of taste buds. We presume that CD44 is involved in local binding, reuptake, and degradation of hyaluronan in the early stages of taste bud formation. CD44 probably does not induce the transformation of epithelial cells into taste bud primordial cells. What is more, CD44 may change its function in the course of developmental events.

Adult↗
PubMed1998

Distribution of N-cadherin and NCAM in neurons and endocrine cells of the human embryonic and fetal gastroenteropancreatic system.

For the first time, the distribution of N-cadherin and neural cell adhesion molecule (NCAM) as well as some neuropeptides in nerve cells and endocrine cells of the human embryonic and fetal gastroenteropancreatic system has been detected in early stages (from the 6th postovulatory week onwards). Epithelial cells of the stomach and small intestine contained gastrin and somatostatin and the epithelium of the small intestine also bombesin-positive cells. Myenteric ganglionic cells showed both bombesin and VIP and were NCAM- and N-cadherin-positive at all ages studied. Some basally granulated epithelial cells of stomach, duodenum and the upper part of jejunum contained N-cadherin. The number of these cells increased from 6th to 10th postovulatory weeks. Nerve cells and the cytoplasm of individual epithelial cells of pancreatic ducts were immunoreactive for NCAM and N-cadherin. NCAM- and N-cadherin-positive cells also appeared in Langerhans islets (> 10 weeks), mainly in their peripheral part. NCAM- and N-cadherin-positive endocrine cells were less numerous than endocrine cells producing somatostatin, bombesin, and VIP, probably reflecting the features of embryonic/fetal histogenesis of Langerhans islets from epithelial endocrine cells of pancreatic ducts. NCAM and N-cadherin were localized on the surface of endocrine islets cells as well as in the cytoplasm of single islet cells. This suggests the involvement of both membrane and soluble forms of adhesion proteins in embryonic/fetal histogenesis of human pancreatic islets. The early occurrence of N-cadherin (6th postovulatory week) in enteroendocrine cells supports the existence of a common precursor. The expression of NCAM and N-cadherin in nerve cells and endocrine cells of the human fetal gastroenteropancreatic system may indicate the involvement of neuronal adhesion mechanisms in the development of neuro-endocrine complexes of fetal stomach, small intestine and pancreas.

Cadherins↗
PubMed1998

Hints of a functional connection between the neuropeptidergic innervation of arteriovenous anastomoses and the appearance of epithelioid cells in the rabbit ear.

Peripheral blood flow can be regulated by specialized vessel segments, the arteriovenous anastomoses. Their wall consists of a relatively thick layer of smooth muscle cells and so-called epithelioid cells. The epithelioid cell is a specialized myogenic cell phenotype expressing nitric oxide synthase. We studied the innervation of the different segments of arteriovenous anastomoses in the rabbit ear using antisera against neuropeptide Y, tyrosine hydroxylase, calcitonin gene-related peptide and substance P, as well as neuron-specific enolase, calbindin D and neurotubulin. The participation was especially examined of neuropeptidergic innervation and a possible morphological connection to the occurrence of epithelioid cells and a paracrine function. The NADPH diaphorase reaction and alpha-smooth muscle actin immunoelectron microscopy served to distinguish epithelioid cells from smooth muscle cells. Using conventional fluorescence microscopy and confocal laser scanning microscopy, we found the most dense innervation pattern of pan-neuronal markers (neurotubulin, neuron-specific enolase), tyrosine hydroxylase-immunoreactive nerve fibres and neuropeptidergic nerve fibres (neuropeptide Y, calcitonin gene-related peptide, substance P) around the intermediate segment in arteriovenous anastomoses, whereas the venous segment was barely marked. Single nerve fibres penetrated into the medial layer and reached the epithelioid cells. Using immunoelectron microscopy, we found intercellular contacts between epithelioid cells, but not the gap junction protein connexin 43. Here, we report for the first time a correlation of the innervation pattern with epithelioid cell type in arteriovenous anastomoses. Our findings suggest that epithelioid cells of the arteriovenous anastomoses are controlled by a dense network of neuropeptidergic nerve fibres in functional connection to their paracrine role as a nitric oxide producer.

Animals↗
PubMed1998

Application of microsatellite markers for monitoring of reconstitution pattern in patients after bone marrow transplantation/peripheral blood progenitor cell transplantation.

We present data on application of microsatellite markers for detection of complete vs. mixed chimerism in individuals after bone marrow transplantation (BMT) and peripheral blood progenitor cell transplantation (PBPCT). Genotypes of the peripheral blood or bone marrow cells of three donor/recipient pairs were studied at several microsatellite loci. In two cases we identified complete chimerism in recipients' peripheral blood and bone marrow after transplantation. These two patients were alive with no symptoms of disease relapse and/or transplant rejection long time after BMT or PBPCT. In the third patient we observed mixed chimerism with the prevalence of donor's genotype while she was alive and well and the prevalence of recipient's genotype when she relapsed and required interferonotherapy. Results of our molecular studies correlated with the patients' clinical outcome. We show that molecular detection of mixed chimerism may be used for prediction of relapse in patients after BMT or PBPCT.

Adolescent↗
PubMed1997

Immunohistochemical detection of human skin nerve fibers.

The autonomic nervous system is involved in different functions such as transduction of afferent sensory inputs, trophic actions, modulation of immunologic events and thermoregulation. In the present investigation, we studied the pattern of human autonomic skin innervation with special reference to its relation to blood vessels, hair follicles, sweat glands and sensory receptors. For the first time, two clinically important areas have been compared: the skin of the forearm and of the face. Using indirect immunohistochemistry, we analyzed the distribution of calretinin (CR), calcitonin gene-related peptide (CGRP), neuropeptide Y (NPY), substance P (SP), neurokinin A (NKA), vasoactive intestinal peptide (VIP), nitric oxide synthase (NOS), tyrosine hydroxylase (TH), histamine, serotonin, enkephalin, and, enzyme histochemically, NADPH-diaphorase (NADPH-d). In the epidermis, we found nerve fibers containing SP, NKA and CGRP. In the dermis, SP-, CR-, VIP-, CGRP- and NKA-positive nerve fibers were detected. Particularly the large nerve fibers contained CR. VIP-positive fibers occurred especially around hair follicles and sweat glands. CGRP-positive nerve fibers were located close to the epidermal basal membrane, in the wall of blood vessels, and to a lesser extent around hair follicles. Immunoreactivity for SP and NKA in the dermis was observed predominantly in the papillary layer near the epidermal basal membrane. All neuropeptides tested in this study were also detected in the nerve fibers of the subcutis. Most of them were CGRP- and VIP-positive. They occurred in association with sweat glands and large arteries. NPY-positive nerve fibers are predominant in the wall of arteries, arterioles and veins. Nerve fibers containing NKA and SP were less common and identified only in the walls of large arteries in deeper dermal layers. In double-staining experiments, the NADPH-d reaction and reactivity to tubulin revealed a partial co-localization in nerve fibers, blood vessel walls, around glands and ganglionic cells. VIP-positive fibers were more common in the face skin than in the forearm. However, in forearm we detected more NPY-, CGRP-, NKA- and SP-positive nerve fibers than in face skin. These findings are important for future studies on skin disorders, such as sensory neuropathies, inflammatory reactions or allergic responses of human skin.

Blood Vessels↗
PubMed1997

Scanning electron microscopical studies of developing gustatory papillae in humans.

Development and morphological changes of human gustatory papillae during postovulatory weeks 6-15 have been studied using scanning and transmission electron microscopy. The first papilla of the tongue appears around postovulatory week 6 in its caudal midline near the foramen caecum. In contrast, the dorsal epithelium of the anterior part of the tongue shows only small hillock- or papilla-like elevations from week 6 on, which comprise an aggregation of 5-20 epithelial cells. From week 7 on, most prominent fungiform papillae develop near the median sulcus and at the margins of the anterior part of the tongue. At their tops, the first primitive taste pores are found around week 10; these are often covered with processes of adjacent epithelial cells. Most pores, however, develop around weeks 14-15. The maturation of taste buds does not coincide with the appearance of taste pores, since taste bud cells are not fully differentiated in the observed period of time. Fungiform papillae are developed before filiform papillae, which do not occur within the first 15 weeks of gestation. Fungiform papillae tend to grow between weeks 8 and 15 of gestation, whereas the size of vallate papillae seems to be constant during this period.

Humans↗
PubMed1997

Repeated measures estimation of measurement bias for self-reported drug use with applications to the National Household Survey on Drug Abuse.

Direct estimates of response bias for self-reports of drug use in surveys require that essentially error free determinations of drug use be obtained for a subsample of survey respondents. The difficulty of obtaining determinations which are accurate enough for estimating validity is well-documented in the literature. Methods such as specimen (e.g., hair, urine) analysis, proxy reports, and the use of highly private and anonymous modes of interview all have to contend with error rates which may only be marginally lower than those of the parent survey. Thus, any methodology for direct validity estimation must rely to some extent on approximations and questionable assumptions. In this chapter, the authors consider a number of methods that rely solely on repeated measures data to assess response bias. Since the assumptions associated with these approaches do not require highly accurate second determinations they may be more easily satisfied in practice. One such method for bias estimation for dichotomous variables that is considered in some detail provides estimates of misclassification probabilities in the initial measurement without requiring that the second measure be accurate or even better than the first. This methodology does require, however, that two subpopulations exist which have different rates of prevalence but whose probabilities of false positive and false negative error are the same. The applicability of these methods for self-reported drug use are described and illustrated using data from the National Household Survey on Drug Abuse. In the discussion of the results, the importance of these methods for assessing the validity of self-reported drug use are examined.

Bias↗
PubMed1997

[Chylothorax in the course of squamous cell lung cancer].

Chylothorax is a rare condition which might be caused by trauma of the chest, malignant tumors, congenital malformation, or inflammation of lymphatic nodes. It occurs as a complication of malignancy in about 50% of all cases, either as the first and the only symptom or at a very late stage of the disease. We presented a case of the 62 year old male with squamous cell cancer of the right lung, treated by chemo- and radiotherapy. Leakage of lymph to the pleura complicated the clinical status of the patient. Due to the primary disease the patient was treated symptomatically, in the hospital and later on in the out-patient clinic. Actually, eleven years after being diagnosed, despite massive fibrosis of the right lung the patient is doing relativity well, working part time.

Antineoplastic Combined Chemotherapy Protocols↗