Eye Muscle Tissue Research Articles

On the gamma radiation response of commercially available 3D printing materials for medical dosimetry

3D printing technology has rapidly spread for decades, allowing the fabrication of medical implants and human phantoms and revolutionizing healthcare. The objective of this study is to evaluate some radiological properties of commercially available 3D printing materials as potential tissue mimicking materials. Among fifteen materials, we compared their properties with nine human tissues. In all materials and tissues, exposure and energy absorption buildup factors were calculated for photon energies between 0.015 and 15 MeV and penetration depths up to 40 mean free path. Furthermore, the Geant4 Monte Carlo toolkit (version 10.5) was used to simulate their percentage depth dose distributions. In addition, equivalent atomic numbers, effective atomic numbers, attenuation coefficients, and CT numbers have been examined. All parameters were considered in calculating the average relative error (σ), which was used as a statistical comparison tool. With σ between 6 and 7, we found that Polylactic Acid (PLA) was capable of simulating eye lenses, blood, soft tissue, lung, muscle, and brain tissues. Moreover, Polymethacrylic Acid (PMAA) material has a σ value of 4 when modeling adipose and breast tissues, respectively. Aside from that, variations in 3D printing materials’ infilling percentage can affect their CT numbers. We therefore suggest the PLA for mimicking soft tissue, muscle, brain, eye lens, lung and blood tissues, with an infill of between 92.7 and 94.3 percent. We also suggest an 89 percent infill when simulating breast tissue. Furthermore, with a 96.7 percent infill, the PMAA faithfully replicates adipose tissue. Additionally, we found that a 59 percent infill of Fe-PLA material is comparable to cortical bone. Due to the benefits of creating individualized medical phantoms and equipment, the results might be seen as an added value for both patients and clinicians.

RETRACTED ARTICLE: Relationship between smoking and serum levels of eye muscle and orbital connective tissue antibodies in patients with Graves ophthalmopathy

Over the past three decades, several studies have quantified the risk of smoking in the development of ophthalmopathy in patients with Graves' hyperthyroidism, with an overall odds ratio of approximately 3.0. Smokers also have a greater risk of more advanced ophthalmopathy than non-smokers. We studied 30 patients with Graves' ophthalmopathy (GO) and 10 patients with upper eyelid signs as the only manifestation of ophthalmopathy, whose eye signs were assessed using the clinical activity score (CAS), NOSPECS classes and upper eyelid retraction (UER) score, half of whom were smokers and half of whom were non-smokers. Serum levels of eye muscle (CSQ, Fp2, G2s) and orbital connective tissue type XIII collagen (Coll XIII) antibodies are valuable markers of ophthalmopathy in patients with Graves' disease. Still, their relationship to smoking has not been investigated. These antibodies were measured by enzyme-linked immunosorbent assay (ELISA) in all patients as a component of their clinical management. Mean serum antibody levels of all four antibodies were significantly greater in smokers than in non-smokers in patients with ophthalmopathy but not in those with upper eyelid signs only. As determined using one-way ANOVA and Spearman's correlation test, there was a significant correlation between smoking severity, assessed as pack-years, with mean Coll XIII antibody level, but not with levels of the 3 eye muscle antibodies. These results suggest that in patients with Graves' hyperthyroidism who smoke, the orbital inflammatory reactions are more advanced than in those with Graves' hyperthyroidism who do not smoke. The mechanism of this enhanced Autoimmunity against orbital antigens in smokers is unclear and worthy of further study.

Исследование качества и безопасности мяса диких животных, полученного в условиях биоценоза Белоосиповского ртутного месторождения

Introduction. Mercury contamination is one of the most common environmental problems. The research objective was to study the qualitative composition and physicochemical properties of raw game meat obtained from the area near the Beloosipovo mercury deposit in order to define any possible contamination w ith xenobiotics.
 Study objects and methods. The research featured rib eye muscle tissue and soft flesh of elks shot on the hunting farms of the Kemerovo Region aka Kuzbass.
 Results and discussion. A complex set of experiments revealed the chemical composition of elk muscle tissue and flesh, as well as the mineral composition of elk muscle tissue. The samples were obtained from different parts of carcasses. The amino acid and fatty acid composition of elk muscle tissue made it possible to describe the biological value, mineral composition, and vitamin profile of elk meat. The physicochemical analysis included toughness, cooking losses, and moisture-retaining capacity, i.e. the properties that ensure juiciness. The research also featured the accumulation of xenobiotics in elk meat samples obtained from the biosinosis near the Beloosipovo merc ury deposit.
 Conclusion. The slaughter yield of elk meat was 51–53%, which exceeds the average yield of farm cattle meat by 4–6%. The moisture content was 73–78%, while the content of protein was between 20–24% and depended on the anatomical location of the muscle sample; the fat content reached 0.75–1.75%. The mercury accumulation at different storage temperature conditions ranged from 0.004 ± 0.001 to 0.009 ± 0.001 mg/kg, while the max imum allowable concentration of mercury is 0.03 mg/kg.

Antibodies against human eye muscle tissue fractions bind to type 2 5'deiodinase and inhibit the lymphocyte MAPK activation derived from hyperthyroid patients with Graves' disease

Searchable abstracts of presentations at key conferences in endocrinology ISSN 1470-3947 (print) | ISSN 1479-6848 (online)

Anti-eye Muscle IgG and IgM Antibodies are Associated with Eye Muscle Type 2 Deiodinase Activities in Hyperthyroid Graves Ophthalmopathy

Objective: Antibodies to eye muscle tissue are demonstrated in hyperthyroid Graves’ disease but their pathognomonic roles have not been clarified yet. We suggest that type 2 deiodinase (DIO2) enzyme can be a target between orbital and thyroid diseases in hyperthyroid Graves’ ophthalmopathy. In this study, the relationship between anti-eye muscle membrane (EyeM) and cytosol (EyeC) IgG, IgA and IgM antibodies and eye muscle DIO2 activity was investigated in hyperthyroid Graves’ disease. Methods: Thirty-two patients with hyperthyroid Graves’ disease (20 had ophthalmopathy), mean age 36 ± 13 years, 27 females and 5 males formed the patient group. Thyroid hormone levels were measured with chemiluminescence immunoassay, antithyroid antibody and anti-eye muscle antibody levels with enzyme-linked immunosorbent assay, but TSH receptor antibodies with radioimmunoassay. Eye muscle DIO2 activity was measured using 125I-T4. Results: EyeM and EyeC IgG antibodies were associated with increased eye muscle DIO2 activity compared to those who were negative for these antibodies (2.79 ± 2.53 vs 17.88 ± 20.06 pmol/mg/min, P<0.015 for EyeM-IgG and 6.88 ± 9.62 vs 26.76 ± 23.86 pmol/mg/min, P<0.008 for EyeC-IgG). The difference in EyeC-IgM antibodies was significant between patients with and without ophthalmopathy (10.1 ± 4.73 vs 3.46 ± 3.18 pmol/mg/min, P<0.023). The presence of EyeM-IgM antibodies was associated with lower eye muscle DIO2 activities with significantly decreased eye muscle thicknesses (8.51 ± 15.38 vs 2.6 ± 0.1 pmol/mg/min and 4.71 ± 1.23 vs 3.9 ± 0.14 mm, P<0.04). Conclusion: Anti-eye muscle antibodies were associated with increased or decreased eye muscle DIO2 activities affecting eye muscle thicknesses in hyperthyroid Graves’ ophthalmopathy. DIO2 enzyme can be a new target between orbital and thyroid diseases in hyperthyroid Graves’ ophthalmopathy.

Essential roles of LEM-domain protein MAN1 during organogenesis in Xenopus laevis and overlapping functions of emerin

Mutations in nuclear envelope proteins are linked to an increasing number of human diseases, called envelopathies. Mutations in the inner nuclear membrane protein emerin lead to X-linked Emery–Dreifuss muscular dystrophy, characterized by muscle weakness or wasting. Conversely, mutations in nuclear envelope protein MAN1 are linked to bone and skin disorders. Both proteins share a highly conserved domain, called LEM-domain. LEM proteins are known to interact with Barrier-to-autointegration factor and several transcription factors. Most envelopathies are tissue-specific, but knowledge on the physiological roles of related LEM proteins is still unclear. For this reason, we investigated the roles of MAN1 and emerin during Xenopus laevis organogenesis. Morpholino-mediated knockdown of MAN1 revealed that MAN1 is essential for the formation of eye, skeletal and cardiac muscle tissues. The MAN1 knockdown could be compensated by ectopic expression of emerin, leading to a proper organ development. Further investigations revealed that MAN1 is involved in regulation of genes essential for organ development and tissue homeostasis. Thereby our work supports that LEM proteins might be involved in signalling essential for organ development during early embryogenesis and suggests that loss of MAN1 may cause muscle and retina specific diseases.

Expression pattern of zinc-finger transcription factor Odd-skipped related 2 in murine development and neonatal stage

The Odd-skipped gene, first identified as a Drosophila pair-rule zinc-finger transcription factor, plays an important role in Drosophila development. The mammalian homolog, Odd-skipped related 2 (Osr2), regulates limb, tooth, and kidney development in mouse embryos. However, the detailed expression pattern of Osr2 during neonatal development remains unclear. In this study, we investigated Osr2 expression patterns in mouse neonatal and embryo tissues using qPCR and in situ hybridization methods. First, we examined the tissue distribution of Osr2 by qPCR, and found it to be highly expressed in the uterus and moderately in the testes, small intestine, and prostate. That expression was also found in eye, kidney, placenta, lung, thymus, lymph node, stomach, and skeletal muscle tissues, and in all embryonic stages. On the other hand, Osr2 was not expressed in brain, heart, liver, or spleen samples. Next, we examined the tissue localization of Osr2 using in situ hybridization. Osr2 was found in the craniofacial region on E13.5, with notable expression in dental germ mesenchyme as well as the renal corpuscle on E17.5. As for neonatal tissues, Osr2 was expressed in the dental papilla, dental follicle, Harderian gland, nasal bone, eyelid dermis, synovial joint, and tibial subcutis. Our findings suggest that Osr2 functions in reproductive system organs, such as the uterus, testes, prostate, placenta, and ovaries. Furthermore, based on its expression in kidney, Harderian gland, eyelid dermis, and tibial subcutis tissues, this transcription factor may be involved in hormone synthesis and function. Together, our results demonstrate the role of Osr2 in postnatal development and embryogenesis.

Performance of growing pigs fed diets based on by-products of maize and wheat processing

Forty-eight crossbred Landrace × Large white pigs with an average body weight of 8.5 ± 0.4 kg and aged 2 months were used to evaluate performance and carcass characteristics of pigs fed two diets in which bran from maize or wheat was used as the energy source. A third diet based on whole maize grain was used as a control. Animals in groups of four, balanced for litter, sex and weight were allotted to dietary treatments in a completely randomised design with four replications. Data were collected on feed intake and weight gain for a period of 4 months. A digestibility trial was carried at the end of the feeding trial using two male pigs per treatment while six pigs per treatment were randomly selected for slaughter to determine carcass characteristics. Daily gain averaged 0.23, 0.31 and 0.13 kg/day, for pigs fed maize bran, wheat bran and whole maize diets, respectively. Average feed intake and final body weight were significantly (P < 0.05) affected by diets. Digestibility of dry matter, crude fibre and calcium were higher (P < 0.05) for pigs fed maize bran while crude protein digestibility was highest (P < 0.05) for the pigs fed wheat bran. There were significant differences (P < 0.05) between the diets in carcass yield, ham, head, trotters and heart weights and also for rib weight, rib eye muscle and other tissues. This study indicates that comparable performance of growing pigs fed cereal bran and full-grain diets are real.

Pathogenesis of thyroid-associated ophthalmopathy: does autoimmunity against calsequestrin and collagen XIII play a role?

Thyroid-associated ophthalmopathy (TAO), or thyroid eye disease, is a complex inflammatory disorder of the eye that, as its name implies, is associated with thyroid disease. TAO can be divided into three subtypes: ocular myopathy, congestive myopathy and mixed congestive and myopathic ophthalmopathy. Although the precise pathophysiology of TAO remains unclear it is likely to reflect an autoimmune reaction involving sensitized T-cells and autoantibodies directed against a thyroid and orbital tissue shared antigen. One well studied candidate in this immune reaction is the thyroid-stimulating hormone receptor (TSH-r), expressed in the orbital fibroblast and pre adipocyte. In our studies of TAO, we have investigated the nature and significance of antibodies targeting other eye muscle and orbital connective tissue (OCT) antigens. Our findings suggest that autoimmunity against the eye muscle antigen calsequestrin and the OCT antigen collagen XIII plays a role in the pathogenesis of TAO. We propose that ocular myopathy and chronic eyelid retraction are due to autoimmunity against skeletal muscle calsequestrin in the extraocular and eyelid muscles, respectively. This may be initiated in the thyroid where calsequestrin expression is upregulated, possibly due to a stimulatory effect of TSH-r antibodies. We also propose that congestive ophthalmopathy results from a reaction against the TSH-r or collagen XIII in orbital fibroblast cell membranes. Further insight into the role of eye muscle and OCT antigens in the pathogenesis of TAO may allow for the development of new therapies to treat the eye disorder and reduce patient morbidity.

Ocular manifestations in children and adolescents with thyrotoxicosis.

Thyrotoxicosis is a rare disorder in childhood and adolescence. The frequency ranges from 0.1 in 100000 in young children to 3.0 in 100000 in adolescents and the most frequent cause is Graves' disease. Few children and adolescents suffer from thyrotoxicosis during the course of autoimmune thyroiditis and non-autoimmune hyperthyroidism due to constitutively active mutations of the TSH-receptor have been reported in single patients with neonatal hyperthyroidism and rare familial cases. From a survey of the literature it is apparent that infiltrative ophthalmopathy is very rare in children and adolescents. Milder ocular manifestations as retraction of the upper eyelid, upper lid lag and "staring eyes" are reported in varying frequencies ranging from 25 to 60% of all patients. In our own series of 43 children and adolescents (34 females, 9 males) with thyrotoxicosis only 16 (37%) had clear ocular manifestations of upper lid retraction, lid lag and slight protrusio bulbi. However, only three of of them had soft tissue involvement and significant exophtalmus. In none of the patient corneal or optic nerve involvement or paralysis of the extraorbital muscles was observed. Newborns with congenital hyperthyroidism due to maternal Graves' disease seem to present significant transitory ocular manifestations more frequently, but again no infiltrative ophtalmopathy requiring specific therapy is present. Recently at least four different newborns with congenital thyrotoxicosis due to gain-of-function mutations of the TSH-receptor have been reported, who also had significant ocular manifestations. The presence of staring eyes, slight proptosis and lid retraction in these newborns without any autoimmune or inflammatory process supports the hypothesis that these ocular manifestations are caused by the increased thyroid hormone actions. The involvement of the eyes is usually transitory in newborns as well as in children and adolescents and the symptoms usually disappear when euthyroidism is restored. The administration of steroids therefore is restricted to the rare severe cases with eye muscle or soft tissue involvement. There are no reports on surgical decompression or orbital irradiation therapy of ophtalmopathy in children and adolescents, which again indicates that ocular involvement in thyrotoxicosis in children and adolescents is less frequent and much less severe than in adults.

Immunohistochemical Studies Using Immunized Guinea Pig Sera with Features of Anti-Human Thyroid, Eye and Skeletal Antibody and Graves’ Sera

Type 2 5' deiodinase enzyme was observed in both thyroid and eye muscle tissues, highlighting its possible role as a common antigen in thyroid-associated ophthalmopathy. Sera of 105 Graves' patients and 40 controls, and immunized guinea pig sera against TCSS peptide, showing homology to the amino acid sequence from 132 to 152 of type 2 5' deiodinase, were investigated to demonstrate the binding effects to human thyroid, eye and skeletal muscle tissues. Twenty-two Graves' patients were positive for anti-TCSS peptide antibodies, of whom 18 cases had ophthalmopathy. The levels of anti-TCSS peptide antibodies were higher not only in Graves' patients with (P<0.0001) and without (P<0.036) eye symptoms compared to controls but also the difference was significant between patients with and without ophthalmopathy (P<0.049). In Western blot, immunized sera showed binding reactions to the supernatant fractions of human thyroid, eye and skeletal muscle tissues at the range of 29 kDa. Patient sera with Graves' ophthalmopathy resulted in positive reactions directed to membrane areas in thyroid follicular cells, and to fibers in eye and skeletal muscles using immunohistochemical method, while no positive staining was present after adding control sera. The binding features of immunized guinea pig sera exhibited similar staining in all human tissues but could be blocked with Graves' sera. Our results suggest that type 2 5' deiodinase enzyme protein could play a role as an antigen in Graves' disease. Immunized guinea pig sera against TCSS peptide exhibited similar binding reactions and stainings to human thyroid, eye and skeletal muscle tissues as patient sera with Graves' ophthalmopathy.

Role of cytokines in the pathogenesis of thyroid-associated ophthalmopathy.

Thyroid-associated ophthalmopathy (TAO) is generally considered to be an autoimmune disorder associated with Graves' disease. However, the nature of autoantigen or mechanism of the development of ophthalmopathy remains unclear. In the present review we focus the accumulating evidence on roles of cytokines in the orbital tissues from patients with TAO and animal models. From the analysis of T-cell clones, T helper 1 (T(H)1)-like clones were predominant in cultures from patients with recent onset hyperthyroidism and T(H)2-like clones were predominant in culture form patients with more remote onset hyperthyroidism. T(H)1-like cytokine profiles are predominant in eye muscle tissue and related to the eye muscle enlargement, while T(H)2-like cytokine profiles are predominant in orbital fat tissue from patients with TAO and negatively related to orbital volume. Therefore, T(H)1-like cytokines, proinflammatory cytokines, may play a role on the development of eye muscle component of TAO in the acute stage. T(H)2-like cytokines, anti-inflammatory cytokines, may play protective role in the chronic stage of TAO. The studies using animal models suggest the genetic background is involved in the pathogenesis of TAO. The studies on polymorphism of the cytokine genes support the proinflammatory role of T(H)1-like cytokines and protective role of T(H)2-like cytokines.

Genetic Variation in White Bass

A protein electrophoretic survey of white bass Morone chrysops was conducted to evaluate the extent and pattern of extant genetic variation and population structuring. Forty-four presumptive structural loci were surveyed from eye, liver, and skeletal muscle tissue from 456 white bass in 19 samples. The level of allelic variation observed within white bass populations was low relative to many other species of fish, but it was consistent with levels of variation observed in the congeneric striped bass M. saxatilis and may represent a feature common to all species in the genus. Significant interpopulation genetic heterogeneity was observed; however, the overall level of differentiation among populations was quite low. Widespread stocking of white bass since 1900 and rapid range expansion could, in part, explain the absence of population structuring that is consistent with geographic proximity or drainage. The high frequencies of an mIDHP* allele in Lake Erie collections that was absent in those from Mississippi drainages is consistent with distinct postglacial origins of these two groups.

Cytokine profiles in eye muscle tissue and orbital fat tissue from patients with thyroid-associated ophthalmopathy.

Eye muscle (EM) and retroorbital fat tissue are two major sites of involvement in thyroid-associated ophthalmopathy (TAO). Lymphocytic infiltration in these tissues is a prominent histological feature of TAO. We have investigated the cytokine gene profiles in EM and orbital fat (OF) tissues from patients with TAO. Total RNA was isolated from EM tissue of 14 patients and from OF tissues of 29 patients with TAO. Cytokine gene expression was assessed by RT-PCR using paired primers for interferon gamma (IFNgamma), tumor necrosis factor alpha (TNFalpha), interleukin (IL)-1beta, IL-2, IL-4, IL-6, IL-10, CD4, CD8, and glyceraldehyde-3-phosphate dehydrogenase. IFNgamma, TNFalpha, IL-1beta, and IL-6 messenger RNA (mRNA) were mainly detected in EM tissue, whereas IL-4 and IL-10 mRNA were detected in only one patient. On the other hand, in OF tissue, IL-4 and IL-10 mRNA were detected in 24% and 38% of the patients, respectively, and IFNgamma, IL-1beta, and IL-6 mRNA were less often detected compared with EM tissue. The enlargement of EM tissue as assessed by computed tomography correlated significantly with TNFalpha mRNA expression in EM tissue. The orbital volume was positively correlated with IL-6 mRNA expression and negatively correlated with IL-4 mRNA and IL-10 mRNA expression in OF tissue. These results suggest that T helper (Th) 1-like cytokines predominate in EM tissue in most patients and that the predominant cytokine profile in OF tissue varies from patient to patient. Both Th1-like and Th2-like immune responses may play roles in the development of two components of ophthalmopathy.

Cytokine Profiles in Eye Muscle Tissue and Orbital Fat Tissue from Patients with Thyroid-Associated Ophthalmopathy

Cytokine Profiles in Eye Muscle Tissue and Orbital Fat Tissue from Patients with Thyroid-Associated Ophthalmopathy

Thyroid-associated ophthalmopathy in black South Africans with Graves' disease: relationship to serum antibodies reactive against eye muscle and orbital connective tissue autoantigens.

The prevalence of hyperthyroidism owing to Graves' disease is increasing among urban black South Africans. Thyroid-associated ophthalmopathy is often observed in this context, but its pathogenesis remains unclear. No close relationship has been noted between antiflavoprotein (Fp) antibodies or thyrotropin receptor antibodies and ocular involvement in black patients. We measured serum antibodies against eye muscle and orbital connective tissue antigens in black patients with Graves' disease, correlating them with eye signs. Of 11 patients with clinical ophthalmopathy, 2 (18%) had antibodies against collagen type XIII, 3 (27%) against flavine adenine dinucleotide (FAD), 1 (9%) against Fp, and 4 (35%) against G2s. Antibody prevalences in eight patients without clinical ophthalmopathy were 12.5% for collagen XIII, 12.5% for FAD, 25% for Fp and 0% for G2s. These differences were not statistically significant. None of the individual mean antibody levels were significantly different between the two subgroups of thyrotoxic patients. Serum antibody levels were negative in 10 black South African controls. In summary, eye muscle and orbital connective tissue antibodies were found in small proportions of patients with Graves' disease with no close relationship of any antibody to eye signs. Thus, a substantial proportion of black South Africans with overt clinical ophthalmopathy remains in whom currently availabe serologic tests are unhelpful for screening and laboratory confirmation.

Thyroid-Associated Ophthalmopathy: Clinical Features, Pathogenesis, and Management

Thyroid-associated ophthalmopathy (TAO) is a progressive eye disorder characterized by immune-mediated inflammation of the extraocular muscles and orbital connective tissue. TAO is linked, in a unique way, with thyroid autoimmunity, in particular Graves’ hyperthyroidism. Our working hypothesis for the pathogenesis of TAO is that recognition of a thyrotropin receptor (TSHR)-like protein in the orbital preadipocytes by antibodies may be the initial event leading to homing of lymphocytes into the orbital tissues. In the course of thyroid inflammation, antibodies and T cells reactive against G2s expressed in thyroid membranes cross-react with the protein in the eye muscle fiber, leading to eye muscle damage and dysfunction. Those patients with anti-G2s antibodies develop ocular myopathy. Antibodies against flavoprotein, the 64-kDa protein, which are produced in the context of eye muscle fiber damage and mitochondrial rupture, are sensitive markers of immune-mediated fiber necrosis in patients with ophthalmopathy but do not directly damage the eye muscle. Antibodies against type XIII collagen, which is localized in the plasma membranes of orbital fibroblast, may be a new marker for the congestive ophthalmopathy subtype of TAO. The measurement of antibodies against key eye muscle and orbital connective tissue autoantigens may have a role in the management of active ophthalmopathy and its prediction in patients with Graves’ hyperthyroidism.

Effects of heparin on vascular dysfunction in diabetic rats.

1. Treatment with heparin has beneficial effects in diabetic nephropathy. The occurrence of increased urinary albumin excretion in diabetic patients reflects general vascular dysfunction, including increased transcapillary permeability of macromolecules. The aim of the present study was to evaluate the effects of heparin on vascular dysfunction in diabetic rats. 2. Male Sprague-Dawley rats were used in two studies. Diabetes was induced by 65 mg/kg, i.v., streptozotocin. In one study, diabetic rats were dosed subcutaneously with different heparin fractions for 8 months and the transcapillary escape rate of albumin (TERalb) was measured in anaesthetized animals. In the other study, heparin was given for 6 weeks, followed by tissue albumin clearance measurements in awake rats. Normal and diabetic rats receiving saline served as controls. 3. Blood glucose did not differ among the diabetic groups and ranged from 22 to 26 mmol/L. The mean (+/- SD) TERalb was increased by diabetes compared with values in normal rats (17.5 +/- 3 vs 14.1 +/- 3.3%/h, respectively). Neither unfractionated nor low molecular weight heparin significantly affected this increase. [131I]-Albumin clearance was significantly increased in diabetic rats in the eye, skin and skeletal muscle tissues compared with normal rats (0.17-0.40 vs 0.1-0.23 microL plasma/g per min). Low molecular weight heparin treatment did not affect the increased organ albumin clearance. 4. In conclusion, heparin treatment does not affect diabetes-induced vascular dysfunction as expressed by increased TERalb and clearance of albumin in rats.

Demonstration of thyrotropin receptor mRNA in orbital fat and eye muscle tissues from patients with Graves' ophthalmopathy by in situ hybridization.

There is a controversy regarding whether there are thyrotropin (TSH) receptors in orbital fat and eye muscle tissues that may play a role in the pathogenesis of Graves' ophthalmopathy. To elucidate whether there are TSH receptors in orbital fat and eye muscle tissues in patients with Graves' ophthalmopathy, we applied the method of in situ hybridization in orbital fat and eye muscle tissues obtained during the operation for patients with Graves' ophthalmopathy, to directly detect TSH receptor mRNA. To identify whether the cells with positive TSH receptor mRNA are fibroblasts, we also did vimentin immunoreactivity study. To further prove the transcript does have a full length of TSH receptor, the samples of total RNA preparations, extracted from orbital fat and eye muscle tissues, were used as a template for reverse transcriptase polymerase chain reaction (RT-PCR) using three primer sets to generate cDNA fragments and cloned for sequencing. The results showed that the expression of TSH receptor mRNA was demonstrated in adipocytes and fibroblasts of orbital fat, and perimysial fibroblasts within eye muscle tissues by in situ hybridization and vimentin immunoreactivity study. Also, by using the RT-PCR, cloning and sequencing, we further proved that the transcript does have a full length of TSH receptor. The present study suggested that there are TSH receptors expressed in orbital fat and eye muscle tissues.

Serological reactivity of recombinant 1D autoantigen and its expression in human thyroid and eye muscle tissue: a possible autoantigenic link in Graves' patients.

Thyroid-associated ophthalmopathy (TAO) is a potentially severe autoimmune disease, in and around the orbit, usually accompanied by Graves' disease. It was the goal of this study to develop a serological indicator for TAO and to characterize its expression in human thyroid and eye muscle tissue. Thus, we have recloned the full-length 1D-complementary DNA and assessed its expression levels in 90 healthy and diseased human thyroids. Only Graves' patients suffering from TAO (n = 29) displayed a significant, 2.1-fold increase of 1D expression levels (P = 0.029), compared with normal controls (n = 9), as assessed using the Mann-Whitney U-test for paired, nonnormally distributed samples. In contrast, a decrease of 1D expression (to 40% of control normal values) was confined to thyroid autonomy (n = 19, P = 0.032). In all other diseased human thyroids, including Graves' thyroids from patients not suffering from clinically overt TAO (n = 9), 1D expression levels were not different from the healthy controls. 1D gene expression was demonstrated in both healthy (n = 10) and diseased (n = 10) eye muscle tissues. Furthermore, a recombinant protein derived from baculovirus-infected Sf9 insect cells was purified under both nondenaturing and denaturing conditions. While under nondenaturing conditions, the molecular mass of recombinant 1D was determined to be 85 kDa; denaturing isolation yielded the expected 64-kDa protein. Autoantibodies against denatured 1D protein were not detectable in sera of diseased or healthy subjects. Immunoreactivity against the 85-kDa, nondenatured protein, evaluated in a panel of 222 different human sera, showed that 82% of Graves' patients suffering from TAO had autoantibodies against recombinant 1D, whereas only 5% of the healthy controls were positive for antibodies against 1D. Taken together, our results demonstrate a high disease sensitivity and specificity of recombinant, nondenatured 1D, to distinguish Graves' disease with or without TAO from other forms of thyroid and/or eye disease. Prospective studies will have to show whether autoantibodies against 1D can also be used as a prognosticator of TAO.