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Molecular biology of terminal transferase.

Terminal transferase is an unusual deoxynucleotide polymerizing enzyme found only in prelymphocytes. The protein was purified to homogeneity from calf thymus glands in 1971 as a 32 kDa protein with a two peptide structure. Subsequent biochemical and immunological analyses of terminal transferase protein in crude extracts from a number of animal species showed a single peptide with a molecular weight of about 58,000. The two peptide structure found earlier was caused by proteolysis. Homogeneous 58 kDa terminal transferase has now been produced from human lymphoblastoid cells and calf thymus glands by immunoaffinity chromatography. In vitro phosphorylation studies showed that the terminal transferase protein contains one phosphorylation site near one end of the polypeptide chain, and the phosphorylation of the enzyme has been confirmed by in vivo labeling experiments. Unambiguous demonstration of the molecular weight of the human terminal transferase was obtained by translation of the cloned human terminal transferase DNA sequence to a 58,308 Da protein. The translated amino acid sequence also provided a possible phosphorylation site near the amino-terminus of the protein. Preliminary analysis of the genomic structure shows a simple intron/exon pattern with the total human terminal transferase gene spanning at least 65 Kb.

Amino Acid Sequence↗

Characterization of thymic weight and thymic peptide thymosin-beta 4: effects of hypophysectomy, sex, and neonatal sexual differentiation.

An initial experiment was conducted to analyze the effects of hypophysectomy (Days 5-30 post hypophysectomy) on thymic weight and thymosin-beta 4 concentrations. After hypophysectomy, thymic weight and thymosin-beta 4 concentrations decreased with time (5, 10, 15, and 30 days post hypophysectomy) and by 30 days post hypophysectomy were reduced by 50%. Exp. II investigated the effects of early sexual differentiation on thymic weight and thymosin-beta 4 concentrations in pituitary intact and hypophysectomized male, female, nonmasculinized rats (via castration of males at Day 2 or 11 of age and androgenized female (at Day 3 of age) rats. Rats were hypophysectomized at 30 days of age and, at 17 days post hypophysectomy, the thymus gland was dissected and weighed. Serum was collected for thymosin-beta 4 analysis. At 47 days of age, males have a greater thymic weight when compared to females (p less than 0.01) but hypophysectomy eliminated any sex differences and significantly reduced thymic weights (p less than 0.05). Hypophysectomy did produce increased thymosin-beta 4 concentrations across all treatments (p less than 0.01) and is considered to be a combination response of decreased gonadal function via lack of pituitary hormonal stimulation (luteinizing hormone and follicle stimulating hormone) and lack of gonadotrophic effects on the thymus gland and its secretory capacity. Hypophysectomized males and Day-2 castrate males had increased thymosin-beta 4 concentrations when compared to hypophysectomized females and neonatally androgenized females (p less than 0.05). Differences of thymic weights between male and female rats during prepubertal development are probably related more to presence or absence of gonads, gonadal steroids, and response of the thymus to pituitary secretagogues rather than to neonatal sexual differentiation of thymic function and secretion of thymosin-beta 4.

Animals↗

[Characteristics of the lymphoid cells of rabbits].

By means of scanning electron microscopy it has been stated that the thymus gland, spleen, paratracheal lymph node, Peyer's patches and appendix in an intact rabbit of "Shinshilla" strain have T- and B-lymphocytes in different proportion. T-lymphocytes are seen as spherical cells with rough surface, with a few microprocesses, have 3.0 +/- 0.1 mcm--4.7 +/- 0.2 mcm in diameter depending on their location. B-lymphocytes are seen as spherical cells with a considerable amount of microvilli on their surface, have 2.9 +/- 0.1 mcm--4.4 +/- 0.1 mcm in diameter depending also on their location. In the thymus gland, cells with rough surface, a few microprocesses and crater-like hollows prevail. In the spleen and lymph node, most of the cells are of similar diameter and have microvilli on the surface. Lymphoid population in the Peyer's patch and appendix is represented by large cells with microprocesses.

Animals↗

[Pseudoparalytic myasthenia gravis. Diagnostic and therapeutic aspects in 60 separate cases].

Based on 60 of our own cases and on the medical literature the authors discuss the diagnostic, pathophysiological and therapeutic aspects of myasthenia gravis. Myasthenia is suspected in cases of motor weakness of changing intensity, diminishing by rest. The weak muscles are innervated by different peripheral nerves. At the beginning a weakness of upperlid-muscles, external eye muscles and bulbar muscles is particularly frequent. There is no sensory loss or other neurological symptoms. A transitory disappearance of motor weakness after an intravenous dose of Edrophonium (Tensilon) is a typical diagnostic sign. The effect is less evident with eye-muscle weakness. A typical appearance of potentials after repetitive stimulation of peripheral nerves as well as other characteristics in electrophysiological testing of muscles are of high diagnostic value. This allows differentiation from other types of muscle weakness. In the pathogenesis of myasthenia an autoimmune process related to a persistent thymus gland plays an important part. This leads to an ultrastructural change in the postsynaptic membrane of the muscle fibre. The postsynaptic membrane no longer reacts in a normal way to acetylcholine as a transmitter substance at the level of the motor endplate. Therefore the first step in the treatment of myasthenia consists of cholinesterase-inhibitors, specially Neostigmin (Prostigmin) and Pyridostigmin (Mestinon). Thymectomy is advised in all cases of myasthenia with the exception of the pure ocular form and of myasthenia in patients older than 60 years. The thymus gland is practically always persistent or hypertrophic in myasthenia. The suprasternal access is recommended. A thymoma should always be operated upon because of the danger of malignancy. In cases where thymectomy is not performed or not successful and if cholinesterase-inhibitors are not sufficiently efficient, treatment with corticosteroids or ACTH is recommended.

Adolescent↗

The thymolytic effect of cocaine and monoaminergic drugs in the mouse.

Cocaine abuse is known to be associated with suppression of the immune system. In this experiment, animals treated with cocaine (CC) in their drinking water (200 mg/L) for 10 days resulted in a significant decrease in thymus weight in HSD-NDA male Swiss mice. A subsequent in-vitro experiment was designed to investigate the direct effect of cocaine and other dopaminergic agents on thymocyte proliferation. Thymus glands were aseptically removed from mice and thymocytes were isolated and then incubated in microtiter plates with various concentrations (10[-8]-10[-4] M) of CC, apomorphine (AM), haloperidol (HP), dopamine (DA) and epinephrine (EN) for 18 hrs. The results revealed that cocaine inhibits [3H]-thymidine uptake into DNA in a dose-dependent manner. Apomorphine, haloperidol and dopamine also exhibited a similar dose dependent inhibition of thymocyte proliferation. The IC25s for DNA synthesis inhibition were 8.2 x 10(-6) M, 4.3 x 10(-7) M, 2.5 x 10(-7) M, and 1.1 x 10(-7) M for CC, HP, DA and AM, respectively. EN was found not to have any significant effect on DNA synthesis. The results suggest that thymus gland atrophy, associated with use of CC may be related to the inhibition of thymocyte proliferation.

Animals↗

The growth of nonlymphoid thymic components in vitro: age-related differences during development.

An explant culture procedure has been developed that makes it possible to measure the relative growth capacity of the epithelial and mesenchymal cells of the canine thymus gland. Standardized growth conditions were obtained by size-grading thymic fragments and counting to allow uniform fragment density during culture. After 6 d in culture, outgrowth from the fragments formed colonies that could be classified into epithelial, mixed, or spindle cell type. Uniform fragment size and number in each flask allowed calculation of the total plating efficiency, relative distribution of colony types, and mean colony diameters for thymic fragments collected from fetuses (50 d of gestation), neonates (0 d postpartum), and juveniles (70 d postpartum). Data show age-related changes in the proliferative capacity of the cells in all three colony types. The most significant difference was seen in the epithelium, which showed a 30% reduction in mean colony diameter over the 2 wk between fetal and neonatal ages and a 23% reduction over the postnatal period of 70 d. Significant reductions were seen in the other colony types as well. Because the severity of the effect of many injurious agents is proportional to the rate of growth of the target cells, these data suggest that the thymus gland of the fetus may be more sensitive to physical or chemical injury than is the neonate or adult.

Acid Phosphatase↗

Altered major histocompatibility complex-restricted antigen recognition by T cells from elderly humans.

Positive selection of T cells within the thymus gland leads to major histocompatibility complex (MHC)-restricted recognition of antigen by T lymphocytes. As the thymus gland involutes with age, altered MHC-restricted antigen recognition by T cells from elderly humans would be expected. We have tested this hypothesis by comparing the proliferative response of T cells and T cell clones from aged and young subjects to influenza determinants presented by autologous or allogeneic antigen-presenting cells (APC). Under conditions in which the allogeneic mixed lymphocyte reaction was minimal, T cells from six of seven aged donors but only one of seven young donors were stimulated by influenza vaccine presented by allogeneic APC. More importantly, one-half of the influenza-specific T cell clones derived from aged donors, but none of the clones derived from young donors, were activated by influenza vaccine presented by allogeneic APC. While 80% of the MHC-nonrestricted influenza-specific T cell clones expressed the gamma/delta T cell receptor, 20% of these clones expressed the alpha/beta T cell receptor. Thus, changes in MHC-restricted antigen recognition by T cells and in altered distribution of alpha/beta versus the gamma/delta T cell receptor bearing antigen-specific T cell clones occur with aging.

Adult↗

Assessment of pulmonary and intrathymic hemosiderin deposition in sudden infant death syndrome.

The aim of this study was to stain lung and thymus gland sections that had been taken from infants who had died of sudden infant death syndrome (SIDS) for interstitial hemosiderin and to compare the results with those obtained for controls. There were two groups of SIDS infants, one with, and a second group without, histories of apparent life-threatening events (ALTEs). No significant difference in numbers of cases with interstitial hemosiderin deposition was found between SIDS infants with histories of ALTEs (n = 4 of 12, 33.3%), SIDS infants without histories of ALTEs (n = 4 of 22, 18.2%), and controls (n = 4 of 24, 16.7%). However, if four of the control cases with histories of previous chest trauma were excluded, there was a significantly greater number of cases with pulmonary interstitial hemosiderin in the SIDS infants with histories of ALTEs compared with the subgroup of control infants with no previous chest trauma (n = 1 of 20, 5%) (P < .05). No relationship could be established between the timing of the ALTEs, the type of resuscitation or age of the infant at death, and the presence of hemosiderin. None of the sections of thymus gland stained positively for hemosiderin. Positive staining for pulmonary interstitial hemosiderin, therefore, differentiated a group of SIDS infants with histories of previous ALTEs from a subgroup of control infants with no histories of previous chest trauma. However, pulmonary interstitial hemosiderin staining could not be used with certainly to confirm or exclude previous ALTEs in individual SIDS cases as not every SIDS case with a history of an ALTE stained for pulmonary interstitial hemosiderin. In addition, positive staining occurred for SIDS infants without histories of ALTEs and also for control infants who died of other causes.

Case-Control Studies↗

Tolerance to the effect of delta1-tetrahydrocannabinol on corticosterone levels in mouse plasma produced by repeated administration of cannabis extract or delta1-tetrahydrocannabinol.

1 Three injections of cannabis extract (500 mg/kg s.c. given over 3 or 5 days) diminished thymus gland weight but not the weights of spleen or liver in weanling female and adult male mice kept at room temperature.2 Both cannabis extract (500 mg/kg s.c.) and Delta(1)-tetrahydrocannabinol (Delta(1)-THC; 10 and 20 mg/kg i.p.) elevated corticosterone levels in mouse plasma.3 A pretreatment that consisted of three daily subcutaneous injections of 500 mg/kg of cannabis extract and that was shown to produce tolerance to the ;cataleptic' effect of Delta(1)-THC (2 mg/kg i.v.) in mice, also produced tolerance to the effect of Delta(1)-THC (10 mg/kg i.p.) on corticosterone levels in mouse plasma. However, this pretreatment did not reduce the rise in plasma corticosterone concentration produced by immobilization.4 Tolerance to the effect of Delta(1)-THC (10 mg/kg i.p.) on corticosterone levels in mouse plasma was also produced by the pretreatment of mice with a single injection of Delta(1)-THC (10 mg/kg s.c.). Three daily injections of Delta(1)-THC (10 or 30 mg/kg s.c.) also produced tolerance.5 In a thermoneutral environment (30-32 degrees C) in which cannabis extract does not produce hypothermia, the drug no longer reduced thymus gland weight. However the effect of cannabis extract and of Delta(1)-THC on corticosterone plasma levels was the same at room temperature as at 30-32 degrees C. Tolerance to the latter effect of Delta(1)-THC was also produced equally readily under the two conditions.6 It is concluded that pretreatment with cannabis extract or Delta(1)-THC can produce tolerance to the effect of Delta(1)-THC on corticosterone levels in mouse plasma and does so without impairing the effect of immobilization stress on corticosterone release. In addition, both the rise in corticosterone plasma levels produced by cannabis or Delta(1)-THC and the development of tolerance to this effect can still take place in the absence of hypothermia.

Animals↗

The binding properties, with blood proteins, and tissue distribution of 22-oxa-1 alpha,25-dihydroxyvitamin D3, a noncalcemic analogue of 1 alpha, 25-dihydroxyvitamin D3, in rats.

The binding properties, with blood proteins, and tissue distribution of 22-oxa-1 alpha,25-dihydroxyvitamin (22-oxacalcitriol; OCT), a noncalcemic analogue of 1 alpha,25-dihydroxyvitamin D3 [1,25(OH)2D3], in rats were investigated. The binding affinity of OCT to plasma vitamin D binding protein (DBP) is extremely low and OCT mainly circulates in the blood as an intact form nonspecifically bound to lipoproteins especially to chylomicrons and low density lipoprotein (LDL). OCT intravenously injected into normal rats rats rapidly disappeared from the blood, and rapidly appeared in the bile as glucuronides of intact OCT and 1 alpha, 3 beta,20(S)-trihydroxy-9,10-secopregna-5,7,10(19)-triene (23,24,25,26, 27-pentanorOCT; pentanorOCT) as an OCT metabolite. When OCT or 1,25(OH)2D3 was injected into normal rats, significant amounts of OCT and 1,25(OH)2D3 were quickly detected in the thyroid and parathyroid glands, thymus, adrenals, liver, plasma, small intestine, kidneys, and calvaria. The detected amounts of OCT in the parathyroid glands, thymus, adrenals, liver, small intestine, and kidneys were significantly higher than the respective values for 1,25(OH)2D3 2 and/or 10 min after injection, while those of OCT in the plasma and calvaria were significantly lower than those of 1,25(OH)2D3. The in vivo rapid turn-over, nonspecific transportation, and incorporation of detectable amounts into the tissues are typical characteristics of OCT which may account for its specific activities.

Animals↗

KIT (CD117) is frequently overexpressed in thymic carcinomas but is absent in thymomas.

KIT (CD117), a tyrosine kinase receptor, has not been widely studied in epithelial tumours. In a systematic immunohistochemical survey of KIT expression on tissue arrays incorporating 671 cases, it was found that thymic carcinomas frequently express KIT. Twenty-two thymic carcinomas, 110 thymomas, and 16 non-neoplastic thymus glands were retrieved for further analyses. Immunohistochemically, 19 (86%) thymic carcinomas revealed heterogeneous to diffuse membranous positivity, whereas no thymomas or normal thymus glands contained positive epithelial cells. Using reverse transcriptase-polymerase chain reaction (RT-PCR), c-kit transcripts could be demonstrated in all immunohistochemically positive cases. PCR amplification and direct sequencing of the c-kit juxtamembrane domains (exons 9 and 11) and tyrosine kinase domain (exons 13 and 17) were also performed on the thymic carcinomas but mutations were not found. Some non-thymic epithelial tumours showed frequent KIT expression including adenoid cystic carcinomas of the salivary gland (100% positive), chromophobe renal cell carcinomas (94%), renal oncocytomas (67%), and neuroendocrine tumours (34%). Other carcinomas were infrequently immunoreactive for KIT. The findings of this study suggest that KIT is involved in the pathogenesis of thymic carcinomas. The overexpression of KIT in thymic carcinomas has potential diagnostic utility in differentiating these tumours from thymomas and carcinomas arising from other sites, which express KIT infrequently.

Carcinoma↗

[Castleman's disease: an unusual neck neoplasm].

Castleman's tumour of unusual location on the neck of 12 years old girl has been described. From two forms of Castleman's disease which exist--the hyaline vascular kind of disease. The authors presented their opinion on the affection arising and differential diagnosis Castleman's tumour from tumours of thymus gland origin.

Castleman Disease↗

[Immunostimulating effect of synthetic surface-active agents in experimental tuberculosis].

The influence of synthetic surfactants of different classes (BX-14, O-14, B-14) on an immunologic reactivity of mice with evolving experimental tuberculosis was studied. The use of the substances leads to an increase of the E-rosette forming cells (E-RFC) in the mice thymus gland and spleen, and stimulates the inhibited function of spleen natural killers, without modifying (except the agent O-14) the cytotoxicity level of thymus gland natural killers. The above substances can enhance the absorbing capacity of the spleen macrophages as well as their ability to reduce nitro blue tetrazolium. Attention is drawn to the possibility of using immunostimulators belonging to a group of surfactants to correct immunologic reactivity impairments caused by tuberculosis.

Adjuvants, Immunologic↗

Myasthenia gravis: influence of thymic changes on clinical parameters. A study on the basis of 364 thymectomies.

In 364 patients with myasthenia gravis the morphology of thymus glands, thymomas and muscle samples was studied and compared to different clinical, therapeutic and prognostic parameters of the disease. The role of active thymus as the most outstanding finding, was considered for the result of thymectomy and for the prognosis to be positive: The more active the thymus the better the outcome of thymectomy and the final and lasting prognosis of the disease.

Adolescent↗

The thymus-pituitary axis and its changes during aging.

The pituitary-thymic axis constitutes a bidirectional circuit where the ascending feedback loop is effected by thymic factors of epithelial origin. The aim of the present article is to review the evidence demonstrating that aging brings about a progressive disruption in the integration of this network. In doing so, we briefly review the experimental evidence supporting the view that immune and neuroendocrine aging are interdependent processes. The advantages and limits of the nude mouse as a model of thymus-dependent accelerated aging is also discussed. Next, we review a number of studies which show that the endocrine thymus produces several bioactive molecules, generally called thymic hormones, which in addition to possessing immunoregulatory properties are also active on nervous and endocrine circuits. In particular, the reported activities of thymosin fraction 5 (TF5), thymosin alpha-1 and thymosin beta-4 on beta-endorphin, ACTH, glucocorticoids, LHRH and LH secretion in different animal and cell models are reviewed. The known hypophysiotropic actions of other thymic hormones like thymulin, homeostatic thymus hormone (HTH) and thymus factor are summarized. Aging has a significant impact on pituitary responsiveness to thymic hormones. Thus, it has been reported that TF5 and HTH have thyrotropin-inhibiting activity in young but not in old rats. Furthermore, intravenous administration of HTH was also able to reduce plasma GH and increase corticosterone levels in both young and old rats, although these responses were much weaker in the old animals. Further evidence on this topic is discussed. It is proposed that in addition to its central role in the regulation of the immune function, the thymus gland may extend its influence to nonimmunologic components of the body, including the neuroendocrine system. The early onset of thymus involution might therefore act as a triggering event which would initiate the gradual decline in homeostatic potential that characterizes the aging process.

Aging↗

Myasthenia gravis: An embryologic and phylogenetic perspective.

Myasthenia gravis is a rare disease with unique clinical features. These include (1) progressive fatigue on exertion, most evident clinically in the muscles of the head and neck (2) a definite relationship of the disease to the thyroid, parathyroid and thymus glands. A hypothesis is advanced with correlates these facts on the basis of a common embryologic origin of these glands and the branchial arch muscles. A corollary is that the muscles of the head and neck differ from the somatic musculature in some as-yet-unidentified manner, for example, pharmacologically. A plea is made for comparative studies between the two.

Biological Evolution↗

Studies of patients' thymi aid in the discovery and characterization of immunodeficiency in humans.

Studying the molecular and genetic bases of primary immunodeficiency is valuable at several levels. First, such information directly benefits patients in both short- and long-term management. Sophisticated diagnostic tools based on these studies can be used early and lead to appropriate treatment before potentially fatal infections and complications arise. Genotyping is also critical for future development and implementation of gene therapy. Secondly, investigating primary immunodeficiency helps understand the normal immune system in humans. As described in this report, the roles of zeta-associated protein of 70 kDa (Zap-70), CD25, and CD3delta are substantially different in humans when compared with the roles of homologous molecules in other species. Last, information obtained from these studies can be applied to other fields of investigation. Prominent examples for such applications include the intensive effort to design and produce specific inhibitors of Zap-70 and Janus kinase 3 as specific immunosuppressive agents. Most types of primary immunodeficiency in general and severe combined immunodeficiency in particular are rare and therefore cannot be easily studied by using traditional genetic methodology. Instead, biochemical methods were used to explore for candidate genes as was the case in the discovery of Zap-70 deficiency. Critical to the success of these discoveries was the careful analysis of patients' thymus glands. Detection of abnormalities in the thymus in these patients, which preceded identification of the genetic defect, aided in the assessment of the severity and nature of the immune disorder (primary versus secondary). Such assessment is critical before high-risk bone marrow transplantation. Equally important was the contribution of studies of the thymus to the description of novel phenotype of immunodeficiency as clearly demonstrated in defining CD8 lymphocytopenia, Zap-70 deficiency, and CD25 deficiency. Indeed, analysis of the thymus directly pointed to CD25 as candidate gene. Recently, the study of thymocyte-derived transcripts using DNA microarrays was key to discovering CD3delta deficiency. Finally, immunohistochemical analysis of the thymus was critical in pinpointing the roles of Zap-70, CD25, and CD3delta in the development of human T cells.

CD3 Complex↗