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N Wohllk

Publications and source records attributed to N Wohllk.

18 recordsLinked to original sources
PubMed2001

[Germline mutations of the ret proto-oncogene in Chilean patients with hereditary and sporadic medullary thyroid carcinoma].

BACKGROUND: Medullary thyroid carcinoma (MTC) may occur either as a sporadic or familial disease. Multiple endocrine neoplasia (MEN) type 2, inherited as an autosomal dominant disease, is characterized by MTC only (FMTC) or coexistence of MTC with other endocrine neoplasia (NEM 2A, 2B). Germline mutations of the RET proto-oncogene (cRet) are found in the inherited forms and in some apparently sporadic MTC cases. AIM: To study RET mutations in 8 families with MEN 2. MATERIAL AND METHODS: RET mutations were screened in peripheral blood DNA from 18 patients and 87 high risk carriers belonging to 8 MEN 2 families and 52 sporadic MTC. Exons 10, 11, 13, 14, 15 and 16 of the c-Ret were amplified by polymerase chain reaction (PCR) and examined by direct sequencing of PCR products and/or restriction enzyme analysis. RESULTS: Five MEN 2A and one FMTC families with a germline mutation at codon 634, one MEN 2A and one FMTC family carrying a mutation at codon 620 were identified. Mutations were found in 23 out of 87 high risk carriers. In addition, we detected a S891A (exon 15) germline mutation in a sporadic MTC patient and in one out of her three sons and V804M (exon 14) in another sporadic MTC case and in one out of his six relatives, indicating in both cases the presence of a sporadic misclassified familial disease. CONCLUSIONS: These results underscore the importance of routine application of c-Ret testing in all cases of MTC either familial or sporadic.

Adolescent↗
PubMed2000

[Treatment of diffuse hyperthyroid goiter with radioiodine: influence of propylthiouracil pretreatment].

BACKGROUND: To stabilize Graves disease and deplete the preformed hormone, the use of antithyroid drugs prior 131I therapy has been suggested, specially in those patients with severe thyrotoxicosis and in the elderly. However, PTU may reduce the effectiveness of 131I. AIM: To study the effects of PTU pretreatment before 131I administration. SUBJETS AND METHODS: A retrospective analysis of the medical records of patients with Graves disease treated with 131I from 1989 to 1997 was made. Of 244 patients with adequate follow-up for at least 12 months after 131I treatment, 142 had not been pretreated and 102 had received PTU prior to 131I therapy. Pretreated patients were distributed according to the number of days that PTU was discontinued before receiving 131I, forming four groups (a = 5 d, b = 6-14 d, c = 15-30 d and d = 31-60 d). Radioiodine was delivered according to our protocol of 120 microCi per gram of thyroid tissue, as estimated by clinical examination. Therapy was considered successful when laboratory evidence of euthyroidism or hypothyroidism after one year of treatment was obtained and as a failure when undetectable TSH values persisted after 12 months of treatment with 131I. RESULTS: All groups were comparable as to age, gender, goiter size, and 24 h radioiodine uptake. Control of hyperthyroidism was achieved in 76% of the non pretreated group. A similar percentage was observed in groups (b), (c) and (d). However, the disease was controlled in only 50% of group (a) patients (p < 0.003). CONCLUSIONS: The therapeutic efficacy of 131I is significantly reduced when the PTU is stopped for only a few days prior to the use of radioiodine. We postulate that PTU has to be discontinued for at least 10 days before radioiodine administration.

Adult↗
PubMed2000

[Multiple endocrine neoplasia: a clinical model for applying molecular genetic techniques].

Multiple endocrine neoplasias (MEN) are syndromes inherited as autosomal dominant. The application of the techniques of molecular biology has made possible the identification of the genes causing MEN 1 and 2. The gene responsible for MEN 1 belongs to the family of tumor suppressor genes and encodes for a protein named MENIN whose function remains to be elucidated. The identification of mutant MEN 1 gene carriers who are at risk of developing this syndrome requires frequent biochemical screening for the development of endocrine tumors. MEN 2 is a consequence of mutations in the Ret proto-oncogene (c-Ret). This gene encodes for a tyrosine kinase receptor thought to play a role in the development of neural crest-derived tissue. Members of kindred with either MEN 2A or MEN 2B should be screened by direct DNA testing early in life for mutations in c-Ret. Those with the mutation should be advised to have thyroidectomy at five years of age in children with MEN 2A and earlier in children with MEN 2B. Some cases of sporadic MTC are actually MEN 2A or Familial MTC after c-Ret testing is done, therefore routine application of this test is recommended in all cases of apparent sporadic MTC.

Adult↗
PubMed1998

A human succinate-ubiquinone oxidoreductase CII-3 subunit gene ending in a polymorphic dinucleotide repeat is located within the sulfonylurea receptor (SUR) gene.

We report the cloning of two variant genes encoding the CII-3 subunit of succinate-ubiquinone oxidoreductase complex II. One gene is located within intron 10 of the human sulfonylurea receptor gene. The 3' boundary of this gene ends in a polymorphic dinucleotide repeat. The second gene CII-3b is expressed at a low level and contains a 102-bp internal deletion compared to CII-3 cDNA. These genes should prove valuable in the characterization of Complex II disorders.

ATP-Binding Cassette Transporters↗
PubMed1996

Application of genetic screening information to the management of medullary thyroid carcinoma and multiple endocrine neoplasia type 2.

Application of RET proto-oncogene mutation analysis to the clinical management of MEN 2 and FMTC has simplified and enhanced the power of earlier used screening and treatment efforts for hereditary MTC. The approaches outlined herein are cost-effective, have improved diagnostic accuracy, and hold the promise of improved cure rates for this neoplasm. Further studies to elucidate the mechanism by which these activating mutations cause transformation may lead to other strategies for prevention or treatment of this neoplasm.

Base Sequence↗
PubMed1996

Relevance of RET proto-oncogene mutations in sporadic medullary thyroid carcinoma.

Analysis of peripheral blood or tumor DNA samples from 101 patients with apparent sporadic medullary thyroid carcinoma (MTC) was performed to assess the frequency of RET proto-oncogene mutations in this patient population. Peripheral blood and/or tumor DNA was amplified by polymerase chain reaction. DNA sequence or restriction enzyme analysis was performed to detect mutations of RET proto-oncogene codons 609, 611, 618, 620, 634, 768, and 918. Six of 101 patients with apparent sporadic MTC had peripheral blood DNA mutations more commonly associated with hereditary MTC. In 4 patients, these mutations led to the identification of previously unrecognized kindreds. The remaining 2 patients were examples of de novo mutations. A codon 918 mutation was found in 14 of 57 (approximately 25%) tumor DNA samples. Mutations were not identified in the remaining patients. In this large cancer center population, approximately 6% of patients with sporadic MTC carry peripheral blood DNA mutations, either inherited or de novo, more commonly associated with MEN 2A or familial MTC. Seven additional gene carriers were identified as a direct result of these studies, a 2-fold multiplying effect. We conclude routine application of RET proto-oncogene testing should be included in all cases of apparent sporadic MTC.

Base Sequence↗
PubMed1996

Inactivation of the first nucleotide-binding fold of the sulfonylurea receptor, and familial persistent hyperinsulinemic hypoglycemia of infancy.

Familial persistent hyperinsulinemic hypoglycemia of infancy is a disorder of glucose homeostasis and is characterized by unregulated insulin secretion and profound hypoglycemia. Loss-of-function mutations in the second nucleotide-binding fold of the sulfonylurea receptor, a subunit of the pancreatic-islet beta-cell ATP-dependent potassium channel, has been demonstrated to be causative for persistent hyperinsulinemic hypoglycemia of infancy. We now describe three additional mutations in the first nucleotide-binding fold of the sulfonylurea-receptor gene. One point mutation disrupts the highly conserved Walker A motif of the first nucleotide-binding-fold region. The other two mutations occur in noncoding sequences required for RNA processing and are predicted to disrupt the normal splicing pathway of the sulfonylurea-receptor mRNA precursor. These data suggest that both nucleotide-binding-fold regions of the sulfonylurea receptor are required for normal regulation of beta-cell ATP-dependent potassium channel activity and insulin secretion.

ATP-Binding Cassette Transporters↗
PubMed1996

The molecular basis for familial persistent hyperinsulinemic hypoglycemia of infancy.

Familial persistent hyperinsulinemic hypoglycemia of infancy (PHHI) is a glucose metabolism disorder in neonates characterized by inappropriate insulin secretion in the presence of profound hypoglycemia. Loss of function mutations in the sulfonylurea receptor (SUR) gene recently have been implicated as a cause for familial PHHI in nine independent families. This review will describe the combined positional cloning and candidate gene strategy used to identify the SUR gene as the one responsible for PHHI. Potential roles for SUR in other disorders of insulin secretion remains to be determined.

ATP-Binding Cassette Transporters↗
PubMed1995

Mutations in the sulfonylurea receptor gene in familial persistent hyperinsulinemic hypoglycemia of infancy.

Familial persistent hyperinsulinemic hypoglycemia of infancy (PHHI), an autosomal recessive disorder characterized by unregulated insulin secretion, is linked to chromosome 11p14-15.1. The newly cloned high-affinity sulfonylurea receptor (SUR) gene, a regulator of insulin secretion, was mapped to 11p15.1 by means of fluorescence in situ hybridization. Two separate SUR gene splice site mutations, which segregated with disease phenotype, were identified in affected individuals from nine different families. Both mutations resulted in aberrant processing of the RNA sequence and disruption of the putative second nucleotide binding domain of the SUR protein. Abnormal insulin secretion in PHHI appears to be caused by mutations in the SUR gene.

ATP-Binding Cassette Transporters↗
PubMed1995

RET proto-oncogene mutations in multiple endocrine neoplasia type 2 and medullary thyroid carcinoma.

The identification of RET proto-oncogene mutations in patients with MEN2 2 years ago was a watershed event in the management of this genetic cancer syndrome. The identification of a finite number of mutations that together causes more than 95% of hereditary and 15-25% of sporadic MTC has made it possible to develop simple and definitive tests to screen individuals at risk for this tumour syndrome. The impact of this technology is enormous. It is now possible to reassure 50% of family members at risk that they, and their children, do not have to worry about developing MTC. In the other 50% who are gene carriers, it is now possible to approach clinical management with greater certainty and plot strategies that are likely to result in a greater percentage of curative therapy. It seems likely that this technology will also have an impact on the management of sporadic MTC, although it is still too early to define a specific role for mutational analysis in these patients, except to exclude hereditary disease. The identification of specific mutations causative for MTC makes it possible to conceive future strategies for the treatment or prevention of MTC and to further extend the impact of these exciting findings.

Carcinoma, Medullary↗
PubMed1994

[Determination of antibody antireceptor of TSH, experience in patients with thyroid disorders and controls].

In order to measure TSH receptor antibodies (TRAb) we tried to set up a radioreceptor assay using human thyroid membranes. Due to lack of appropriate binding activity of the material obtained, we decided to use a kit which provides solubilized porcine membrane-receptors to TSH instead of human membranes, as well as calibrators that have been standardized in a receptor assay against MRC LATS std B. With these reactives we have measured TRAb in sera from 7 normal controls (C), 54 thyrotoxic patients (43 diffuse goiters [BDH], 8 multinodular goiters [BHM] and 3 Subacute Thyroiditis [TSA]), 3 patients with Hashimoto's Thyroiditis (TH) and in 6 non-hyperthyroid Graves ophthalmopathy patients. Measurement were initially performed using calibrators and the results expressed as U/L; since a very good correlation between the expression U/L and the calculated Inhibition Index (I.I.) was found (r = 0.99, n = 15, p < 0.001), results are shown using latter. In C mean +/- SD value for I.I. was 3.4 +/- 2.37%, so we decided to use, as cut off criteria for differentiating between normal and abnormal results, the figure 11%, which represents the mean +/- 3 SD. According to this, 93% of BDH had elevated TRAb activity while only slightly more than one third of MBH had elevated values, this difference being highly significant (p < 0.0001); both TSA and TH patients showed low TRAb activity while all Graves ophthalmopathy pts had elevated values, thus suggesting that they had a latent disease. We concluded that the methodology that is adequate and practical for clinical purposes.(ABSTRACT TRUNCATED AT 250 WORDS)

Antibodies↗
PubMed1993

[Lack of in vitro effect of antithyroid drugs upon peroxidase antigen expression in autoimmune thyroid disease].

The aim of this study is to determine whether antithyroid drugs (ATD) act via inhibiting thyroid hormone synthesis or by interfering with the immune process which leads to autoimmune disease. Previously we had demonstrated that ATD do not affect HLA-DR and TPO antigen expressions induced by appropriate stimulus in normal thyrocytes, so we decided to study what happens when the same experiments are performed using autoimmune thyrocytes. Cultured thyroid tissue from patients operated on for Graves' Disease or Hashimoto's Thyroiditis were stimulated with TSH or TBII alone or associated with ATD; TPO antigen expression was evaluated by the Cytotoxicity Assay using human monoclonal antiTPO. When autoimmune thyrocytes were cultured and no stimulus was used or with the addition of MMI or PTU alone, very low values of TPO expression were noted (8.6 +/- 6.7, 5.0 +/- 7.1 and 4.2% +/- 2.3% respectively); if they were stimulated with TSH or TBII, a sharp rise of TPO antigen expression was detected (54.4 +/- 23.3 and 62.6 +/- 16.5%), these figures being significantly different from unstimulated cells (p < 0.001). If both stimulus were used associated with ATD, the high TPO antigen expression was unaffected. It is concluded that, at least in vitro, ATD have no effect upon induced-antigen expression in autoimmune thyrocytes. Since they do not alter antigen presentation, a primary step in the immune process, it is difficult to accept that their mechanism of action is through interference with the immune response.

Antithyroid Agents↗
PubMed1993

[Experimental evidence that endogenous TSH modulates microsomal antigen expression: clinical implications].

The purpose of this work was to study if TSH has a role in TPO antigen expression in vivo. Using the cytotoxicity assay we measured TPO expression and correlated it with TSH serum levels in 3 groups of rats: control, hypothyroid and hypothyroid supplemented with thyroxine. For comparative purposes, in the cytotoxicity assay we used rat monoclonal antiTPO or human sera with high titles for antiTPO antibodies. Hypothyroid rats showed marked elevations of TSH serum levels and TPO antigen expression in their thyrocytes when compared to the control and supplemented group. A positive correlation between TPO antigen and TSH levels was observed (r = 0.69, p < 0.001). There was an excellent correlation between TPO results using rat monoclonal or human sera antibodies (r = 0.94 p < 0.0001). It is concluded that TSH modulates TPO antigen expression. These data are of clinical relevance considering that TSH modulates the expression of other antigens that can maintain the immune response and perpetuate the immune disease in patients with Graves disease treated with antithyroid drugs. Thus, the avoidance of TSH hypersecretion with administration of thyroxine could be useful to treat these patients.

Analysis of Variance↗
PubMed1993

[Importance of gamma-interferon and DR expression in autoimmune thyroid disease].

The purpose of this paper is to study some of the mechanisms by which antigen presentation, one of the first steps in the immune process, is modulated. Three different experiments are performed: a) Using thyroid tissue from patients operated on for Graves' Disease, spontaneous HLA-DR and TPO antigen expressions were measured through the Cytotoxicity Assay; these findings were correlated with the presence of antiTPO in the sera of these patients. It was found that only patients whose thyroid tissue spontaneously expressed both antigens had circulating antiTPO in their sera, thus demonstrating that dual expression is basic for antibody production. b) Blood samples from other Graves' patients were obtained; peripheral lymphocytes were isolated and cultured in complete media for 5 days, then supernatant was separated and IFN-g concentration was measured by a sandwich type RIA; the same procedure was done in 12 normal controls in order to compare the results. It was found that lymphocytes from Graves' patients secreted significantly more IFN-g than normal controls (20.9 +/- 13.54 U/ml vs 3.7 +/- 3.22 U/ml respectively, p < 0.001) confirming that they were sensitized, so this determination could be used as a marker of immune process, if other infectious conditions are excluded; also it was found that IFN-g hypersecretion persists once hyperthyroidism has been treated, pointing out that the immune abnormality is still present.(ABSTRACT TRUNCATED AT 250 WORDS)

Antigen Presentation↗
PubMed1993

[Thyroid profile in normal pregnancy].

The aim of this study was to update the thyroid hormone profile in normal pregnant women with adequate iodine nutrition, to analyze the physiological changes that occur during pregnancy and to know the role that TBG and bHCG exert on these changes. One hundred six pregnant women without goiter, former thyroid diseases or positive antimicrosomal antibodies were studied. Fifty three of them were prospectively followed during the gestational period. Thirty age matched non pregnant women were studied as a control group. Serum T3t, T4t, T41, conventional and IRMA TSH, rT3, TBG, bHCG, antimicrosomal antibodies and urinary iodine content were measured. Median urinary iodine content was 18.9 ug/ml in pregnant women, discarding iodine deficiency, the main observed changes occurred between weeks 6 and 14 with significant elevations of T3t, T4t, T41, rT3, TBG and bHCG and TSH decrease. There was a positive correlation between TBG and T3t and T4t indicating a causal relationship. There was a negative correlation between T41 and TSH and between TSH and bHCG and a positive correlation between T41 and bHCG, suggesting a thyroid stimulator effect of bHCG which would raise T41 and thus inhibit TSH secretion.

Adolescent↗
PubMed1993

[Treatment of hyperthyroidism during pregnancy: experience with 19 cases].

Untreated hyperthyroidism in pregnant women is associated with a high incidence of maternal and fetal complications. Thus, its treatment is mandatory, ideally using PTU because it has lesser transplacental passage. From 1987 and 1991 we have attended 19 hyperthyroid pregnant women. Of these, 18 had diffuse and 1 nodular goiter and in 10, thyrotoxicosis preceded pregnancy. PTU was used in 17 women (7 received it along the whole pregnancy), five had to be operated due to poor response, one received propranolol and one patient was not treated due to lack of attendance. Cesarean section was performed in 12 women, 5 had vaginal delivery, one had a miscarriage at the 20th week of pregnancy due to a neurological malformation and one patient was lost from control before delivery. The newborn of the untreated woman had a neonatal thyrotoxicosis and the resting 16 did not show evidence of thyroid disfunction. Newborns from mothers receiving PTU until delivery had significantly lower rT3 levels and non significant changes in T4 and T3. At the end of the observation period, 8 patients were euthyroid, 3 hypothyroid (2 after 131-l and 1 after surgery), 4 continued on PTU and 4 were lost from control. It is concluded that the outcome of pregnancy may be uneventful in hyperthyroid women provided that there is a close and adequate follow up.

Adult↗
PubMed1991

[Spontaneous expression of class II (HLA-DR) molecules in thyroid pathology].

The spontaneous expression of Class II molecules (HLA-DR) was studied in cultured thyrocytes obtained from patients with Graves Disease (n = 7), Hashimoto's Thyroiditis (n = 5), euthyroid nodular goiter, (n = 12), papillary carcinoma (n = 5), and laryngeal carcinoma (3 normal thyroid glands). If nodular goiters and papillary carcinoma are of autoimmune origin, as Graves Disease and Hashimoto's Thyroiditis, they should spontaneously express HLA-DR antigen on their cell surface as this has been considered one of the initial steps of autoimmunity. The study was performed using the cytotoxicity assay. Immediately after the thyroid glands were obtained, thyrocytes were labelled with 51-Cr and incubated overnight; the cells were destroyed by adding monoclonal antiHLA-DR antibody and rabbit complement. The cytotoxicity index (CI% + SD) which reflects 51-Cr release from lyzed cells was used to measure antigen expression. While Graves Disease's and Hashimoto's Disease's thyrocytes expressed HLA-DR in high proportion, normal thyrocytes and thyroid cells from other diseases did so in minimal proportion (28.12 +/- 10.71 vs 2.26 +/- 2.32, p < 0.001). These findings strongly suggest that nodular goiters and papillary carcinoma are not of autoimmune origin since they are unable to express HLA-DR on their cell surface. It is postulated that HLA-DR expression is the result of the influence of T lymphocytes previously sensitized to thyroid antigens.

Autoimmune Diseases↗