Biomedical subjects

Y Malthièry

Publications and source records attributed to Y Malthièry.

10 recordsLinked to original sources

PubMed1997

Male infertility associated with multiple mitochondrial DNA rearrangements.

Male sterility results from a number of characterized exogenous or genetic dysfunctions preventing normal differentiation into mobile spermatozoa. This may now be overcome by intra cytoplasmic sperm injection (ICSI). This practice does not require mobile, or even mature spermatozoa for in vitro fecondation. However, a functional respiratory chain, partly encoded by the mitochondrial DNA (mtDNA), is required for the mobility of the spermatozoa. We report the case of an infertile patient who wished to procreate. ICSI was proposed but he displayed multiple mtDNA deletions of possible nuclear origin in the spermatozoa and in the deltoid muscle. Even though mtDNA is maternally inherited, the possibility of a nuclear-driven mutation affecting the integrity of the mtDNA should be taken into account when ICSI is to be performed. Together with recent genetic in vitro manipulations in mammals, our data point to the importance of studying the mtDNA structure in human spermatozoa, and the potential risks of these non-natural practices for procreation.

Adult↗

PubMed1997

Oligoasthenospermia associated with multiple mitochondrial DNA rearrangements.

A patient who wished to be treated for infertility by intracytoplasmic sperm injection (ICSI) was referred to our group for assessment. Upon clinical examination, a ptosis (partial closure of the eyelid) was noted, and histology revealed ragged red fibres in the skeletal muscle. Southern blot analysis of spermatozoa and skeletal muscle revealed the presence of multiple mitochondrial DNA deletions. This kind of rearrangement may be of nuclear origin since three nuclear loci have been ascribed to multiple mitochondrial DNA deletions in humans. Since mitochondrial DNA is maternally transmitted, the use of ICSI was feasible. However, an alteration of nuclear gene product affecting the integrity of mitochondrial DNA, and thus sperm mobility, might be transmitted to the offspring with the risk of developing a mitochondrial DNA disease.

Adult↗

PubMed1994

Association of deletion and homoplasmic point mutation of the mitochondrial DNA in an ocular myopathy.

The mitochondrial DNA of a 41 year old patient with ocular myopathy was explored. We found a deletion of 3540 base pair in about 50% of the mitochondrial genomes associated with a homoplasmic point mutation. The mutation at nucleotide pair 7444 converts stop codon AGA into lysine codon AAA (human mitochondrial genetic code). The synergistic effect between two point mutations has already been described in mitochondrial pathology but this is the first time that an association between a deletion and a point mutation is shown.

Adult↗

PubMed1993

Epitope mapping of human thyroid peroxidase defined seven epitopes recognized by sera from patients with thyroid pathologies.

Human thyroid peroxidase (hTPO) is the major component of the microsomal antigen. In almost cases, antibodies against this protein are found in sera from patients with autoimmune thyroid diseases. Overlapping cDNAs which correspond to the complete hTPO mRNA obtained from a thyroid library or by polymerase chain reaction were cloned and expressed as fusion proteins in a prokaryotic vector. Seven antigenic determinants between 21 and 49 amino acids were defined by cloning, subcloning of the immunoreactive regions and screenings with the two rabbit polyclonal anti hTPO antibodies. This study confirms the antigenic nature of the sequences 70-160 and 590-675 but above all refines the localization of three shorter distinct antigenic peptides corresponding to the sequences 68-105, 106-126 and 574-621. Moreover, four other determinants were characterized on the sequences 233-277, 467-515, 641-685 and 701-730. Analysis of sera from patients with autoimmune thyroid diseases (AITD) against the seven immunoreactive peptides confirms the heterogeneous nature of autoantibodies to hTPO.

Autoantibodies↗

PubMed1992

A major human thyroglobulin epitope defined with monoclonal antibodies is mainly recognized by human autoantibodies.

The antigenic nature of 15 anti-human thyroglobulin (hTg) monoclonal antibody (mAb) epitopes was studied by two different approaches. First, we tested two successive protease-digest products of hTg. Only four mAb from the same cluster of reactivity recognized a low-molecular weight peptide, the other mAb only bound native hTg or high-molecular weight digest fractions. Second, these 15 mAb were used to immunoscreen hTg expression libraries. Only the same four mAb revealed immunoreactive clones corresponding to region 1149-1295 on the hTg primary sequence. After subcloning, this antigenic determinant was reduced to a 102-amino acid peptide (hTg region 1149-1250). The two different methodologies were coherent and complementary, and demonstrated that hTg sequence 1149-1250 is the target for this cluster of four mAb. Moreover, anti-hTg autoantibodies which cross-reacted with these mAb bound the 102-amino acid peptide. This epitope was the one most frequently detected by sera from autoimmune thyroid disease. The data confirm the presence of an immunodominant domain in the central part of the hTg molecule and suggest that this mAb epitope may be a powerful probe for the diagnosis of autoimmune thyroid disorders.

Amino Acid Sequence↗

PubMed1991

Epitope mapping of human thyroglobulin reveals a central immunodominant region.

Thyroglobulin is the thyroid hormone precursor and the major antigen frequently involved in autoimmune diseases. The primary structure of human thyroglobulin is known but the spatial structure remains largely undetermined. By using fusion protein produced in prokaryotic system we have characterized seven short immunoreactive peptides carrying at least one epitope. None of them includes hormonogenic sites, but five are concentrated in the central part of the monomeric molecule, which thus emerges as the major immunogenic region of this protein.

Epitopes↗

PubMed1990

Epitope mapping of human thyroglobulin. Heterogeneous recognition by thyroid pathologic sera.

Thyroglobulin is the major Ag of the thyroid gland involved in autoimmune pathologies. Epitope mapping was carried out with a rabbit polyclonal immune serum against fusion proteins expressed in prokaryotic cells. After screening of an initial human thyroglobulin cDNA library and subcloning of immunoreactive clones, seven epitopes were characterized and localized on the human thyroglobulin monomeric molecule. One was close to each extremity of the molecule, and five others were concentrated in the middle, covering a sixth of this 2748-amino-acid chain. The immunoreactivities of 18 autoimmune sera from different thyroid pathologies were tested against the seven previously characterized epitopes. Those from Hashimoto's thyroiditis were the most immunoreactive. Immune responses were heterogeneous for sera from different pathologies as well as for those from the same pathology. The central epitopes and the near-C-terminal epitope, however, were the epitopes most often recognized by the immune sera. These findings show that some autoepitopes overlap accurately with some heteroepitopes characterized by a polyclonal immune serum directed against the mature protein.

Amino Acid Sequence↗

PubMed1990

Evidence for an alternate splicing in the thyroperoxidase messenger from patients with Graves' disease.

An initial lambda gt 11 cDNA library constructed from a human Graves' patient thyroid was screened with an immunopurified rabbit anti-human thyroperoxidase (hTPO) polyclonal antibody. A 869 bp clone was obtained. It presents a 130 bp deletion as compared to the published sequence and a 77 bp insertion in the 3' non-coding region. Screening of a pUC cDNA library from another Graves' patient thyroid exhibited the same 130 bp deletion in two other cDNA clones. PCR analysis of mRNA transcripts confirmed the presence of the two messengers in two other Graves' thyroid tissues. In all the cases, this new spliced mRNA species represents between 40% and 50% of the total hTPO mRNAs. With respect to the structure of the hTPO gene, the present deletion suggests an alternate splicing of exon 16. The juxtaposition of exon 17 to exon 15 encoding the transmembrane domain leads to a shift in the reading frame. By the use of a different stop codon, the spliced mRNA generates a modified 56 - COOH terminal aminoacids (aa) sequence.

Amino Acid Sequence↗

PubMed1989

Thyroglobulin structure and function: recent advances.

Thyroglobulin is a large-size iodoglycoprotein specific to thyroid tissue and is the substrate for the synthesis of thyroid hormones, thyroxine and 3,5,3'-triiodothyronine. Recent studies, which greatly benefited from recombinant DNA methodologies, improved the knowledge of several structural features of this dimeric protein and permitted insights into some structure-function relationships. Analysis-function of the primary structure of the human thyroglobulin monomer revealed several main characteristics: 1) 3 types of internal homologies; 2) extensive homology with the bovine thyroglobulin monomer and known partial sequences in the thyroglobulins of other mammalian species; 3) significant homologies with 2 other non-thyroid proteins (acetylcholinesterase and the invariant chain of the Ia class II histocompatibility antigen); 4) a terminal localization of the hormonogenic sites at both ends of the monomer. Current studies aim at determining conformational characteristics, understanding the molecular mechanisms of thyroid hormone formation and unraveling those interactions which in the thyroid cell and the thyroid follicle will permit this large pro-hormone to synthesize and release a few small thyroid hormone molecules. A more precise knowledge of this molecule in higher vertebrates and during evolution would impart valuable information concerning thyroid pathology, since thyroglobulin has been implicated in some genetic and in autoimmune thyroid diseases.

Amino Acid Sequence↗

PubMed1989

[Structural heterogeneity of human thyroglobulin in relation to its messenger RNA maturation].

One human thyroglobulin cDNA clone present 192 nucleotides deletion which correspond to 64 amino-acid deletion on the protein molecule. We show that this fact figure the exon loss on the mRNA molecule using as template during cDNA synthesis by comparison between mRNA and genomic DNA nucleotide sequences. Different possibility can explain this fact. We retain mainly mistake in mRNA maturation fidelity. This type of event is not an exception and can be an explanation for some pathologies.

Chromosome Deletion↗