[The therapeutic purpose of plastic surgery]. / La finalité thérapeutique en chirurgie plastique.

The acts of plastic surgery consist in providing care to patients and pursue a therapeutic purpose. They benefit from VAT exemption in accordance with European legislation. They appear in the Common Classification of Medical Acts as therapeutic acts, whether or not they are covered by health insurance. To assist the plastic surgeon during the consultation in his role as a medico-surgical expert, we propose a global assessment scale for physical disgraces. This scale specifies the importance of disharmony, its etiology, and considers the patient in his entirety, both biologically and psychosocially. All repercussions are analyzed: physiopathological impact, pain, functional impact on daily life activities, pleasure and sexual activities, psychological, social, and professional impact. Each item is independently rated. Depending on the overall score obtained, the therapeutic nature of the act can be confirmed. The proposed scale is a simple tool that easily and seriously supports the therapeutic nature of our acts. It ensures argued expertise, a reliable and indisputable synthesis. We are, above all, doctors, caregivers, contributing to health as defined by the WHO: "a state of complete physical, mental, and social well-being." We alone are able to judge the therapeutic nature of a service, in agreement with our patients.

[Three recurrent episodes of Tako-Tsubo cardiomyopathy precipitated by an emotional stressful event: a case report]. / Trois épisodes récurrents de Tako-Tsubo secondaires à un choc émotionnel : à propos d'un cas clinique.

Tako-Tsubo cardiomyopathy, first described in 1990 by Sato in Japan, has recently gained increasing consideration when reported in non-Japanese patients, including the United States and Europe. Typical presentation mimics acute coronary syndrome, with acute chest pain and/or dyspnoea, associated to electrocardiographic changes and moderate cardiac biomarkers release, but in which coronary angiography reveals no coronary arteries lesions and echocardiography or left ventriculography shows a reversible left ventricle systolic dysfunction. Prognosis is good, in contrast to acute coronary syndrome, provided that the patients survive the possible life-threatening acute presentation, with correction of the left ventricle systolic dysfunction within several days or weeks. As noted in several reviews, 3.5% to 10% of the patients have a recurrence during the first few years after the initial presentation. Here, we described a case of a 60-year-old female who had three episodes of Tako-Tsubo always preceded by severe emotional stress suggesting a potential common etiopathogenesis.

[Perilunate dislocations and fracture-dislocations of the wrist, a review of 14 cases]. / Luxations et fractures-luxations périlunaires du carpe, étude rétrospective d'une série de 14 cas.

OBJECTIVES: We report a retrospective series of 14 dislocations or perilunate fracture-dislocations. The results of our series are compared with the data of the literature and we discuss epidemiology, types of lesions, surgical treatment, complications and prognosis of this pathology. METHODS: The series included seven pure dislocations and seven fracture-dislocations including three trans-scapho-lunate forms (including one Fenton's syndrome). The displacement of all these lesions was posterior. The mean age was 35 years. Sixty-four percent were manual workers. All 14 patients had undergone surgical treatment through a dorsal approach in the first seven days following the injury. They were reviewed clinically and radiologically with a mean follow-up of 25 months. RESULTS: The average Cooney functional score was 72/100 with two excellent, six good, four fair and two poor results. Average flexion-extension motion arc was 74%, the grip strength was 77% compared to the other wrist. Persistent wrist pain was almost constant. One carpal instability was observed and one patient required a four-corner arthrodesis for SLAC wrist. Eighty-five percent of all patients were employed at least. CONCLUSIONS: Early diagnosis and anatomical reduction can provide satisfactory functional results. Emergency surgical treatment is required. We prefer a dorsal approach and we do not perform primary closed reductions.

[Heterotopic digital transplantation in an 80-year-old man]. / Replantation digitale hétérotopique chez un homme de 80 ans.

The authors report a case of amputation of the first, second and third fingers of the left hand in an 80-year old man. As the thumb was not replantable, a pollicization by hetero-replantation of the index was performed as an emergency. With a 1-year follow-up, the functional result was satisfactory. Analysis of the case report again shows that in the case of thumb amputation, hetero-replantation according to the principles of "finger-bank" is the solution of choice and demonstrates its feasibility in the elderly. The authors discuss the recovery of sensation.

DNA-interacting proteins in the spermiogenesis of the mollusc Murex brandaris.

Sperm chromatin of Murex brandaris (a neogastropod mollusc) undergoes a series of structural transitions during spermiogenesis. The DNA-interacting proteins responsible for these changes as well as the mature protamines present in the ripe sperm nucleus have been characterized. The results reveal that spermiogenic nuclear proteins are protamine precursors that are subjected to a substantial number of small N-terminal deletions that gradually modify their overall charge. The composition of mature protamines is remarkably simple in turn, promoting an efficient and extremely tight packaging of DNA. The pattern of spermiogenic chromatin condensation in M. brandaris clearly departs from that corresponding to vertebrate chromatin.

New toxins acting on sodium channels from the scorpion Leiurus quinquestriatus hebraeus suggest a clue to mammalian vs insect selectivity.

Two new toxins were purified from Leiurus quinquestriatus hebraeus (Lqh) scorpion venom, Lqh II and Lqh III. Lqh II sequence reveals only two substitutions, as compared to AaH II, the most active scorpion alpha-toxin on mammals from Androctounus australis Hector. Lqh III shares 80% sequence identity with the alpha-like toxin Bom III from Buthus occitanus mardochei. Using bioassays on mice and cockroach coupled with competitive binding studies with 125I-labeled scorpion alpha-toxins on rat brain and cockroach synaptosomes, the animal selectivity was examined. Lqh II has comparable activity to mammals as AaH II, but reveals significantly higher activity to insects attributed to its C-terminal substitution, and competes at low concentration for binding on both mammalian and cockroach sodium channels. Lqh II thus binds to receptor site 3 on sodium channels. Lqh III is active on both insects and mammals but competes for binding only on cockroach. The latter indicates that Lqh III binds to a distinct receptor site. Thus, Lqh II and Lqh III represent two different scorpion toxin groups, the alpha- and alpha-like toxins, respectively, according to the structural and pharmacological criteria. These new toxins may serve as a lead for clarification of the structural basis for insect vs mammal selectivity of scorpion toxins.

Nuclear basic proteins in spermiogenesis.

In animal species, spermiogenesis, the late stage of spermatogenesis, is characterized by a dramatic remodelling of chromatin which involves morphological changes and various modifications in the nature of the nuclear basic proteins. According to the evolution of species, three situations can be observed: a) persistence of somatic histones or appearance of sperm-specific histones; b) direct replacement of histones by generally smaller and more basic proteins called protamines; and c) occurrence of a double nuclear basic protein transition: histones are not directly replaced by protamines but by intermediate basic proteins which are themselves replaced by one or several protamines. However, in some species, two kinds of intermediate basic proteins can be distinguished in spermatid nuclei: transition proteins and protamine precursors. Whereas transition proteins are not structurally related either to histones or to protamines, protamine precursors are further processed at the end of spermiogenesis to give rise to the mature protamine. The molecular characteristics of the protamines as well as number of protamine types present in the spermatozoon vary from species to species. In some cases, protamine-encoding genes, although present, are not expressed to a significant level. The diversity and the precise function of intermediate basic proteins remain open to discussion. Some of them are the precursors of protamines but the mechanism, sequential or not, as well as the enzyme(s) involved in the proteolytic processing, remain to be discovered.

Spermatogenesis of the lizard Lacerta vivipara: histological studies and amino acid sequence of a protamine lacertine 1.

The lizard Lacerta vivipara is a seasonal breeder with a well characterized reproductive cycle. An histological study of the lizard testis has been performed at different stages of spermatogenesis and the nuclear basic proteins content was assessed by electrophoretical analysis. Two protamines, lacertines 1 and 2, are present in spermatozoa in April and May. We have isolated lacertine1 and characterized a protamine with a mass of 4,963.7 Da. Amino acid sequence of this protamine (41 residues) was established from data provided by automated Edman degradation. It is characterized by a basic amino acid stretch in the N- and C-terminal regions and by a central part which only consists of 3 different intermingled amino acids. This protamine presents 62% homology with scylliorhinine Z3 from dog-fish Scylliorhinus caniculus and 58% homology with quail protamine. The reported lizard protamine sequence is the first reptilian protamine sequence available so far.

Sequence analysis and structural features of the largest known protamine isolated from the sperm of the archaeogastropod Monodonta turbinata.

Protamine of the archaeogastropod mollusc Monodonta turbinata has been isolated and characterized. With a mass of 13,476 Da, it is the largest known protamine. Amino acid sequence of this protamine (106 residues) was established from data provided by automated sequence analysis and mass spectrometry of the protein and of its fragments. The primary structure of the NH2-terminal region exhibits repetitive sequence motifs "Basic-Ser" (mainly R-S) and both central and COOH-terminal regions are composed by arginine clusters. The amino acid sequence of Monodonta turbinata protamine shows structural similarities with other protamines from invertebrates and from birds and mammals.

Persistence of protamine precursors in mature sperm nuclei of the mouse.

During mouse spermiogenesis, two protamines, mP1 and mP2, are synthesized in replacement of histones. One of them (protamine mP2, 63 residues) appears at first in elongating spermatid nuclei as a protamine of 106 residues (pmP2) with an amino-terminal extension that is progressively excised. The two protamines were previously described as the only proteins associated with DNA in sperm chromatin. This paper shows that the nuclear proteins of mouse spermatozoa are indeed heterogeneous: at least six minor polypeptides in addition to protamines can be identified. The primary structure of four of them has been established. They are intermediate in the maturation of the precursor of protamine mP2 and correspond to polypeptides pmP2/11, pmP2/16, pmP2/20, and pmP2/32, characterized previously in mouse testis. Therefore, these intermediates of proteolysis generated from pmP2 inside spermatid nuclei persist in mature sperm, whereas the largest precursors, pmP2 and pmP2/5, disappear. These findings clearly indicate that limited proteolysis events still occur outside of the testis.

Electroblotting proteins onto carboxymethylcellulose membranes for sequencing.

Proteins and peptides separated by polyacrylamide gel electrophoresis either in acetic acid-urea or in sodium dodecyl sulfate (SDS)-containing gels were electroblotted onto carboxymethylcellulose membrane and eluted from the membrane by dilute acid. Polypeptides thus recovered with a high yield can be used to get structural or biological data.

Phosphorylation of human sperm protamines HP1 and HP2: identification of phosphorylation sites.

Human sperm is characterized by a high heterogeneity of its basic nuclear protein complement of pro-protamines, protamines and histones. This heterogeneity is increased by the persistence of phosphorylated protamines in mature spermatozoa. Alkaline phosphatase treatment of whole protein indicated that protamines HP1 and HP2 were phosphorylated to various degrees. Presence of non-phosphorylated and phosphorylated protamines HP1 and HP2 was further demonstrated by electrospray mass spectrometry. Phosphorylation sites of mono- and di-phosphorylated protamine HP1 were identified by automatic Edman degradation of the protein after phosphoserine derivatization to S-ethylcysteine. In both phosphorylated forms, Ser-10 was found phosphorylated; in the di-phosphorylated form, Ser-8 was identified as the second site of phosphorylation. In protamine HP2, the unique site of phosphorylation (Ser-14) was located after limited acid hydrolysis of enzymic peptides and thin-layer electrophoresis.

Post-translational chemical modifications of proteins--III. Current developments in analytical procedures of identification and quantitation of post-translational chemically modified amino acid(s) and its derivatives.

1. The Chemical modifications of amino acids and their derivatives are mainly due to different post-translational enzymatic reactions. 2. The enzymatic reactions resulting in amino acids such as acetylation-, formylation, methylation-phosphorylation-, sulfation-, hydroxylation, ADP ribosylation-, carboxylation-, amidation-, adenylylation-, glycosylation-, ubiquitination-, prenylation and acylation are listed and analytical methods are reported and extensively reviewed. 3. The post-translationally modified cross-linking molecules after maturations such as desmosines, allo-desmosine, hydroxy-, lysylpyridinoline, 3-hydroxypyridinium derivatives, cyclopentenosine recently found in matured elastin, and in collagen, and pulcherosine a novel tyrosine-derived found in fertilization envelope of Sea Urchin embryo, di-tyrosine in resilin, gamma-glutamyl-lysine isopeptide cross-linking molecule etc. are listed and both physico-chemical and analytical methods are extensively reviewed and discussed. 4. Other consequences of post-translational modifications encountered in the analytical procedure such as N-terminal step-wise Edman degradation of glycosylated site(s), phosphorylated-site(s) and or sulfated-site(s) were also reported by us.

Amino acid sequence of the human intermediate basic protein 2 (HPI2) from sperm nuclei. Structural relationship with protamine P2.

Human intermediate basic protein 2 (HPI2) is a low-molecular-mass basic protein present in small amounts in human sperm nuclei. The amino acid composition of the protein, its N-terminal amino acid sequence and peptide maps obtained after digestion with endoproteinases Lys-C and Glu-C, reveal that HPI2 is structurally related to human protamine species P2 (HP2), which is rich in Arg, His and Cys residues. Compared to HP2, which is one of the two major sperm protamines, HPI2 has an N-terminal extension of 24 residues which includes six acidic residues and does not possess any Arg residues. The amino acid sequence of HPI2 (81 residues) is identical to the sequence of the C-terminal region of another minor sperm nuclear protein, human intermediate basic protein 1 (HPI1, 101 residues), which was sequenced previously [Martinage, A., Arkhis, A., Alimi, E., Sautière, P. & Chevaillier, P. (1990) Eur. J. Biochem. 191, 449-451]. Due to this structural similarity, HPI2 must be considered as an intermediate in the maturation of proprotamine HPI1 limited proteolysis.

Possible relationship between coding recognition amino acid sequence motif or residue(s) and post-translational chemical modification of proteins.

1. The "code-sequence" of N-glycosylation site(s), the amino acids located around O-glycosylation site(s), the sequence motifs of several kinases, the sequence motifs of--sulfation, amidation, isoprenylation, myristoylation, palmitoylation and N-acetylation, Aspartic and Asparagine hydroxylation-site, gamma-carboxyglutamate domain, phosphopantetheine attachment site etc. are extensively listed, compared to those reported by "PROSITE" Computer Screen Center and discussed. 2. The structural aspects of protein-DNA recognition are quoted as discussion and conclusion.

Molecular characterization of six intermediate proteins in the processing of mouse protamine P2 precursor.

In mouse spermatozoa, DNA is compacted by two protamines mP1 and mP2. Protamine mP2 (63 residues) is synthesized in spermatid nuclei as a precursor pmP2 (106 residues) which is subsequently processed at the end of spermiogenesis [Yelick, P.C., Balhorn, R., Johnson, P.A., Corzett, M., Mazrimas, J.A., Kleene, K.C. & Hecht, N.B. (1987) Mol. Cell. Biol. 7, 2173-2179]. Six proteins, three of which were described earlier [Chauvière, M., Martinage, A., Debarle, M., Alimi, E., Sautière, P. & Chevaillier, Ph. (1991) C.R. Acad. Sci. 313, 107-112], have molecular and electrophoretic properties similar to those of pmP2. They were isolated from purified testis nuclei and characterized by amino acid composition, N-terminal sequence and peptide mapping. From the amino acid compositions, it appears that all six proteins are rich in arginine, cysteine and histidine and are closely related to pmP2 and mP2. The N-terminal sequence of each protein overlaps a distinct region of the N-terminal part of pmP2. The C-terminal part of protamine mP2 starting at arginine 15 is common to all proteins as assessed by amino acid compositions and peptide maps. All these structural data demonstrate that the six isolated proteins are products of pmP2 precursor processing. The six intermediate proteins pmP2/5, pmP2/11, pmP2/16, pmP2/20, pmP2/26 and pmP2/32 which contain 102, 96, 91, 87, 81 and 75 residues, respectively, are generated from the pmP2 precursor after N-terminal excision of 4, 10, 15, 19, 25 and 31 residues, respectively. The C-terminal sequence of protamine mP2 is strictly identical to that of its precursor; therefore, no maturation occurs in this part of the molecule. At the present time, the proteolytic pathway involved in the amino-terminal processing leading to the mature form of the protamine mP2 (63 residues) has not been elucidated. However, the different representation of six intermediates in the testis suggests that some stages of processing are faster than others or that some cleavage sites are preferred. The proteins described in this paper could result either from stepwise excision of N-terminal residues or from non-sequential cleavages.

Post-translational chemical modification(s) of proteins.

1. The role played by the modification of protein in determining its fate is reported by us. 2. Post-translational modifications such as acetylation, phosphorylation, sulfation, methylation, hydroxylation, ADP-ribosylation, maturation, amidation, carboxylation, adenylylation, glycosylation, ubiquitination, and prenylation are extensively reviewed. 3. Each post-translational modification's significance and its role played in biological function(s) is summarized in the general discussion and the conclusion's remark is directed at the problems left to solve (e.g. post-translational modification reactions in recombinant protein in modern genetic engineering).

Phosphorylation of Toxoplasma gondii major surface antigens.

The phosphorylation of the major surface proteins of Toxoplasma gondii tachyzoites was investigated. Metabolic labeling of intracellular tachyzoites with [32P]-orthophosphate followed by immunoprecipitation with specific monoclonal antibodies showed that all five major surface proteins were labeled as expected for GPI-anchored proteins. More detailed analysis of the phosphorylated sites revealed that surface proteins P30 and P22 contained phosphoserine residues in addition to the phosphorylated molecules that are presumably localized in the GPI-membrane anchors.

Molecular structure of human protamine P4 (HP4), a minor basic protein of human sperm nuclei.

Protamine HP4 is a minor protein which was purified from human sperm nuclei. It was characterized by its amino acid composition, peptide mapping after digestion with highly specific endoproteinases and finally by its amino acid sequence. Protamine HP4 contains high amounts of arginine, cysteine and histidine. The primary structure of the protein was established by sequence analysis of intact protamine and of its fragments. HP4 is a P2-type protamine of 58 residues (Mr 7783) structurally related to human protamines HP2 and HP3 from which it only differs by an amino-terminal extension of one and four residues, respectively. These three protamines exhibit a close structural relationship with mouse protamine mP2. The heterogeneity of protamines in human sperm nuclei is discussed.

Nuclear transition protein 1 from ram elongating spermatids. Mass spectrometric characterization, primary structure and phosphorylation sites of two variants.

The ram transition protein 1 (TP1) is present in spermatid cell nuclei in the nonphosphorylated, monophosphorylated and diphosphorylated forms. Its primary structure was determined by automated Edman degradation of S-carboxamidomethylated protein and of peptides generated by cleavage with thermolysin and endoproteinase Lys-C. The ram TP1 is a small basic protein of 54 residues and structurally very close to other mammalian TP1. The mass spectrometric data obtained from the protein and its fragments reveal that ram TP1 is indeed a mixture (approximately 5:1) of two structural variants (Mr 6346 and 6300). These variants differ only by the nature of the residue at position 27 (Cys in the major variant and Gly in the minor variant). The study of phosphorylation sites has shown that four different serine residues could be phosphorylated in the monophosphorylated TP1, at positions 8, 35, 36 or 39. From previous physical studies, it has been postulated that the Tyr32 surrounded by two highly conserved basic clusters was responsible for the destabilization of chromatin by intercalation of its phenol ring between the bases of double-stranded DNA. The presence of three phosphorylatable serine residues in the very conserved sequence 29-42 is another argument for the involvement of this region in the interaction with DNA.