Death of an apprentice bodybuilder following 2,4-dinitrophenol and clenbuterol intake.

We present the case of a 17-year-old man, who died after 2,4-dinitrophenol (DNP) and clenbuterol consumption, which he likely took for physical enhancement. Forensic post-mortem examination revealed a yellowish skin colour and nonspecific signs of asphyxia. Analytical confirmation of the intoxication was obtained in blood and urine, with high levels of DNP and clenbuterol. Both of these substances are used by bodybuilders as DNP enhance lipolysis and clenbuterol has anabolic properties, but their toxicity is underestimated. DNP uncouples oxidative phosphorylation, leading to thermogenesis and even relatively small doses can cause fatal hyperthermia. Clenbuterol is a ß2 agonist that causes electrolyte disturbances (hypokalemia and hyperglycemia mostly) and death have been described through coronary vasospasm. Given the circumstances in which the body was found and toxicological results, we believe the cause of death to be fatal hyperthermia from DNP intake. These substances are illegal in many countries, but easily bought online. Through this availability, the last decades have seen an increase of fatal intoxications. Websites selling them are regularly closed by French public authorities and Interpol, but unfortunately it seems insufficient.

Is the pterygopalatomaxillary suture (sutura sphenomaxillaris) a growing suture in the fetus?

The pterygopalatomaxillary suture is considered as having an important role in the posteroanterior growing of the maxilla. To determine whether this suture is a growing suture in the fetus, we performed a histological study of this suture in a fetus aged of 16 weeks of amenorrhea. Serial sections (5 microm) of the pterygopalatomaxillary suture area have been performed. Fibrous sutures are separating four pieces of ossification (maxilla, palatine bone, lateral and medial plates of the pterygoid process). A fibroblastic growing site has been observed on the dorsal aspect of the pterygopalatomaxillary suture, in contact to the anterior border of the lateral plate of the pterygoid process. The posteroanterior growing of maxilla is dependent on a growing suture located on the anterior border of the pterygoid process. The pterygoid process (via its lateral plate) makes the junction between the maxilla and both the cranial base and the condylar mandibular site of growth.

Immunocytochemical characterisation of two generations of fibers during the development of the human quadriceps muscle.

We have carried out a comprehensive study of the formation of muscle fibers in the human quadriceps in a large series of well dated human foetuses and children. Our results demonstrate that a first generation of muscle fibers forms between 8-10 weeks. These fibers all express slow twitch myosin heavy chain (MHC) in addition to embryonic and foetal MHCs, vimentin and desmin. Between 10-11 weeks, a subpopulation of these fibers express slow tonic MHC, being the first primordia of muscle spindles. Extrafusal fibers of a second generation form progressively and asynchronously around the primary fibers between 10-18 weeks, giving the muscle a very heterogeneous aspect due to different degrees of organization of their proteins. By 20 weeks, these second generation fibers become homogeneous and thereafter undergo a process of maturation and differentiation when they eliminate vimentin, embryonic and foetal MHCs to express either slow twitch or fast MHC. The differentiation of these second generation fibers into slow and fast depends upon different factors, such as motor innervation or level of thyroid hormone. Around the intrafusal first generation fibers, additional subsequent generations of fibers are also progressively formed. Some differ from the extrafusal second generation fibers by expressing slow tonic MHC, others by continuous expression of foetal MHC. The differentiation of intrafusal fibers is probably under the influence of both sensory and motor innervation.

Replicative potential and telomere length in human skeletal muscle: implications for satellite cell-mediated gene therapy.

In this study, we have evaluated the ability of human satellite cells isolated from subjects aged from 5 days to 86 years to proliferate in culture. Cells were cultivated until they became senescent. The number of cell divisions was calculated by counting the number of cells plated in culture compared to the number of cells removed following proliferation. Telomere length, which is known to decrease during each round of cell division, has been used to analyze the in vitro replicative capacity and in vivo replicative history of human satellite cells at isolation. The rate of telomere shortening in myonuclei of these muscle biopsies was also examined. Our results show that both proliferative capacity and telomere length of satellite cells decreases with age during the first two decades but that the myonuclei of human skeletal muscle are remarkably stable because telomere length in these myonuclei remains constant from birth to 86 years. The lack of shortening of mean terminal restriction fragments (TRF) in vivo confirms that skeletal muscle is a stable tissue with little nuclear turnover and therefore an ideal target for cell-mediated gene therapy. Moreover, our results show that it is important to consider donor age as a limiting factor to obtain an optimal number of cells.

Plasticity of human satellite cells.

Satellite cells were isolated from human quadriceps and masseter muscles and the phenotype of these cells examined in vitro. The expression of the different isoforms of the myosin heavy chains (embryonic, fetal, fast and slow) and light chain isoforms was used to assay myotube diversification. We found that fused cultures of human satellite cells express adult fast and slow MHCs in addition to the embryonic and fetal isoforms. Only the four fast light chains (MLC1emb, MLC1F, MLC2F and MLC3F) were synthesized. No slow MLCs were ever detected in these cultures. In order to determine if the human satellite cells were committed to distinct fast and slow myogenic lineages, a clonal analysis was carried out on both cell populations. All myogenic clones expressed fast and slow MHCs, suggesting that there is no evidence for different fast and slow satellite cell lineages in human skeletal muscle.

Shorter telomeres in dystrophic muscle consistent with extensive regeneration in young children.

Muscular dystrophies are characterised by continuous cycles of degeneration and regeneration resulting in an eventual diminution of the muscle mass and extensive fibrosis. In somatic cells chromosomal telomeres shorten with each round of cell division and telomere length is considered to be a biomarker of the replicative history of the cell. We have previously shown that human myoblasts have a limited proliferative capacity, and that normal skeletal muscle has a very low level of nuclear turnover. However, in patients suffering from muscular dystrophy the satellite cells will be forced to make repeated rounds of cell division, driving the cells towards senescence. In this study we have used the telomere length to quantify the intensity of the muscle cell turnover in biopsies from dystrophic patients of different ages. Our results show that as soon as the first clinical symptoms become apparent the muscle has already undergone extensive regeneration and the rate of telomere loss is 14 times greater than that observed in controls. This confirms that the decline in regenerative capacity is due to the premature senescence of the satellite cells induced by their excessive proliferation during muscle repair.

45,X/46,XY mosaicism: report of 27 cases.

OBJECTIVES: There exist substantial differences between prenatally and postnatally diagnosed cases of 45,X/46,XY mosaicism. Ninety percent of prenatally diagnosed cases show a normal male phenotype, whereas the postnatally diagnosed cases show a wide spectrum of phenotypes. This 10% risk of an abnormal outcome in prenatally diagnosed cases requires further attention. The purpose of the present study is to provide more information on the postnatally diagnosed 45,X/46,XY mosaicism cases. To date, only a few series have been reported. An accurate diagnosis in these patients is essential not only to their follow-up, but also to providing appropriate genetic counselling and subsequent prenatal diagnosis to their parents. METHODS: The clinical, cytogenetic, endocrinologic, histologic and molecular biological findings of 27 patients with 45, X/46,XY mosaicism are analyzed. RESULTS: The reported cases showed a wide spectrum of phenotypes as Turner syndrome, mixed gonadal dysgenesis (MGD), male pseudohermaphroditism (MPH) and apparently normal male. However, Ulrich-Turner stigmata were the most common features found in this series. Patients with MGD or MPH presented with various degrees of sex reversal such as hypospadias and/or abnormal internal genitalia. No correlation between the proportion of the 45,X/46,XY cell lines in the blood or the fibroblasts and the phenotype was found. Mild mental retardation was present in 4 of the patients and 2 patients showed signs of autism. CONCLUSIONS: Two major points are emphasized in this series: 1) the presence in 7 histologically analyzed streak gonads of a homogeneous 45,X chromosomal complement suggests that the invasion of the primitive genital ridge by a such a cell line may induce abnormal gonadal development; 2) 3 males, apparently normal at birth, developed late onset abnormalities such as dysgenetic testes leading to infertility, Ulrich-Turner stigmata, dysmorphic features, and mild mental retardation. These data indicate the importance of an accurate clinical and histologic evaluation of any patient presenting with 45, X/46,XY mosaicism.

Intestinal lesions containing coronavirus-like particles in neonatal necrotizing enterocolitis: an ultrastructural analysis.

Since the outbreaks of neonatal necrotizing enterocolitis occurring in maternity hospitals of Paris and suburbs in 1979-1980, it has been possible to examine by light and electron microscopy gut specimens from ten newborns with this illness. Coronavirus-like particles, enclosed in intracytoplasmic vesicles of damaged epithelial cells of the intestinal mucosa, were observed in the small intestine, appendix, and colon. The ultrastructural study, supported by bacteriologic findings, suggests the role of coronavirus-like particles in the appearance of the lesions. Secondary proliferation of mainly anaerobic bacteria, probably responsible for pneumatosis, may aggravate the disease.

Breathing dry air causes acute epithelial damage and inflammation of the guinea pig trachea.

Drying and cooling of the airways mucosa caused by respiratory water loss may be responsible for exercise- and hyperventilation-induced asthma. Therefore we designed this study to investigate whether breathing dry air is capable of causing structural changes of the airways mucosa. Anesthetized guinea pigs breathed spontaneously through a tracheostomy either dry (n = 15) or water-saturated (n = 12) air at approximately 38 degrees C for 30 or 60 min, during which time total pulmonary resistance (TPR) was measured. Immediately afterward, the animals were killed and the lungs and airways were prepared for histological examination (light microscopy, transmission electron microscopy, and scanning electron microscopy). With dry as well as humid air, there was no change in TPR or in the structure of the main bronchi and lung parenchyma. With humid air the tracheal mucosa was normal in six guinea pigs and exhibited minor changes of the ciliae in eight and localized epithelial damage on light microscopy in the remaining animal. With dry air we found widespread loss of the ciliae on scanning electron microscopy in 10 of 12 animals, associated with detachment or sloughing of the epithelium, subepithelial vascular congestion, edema, and cellular infiltration on light microscopy. Our data demonstrate that a short exposure of the trachea to dry air causes marked epithelial lesions and local inflammation.

Immaturity of muscle fibers in the congenital form of myotonic dystrophy: its consequences and its origin.

Skeletal muscle maturation is impaired in children with congenital myotonic dystrophy. This immaturity is characterized at the light microscopy level by an abnormal presence of myotubes, small fascicles of muscle fibers, thin myofibers, and delayed muscle fiber type differentiation with a peripheral halo lacking mitochondrial oxidative enzyme activity. At an ultrastructural level, the characteristics are a paucity of myofibrils with a peripheral rim devoid of mitochondria and myofibrils in the fibers. In time the muscle is able to gain a certain degree of maturity as shown in one of our cases who had two successive muscle biopsies. The muscle, however, never becomes normal but retains discrepancies in fiber size and fiber type distribution and shows some fiber necrosis. Maturation of the motoneurons is normal, which may explain necrosis of immature muscle fibers. In an experimental study carried out to look for evidence of a circulatory factor in mothers of children with congenital myotonic dystrophy, it was found that sera from these mothers administered intra-peritoneally to newborn rats does in fact impair muscle maturation, whereas rats injected similarly with sera from control women showed normal muscle maturation.

Myosin heavy and light chain expression during human skeletal muscle development and precocious muscle maturation induced by thyroid hormone.

It has now been well established that during mammalian muscle development there is a sequential transition of the myosin isoforms, with the developmental isoforms being replaced just before or just after birth by the adult isozymes. In a previous study of human fetal muscle, we demonstrated the differentiation of two fiber populations as early as 15 weeks: one population of large diameter fibers containing predominantly slow myosin heavy and light chains, and another population which stained homogeneously for fetal myosin heavy chain and corresponded to histochemical type IIC fibers. We have carried out an immunocytochemical and biochemical study of human fetal quadriceps between 7 and 40 weeks. A chronology of the changes which occur in the expression of the myosin heavy and light chains is correlated with the results obtained by enzyme histochemistry. Evidence is also presented that in man excessive amounts of thyroid hormone act directly on the muscle, and result in a precocious accumulation of the adult myosin heavy chains and a precocious maturation of the muscle.

Computerized tomographical study of dorsal neck muscles for insertion of EMG wire electrodes.

A computerized tomographical (CT) study of the main dorsal neck muscles was performed on 60 subjects (30 males and 30 females) in order to quantify individual anatomical variations and to describe valid means of accurately inserting intramuscular wire electrodes in these deep and superimposed muscle layers. Depth and thickness of transverso-spinalis (TS), semispinalis capitis (SSC) and splenius capitis (SPL) muscles were measured on a cross-sectional image of the neck at the level of the fith cervical vertebrae. Thickness and depth were significantly greater in males than in females. SSC and SPL thicknesses were less than 10 mm in most subjects and their depths were highly variable depending on the subject's morphology. TS was thicker but its depth was also variable. This CT study clearly demonstrated that the interindividual variations of dorsal neck muscles are important and CT may provide a valid way to position wire electrodes accurately in most cervical muscles.

Mandibulo-facial dysostosis: comparison study of a neonate with mandibulo-facial dysostosis and a normal neonate.

Mandibulo-facial dysostosis (MFD) is a malformative syndrome with autosomal dominant transmission and variable expressivity that mainly affects derivatives of the first and second branchial arches. The subsurface anatomy of this condition is still partly unexplored since there have been only four reported dissections of MFD. A detailed dissection of the head and neck of a neonate with MFD is described and compared with a normal neonate. Theories of the pathogenesis are discussed on the basis of these observations.

Improvement of dysphagia following cricopharyngeal myotomy in a group of elderly patients. Histochemical and biochemical assessment of the cricopharyngeal muscle.

Cricopharyngeal myotomy is not effective in all cases of dysphagia. However, it should be the specific treatment in cases of dysphagia caused by a primary cricopharyngeal muscle dysfunction. Of a group of 10 patients with swallowing disorders in the absence of any defined cause, 7 (mean age, 81.6 years) were improved by a myotomy and 3 were not. The cricopharyngeal muscle was studied histologically and biochemically and compared to muscle obtained from nondysphagic subjects. In the muscle of the 7 improved patients, homogeneous histologic abnormalities were demonstrated: connective tissue infiltration, inflammatory cell infiltration, and degenerative changes of the muscle fibers. Conversely, muscles of the nonimproved patients and of the controls did not present the same degree of histologic lesions.

Fetal development of the urethral sphincter.

A morphological study was carried out on 25 human embryos (3-60 mm CR) and 20 human fetuses (15-40 weeks). The anatomical analysis was completed by immunocytochemical studies of different markers of muscle differentiation (vimentin, desmin, titin and isoforms of the myosin heavy chains). A mesenchymal condensation forms around the urethra after the division of the cloaca (E.H. 12-15 mm CR). The m. pubo-rectalis appears in 20-30 mm CR embryos, following the opening of the anal membrane. Striated muscle fibers can be clearly differentiated at 15 weeks. At this time, the smooth muscle layer also becomes thicker at the level of the bladder neck and forms the inner part of the urethral musculature. The urethral sphincter is a functional unit composed of central smooth muscle fibers and peripheral striated muscle fibers. In the human fetus, this musculature mainly develops in the anterior wall of the urethra. We analysed the expression of smooth and skeletal markers as an original approach to the muscular development in this region.

Anatomical and sonographical studies on the development of fecal continence and sphincter development in human fetuses.

The aim of this study was to specify the sonographic, anatomical and morphological aspects of the fetal anal sphincter and to compare them with pathological and physiological findings. The sphincter was examined by serial sectioning and staining of embryo and fetal tissue and by real-time ultrasound. Its function was analysed using amniotic fluid digestive enzyme assays in cases of anorectal atresia and cystic fibrosis. Morphological findings indicate that the functional components of the anal sphincter do not differentiate before 30 weeks and therefore do not account for the observed anal continence at 22 weeks. Ultrasound measurements of the sphincter indicate three developmental phases: 1) slow growth from 14 to 19 weeks; 2) rapid growth from 19 to 30 weeks; 3) subsequently, no further increase, but contractions indicative of peristaltism. Amniotic fluid digestive enzyme assays indicate that anal sphincter maturation begins with perforation of the anal membrane at 12 weeks. Comparison of pathological cases (anorectal atresia and cystic fibrosis) suggests two possible explanations of fetal anal obstruction: increasing viscosity of digestive secretion or the presence of the three anal sphincter muscles, even if still immature. Our results clarify the evacuation and retention of meconium during fetal life and the role of the terminal part of the digestive tract, notably the anal sphincter. Prenatal diagnosis of anorectal atresia is therefore possible before 20 weeks of gestation by measurement of amniotic fluid digestive enzymes and ultrasonography, thus enabling better neonatal management.

Management of ovarian cyst detected by prenatal ultrasounds.

As a result of refinements in Prenatal Ultrasonography (US), neonatal ovarian cysts are more frequently encountered than in the past. Between January 1981 and December 1990, 21 consecutive fetuses with ovarian cysts were followed up by ultrasonography. 23 ovarian cysts were diagnosed between 28 and 38 weeks gestation. 18 cysts were initially large cysts (more than 50 mm in diameter). In 9 cases, an anechoic cyst was observed and a US-guided needle aspiration of the cyst was performed (2 antenatally and 7 postnatally). Ultrasonographic patterns of complicated cyst were observed in 3 fetuses. After a postnatal ultrasound control, 11 infants were operated on: 8 surgical interventions were required for complicated cysts (torsion: 4, hemorrhage: 4), 3 cases irrespective of their sonographic appearance in our initial experience. In the remaining cases, spontaneous resolution was followed by repeated ultrasonographic examination. Prenatal ultrasonography today allows diagnosis of ovarian cysts and may suggest antenatal complications. To preserve as much ovarian tissue as possible, cysts greater than 50 mm in diameter should be candidates for percutaneous aspiration and complex cystic masses should be operated on.

[Rhabdomyosarcomas discovered in newborn infants. Study of markers of muscle differentiation]. / Rhabdomyosarcomes de découverte néonatale. Etude des marqueurs de différenciation musculaire.

Rhabdomyosarcomas were observed at birth in two premature infants. The evolution of these tumors was both rapid and fatal. One case was a botryoid sarcoma of the bladder and the second was a cervical alveolar sarcoma of Riopelle and Thériault. A cytological differentiation of the tumor was observed in the first case spontaneously and in the second following chemotherapy. In the second case histology, immunocytochemistry and a novel biochemical analysis of the contractile proteins demonstrated this differentiation of the tumor. The cellular reactivity varies according to the degree of muscular differentiation that can be defined either by studying the type of intermediate filaments present or the expression of specific muscle contractile proteins.

[Association of mixed gonadal dysgenesis and non-classic 21-hydroxylase deficiency]. / Association d'une dysgénésie gonadique mixte et d'une forme non classique de bloc de la 21-hydroxylase.

BACKGROUND: The rare association of mixed gonadal dysgenesis and non classical congenital hyperplasia by 21-hydroxylase deficiency poses the problem of their respective responsibility in the development of sexual ambiguity. CASE REPORT: In a newborn with ambiguous genitalia, blood 17-OH progesterone was moderately elevated (3.9 to 14.1 ng/mL) leading to the diagnosis of non-classical 21 hydroxylase deficiency, Molecular studies later confirmed this diagnosis. However, the presence of a palpable gonad and the karyotype (45 X/46 XY mosaicism) indicated a mixed gonadal dysgenesis as the cause of sexual ambiguity. Histological examination revealed the presence of a testis and a streak gonad. CONCLUSION: This observation emphasizes the need for a complete clinical and biological analysis in all newborns with sexual ambiguity.

[Limb-girdle dystrophy and pregnancy: a case report]. / Myopathie des ceintures et grossesse: un cas.

A case of pregnancy with Limb-girdle dystrophy is presented. The course of pregnancy remained uncomplicated, the patient has been delivered at 42 week's gestation by cesarean section, performed for dystocia and fetal distress. Histological and immunohistochemical examinations of the myometrium were unable to show its involvement in the muscular disease. The influence of the muscular disease and the route of delivery for such patients are discussed.