Molecular and cytogenetic evidence for cryptic speciation within a rare endemic Malagasy lemur, the Northern Sportive Lemur (Lepilemur septentrionalis).

Evolutionary relationships of different populations of the threatened malagasy lemur Lepilemur septentrionalis were assessed by sequence analysis of mitochondrial DNA (D-loop region and partial Cyt b gene). One hundred and fifty nine samples were collected from five main different localities in the northern part of Madagascar. We applied the phylogenetic species concept based on fixed diagnostic differences to determine the status of different geographical populations. No nucleotide site diagnoses Ankarana from Andrafiamena or Analamera. However, numerous fixed differences separate Sahafary from all other populations. These results were corroborated by phylogenetic trees. As previous cytogenetic studies, our molecular data suggest that two cryptic species of Lepilemur occur in the extreme north of Madagascar. This speciation is probably caused by chromosomal rearrangements in at least one of the evolutionary lineages. Our study comprises another striking example of how molecular genetic assay can detect phylogenetic discontinuities that are not reflected in traditional morphologically based taxonomies. Our study indicates that the Sahafary population is a hitherto undescribed endangered endemic species which urgently needs conservation efforts.

Taxonomic relationships and sampling effects among Lepilemuridae and Lemuridae using a partial cytochrome b gene.

Partial cytochrome b sequences were used to study relationships between three Lepilemuridae species (Lepilemur dorsalis, L. septentrionalis and L. leucopus) and other Lemuridae species. L. dorsalis were subdivided into two sub-groups, according to their capture area (Nosy-Be island and Sahamalaza peninsula). Relationships deduced from phylogenetic trees as well as genetic distances lead to the classification of the Lepilemurs analysed here into separate species. These Lepilemurs form a monophyletic clade which is the sister clade of all other Lemurs used in this study. Reconstructions using randomly chosen sequences and step by step addition of sequences indicate that phylogenetic results for closely related species need to be analysed with caution, if only a small number of sequences are used to obtain them.

[Osteo-articular ultrasonography of the shoulder]. / Echographie ostéo-articulaire de l'épaule.

The objective of this course is to present the normal and pathological aspects of the various periarticular structures of the shoulder, and the practical modalities of their analysis. The anatomy and pitfalls of tendon imaging is recalled, as well as the interest to detect intra and peri articular effusions. The semiological aspects of complete and partial tears of the rotator cuff and the various impingement syndromes, well demonstrated with dynamic sonography, are then studied. The examination requires time and knowledge but the diagnostic and therapeutic impact is very important regarding the low cost of this technique. The standardisation of the procedure and the production of normal reference images seems to guarantee a global increase in quality of the sonographic examinations.

MFASAT: a new alphoid DNA sequence isolated from Macaca fascicularis (Cercopithecidae, Primates).

A new highly repeated DNA fragment isolated from Macaca fascicularis (MFASAT) is described. Our findings obtained by sequencing, Southern blot analysis, and fluorescent in situ hybridization (FISH) on metaphasic chromosomes strongly suggest that MFASAT can be considered as a member of the alphoid DNA family characteristic of Old World monkeys. The chromosomal localization of MFASAT, obtained by FISH, showed that this alphoid DNA is present in the peri-centromeric area of all the chromosomes. MFASAT showed a high degree of conservation when compared, by sequence alignment, to other Macaca species and Papio papio as expected for species with considerable genome conservation. A low degree of homology has been found comparing M. fascicularis alphoid DNA with a more distantly related Cercopithecidae species such as Cercopithecus aethiops.

Identification of Mycobacterium DNA in an Egyptian Pott's disease of 5,400 years old.

The antiquity of tuberculosis in the Old World is controversial because the morphology of the lesion in skeletal remains is non-specific. We report the recovery of a DNA fragment from a 5,400-year-old Predynastic Egyptian skeleton that exhibits a kyphotic, 'hunchback' spinal deformity consistent with Pott's disease and suggestive of tuberculous vertebral involvement. The recovered DNA fragment was sequenced and is consistent with an original Mycobacterium sequence. We cannot prove that it is M. tuberculosis, M. bovis or an ancient mycobacteria resembling the two current forms because the observed modifications in the sequence could be attributed to the antiquity of Mycobacterium and/or to the effects of Taq polymerase. This provides the most specific evidence for the antiquity of human Mycobacterium disease in the world.

Phylogenetic relationships among Indriidae (Primates, Strepsirhini) inferred from highly repeated DNA band patterns.

Comparative studies of highly repeated DNA from different species of Indriidae (Primates, Strepsirhini) allowed confirmation of the specific status of Avahi occidentalis, A. laniger and Propithecus tattersalli. The comparison of their band patterns revealed the existence of specific and common bands from which a cladogram of the family is inferred. This cladogram shows that Avahi clade is the sister-group of Indri and Propithecus clade, and that P. verreauxi is related to P. diadema. These results were discussed in view of those obtained from cytogenetic, morphological and molecular data (mitochondrial DNA). This study shows the capacity of the repeated sequence pattern comparison to be used as a tool for confirming taxa status, (taxinomic classification is a primary determinant of management priorities for endangered species, neglect of distinct taxa may lead to their extinction), and for inferring phylogenetic relationships among related species.

First isolation of tandemly repeated DNA sequences in New World vultures and phylogenetic implications.

A highly repeated DNA sequence composed of closely related subunits that ranged from 171 to 176 base pairs has been cloned and characterized in the king vulture (Sarcoramphus papa). Related sequences were also isolated in the black vulture (Coragyps atratus). This new family of avian repetitive DNA elements is here termed the "HaeIII family." Genomic DNAs from a number of avian species were probed with one of the king vulture restriction fragments. In the cathartids, the hybridization patterns showed no individual or sexual variations. A strong HaeIII ladder was present in the two aforementioned species as well as in the Andean condor (Vultur gryphus), but in the black vulture the bands of the ladder alternated in intensity. Weaker hybridization signals were obtained in two ciconids, the jabiru stork (Jabiru mycteria) and the white stork (Ciconia ciconia). The HaeIII repeat was not detected in accipitrid birds of prey, a Polyborinae falconid, pelecanids, and psittacids.

Confirmation of the taxonomic position of Callimico goeldi (Primates, Platyrrhini) on the basis of its highly repeated DNA patterns.

The systematic position of Callimico goeldi has been analyzed by comparison of its highly repeated DNA fragments with those of three species of Callithricidae and two species of Cebidae. Our data, in agreement with those obtained by other authors by means of cytogenetic and molecular biology techniques, are in favour of the classification of C. goeldi in the family Callithricidae.

Comparison of highly repeated DNA sequences in some Lemuridae and taxonomic implications.

Highly repeated DNA sequences of Eulemur fulvus mayottensis, E. coronatus, Lemur catta, and Hapalemur griseus griseus have been identified and compared. Sequence analysis of highly repeated DNA fragments isolated from L. catta and Hapalemur showed a high percentage of similarity (nearly 95%), as did fragments isolated from the two very close Eulemur species, whereas comparison of the DNA fragments isolated from the two Eulemur species and the L. catta/Hapalemur group showed a very low percentage (approximately 40%) of identity, as might be expected for distant species. These results confirm our previous data, obtained by Southern blot hybridization techniques on the same species, and strongly support the existence of a common trunk between L. catta and Hapalemur, but different from the leading to the Eulemur species.

[Electrophoretic study of the protein composition of human seminal vesicle secretion]. / Etude électrophorétique de la composition protéique de la sécrétion vésiculaire humaine.

The SDS-PAGE and two-dimensional reductived electrophoretic study of the liquid aspirated from human seminal vesicles showed that this medium contains high molecular weight proteins (greater than 150 kDa). In addition to these high molecular weight proteins, native seminal vesicle secretion is characterised by the presence of a large 47.4 kDa protein band corresponding to seminogelin, described by Lilja. The high molecular weight proteins actually correspond to associations of three types of protein subunits with molecular weights of 75-76, 63-64 and 47-48 kDa, which represent the structural elements of the coagulum; one of these subunits consists of seminogelin. Seminal vesicle secretion contains very little or no proteolytic enzymes, so that its protein composition remains stable even after incubation for 24 hours.

Comparative study of semen in diabetic and healthy men.

This study examines the consequences of diabetes on male reproductive function. We have studied a group of 18 diabetics (15 type I, 3 type II) age 24 to 40 years (mean 34 +/- 5.3) with duration of diabetes from 2 to 30 years (mean 11.8 +/- 8.3). This group was compared to 20 semen donors. In diabetics higher sperm (p = 0.0063) and round cell (p = 0.031) concentrations/ml as well as a higher total spermatozoa count (p = 0.023) were observed. The percentage of abnormal sperms were also higher. None of the couples had consulted for sterility and they had a mean of 2 children. In this group of patients, diabetes was not a cause of subfertility. Specific spermiological alteration in diabetics might reflect a testicular damage secondary to their diabetes or disease.

Secretory proteins of human seminal vesicles and their relationship to lipids and sugars.

Proteins secreted by human seminal vesicles are strongly positively charged. Cellulose acetate electrophoresis show how the presence of two protein bands of vesicular origin (N3 and N4). When studied on SDS-PAGE there are three main bands of molecular weight 67, 45 and 40 kDa, respectively. These proteins may be separated by a chromatographic process using gel filtration on Sephadex G 25 M and ion exchange chromatography on CM and SP Sephadex C 50. Fructose, secreted by seminal vesicles, is excreted with specific proteins and these complexes take part to coagulum formation. During liquefaction glucose appears progressively and fructose is released from complexes with proteins. Interconversion processes that transform fructose into glucose, originate from prostatic secretion. In man, the liquefaction process seems to be not due to proteolysis, but by the way of other mechanisms that transform vesicular proteins of very high molecular weight into sub-units with lower molecular weights in the first minutes after ejaculation. In species other than man, i.e. lemurian, fructose takes part in the coagulation process with other components (albumin, ions, -SH groups). Cholesterol appears to be in relation with proteins which have high molecular weights. Half of phospholipid content of seminal plasma is probably free. Incubation of seminal plasma with spermatozoa show that these cells use triglycerides for their metabolism. The ratio between cholesterol and phospholipids is an important marker for the capacitation-decapacitation process.

[Peroperative spinal cord ultrasonography in neurosurgery]. / L'échographie médullaire per-opératoire en neurochirurgie.

Intraoperative spinal sonography is a new method of investigation. We are reporting 14 observations. The results of these explorations permit to describe the normal spinal sonography and a spinal ultrasonic semeiology. We underline the advantage of this technic in the neurospinal surgery particularly to locate the tumors and to facilitate the removal.