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L’hypoxemie, si elle n’est pas reconnue, entraine a court ou moyen termes et suivant sa gravite un syndrome de defaillance multiviscerale qui peut etre letal. Sa recherche est primordiale et systematique pour tous les patients vus dans le cadre de l’urgence. Suivant sa gravite et son etiologie, la methode d’oxygenation varie et doit etre reevaluee rapidement afin de l’optimiser. Quand les methodes d’oxygenation non invasives sont depassees, elles sont relayees par des techniques invasives qui necessitent une autre orientation que celle des urgences.

Acetabular cup orientation (inclination and anteversion) is a fundamental topic in orthopaedics and depends on pelvis tilt (positional parameter) emphasising the notion of a safe range of pelvis tilt. The hypothesis was that pelvic incidence (morphologic parameter) could yield a more accurate and reliable assessment than pelvis tilt. The aim was to find out a predictive equation of acetabular 3D orientation parameters which were determined by pelvic incidence to include in the model. The second aim was to consider the asymmetry between the right and left acetabulae. Twelve pelvic anatomic specimens were measured with an electromagnetic Fastrak system (Polhemus Society) providing 3D position of anatomical landmarks to allow measurement of acetabular and pelvic parameters. Acetabulum and pelvis data were correlated by a Spearman matrix. A robust linear regression analysis provided prediction of acetabulum axes. The orientation of each acetabulum could be predicted by the incidence. The incidence is correlated with the morphology of acetabula. The asymmetry of the acetabular roof was correlated with pelvic incidence. This study allowed analysis of relationships of acetabular orientation and pelvic incidence. Pelvic incidence (morphologic parameter) could determine the safe range of pelvis tilt (positional parameter) for an individual and not a group.

The goal in this paper was to rebuild a three dimensional (3D) reconstruction of the dorsal and ventral pancreatic buds, in the human embryos, at Carnegie stages 15-23.The early development of the pancreas is studied by tissue observation and reconstruction by a computer-assisted method, using a light micrograph images from consecutive serial sagittal sections (diameter 7 microm) of ten human embryos ranging from Carnegie stages 15-23, CRL 7-27 mm, fixed, dehydrated and embedded in paraffin, were stained alternately with haematoxylin-eosin or Heindenhain'Azan. The images were digitalized by Canon Camera 350 EOS D. The serial views were aligned automatically by software, manual alignment was performed, the data were analysed following segmentation and threshold.The two buds were clearly identified at stage 15. In stage 16, both pancreatic buds were in final position, and begin to merge in stage 17. From stage 18 to the stage 23, surrounding connective tissue differentiated. In the stage 23, the morphology of the pancreas was definitive. The superior portion of the anterior face of the pancreas's head was arising from the dorsal bud. The rest of the head including the uncinate process emanated from the ventral bud.The 3D computer-assisted reconstruction of the human pancreas visualized the relationships between the two pancreatic buds. This explains the disposition and the modality of the components fusion. This embryologic development permits a better understanding of congenital abnormalities.

Le but de ce travail etait d’evaluer la valeur de l’angle d’incidence pelvienne (IP) comme facteur predictif de descente perineale. Ce facteur morphologique osseux, acquis a la fin de la croissance, n’a jamais ete correle a un trouble de la statique pelvienne. Dans une etude retrospective de 197 femmes, la descente du perinee au repos et durant la poussee est evaluee a partir de defecographies. L’angle d’IP (53° ± 9 °C, independant de la position du sujet) est defini comme l’angle entre la droite perpendiculaire au milieu du plateau superieur du sacrum et la droite reliant ce dernier avec le milieu de l’axe bicoxofemoral. L’angle d’IP etait correle a la descente perineale. L’angle d’IP etait statistiquement plus grand lors de la descente du perinee au repos (64 versus 53 °C; p 62 °C) avait une sensibilite (73 %), une specificite (82 %), une valeur predictive positive (81 %) et une valeur predictive negative (75 %). Une grande IP (> 62 °C) est un facteur predictif de la descente perineale au repos avant l’apparition des autres facteurs de risque lies a l’âge. Une grande IP (> 62 °C) s’accompagne d’un large porte-a-faux et d’une horizontalisation du perinee, provoquant une augmentation des contraintes entre les insertions de celui-ci. Une simple radiographie du bassin de profil pourrait permettre de depister un pelvis a risque de trouble de la statique pelvienne.

Le diagnostic de syndrome canalaire du nerf pudendal est un diagnostic avant tout clinique. Il n’y a aucun marqueur clinique ou paraclinique specifique de cette pathologie. Un faisceau d’arguments permet cependant d’evoquer le diagnostic. Un groupe d’experts a valide des criteres diagnostiques simples (criteres de Nantes). Les criteres indispensables au diagnostic sont: l’existence d’une douleur situee dans le territoire anatomique du nerf pudendal, aggravee en station assise, ne reveillant pas la nuit, sans hypoesthesie objective a l’examen clinique mais avec un bloc anesthesique du tronc du nerf pudendal positif. D’autres elements cliniques pourront apporter des arguments supplementaires au diagnostic de nevralgie pudendale. En revanche, nous proposons des criteres d’exclusion: douleurs purement coccygienne, fessiere ou hypogastriques, douleurs uniquement paroxystiques, prurit, presence d’anomalies d’imagerie susceptibles d’expliquer la symptomatologie. La nevralgie pudendale peut etre typique mais egalement deroutante en raison de signes associes qui temoignent de reactions d’hypersensibilisation peripherique et centrale a expression neuropathique, musculaire ou vegetative.

Une compression chronique du nerf pudendal dans un site d’etroitesse anatomique (syndrome canalaire) peut etre a l’origine de douleurs perineales invalidantes. Ce type d’atteinte doit etre diagnostique de facon specifique, car cela peut constituer une indication de neurolyse chirurgicale. Dans ce cadre, il est usuel de demander un examen electroneuromyographique (ENMG) du perinee, qui sera base sur l’etude de l’activite electromyographique de muscles perineaux, des reflexes sacres et des conductions motrices du nerf pudendal. Differentes considerations physiopathologiques et techniques expliquent certaines limites de l’ENMG qu’il faut connaitre. C’est ainsi que les methodes utilisees n’evaluent pas les anomalies fonctionnelles a l’origine des douleurs, mais plutot les alterations structurelles du nerf pudendal (demyelinisation ou perte axonale). De plus, seule l’innervation motrice directe ou reflexe est evaluee, alors que l’etude specifique des conductions sensitives serait sans doute plus sensible a objectiver une compression nerveuse. Enfin, il n’est pas possible de distinguer l’atteinte compressive des nombreuses autres causes de lesion nerveuse pudendale (chirurgicales, obstetricales, liees a une constipation chronique…). Ainsi, l’ENMG perineal a une sensibilite et une specificite limitees dans le diagnostic de syndrome canalaire pudendal et ne renseigne pas directement sur le phenomene douloureux. Le diagnostic de nevralgie pudendale repond en fait a des criteres cliniques precis et l’ENMG ne peut que donner des arguments supplementaires, mais non formels en faveur de ce diagnostic. L’ENMG perineal permet surtout de faire un « etat des lieux » de l’innervation perineale en prevision d’un geste chirurgical de decompression, et pourrait eventuellement fournir certains elements predictifs de l’interet de l’intervention. En revanche, l’ENMG ne permet generalement pas de localiser precisement le site de compression et n’a, dans tous les cas, aucune utilite dans la surveillance peroperatoire.

Reliability and reproducibility of two radiological pelvic parameters are tested: thickness (length of the segment defined by the middle of the upper endplate of the sacrum to the middle of the bi-coxo-femoral axis) and pelvic incidence (angle defined by the perpendicular line to the centre of the upper endplate of the sacrum and the thickness line). These two parameters provide a pelvis description and assess the relation between pelvis and spinal curves. The anatomical reliability of these radiological parameters was not achieved. The values of these two parameters from X-ray versus direct measurement on 12 anatomical specimens are compared. The direct measurement was performed by means of an electromagnetic Fastrak system (Polhemus society) providing 3D position of anatomical landmarks and allowing to measure the incidence and the thickness. These parameters were also measured from sagittal X-ray. Their values were compared. Incidence: the paired t-test and the variance ratio test were not statistically significant and a highly significant positive correlation existed between anatomical and radiological values (r=0.98; P

A liquid chromatographic method has been developed to separate a series of quinolones which are widely used as veterinary drugs: danofloxacin, difloxacin, enrofloxacin (and its metabolite ciprofloxacin), marbofloxacin, norfloxacin and sarafloxacin. Separation was performed by using an end-capped high-purity silica-based C8 column and a mobile phase consisting of acetonitrile-aqueous oxalic acid buffer solution. In order to establish the optimal conditions for an isocratic separation, a three-factor Doehlert experimental design was applied. Gradient elution was applied which reduced analysis time. Figures of merit of the method proposed, with fluorimetric detection, were evaluated.

ResumenObjetivoEstimar la prevalencia de tuberculosis pulmonar, analizar factores asociados a su positividad y analizar el uso de servicios de salud en tosedores usuarios del primer nivel de atención en zonas de alta y muy alta marginación de la Región Fronteriza de Chiapas, México.MétodosEn 1997 se realizó una búsqueda activa de tosedores (n = 573), entre usuarios (n = 2.203) de una muestra aleatoria de siete unidades del primer nivel de atención, a los que se les solicitaron tres muestras de esputo para la realización de baciloscopías y cultivos.ResultadosSe encontró una tasa de positividad a tuberculosis pulmonar de 11,1 por 100 pacientes estudiados (IC 95% = 6,6–17,2). La positividad a tuberculosis pulmonar se mostró asociada al tamaño y grado de marginación de la localidad de residencia. El 56% de los tosedores acudió a consulta por síntomas no respiratorios.ConclusionesEs necesario reforzar la detección y control de la tuberculosis pulmonar en el primer nivel de atención, así como sensibilizar y brindar educación para la salud a la población en general, con el fin de aumentar la demanda de atención a causa de la tos y reducir el tiempo entre el inicio de la tos y la búsqueda de atención.AbstractObjectiveTo estimate the prevalence of pulmonary tuberculosis and to analyze the utilization of health services, in coughers seeking care at primary care centers, within areas of «severe» and «very severe» poverty, in the Border Region of Chiapas, Mexico.Materials and methodsIn 1997 an active case finding of patients with cough was carried out among all patients aged over 14 years seeking consultation (n = 2.203) in a random sample of seven primary care centers; 573 coughers were found.ResultsA rate of 11.1 positive pulmonary tuberculosis per hundred patients was found (95% CI = 6.6–17.2). The factors associated with positive pulmonary tuberculosis were size and poverty level of the locality of residence. Fifty-six percent of the coughers sought care for non-respiratory symptoms.ConclusionsIt is necessary to improve detection and control of pulmonary tuberculosis at the primary care centers, as well as to develop a health education campaign among the general population, with the aim of increasing the demands for care among coughers and reduce the time elapsed between onset of cough and first medical care.

The dissociation constants of several quinolones in a 7% acetonitrile-water mixture are determined. A model describing the effect of pH on the retention of quinolones by an octadecylsilica stationary phase with a 7% acetonitrile-water mobile phase is proposed. The model uses pH values in the mobile phase instead of pH values in water and takes into account the effect of activity coefficients. Fluorescence of quinolones in the mobile phase is studied, and a chromatographic separation with fluorimetric detection is established. Detection limits for the proposed method range from 3 to 12 μg·L−1.

Resume Notre etude comprenait 75 patientes dont l'âge moyen etait de 55,05 ± 13,27 ans. Ces patientes ont eu une exploration electrophysiologique perineale qui comprenait la detection du muscle sphincter anal externe et la recherche de la latence du reflexe sacre par stimulation du nerf dorsal du clitoris et recueil dans le sphincter anal externe. Des atteintes neurogenes peripheriques isolees du nerf pudendal ou associees a des atteintes musculaires sont correlees avec la presence d'un perinee descendant. Notre etude a tenu compte des antecedents obstetricaux et chirurgicaux dans la survenue de cette atteinte, mais ces derniers n'etaient pas impliques de facon significative.

Background and Purpose A new neurological implant, the Sensor-Reservoir, was developed to provide a relative measurement of ICP, which permits a noninvasive technique to detect and localize occlusions in ventricular drainage systems and, thus, to identify mechanical damage to shunt valves. The “reservoir” of this device can be used to administer medication or a contrast agent, to extract cerebral spinal fluid (CSF), and with the possibility of directly measuring ICP. The Sensor-Reservoir was evaluated to identify possible MRI-related issues at 1.5-T/64-MHz and 3-T/128-MHz. Materials and Methods Standard testing techniques were utilized to evaluate magnetic field interactions (i.e., translational attraction and torque), MRI-related heating, and artifacts at 3-T for the Sensor-Reservoir. In addition, 12 samples of the Sensor-Reservoir underwent testing to determine if the function of these devices was affected by exposures to various MRI conditions at 1.5-T/64-MHz and 3-T/128-MHz. Results Magnetic field interactions for the Sensor-Reservoir were not substantial. The heating results indicated a highest temperature rise of 1.8 °C, which poses no patient risks. Artifacts were relatively small in relation to the size and shape of the Sensor-Reservoir, but may interfere diagnostically if the area of interest is near the device. All devices were unaffected by exposures to MRI conditions at 1.5-T/64-MHz and 3-T/128-MHz. Conclusion When specific guidelines are followed, the Sensor-Reservoir is “MR conditional” for patients undergoing MRI examinations at 3-T or less.

By measuring the pelvic incidence angle, we assessed the relationship between pelvic floor disorders and pelvic morphology, which allowed us to document for the first time the hypothesis that pelvic incidence may be a predictive factor of perineal descent.In a retrospective study of 197 women, the perineal descent at rest and during straining was assessed by defecography. The pelvic incidence angle (53 degrees +/- 9 degrees , independent of the subject position) was defined as the angle between the line perpendicular to the sacral plate at its midpoint and the line connecting this point to the middle of the femoral heads axis. The pelvic incidence angle was correlated with the incidence of pelvic floor descent.In those with pelvic floor descent at rest compared with those without, pelvic incidence angle was significantly larger (64 degrees vs. 53 degrees , P10E-06). As a predictive factor of perineal descent at rest, a great pelvic incidence (62 degrees ) had a sensitivity (73 percent), specificity (82 percent), positive predictive value (81 percent), and negative predictive value (75 percent).A large pelvic incidence (62 degrees ) may be a predictive factor of perineal descent at rest before the apparition of other acquired factors. With pelvic incidence62 degrees , a large overhang between the insertions increases the strains on the perineum, which is rather horizontal.

The aim of this work was to reconstruct, in the rat embryos, stage 12-23, the three dimensional (3D) distribution of the dorsal and ventral pancreatic buds by of a computer assisted method. Ninety-six rat embryos, CRL 3-16 mm, fixed, dehydrated, and paraffin embedded, were submitted to serial histological sections and stained by hematoxylin-eosin and Heidenhain's azan techniques. The images were digitalized by Canon Camera 350 EOS D. The serial views were aligned anatomically by software and the data were analyzed following segmentation and thresholding. The dorsal pancreas developed from the dorsal wall of the duodenum in stage 12, while the ventral pancreas arose from the ventral wall of the hepatic diverticulum in stage 13 and 14. The rotation of ventral pancreas started in stage 15 and was completed in stage 16. The fusion of both buds was evident in stage 17. In stage 23 the limit between dorsal and ventral bud was still marked by the pathway of superior mesenteric vein.

International audience; Entrapment of the pudendal nerve may be at the origin of chronic perineal pain. This syndrome must be diagnosed because this can result in the indication of surgical decompression of the entrapped nerve for pain relief. Electroneuromyographic (ENMG) investigation is often performed in this context, based on needle electromyography and the study of sacral reflex and pudendal nerve motor latencies. The limits of ENMG investigation, owing to various pathophysiological and technical considerations, should be known. The employed techniques do not assess directly the pathophysiological mechanisms of pain but rather correlate to structural alterations of the pudendal nerve (demyelination or axonal loss). In addition, only direct or reflex motor innervation is investigated, whereas sensory nerve conduction studies should be more sensitive to detect nerve compression. Finally, ENMG cannot differentiate entrapment from other causes of pudendal nerve lesion (stretch induced by surgical procedures, obstetrical damage, chronic constipation...). Thus, perineal ENMG has a limited sensitivity and specificity in the diagnosis of pudendal nerve entrapment syndrome and does not give direct information about pain mechanisms. Pudendal neuralgia related to nerve entrapment is mainly suspected on specific clinical features and perineal ENMG examination provides additional, but no definitive clues, for the diagnosis or the localization of the site of compression. In fact, the main value of ENMG is to assess objectively pudendal motor innervation when a surgical decompression is considered. Perineal ENMG might predict the outcome of surgery but is of no value for intraoperative monitoring.

The aim of this study was to assess pelvic asymmetry (i.e. to determine whether the right iliac bone and the right part of the sacrum are mirror images of the left), both quantitatively and qualitatively, using three-dimensional measurements. Pelvic symmetry was described osteologically using a common reference coordinate system for a large sample of pelvises. Landmarks were established on 12 anatomical specimens with an electromagnetic Fastrak system. Seventy-one paired variables were tested with a paired t-test and a non-parametric test (Wilcoxon). A Pearson correlation matrix between the right and left values of the same variable was applied exclusively to values that were significantly asymmetric in order to calculate a dimensionless asymmetry index, ABGi, for each variable. Fifteen variables were significantly asymmetric and correlated with the right vs. left sides for the following anatomical regions: sacrum, iliac blades, iliac width, acetabulum and the superior lunate surface of the acetabulum. ABGi values above a threshold of +/- 4.8% were considered significantly asymmetric in seven variables of the pelvic area. Total asymmetry involving the right and the left pelvis seems to follow a spiral path in the pelvis; in the upper part, the iliac blades rotate clockwise, and in the lower part, the pubic symphysis rotates anticlockwise. Thus, pelvic asymmetry may be evaluated in clinical examinations by measuring iliac crest orientation.

The metabolomic analysis of 11 Ilex species, I. argentina, I. brasiliensis, I. brevicuspis, I. dumosavar. dumosa, I. dumosa var. guaranina, I. integerrima, I. microdonta, I. paraguariensis var. paraguariensis, I. pseudobuxus, I. taubertiana, and I. theezans, was carried out by NMR spectroscopy and multivariate data analysis. The analysis using principal component analysis and classification of the (1)H NMR spectra showed a clear discrimination of those samples based on the metabolites present in the organic and aqueous fractions. The major metabolites that contribute to the discrimination are arbutin, caffeine, phenylpropanoids, and theobromine. Among those metabolites, arbutin, which has not been reported yet as a constituent of Ilex species, was found to be a biomarker for I. argentina,I. brasiliensis, I. brevicuspis, I. integerrima, I. microdonta, I. pseudobuxus, I. taubertiana, and I. theezans. This reliable method based on the determination of a large number of metabolites makes the chemotaxonomical analysis of Ilex species possible.

Pelvis and spinal curves were studied with an angular parameter typical of pelvis morphology: pelvic incidence. A significant chain of correlations between positional pelvic and spinal parameters and incidence is known. This study investigated standards of incidence and a predictive equation of lordosis from selective pelvic and spinal individual parameters. One hundred and forty nine (78 men and 71 women) healthy adults, aged 19-50 years, with no spinal disorders, were included and had a full-spine lateral X-ray in a standardised upright position. Computerised technology was used for the measurement of angular parameters. Mean-deviation section of each parameter and Pearson correlation test were calculated. A multivariate selection algorithm was running with the lordosis (predicted variable) and the other spinal and pelvic parameters (predictor variables), to determine the best sets of predictors to include in the model. A low incidence (44 degrees ) decreased sacral-slope and the lordosis is flattened. A high incidence (62 degrees ) increased sacral-slope and the lordosis is more pronounced. Lordosis predictive equation is based on incidence, kyphosis, sacral-slope and +/-T9 tilt. The confidence limits and the residuals (the difference between measured and predicted lordosis) assessed the predicted lordosis accuracy of the model: respectively, +/-1.65 and 2.41 degrees with the 4-item model; +/-1.73 and 3.62 degrees with the 3-item model. The ability of the functional spine-pelvis unit to search for a sagittal balance depended both on the incidence and on the variation section of the other positional parameters. Incidence gave an adaptation potential at two levels of positional compensation: overlying state (kyphosis, T9 tilt), underlying state (sacral slope, pelvic tilt). The biomechanical and clinical conditions of the standing posture (as in scoliosis, low back pain, spondylisthesis, spine surgery, obesity and postural impairments) can be studied by comparing the measured lordosis with the predicted lordosis.

Our anatomic findings have led us to define conflictual relations that may be encountered in their course by the pudendal n. and its branches. Starting from the clinical study of a group of patients suffering from chronic perineal pain in the seated position, we have defined, beginning with the cadaver, three possible conflictual settings: in the constriction between the sacrotuberal and sacrospinal ligaments; in the pudendal canal of Alcock; and during the straddling of the falciform process of the sacro-tuberal ligament by the pudendal n. and its branches. Consequently, considering so-called idiopathic perineal pain as an entrapment syndrome, the clinical and neurophysiologic arguments and infiltration tests have led us to define a surgical strategy which has currently given 70% of good results in 170 operated patients. Earlier diagnosis should improve on this.

We studied the response of nodal segments of Eucalyptus microtheca F.J. Muell. to salt stress in a tissue culture system. Three clones of Eucalyptus microtheca (37, 42 and 43) were grown in vitro under saline conditions (0-140 mM NaCl) for three months. The survival of all three clones decreased with increasing concentrations of NaCl in the medium, but the presence of up to 70 mM NaCl stimulated rooting in Clones 37 and 42. Shoot elongation of Clone 43 was less affected by salt than that of Clones 37 and 42. Leaf growth, expressed as leaf length, was stimulated 40% in Clone 42 and 33% in Clone 43, but was progressively inhibited up to 27% by 70 mM NaCl in Clone 37. Massive accumulation of Na(+) and Cl(-) occurred, especially in Clones 42 and 43 (4000 and 3000 mmol Na(+) kg(DW) (-1), respectively), as the salinity of the culture solution increased, and this was correlated with inhibition of growth. Because of a reduction in the accumulation of K(+) with increasing salinity, the K(+)/Na(+) ratio decreased from a control value of 4.5 to 0.14 in shoots grown in the presence of 140 mM NaCl. The development of techniques for selecting seedlings by monitoring the physiology of shoots in vitro instead of testing whole plants in vivo will provide a relatively simple method of selection for woody trees.

But Definir une relation entre la descente perineale et la morphologie osseuse du pelvis en calculant l’angle d’incidence pelvienne (AIP). Materiels et Methodes Lors d’une etude retrospective portant sur 197 patientes, la descente perineale au repos et a l’effort a ete evaluee sur la defecographie. L’AIP etait mesure entre la perpendiculaire passant par le centre du plateau de la 1ere vertebre sacree et la droite reliant ce centre aux tetes femorales. L’AIP a ete correle a l’incidence de la descente perineale. Resultats L’AIP etait significativement plus eleve quand le perinee etait descendu (64° vs 59°). Comme facteur predictif de perinee descendu, un AIP > 62° avait une sensibilite de 73,2 %, une specificite de 82 %, une valeur predictive positive de 80,7 % et une valeur predictive negative de 75 %. Discussion Un AIP eleve temoigne d’une horizontalisation et d’une ouverture du pelvis favorisant des troubles de la statique pelvienne. La morphologie osseuse du bassin determine l’orientation des fascias et des muscles pelviens et perineaux, expliquant la possibile survenue de troubles de la statique pelvienne. Conclusion L’angle d’incidence pelvienne est predictif de la descente perineale.

The study was performed using 45 pelvic half section specimens (41 fetal ones and four adults). The macroscopic dissection followed the nerve branches from their spinal roots up to the external anal sphincter. Three nerve branches were found: the anterior ramus arising from the external perineal nerve, the inferior rectal nerve and an independent posterior branch. The anterior and the inferior rectal nerve branches always emerged from the pudendal plexus. The posterior branch arising either from S4 or from the inferior rectal nerve was only found in (31%) of our cases. Five anatomical distributions are described, percentages of every type notified. The fibre content of these nerve bundle branches was evaluated through histological sections using Heidenhain's azan stain and Luxol fast blue. The branches consisted of 2,896 to 2,137 fibres, 20% of them being unmyelinated and 80% containing various myelinated fibres. The nomenclature of these nerve branches has to be debated. The terms of anterior, middle and posterior anal nerves seem more suitable.

Summary Heritable quantitative and qualitative changes can be observed in the nuclear DNA of doubled-haploid (D.H.) plants obtained by pollen culture of Nicotiana sylvestris . D.H. plants resulting from consecutive androgenic cycles contain on the average increasing amounts of total DNA and increasing proportions of highly repeated sequences. All the results suggest that both A-T rich and G-C rich regions are amplified in the D.H. genome. Some hypotheses on the origin of the changes are discussed.

Different lines of Nicotiana sylvestris, a diploid autogamous species (2n =24), were used to initiate protoplast culture from leaf mesophyll cells: The original line (T line) is a pure line from the Institut des Tabacs, Bergerac, France, morphologically very homogeneous (“flat” phenotype). Diploid androgenetic lines were obtained by consecutive pollen cultures from the T line. These lines, theoretically completely homozygous (Doubled-Haploid or DH) show typical alterations of leaf morphology that we previously called “crumpling” (FR: fripe) (De Paepe et al, 1977, De Paepe et Pernes, 1978).

Protoplast cultures are often performed in order to obtain interesting genotypes (coupled with mutagenesis) or to multiply them. But there are few studies on the genetic variability that might be induced by protoplast culture itself (without mutagenesis).