Validation of quantitative polymerase chain reaction methodology for monitoring DNA as a surrogate marker for species material contamination in porcine heparin.

Heparin is a widely used intravenous anticoagulant comprising of a very complex mixture of glycosaminoglycan chains, mainly derived from porcine intestinal mucosa. The species of origin and the absence of contaminants from other species are important determinants of the different physicochemical characteristics of heparin. They also determine the potential for introducing infectious and adventitious agents into heparin batches destined for medicinal use. We perform routine quantitative polymerase chain reaction (Q-PCR) release tests to confirm the quality of all crude heparin batches, including those used for the manufacture of enoxaparin sodium. Here we further demonstrate that the assessment of the DNA content in crude heparin is a good surrogate marker of contamination at the mucosa level. After spiking porcine mucosa with ovine mucosa and processing this material to form crude heparin, we were able to observe similar ratios of species-specific DNA in both the starting and end products. Experiments performed with 3,000 and 1,500 ppm contamination found these concentrations to be well above the detection limit for our assay of heparin batches. Additionally this Q-PCR method can be used to detect contamination in mucosa, thus providing a tool capable of monitoring for contaminants throughout the crude heparin manufacturing process. Q-PCR analysis of industrial crude heparin samples has confirmed over time the value of this method to assess the pure porcine origin of heparin.

Near-field and confocal surface-enhanced resonance Raman spectroscopy at cryogenic temperatures.

For laser spectroscopy at variable temperatures with high spatial resolution a combined scanning near-field optical and confocal microscope was developed. Rhodamine 6G (R6G) dye molecules dispersed on silver nano-particles or nano-clusters were investigated. For optical excitation of the molecules, either an aperture probe or a focused laser spot in confocal arrangement were employed. Raman spectra in the wavenumber range between 300 cm-1 and 3000 cm-1 at room temperatures down to 8.5 K were recorded. Many of the observed Raman lines can be associated with the structure of the adsorbed molecule. Intensity fluctuations in spectral sequences were observed down to 77 K and are indicative of single molecule sensitivity.

[Conservative treatment of grade 3 intra-epithelial cervical neoplasms. Comparative study of CO2 laser vaporization, laser conization and cold scalpel conization]. / Traitements conservateurs des CIN III. Etude comparative des vaporisations au laser CO2, des conisations laser et des conisations au bistouri froid. A propos de 141 cas.

Carbon dioxide laser was used in 79% of the 141 conservative treatments carried out for grade 3 cervical intra-epithelial neoplasia (CIN III) between the years 1982 and 1986 (41% vaporized and 38% coned out by using the laser as against 21% that were treated by scalpel conisation. The mean age of the women treated in this way by the laser was low (28.5 years of age and 32.5 years as against 38.1). Their parity was also low (0.8 and 1.2 as against 2.2). The lesions were very often spread out on the ectocervix and sometimes going into the vagina (26 and 9% as against 26%) or associated with koilocytosis (65.5 and 47.2% as against 41.3%). In one out of two cases vaporisation of the lesion is contra-indicated and the three diagnostic methods that are used (an ecto and endocervical smear, colposcopy and multiple directed small biopsies) do not make it certain that there is no underlying invasion of the tissues. Vaporisation and conisation are easily carried out under colposcopic control and are associated with widespread lesions at the squamo-columnar junction. These treatments sometimes have to be repeated; then they give a cure rate of 92-96% as against 96%. They are more reliable than conventional surgery for widespread lesions and they do make it possible to keep to the morphology of the cervix and thus make it possible for the cervix that is treated to behave more physiologically. It is also much easier and more reliable in these cases to carry out follow-up for carcinoma. This follow-up should be carried out on two occasions. The triple diagnostic method should be carried out again at the third month (2 smears, colposcopy and colposcopically directed small biopsies). This makes it possible to diagnose and treat early the cases where there has been failure of the original treatment. Then ecto and endocervical smears should be repeated at 3-monthly intervals, then at 6-monthly intervals and finally annually to screen for recurrences of these neoplastic conditions, and for koilocytosis which sometimes repeat themselves in an extensive manner.

[Fertilization in vitro with frozen donor sperm after ovocyte stimulation with an LH-RH analog, HMG, and HCG using a short-term protocol]. / Fécondation in vitro avec sperme congelé de donneur après stimulation ovocytaire par analogue Gn-RH, HMG, HCG selon le protocole court.

Frozen cryoprotected donor sperm can be used for IVF.ET. Short administration of Gn.RH analogue with HMG and HCG (Decapeptyl) are used in 39 cycles. Seven clinical pregnancies were obtained, including one which resulted from the transfer of frozen embryos. One spontaneous abortion occurred after 7 weeks amenorrhea. Pregnancy rates are 17.9% per induction cycle, 24.1% per oocyte recovery and 26.9% per embryo transfer. Cumulative pregnancy rate for one year (3 attempts at donor sperm IVF.ET) is 44.7%, but 69.9% if we consider patients who have previously benefited from 12 to 24 AID cycles. The AID success rate is 66% for the 12 months of the first year, 42.3% in the second year and 24.2% in the third year of treatment. These data make it possible to use donor sperm with IVF.ET if tubal disease is associated with severe male factors and if the women fail to conceive after 12, 18 or 24 AID cycles, depending on their age and the existence of hormonal, cervical, tubal or pelvic associated factors.

[Unusual variations of the large arteries of the thigh]. / Seltene Variationen der grossen Oberschenkelarterien.

Three rare variations of the large arteries of the thigh are known: Hypoplasia or aplasia of the superficial femoral artery in combination with persistent sciatic artery which normally is present during embryogenesis for a few days only. Typical finding is a large dorsal artery in the thigh accompanied with a hypoplastic superficial femoral artery. For the first time angiography of this vascular situation is shown in two projections. Complications of persistent sciatic artery include aneurysm (25%) and occlusion (embolic or thrombotic). Duplication of the superficial femoral artery. Absence or duplication of the deep femoral artery caused by separately originating branches. Knowledge of embryogenesis of the vessels helps to understand these artery variations.

[From the pioneer-days of angiography (author's transl)]. / Aus der Pionierzeit der Arteriographie.

The occasion of transferring old archives of angiograms done between 1935 and 1940 to our present department led to the review of techniques and problems of angiography in these days; including a review of contrast agents, initial possibilities for series arteriography, puncture methods and catheter technique.