Distribution of 14 high risk HPV types in cervical intraepithelial neoplasia detected by a non-radioactive general primer PCR mediated enzyme immunoassay.

AIM: To evaluate the presence of high risk human papillomaviruses (HPV) in cervical smears showing intraepithelial neoplasia (CIN). METHODS: The presence of 14 high risk HPV was evaluated in 114 cervical smears with CIN of different degrees, by comparing a non-radioactive polymerase chain reaction (PCR) enzyme immunoassay (EIA) with conventional PCR followed by radioactive Southern blot hybridisation. General primer PCR amplicons detecting low risk and high risk HPV were typed for 14 different high risk HPV types (HPV 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, and 68) by a non-radioactive PCR-EIA. Virus load of HPV 16 positive CIN was assessed using the semiquantitative PCR-EIA. RESULTS: Histological evaluation confirmed CIN I in 49 cases (mean age 29.0 years, range 17 to 52), CIN II in 31 cases (mean age 30.8 years, 18 to 54), and CIN III in 34 cases (mean age 31.1 years, 16 to 57). The non-radioactive PCR-EIA showed an overall agreement rate of 90% (kappa value 0.75) when compared with conventional general primer PCR followed by radioactive Southern blot hybridisation. High risk HPVs were detected in 47% of CIN I, 77% of CIN II, and 97% of CIN III (p < or = 0.02). HPV types 39, 51, 56, and 58 were found in CIN I exclusively (between 2% and 8%). HPV 16 and HPV 31 were detected in 12% and 2% of CIN I, 35% and 21% of CIN II, and 74% and 13% of CIN III, respectively (p < or = 0.03 and p < or = 0.04). The virus load estimated by the semiquantitative PCR-EIA of HPV 16 was similar in CIN I, CIN II, and CIN III. CONCLUSIONS: The PCR-EIA has high clinical sensitivity for detecting CIN II/III (90%). There was a significantly higher prevalence rate of HPV 16 and 31 in CIN III than in CIN I and II.

Laparoscopy-assisted radical vaginal hysterectomy modified according to Schauta-Stoeckel.

In laparoscopy-assisted radical vaginal hysterectomy, laparoscopy is used to develop the paravesical and pararectal spaces. The cardinal ligament is isolated and cut after bipolar coagulation to the level of the deep uterine vein. By the vaginal approach, the ureters are identified before their entry into the bladder pillar. The uterine vessels are pulled down until their laparoscopically coagulated ends become visible. After incision of the vesicocervical reflection, the uterine fundus is grasped and developed (Döderlein maneuver). The lower cardinal and uterosacral ligaments are exposed by pulling the cervix and fundus uteri to the contralateral side. The cardinal and uterosacral ligaments are dissected and ligated, and the specimen is removed. We combined laparoscopic lymphadenectomy with radical vaginal hysterectomy in 33 women with cervical cancer. The mean operating time was 80 minutes for the vaginal phase and 215 minutes for the laparoscopic phase, including paraaortic and pelvic lymphadenectomy and preparation of the cardinal ligaments. Blood transfusions were necessary in four women. Three patients sustained injury to the bladder, one patient to the left ureter, and another patient to the left internal iliac vein. Repair was achieved at primary surgery for all intraoperative complications. No fistula was observed. The patients had fully recuperated after a mean of 28 days. The laparoscopy-assisted Schauta-Stoeckel approach may prove to be a safe alternative to conventional radical abdominal hysterectomy.

The pattern of apolipoprotein B100 containing lipoprotein subclasses produced by the isolated visceral rat yolk sac depends on developmental stage and fatty acid availability.

Explants of visceral rat yolk sacs from gestational days 16, 18 and 22 were used for studying developmental changes of secretion and density distribution of lipoproteins, particularly of those containing apoB. Moreover, the influence of fatty acid supply on the amount and density distribution of secreted apolipoproteins was studied on day 18 of gestation. Active lipoprotein production was observed in yolk sacs taken on days 16 and 18 of gestation. It declined considerably on day 22 of gestation in parallel with the production of total protein, triacylglycerols and cholesterol. On all gestational days, apoB floated mainly in the LDL range ( > or = 70%) with differences in the distribution pattern of LDL subclasses. The lowest density of secreted LDL was found on day 18 of gestation (peak at d = 1.025 g/ml) followed by day 16 (peak at d = 1.035 g/ml) and day 22 of gestation (peak at d = 1.045 g/ml). ApoAIV, apoE and apoAI floated exclusively in the HDL range with a peak at d = 1.089 g/ml independently of the gestational day. After incubation of yolk sacs from the 18th day of gestation with 0.4 mM or 0.8 mM oleate, the density of secreted apoB containing particles was decreased (peaks in the VLDL and IDL density range), whereas palmitate in the same concentrations caused a redistribution of secreted apoB toward higher densities (peaks at d > or = 1.032 g/ml). Taken together, the data provide evidence that the density of LDL subclasses produced by isolated yolk sacs between days 16 and 22 of gestation depended on the gestational stage. Moreover, addition of unsaturated or saturated fatty acids to the organ culture differently affected the secretory rate and the density of lipoproteins delivered by yolk sacs on day 18 of gestation.

[Laparoscopic para-aortic and pelvic lymph node excision--initial experiences and development of a technique]. / Laparoskopische paraaortale und pelvine Lymphonodektomie--erste Erfahrungen und Entwicklung einer Technik.

It was the aim of this study to establish and analyze the laparoscopic technique of para-aortic and pelvic lymphadenectomy. During a one year period (from August 1994 till July 1995) 42 patients underwent pelvic and para-aortic laparoscopic lymphadenectomy. In 29 cases cervical cancer, in 11 cases endometrial cancer and in 2 cases tumors of low malignant potential of the ovary were the indication for lymphadenectomy which was combined with radical vaginal hysterectomy in 19 patients or simple vaginal hysterectomy in 13 patients. During the observation period the mean operating time for para-aortic and pelvic lymphadenectomy decreased and the efficiency of the lymphadenectomy increased significantly: the mean operating time for the first 10 para-aortic lymphadenectomies was 52 minutes and for the pelvic lymphadenectomies 141 minutes, respectively. For the last 10 procedures the para-aortic part took 35 minutes and the pelvic part 110 minutes. Whereas at the beginning of the study a mean of 25 lymph nodes were removed, a mean of 36 lymph nodes were sampled during the last 10 procedures. In 3 patients operative injuries to major vessels were encountered of which two were followed by laparotomy. In 3 other patients laparotomy due to postoperative hemorrhage was necessary. These 6 complications occurred during the first half of the study and were not encountered during the following operations by changing from monopolar to bipolar coagulation and by modifying the regimen for perioperative thrombosis prophylaxis. In the first half of the study in 9 women blood transfusions were necessary and in the second half only 4 patients had to be transfused. In the first 10 patients the mean intraoperative blood loss was 1300 cc, in the last 10 patients 300 cc. After a short learning curve laparoscopic para-aortic and pelvic lymphadenectomy is a safe and effective technique for staging cervical, endometrial, and early ovarian cancers. Though this technique is no standard procedure results of this pilot study warrant prospective studies comparing this technique with conventional procedures.