Human recombinant myeloperoxidase antiviral activity on cytomegalovirus.

In vitro incubation of human cytomegalovirus (Towne strain) with 8 U/ml human recombinant myeloperoxidase plus sodium chloride and glucose nearly abolished viral infectivity. To assay the effect on intracellular infection, cell toxicity of the enzymes was first studied. Even the high dose of 16 U/ml of recombinant myeloperoxidase plus 10 mU/ml glucose oxidase did not decrease MRC5 cell growth. By contrast, this dose reduced proliferation of activated THP1 cells. Even half of the myeloperoxidase dose proved slightly toxic to these cells. Non-cytotoxic concentrations of the reagents were used to monitor their effect on cytomegalovirus infection. In MRC5 cells, even the low dose of 4 U/ml myeloperoxidase plus glucose oxidase inhibited synthesis of cytomegalovirus early antigens, as tested by immunofluorescence. Viral release in the supernatant was decreased by 4 logs. In THP1 cells, which produce endogenously hydrogen peroxide, myeloperoxidase alone (8 U/ml) decreased the formation of early and late antigens by 53 and 44%, respectively.

A human milk factor susceptible to cathepsin D inhibitors enhances human immunodeficiency virus type 1 infectivity and allows virus entry into a mammary epithelial cell line.

Human immunodeficiency virus type 1 (HIV-1) growth in lymphocyte cultures was increased when the virus inoculum was incubated in breast milk. The enhancing effect of milk was abolished by anti-cathepsin D antibody or by pepstatin A, a cathepsin D inhibitor. The cathepsin D-producing CD4-negative MCF7 mammary cells supported the growth of some HIV-1 isolates. An MCF7 line chronically producing HIV-1 IIIb was obtained. Cathepsin D may induce conformational modification of viral gp120, allowing direct interaction with a coreceptor. We demonstrated the presence of CXCR4 mRNA in MCF7 cells.

Management of common bile duct stones in a single operation combining laparoscopic cholecystectomy and perioperative endoscopic sphincterotomy.

BACKGROUND: Laparoscopic cholecystectomy (LC) has become the reference treatment for biliary lithiasis, but the management strategy for common bile duct stones (CBDS) remains a subject of controversy in the absence of an established consensus. While conventional surgery remains the reference treatment for CBDS, minimally invasive techniques are becoming more and more popular. These methods consist of the extraction of the common bile duct stones either exclusively by laparoscopy or by sequential treatment with endoscopic sphincterotomy (ES) followed by LC. The aim of this study was to evaluate the treatment of CBDS in a one-stage operation by laparoscopic cholecystectomy (LC) and perioperative endoscopic sphincterotomy. PATIENTS AND METHODS: Between January 1994 and March 1998, 44 patients, 20 male and 24 female, (sex ratio 1.2) with a median age of 57 years (range 28-84 years) were treated for suspected or confirmed CBDS. The CBDS were uncomplicated in 39 cases (88%) and associated with a complication in 5 cases (12%), namely, cholangitis (2 cases) or acute pancreatitis (3 cases). The perioperative ES was performed immediately after the LC during the same operative time, with perioperative cholangiography being systematically performed (1 failure). In 6 cases, a transcystic drain was left in place (to ensure complete evacuation of the CBDS postoperatively) when there were more than three stones and/or when they were larger than 6 mm. The patient was positioned in the left lateral position in order to perform the ES. RESULTS: Mean operative time for LC was 60 min, range 40-90 min. The general anesthesia was prolonged by 40 min in order to perform an ES (range 30-60 min). The perioperative ES was unsuccessful in one case (2%), due to the impossibility of catheterizing the papilla, the preoperative MR cholangiogram being normal. Immediate clearance of the CBD was achieved in 95% of the cases (42 p). In 2 cases, residual stone was found in the sixth day after cholangiography and was spontaneously evacuated as shown by 21st-day control. There was no mortality or postoperative complications. The duration of the postoperative hospitalization was 4.6 days (range 3-6). CONCLUSIONS: We believe that LC combined with perioperative ES is a quick, reliable, and safe technique for the treatment of CBDS during a single operative procedure, although this approach is limited by the proximity and availability of an endoscopic team.

HIV-1 infectivity and host range modification by cathepsin D present in human vaginal secretions.

OBJECTIVE: To investigate HIV-1 infectivity in the natural environment of vaginal secretions. DESIGN: Vaginal wash samples collected from 14 healthy women were incubated in vitro with various HIV-1 strains for 10 min at 37 degrees C and then assayed for infectivity on primary lymphocyte cultures, or on CEM cells, or on CD4- ME180 cells derived from vaginal epithelium. METHODS: HIV-1 infectivity was measured by early virus growth in the various host cells tested using a quantitative p24 assay and by the Karber procedure. RESULTS: Preincubation of HIV-1(IIIB) with vaginal wash samples or 2 microg/ml cathepsin D increased the ability of the virus to grow in lymphocyte cultures. The vaginal wash effect was abolished by 5 microg/ml pepstatin A, an inhibitor of aspartyl proteases. Presence of precursor and mature forms of cathepsin D in vaginal wash was demonstrated after passage through a pepstatin A-agarose column. Median tissue culture infective doses of HIV-1(IIIB) and HIV-1(JRFL) strains were increased 14.4-fold and 18-fold, respectively, after preincubation in vaginal wash sample, and were increased by pretreatment with 2 microg/ml cathepsin D. When CD4 receptors of CEMss cells were blocked by OKT4a monoclonal antibody, the cells lost susceptibility to HIV-1 (IIIB), but supported the growth of virus pretreated with vaginal wash sample or cathepsin D. These treated viruses were able to initiate infection of CD4-ME180 epithelial cells, which were not receptive to untreated virus. ME180 cells were shown to possess the messenger of CXC-chemokine receptor-4. CONCLUSIONS: Vaginal secretions may help HIV-1 transmission to women by increasing infectivity for CD4+ cells and allowing entrance into some CD4-epithelial cells.

Management of common bile duct stones by laparoscopic cholecystectomy and endoscopic sphincterotomy: pre-, per- or postoperative sphincterotomy?

BACKGROUND/AIMS: The aim of this study was to evaluate the treatment of common bile duct stones (CBDS) by endoscopic sphincterotomy (ES) and laparoscopic cholecystectomy (LC), ES being performed either pre-, per- or postoperatively. METHODS: Between January 1990 and June 1997, 386 patients with a median age of 60 (range 18-92) years were treated for suspected or confirmed CBDS. The CBDS were uncomplicated in 264 cases (70%) but associated with a complication in 122 cases (30%), namely, cholangitis (69 cases) or acute pancreatitis (53 cases). ES combined with LC was carried out in 233 cases (60%): ES was preoperative (sequential treatment in two stages) in 197 cases (51%); peroperative in 30 cases (7%), or postoperative in 6 cases (2%). Laparoscopic extraction was performed in 58 cases (15%) and conventional surgery in 82 cases (21%). RESULTS: With respect to sequential treatment, endoscopic retrograde cholangiography showed the presence of CBDS in 117 cases (60%) and preoperative ES allowed the release of the CBDS in 82% of these cases. The complication rate of sequential treatment was 8% (15 cases) after ES and 7% (13 cases) after LC, with 1 death (0.5%). A peroperative ES performed after LC enabled evacuation of the CBDS in 28 cases (93%) without any complications or mortality. Postoperative ES was successful in 100% of cases with residual lithiasis in 16% (1 case) and a complication rate of 16% (1 case). CONCLUSION: Along with conventional surgery and laparoscopic extraction, ES combined with LC represents an effective alternative in the management of CBDS. Since it can be performed peroperatively, it allows a one-stage, minimally invasive treatment of most uncomplicated CBDS.

Detection and identification of human papilloma viral DNA, types 16, 18, and 33, by a combination of polymerase chain reaction and a colorimetric solid phase capture hybridisation assay.

A colorimetric microplate hybridization assay was developed previously to simplify detection procedures of DNA fragments resulting from polymerase chain reactions (PCR). This format has now been adapted for the simultaneous detection and identification of three human papillomavirus (HPV), types 16, 18 and 33, associated frequently with cervical cancer. This post-PCR detection system uses three type-specific capture oligonucleotides linked covalently to a single microplate well and three type-specific multibiotinylated oligonucleotidic probes for detection. It therefore offers a double specificity; the first is conferred by pairs of primers, specific of each type of virus tested, and the second, by the sets of capture and detection probes which are complementary to internal regions of the amplified DNA fragments. The detection format outperformed agarose gel electrophoresis of amplified DNA products in sensitivity and specificity. The rapidity and simplicity of this hybridisation system would justify its use in routine diagnostic examination of cervical specimens (smears and biopsies).

[Laparoscopic treatment of gastroesophageal reflux using fundoplication. Midterm results]. / Traitement laparoscopique du reflux gastro-oesophagien par fundoplicature. Résultats à moyen terme.

UNLABELLED: The aim of this study was to see whether the results of surgical treatment of gastroesophageal reflux disease (GERD) by laparoscopic fundoplication were satisfactory and stable over time. PATIENTS AND METHODS: From July 1992 to September 1996, 161 patients with medical treatment-dependent GERD were operated. 17 patients were excluded from the study (conversion or immediate laparotomy). The group of 144 patients included consisted of 92 men and 52 women with a mean age of 50 (25-77 years). The preoperative work-up included endoscopy, esophageal manometry and 24-hour pH monitoring. The surgical procedures were complete fundoplication without section of the short vessels (Nissen-Rossetti: 122 cases), with section of the shorts vessels (Nissen: 18 cases), or partial fundoplication of 270 degrees (Toupet: 4 cases). The patients were reviewed clinically 3 months after the operation, with repeat manometry and pH monitoring; yearly survey was performed. RESULTS: There was no postoperative mortality. The morbidity consisted of 2 respiratory complications (1.5%) with good recovery. With a follow-up of 3 and 21 months, the dysphagia rate was 24 vs 2% of patients (p < 0.05), the rate of gas bloat syndrome was 18% vs 21% (n.s.), and the GERD recurrence 6% vs 14% (p < 0.05). The mean time to recurrence was 8 months. 2 patients were reoperated: one for incisional hernia and one for slipped-Nissen. CONCLUSION: The results of laparoscopic treatment of GERD with fundoplication procedures at 2 years follow-up showed an 86% cure rate of GERD control and 94% satisfaction rate for the patients who were investigated.

[Results of fundoplication by laparoscopic approach in the treatment of gastroesophageal reflux. Apropos of 224 cases]. / Résultats de la fundoplicature par voie laparoscopique dans le traitement du reflux gastro-oesophagien. A propos de 224 cas.

STUDY AIM: The aim of this paper is to evaluate prospectively immediate and 2-year results of laparoscopic fundoplicature (LF) for gastroesophageal reflux disease (GERD). PATIENTS AND METHODS: Patients presenting GERD who had been previously submitted to a long-term medical treatment were included in this study. Preoperative workup included upper GI tract endoscopy, esophageal manometry and 24-hour pHmetry. Standard surgical procedure incorporated a Nissen-Rossetti 360 degrees fundoplicature. Short vessels division (Nissen operation) was performed in case of high strength of the wrap and a partial fundoplicature (Toupet 270 degrees) was performed when motility disorders of the esophagus were demonstrated by manometry. Postoperative morbidity and results were evaluated, with a clinical appreciation at 3 and 22 months, and by manometry and pHmetry at 3 months. RESULTS: Two hundred and thirty-five patients were observed, and 224 included in the study (143 men and 92 women). Nissen-Rossetti fundoplication was performed in 169 cases (80%), Nissen in 30 (14%) and Toupet in 13 (6%). In 12 cases (5%). LF was converted to an open Nissen-Rossetti procedure. There was no hospital mortality and complications were noted in three cases (1.5%): pneumonia (n = 2) and gastroplegia (n = 1). With a mean 22-month follow up, among the 103 patients who answered to a questionnaire, the rate of relapse of GERD was 14%, dysphagia was present in 2% and four patients had been reoperated on (one for a slipped Nissen, one for a stenosis of the esogastric junction and two incisional hernias). CONCLUSION: On the basis of this experience, LF for GERD is a safe and efficient operation, with 86% of good results at 2 years.

[The remaining indications for open surgery in common bile duct lithiasis]. / Indicatii reziduale ale chirurgiei deschise în litiaza caii biliare principale.

Once with the continuous development of mini-invasive procedures, the open surgery indications in the treatment of CBD stones are decreasing. Between January 1986 and December 1987, 514 patients with average age 60 years old (18-94) were treated for suspected or confirmed CBDS. Three distinctive periods were studied function of how the conventional surgery were either exclusive (group I), either together with the beginning era of laparoscopy (group II), or together with institutionalised laparoscopy (group III). Group I had 110 cases, group II--207 cases and group III--197 cases. The conventional surgery were 100% performed in group I, 26% (52 cases) performed in group II, 14% (28 cases) performed in group III. If the laparotomy was the only therapeutical option in patients with CBDS from group I, the therapeutical procedures become more various after 1990. It was done endoscopic sphincterotomy in 59% (124 cases) in group II and in 64% (126 cases) in group III. The laparoscopic extraction of CBDS was 12% (cases) in group II and 19% (37 cases) in group III. The overall mortality in conventional surgery was 1.5% with 1.8% in group I and 0% in group II and III. The complications rate was 13% with 8% in group II and 21% in group III. The postoperative hospitalisation time average was 16 days for laparotomy of the 3 groups. The conventional surgery is main indicated when laparoscopic surgery for CBDS extractions or endoscopic sphincterotomy is contraindicated or failed; also it is useful in some complicated forms of CBDS.

[Outcome of surgical treatment of cancer of the stomach. Report of 330 cases]. / Risultati della terapia chirurgica del cancro dello stomaco. A proposito di 330 casi.

The study concerned 330 cases of gastric adenocarcinoma operated from January 1969 to June 1996. Clinical, histological and therapeutic features were analyzed and compared during 3 periods (Group 1: 1969-79, Group 2: 1979-88, Group 3: 1989-96). The aim was to evaluate changes and results occurred in gastric carcinoma. The median age was 65 +/- 11 year (range 25-90), the sex ratio 2. The most common location was in the lower third of the stomach and the cancer was often far advanced (71 percent of stages III and IV). Subtotal gastrectomy was replaced by total gastrectomy and large lymph node dissection was systematically performed in group 3. Curative resection rate increased from 28 to 60 percent between group 1 and group 3. Despite the more radical surgery, post-operative mortality rates decreased respectively 22%, 9%, 8% for the groups 1, 2, 3 (p < 0.01) and global morbidity remained stable while the rate of intra-abdominal infection have changed in group 3 (4.8% vs 2.7% and 2.1% in respectively groups 1 and 2; p = ns). Two year survival rates were 20% for group 1.25% for group 2 and 35% for group 3 (p < 0.01). This results can be considered as encouraging but have to be confirmed at 5 year survival.

[Surgical treatment of adenocarcinoma of the stomach: 1969-1994. Apropos of 261 cases]. / Le traitement chirurgical de l'adénocarcinome de l'estomac: évolution de 1969 à 1994. A propos de 261 cas.

The study concerned 261 cases of gastric adenocarcinoma operated on between 1969 and 1994. Clinical, histological and therapeutic features were analyzed and compared during 3 periods (Group 1: 1969-78, Group 2: 1979-88, Group 3: 1989-94). The aim was to evaluate changes and results occurred in gastric carcinoma. The median age was 65 years (rage 25-90 years), the sex ratio, 1.9. The most common location was in the lower third of the stomach and the cancer was often far-advanced (73 percent of stages III and IV). Subtotal gastrectomy was replaced by total gastrectomy; the number of distal gastrectomy was unchanged. Large lymph node dissection was systematically performed in group 3. Curative resection rate increased from 28 to 51 percent between group 1 and group 3. Despite the more radical surgery, morbidity and post operative mortality rates decreased (respectively: 44.5%, 33%, 25.4%, and 22%, 10%, 7.2% for the groups 1, 2, 3, p < 0.01). 2-year survival rates were 20 percent for group 1, 25 percent for group 2 and 32 percent for group 3 (p < 0.01). This rates can be considered as satisfying in view of the high rate of III and IV stages.

[Retro-duodenal papillary perforation after endoscopic sphincterotomy. Role of surgical treatment by exclusion of the duodenum. Apropos of 3 cases]. / Perforation papillaire rétroduodénale après sphinctérotomie endoscopique. La place du traitement chirurgical par exclusion du duodénum. A propos de 3 cas.

Among the complications of endoscopic sphincterotomy whose rate is about 10% of cases, the retroduodenal papillary perforation represents about 1% of cases. The diagnosis lies on radiological examination which may show during the sphincterotomy the extravasation of the contrast fluid used for the retrograde cholangiography; it can be suggested by the presence of clinical signs of retroperitoneal sepsis or peritonitis. The treatment depends on the severity or peritonitis. The treatment depends on the severity of the clinical symptomatology; it is generally a medical treatment associating nasogastric aspiration and antibiotherapy, and more rarely a surgical one. There is no consensus concerning surgical modalities. We report 3 cases of retroduodenal papillary perforation treated surgical by a duodenal exclusion aiming to transform a complex fistula in a bilio-pancreatic fistula which can be more easily managed by somatostatine-like drugs.

[Virology: diagnosis or screening?]. / Virologie: diagnostic ou dépistage?

André Dachy is a sensitive clinician as well as a microbiologist who knows about hidden meanings of laboratory tests. Pediatrics was enriched by his dual interest in observation of the children and of the microbes which threaten them. As time went by, Dachy's interest for social medicine grew. Thus, it is justified to address him the present consideration on relevancy of virology diagnosis and modalities of screening.

Transitional cell carcinoma of the bladder: evaluation of the role of human papillomaviruses.

OBJECTIVES: The study evaluated the conflicting results of the role of human papillomavirus (HPV) in the development of bladder carcinoma. METHODS: We analyzed the frequency of HPV types 6, 11, 16, 18, and 33 by using polymerase chain reaction on formalin-fixed, paraffin-embedded specimens, from 75 cases of transitional cell carcinoma (TCC) of the bladder. Fifteen samples of normal urothelium adjacent to TCC (10) or from normal bladder obtained at autopsy (5) served as negative controls. RESULTS: HPV type 16 deoxyribonucleic acid (DNA) was detected in 2 (2.7%) of the 75 cases of TCC and in none of the normal urinary bladder cases. The 2 patients with HPV type 16 were immunosuppressed after undergoing renal and cardiac transplantation. CONCLUSIONS: These results strongly suggest that HPVs play a minor role in the development of TCC of the bladder in the general population, although they can act as oncogenic agents in predisposed patients, such as those who are immunosuppressed.

Human papillomavirus type 16 associated with multifocal transitional cell carcinomas of the bladder in two transplanted patients.

This report describes two cases of rapidly progressive, multifocal transitional cell carcinomas of the bladder that developed in two patients after renal and cardiac transplantation, respectively. In both cases human papillomavirus (HPV) type 16 DNA was detected using the polymerase chain reaction DNA amplification method. To our knowledge, this HPV type has not been previously described in multifocal bladder transitional cell carcinoma in transplanted patients. Our findings suggest that HPV may play a major role in the development of rapidly progressive, multifocal transitional cell carcinoma in immunosuppressed patients.

[Treatment of lithiasis of the common bile duct by endoscopic sphincterotomy and laparoscopic cholecystectomy]. / Le traitement de la lithiase de la voie biliaire principale par sphinctérotomie endoscopique et cholécystectomie laparoscopique.

This paper evaluates the treatment of common bile duct stones by endoscopic sphincterotomy (SE) and laparoscopic cholecystectomy (CL). 733 patients presenting with symptomatic cholelithiasis were operated on between March 1990 April 1993; 131 (18%) of them had a preoperative suspicion of common bile duct stones (LVBP): jaundice for 41, biliary acute pancreatitis for 27 and altered liver function tests for 63. 131 retrograde cholangiographies (CPRE) were attempted with an associated SE (113 cases) in the presence of LVBP, biliary pancreatitis, enlargement of common bile duct and appearance of forced papilla. CL was performed 24 to 48 hours later. CPRE +/- SE had no mortality; 1 patient presented a retroduodenal perforation of CBD, requiring surgery. 58 cases (44.2%) of LVBP were diagnosed, without a statistically significant difference according to the clinical pattern. In the group with altered liver function tests only alkaline phosphatase was significantly predictive of LVBP. There was no mortality or morbidity related to CL; conversion rate was 9.8%; 4 of 12 cases of conversion were related to persistence of stones in the common bile duct, without any possibility of laparoscopic extraction. Mean hospital stay was 7.4 days. Efficacy of this sequential method of treatment of LVBP was 91.3%: this method seems satisfactory, not dangerous and minimally invasive, and should be indicated for pre-operative suspected common bile duct stones.

[Cancer of the cervix, papillomavirus, contraception and tobacco]. / Cancer du col utérin, virus du papillome, contraception et tabac.

This survey started in May 1989 and deals with 2,436 patients treated by 10 gynecologists from the Brussels region. In addition to routine cytological analysis in cervical smears, an additional test for Human Papilloma Virus (HPV 16, 18 and 33) was advised for these women. Gynecologists filled in a questionnaire about these patients. In cases with benign cytology, virus prevalence was 2 times greater in woman who took contraceptive pills, over at least 6 months, than in each of the other groups using either diaphragm or male condoms or no contraception, or at menopausal age. It was strange to find that viral prevalence varied according to the pill trade mark, but not according to the pill hormonal content. No confounding factor in population recruitment could be detected. In contrast to viral infection, prevalence of cancer lesions was not increased in the oral contraception group. Squamous intraepithelial lesions of Low Grade and High Grade (SIL LG and HG) were on the contrary 2 times less frequent in women taking the pill, with or without HPV infection. During follow-up of 223 women, the same HPV genotype persisted in SIL lesions but it disappeared within 4 to 8 weeks in 68% of normal cervical smears. However, beyond this period, the proportion of women with persisting virus remained nearly constant, indicating that a subgroup may be less able to mount a defence against the infection. Risk factor of cigarette smoking for SIL LG or HG was 1.6 in women with HPV and 4.0 in those with no virus. This factor was 3.3 in women with oral contraception and 4.5 in the other group.

Demonstration of human papillomavirus type 2 in a verrucous carcinoma of the foot.

This report describes one case of verrucous carcinoma of the foot containing human papillomavirus type 2 DNA which was detected by the polymerase chain reaction DNA amplification method. Our findings suggest that human papillomavirus type 2, which was classically associated with palmoplantar warts, may also play a role in the pathogenesis of peripheral verrucous carcinoma.