Apoptosis and meiotic segregation in ejaculated sperm from Robertsonian translocation carrier patients.

BACKGROUND: To better understand the infertility of patients with Robertsonian translocation, the biochemical and ultrastructural apoptotic characteristics of apoptosis in the sperm of patients and fertile donors were studied. METHODS: Ejaculated sperm samples of seven Robertsonian translocation carriers and seven fertile donors were analyzed after cryopreservation. The proportion of both viable and dead spermatozoa expressing activated caspases was detected by flow cytometry through the use of different specific carboxyfluorescein-labeled caspase inhibitors. Sperm DNA fragmentation was evaluated by the TUNEL method. The percentages of intact spermatozoa or spermatozoa with ultrastructural features of apoptosis, immaturity or necrosis were estimated by electron microscopy. Meiotic segregation analysis was performed by FISH. RESULTS: Significantly lower concentration, forward motility and normal morphology of spermatozoa were found in ejaculated samples of the Robertsonian patients than fertile donors. Compared with the control group, in Robertsonian translocation carriers: (i) the caspase assays showed a significantly increased (P < 0.05) proportion of viable spermatozoa with activated poly-caspases (57.4 versus 25.8%), caspase-3 (43.5 versus 13.4%), caspase-8 (44.4 versus 17.1%) and caspase-9 (42.4 versus 10.0%); (ii) the rate of DNA fragmentation was higher (26.3 versus 12.8%); and (iii) sperm ultrastructural examination highlighted a higher percentage of immature (28.0 versus 10.0%) and apoptotic (24.5 versus 18.5%) spermatozoa. FISH study showed predominant normal/balanced spermatozoa (78.34-85.53%). CONCLUSIONS: These results show a predominant proportion of balanced and normal gametes and higher numbers of spermatozoa showing apoptosis and immaturity features in oligoasthenozoospermic Robertsonian translocation carriers than in fertile donors. This suggests defects in spermatogenesis and especially spermiogenesis of these infertile patients.

Preoperative uterine artery embolization (PUAE) before uterine fibroid myomectomy.

PURPOSE: To evaluate the potential of uterine artery embolization to minimize blood loss and facilitate easier removal of fibroids during subsequent myomectomy. METHODS: This retrospective study included 22 patients (median age 37 years), of whom at least 15 wished to preserve their fertility. They presented with at least one fibroid (mean diameter 85.6 mm) and had undergone preoperative uterine artery embolization (PUAE) with resorbable gelatin sponge. RESULTS: No complication or technical failure of embolization was identified. Myomectomies were performed during laparoscopy (12 cases) and laparotomy (9 cases). One hysterectomy was performed. The following were noted: easier dissection of fibroids (mean 5.6 per patient, range 1-30); mean intervention time 113 min (range 25-210 min); almost bloodless surgery, with a mean peroperative blood loss of 90 ml (range 0-806 ml); mean hemoglobin pretherapeutically 12.3 g/dl (range 5.9-15.2 g/dl) and post-therapeutically 10.3 g/dl (range 5.6-13.3 g/dl), with no blood transfusion needed. Patients were discharged on day 4 on average and the mean sick leave was 1 month. CONCLUSION: Preoperative embolization is associated with minimal intraoperative blood loss. It does not increase the complication rate or impair operative dissection, and improves the chances of performing conservative surgery.

Malignant degeneration of perineal endometriosis in episiotomy scar, case report and review of the literature.

Although malignant degeneration of cutaneous endometriosis is rare at only 0.3-1% in endometriosis surgical scars, diagnosis and management need to be defined. A case of malignant degeneration of perineal endometriosis is reported, with a review of literature. Physiopathology, epidemiological data, diagnostic and therapeutic methods are discussed for malignant degeneration of cutaneous endometriosis. Any scar lesion that evolves in response to the menstrual cycle should be considered endometriosis until proven otherwise, and thus could require surgical resection, with histological analysis. A history of cutaneous endometriosis with frequent recurrences can indicate malignant degeneration. All cases require long-term clinical follow-up because, despite the rarity of this diagnosis, the delay between benign endometriosis and malignant transformation can vary from a few months to over 40 years.

Peginterferon alpha-2b plus ribavirin for treatment of chronic hepatitis C with severe fibrosis: a multicentre randomized controlled trial comparing two doses of peginterferon alpha-2b.

We compared sustained virological response (SVR) in chronic hepatitis C patients with severe fibrosis treated with pegylated interferon (Peg-IFN) alpha-2b 1.5 microg/kg/week or 0.75 microg/kg/week in combination with ribavirin 800 mg/day for 48 weeks. This was a multicentre randomized controlled study. SVR was observed in 44.5% (45/101) of patients treated with the standard dose of Peg-IFN and 37.2% (38/102) of patients treated with the low dose (NS). In patients with genotypes 1, 4 and 5, SVR was observed in 25.0% of patients who received the standard dose and 16.9% of patients who received the low dose of Peg-IFN (P = NS). In patients with genotypes 1, 4 and 5 and low viraemia, SVR was obtained in 27.3% of patients treated with the standard dose and 25.8% of patients treated with the low dose (P = NS). In the high-viraemia subgroup, SVR was obtained in 24.0% and 9.1% of patients, respectively. In patients with genotypes 2 and 3, SVR was similar in both groups (73.2%vs 73.0%). Thus, (1) patients with genotypes 2 and 3 and severe fibrosis can be treated with low dose of Peg-IFN and ribavirin, (2) this study suggests that patients with genotypes 1, 4 and 5 and high viraemia could receive a standard dose of Peg-IFN associated with ribavirin for 48 weeks, (3) side effects limit the efficacy of the treatment with standard dose of Peg-IFN in patients with genotypes 1, 4 and 5 and low viraemia, (4) more studies are needed for patients with genotype 2 or 3 to define the optimal duration (24 or 48 weeks) in patients with severe fibrosis.

[Non Hodgkin's and Hodgkin's lymphomas and HIV: frequency, outcome and immune response under HAART; Clermont-Ferrand University Hospital, 1991-2003]. / Lymphomes non hodgkiniens et hodgkiniens et infection VIH: fréquence, pronostic et reconstitution immune sous trithérapie antirétrovirale; CHU de Clermont-Ferrand, 1991-2003.

OBJECTIVES: The authors had for aim to identify cases of non Hodgkin's (NHL) and Hodgkin's (HL) lymphomas in HIV1-infected patients to assess 1) their incidence, before and after 1996, 2) the clinical features and outcome under treatment together with the survival rate of the patients, 3) the immune reconstitution of lymphoma-free patients under HAART. PATIENTS AND METHODS: A retrospective study was made of HIV1-infected patients managed at the Clermont-Ferrand University Hospital from 1991 to 2003 for the diagnosis and treatment of HIV1-related lymphomas. RESULTS: Forty-one patients were included: 35 NHL and 6 HL giving a cumulative incidence rate estimate from 2.4% between 1991 and 1996 to 3.4% between 1997 and 2003 while other opportunistic diseases were decreasing. A high proportion of aggressive and disseminated disease was observed among NHL cases. Complete remission was achieved in 17 (49%) and 5 (83%) NHL and HL cases respectively. The mean survival was 109+/-54 months and was correlated with CD4 cell count at lymphoma diagnosis (univariate analysis). Among responding patients, 5 died: 3 from opportunistic infections, 1 commited suicide, and 1 from hepatic carcinoma. For responding patients, the mean increase of CD4 cell count under HAART was 58/mm3 over a 2 year-period and 192/mm3 over a 5 year-period of follow-up. CONCLUSIONS: The incidence of lymphomas in HIV-infected patients has not decreased since the introduction of HAART. The immune status assessed by CD4 cell count on diagnosis is correlated with survival. Immune restoration in lymphoma-free patients under HAART is poor.

Guanylyl cyclase C as a reliable immunohistochemical marker and its ligand Escherichia coli heat-stable enterotoxin as a potential protein-delivering vehicle for colorectal cancer cells.

mRNA-based technologies and preclinical research in a variety of animal models have shown that guanylyl cyclase C (GCC) is a highly sensitive and specific molecular marker for the diagnosis of colorectal cancer (CRC). GCC is also a receptor for Escherichia coli (E. coli) heat-stable enterotoxin (STa) and can be used for STa-directed delivery of small-sized imaging agents to human CRC tumours. In this study, we have evaluated GCC as a new immunohistochemical (IHC) marker for CRC tissues and STa as a suitable vector for delivering high-sized protein molecules to CRC cells. Firstly, we have developed a highly sensitive EnVision(+)-based IHC staining method for detecting GCC in serial paraffin-embedded sections of primary and metastatic CRC (38 cases) or non-CRC (14 cases) adenocarcinomas. Carcinoembryonic antigen (CEA) and cytokeratin 20 (CK20) were chosen as controls. Our results indicate that GCC staining was positive in 100% of CRC tumours and was comparable to CEA (95%) or CK20 (92%). In contrast to CEA and CK20, GCC was negative in all of the extra-intestinal non-CRC tumours examined. GCC appears to display higher specificity than either CEA or CK20 while retaining high sensitivity, suggesting that it is a better CRC marker than CEA or CK20. Secondly, STa was genetically coupled to green fluorescent protein (GFP) and the resulting GFP-tagged STa was characterized for expression in E. coli and enterotoxicity in mouse. The binding characteristics of GFP-STa in CRC Caco-2 cells were followed by immunofluorescence microscopy. In this work we show that GFP-tagged STa is biologically active and has retained its ability to internalise into Caco-2 cells making it a potential vehicle for the delivery of anticancer therapeutic protein agents.

Isolated spleen agenesis: a rare cause of thrombocytosis mimicking essential thrombocythemia.

Thrombocytosis is a common feature of myeloproliferative disorders but may also result from various conditions including chronic iron deficiency, hemorrhage, chronic inflammation and splenectomy. We report two cases of secondary thrombocytosis caused by isolated and congenital asplenia, mimicking essential thrombocythemia. These two adult cases of spleen agenesis were unexpected. We conclude that in thrombocytosis without clinical evidence of splenomegaly, attentive screening of blood in search of Howell-Jolly bodies and abdominal ultrasonography should always be performed not only to detect mild spleen enlargement but also to make sure of the presence of this organ.

[Sarcoidosis gastropathy: diagnosis and contribution of echo-endoscopy]. / Gastropathie sarcoïdosique: diagnostic et apport de l'écho-endoscopie.

We report a case of gastric sarcoidosis in a 36-year old woman. The clinical and endoscopic ultrasonography findings initially suggested linitis plastica. Subsequently the discovery of unexpected gastric and then, bronchial granulomas after bronchoscopy and bilateral ocular fixation on gallium scanning, led to the diagnosis of sarcoidosis. Endoscopic ultrasonography showed abnormal hypertrophy of the third hyperechoic layer. These findings usually suggest among the etiologies of giant gastric folds, malignant diseases and particularly linitis plastica.

[Hepatocellular carcinoma in cirrhosis: semeiology and performance of magnetic resonance imaging and lipiodol computed tomography]. / Carcinome hépatocellulaire sur cirrhose: sémiologie et performances de l'imagerie par résonance magnétique et de la tomodensitométrie après artériographie lipiodolée.

OBJECTIVES: a) Describe hepatocellular semiology in magnetic resonance imaging and lipiodol computerized tomography in patients with cirrhosis, who are candidates for surgery; b) Clarify the respective roles of magnetic resonance imaging and lipiodol computerized tomography in hepatocellular detection. METHODS: Twenty four patients with suspected hepatocellular carcinoma underwent successive magnetic resonance imaging and lipiodol computerized tomography. Thirty-four of the 67 lesions seen by lipiodol computerized tomography and 28 of 52 lesions seen by magnetic resonance imaging were confirmed histologically. RESULTS: In lipiodol computerized tomography, 44% of hepatocellular carcinomas had a dense and homogeneous pattern; 24% had a homogeneous but slightly dense pattern. Sixteen distinct deposits were described: 4 were confirmed as hepatocellular carcinoma and 12 were not controlled histologically. In magnetic resonance imaging 57% of hepatocellular carcinomas have a high intensity on T1 and T2 weighted spin echo images, 38% were hyperintense on T2 and hypo or isointense on T1 weighted images. Eighty-six percent of hyperintense T1 and T2 weighted images were hepatocellular carcinoma. When the gold standard was histology, lipiodol computerized tomography sensitivity (81%) was higher than magnetic resonance imaging (68%). When the gold standard was lipiodol computerized tomography, the sensitivity of magnetic resonance imaging was 47 +/- 12%. CONCLUSIONS: a) The sensitivity of lipiodol computerized tomography was better than resonance magnetic imaging; b) the homogeneous and slightly dense pattern corresponded to a hepatocellular carcinoma in 50% of cases; c) on magnetic resonance imaging any lesions with high intensity on T1 and T2 spin echo images strongly suggests hepatocellular carcinoma; d) if surgical resection after ultrasonography is being considered, the second step should be an magnetic resonance imaging.

Fibrogenesis in peritoneal endometriosis. A semi-quantitative analysis of type-I collagen.

OBJECTIVE: To evaluate the degree of fibrosis in the stroma of peritoneal endometriosis, according to the macroscopic appearance. METHODS: Red and black lesions of peritoneal endometriosis were collected and type-I collagen was immunohistochemically identified. The percentage of the stained area in the stroma was measured using a computerized image analysis system. RESULTS: There was a significant difference in the mean percentage of the total stained area between red and black lesions. CONCLUSION: The amount of type-I collagen is significantly higher in the stroma of black peritoneal lesions than in red peritoneal lesions.

Increased mast cell density in peritoneal endometriosis compared with eutopic endometrium with endometriosis.

PROBLEM: Factors regulating fibrosis in peritoneal endometriosis are poorly understood. We hypothesized that mast cell proteases may play a role in the process of fibrosis in peritoneal endometriosis. As the first step in examining the possible contribution of mast cells to fibrosis, we investigated their distribution in peritoneal endometriosis lesions according to their macroscopic appearance. METHOD OF STUDY: Mast cells were identified by immunohistochemistry using monoclonal antibody against mast cell tryptase on formalin-fixed, paraffin-embedded sections. Mast cell density (mean mast cell count per 0.13 mm2 of stroma) was measured using a computerized image analysis system. RESULTS: Mast cell density was significantly increased in black peritoneal lesions compared with red peritoneal lesions in endometriosis. CONCLUSIONS: Mast cells may be involved in the pathogenesis of peritoneal endometriosis.

Angiogenesis in endometriosis.

We evaluated angiogenesis in eutopic endometrium and ectopic endometrium from patients with endometriosis. Microvessels were identified by immunohistochemistry using anti-von-Willebrand factor antibody and vascular parameters were measured using a computerized image analysis system. No relationship was observed between vascular density and type of endometriotic lesion. Heterogeneity in vascular density was observed in different lesions within the same patient. Between red and black peritoneal lesions a significant difference was observed in the frequency distribution of luminal diameter. In black lesions the variability of luminal diameter was decreased and most microvessels were less than 20 mm in diameter. We suggest that the frequency distribution of microvessel luminal diameter of red and black peritoneal lesions may be different and should be important when studying the regulation of angiogenesis in endometriotic implants.

[Segmental absence of colonic muscularis propria. Report of a case in an adult]. / Absence segmentaire de musculeuse colique. A propos d'un cas chez l'adulte.

We report a case of segmental defect of colonic muscularis propria revealed by perendoscopic perforation in a 64-year-old woman. Segmental absence of intestinal musculature is well documented in new-borns and infants and is more frequent in small bowel. It is characterized by localised absence of muscularis propria without fibrous scar. The remaining layers of the bowel wall are intact. The pathogenesis of this lesion is discussed.

Arterial vascularization of the gastrointestinal tract in Macaca fascicularis (cynomologus monkey).

We describe the origin, course, and distribution of the arteries responsible for vascularization of the subdiaphragmatic gastrointestinal tract of Macaca fascicularis as well as the characteristics of the celiac trunk and the superior and inferior mesenteric arteries, studied in a series of 50 animals. Detailed knowledge of these systems is an essential requirement if experimental surgery is to be successfully performed in these laboratory animals.

[Ploidy analysis in a case of ovarian small cell carcinoma with hypercalcemia]. / Analyse de la ploïdie dans un cas de carcinome de l'ovaire à petites cellules avec hypercalcémie.

The authors studied DNA content in a case of small cell carcinoma with hypercalcemia. This tumor exhibited typical clinical, histological, immuno-histochemical, ultrastructural and biological patterns. DNA content was measured both by flow and image cytometry performed on unfixed tumoral samples. The proliferation index was 10%. These results are similar to those of the literature obtained retrospectively from 10% formalin fixed tissues. The DNA content is a clue to distinguish this entity from other small cell carcinomas of the ovary because immunohistochemical findings are not always informative. The diagnosis of small cell carcinoma of hypercalcemic type should be questioned if DNA content is abnormal.

Meckel's cartilage in the human embryo and fetus.

This work studied the development of the ventral part of Meckel's cartilage in a series of human embryos (classified in stages) and fetuses. These stages appeared particularly important: stage 16, appearance of Meckel's cartilage; stage 20, beginning of membranous ossification of mandible; and stage 23, end of the embryonic period (8th week). The primitive bony nodule which develops from the embryonic mesenchyme appears as a double bony layer forming a groove containing the neurovascular bundle, into which the dental lamina is also invaginated. It was concluded that during the fetal period, the cartilage participates in the formation of the body of the mandible in an area close to the mental foramen via endochondral ossification. The cartilage disappears in parallel with the development of ossification by the sixth month.