Presentation of a patient with in situ amelanotic melanoma of the vulva.

Amelanotic malignant melanoma of the vulva is extremely rare. The authors describe here a case of amelanotic malignant melanoma of the vulva, occurring in a 71-year-old woman without any clinical symptoms. The woman had a small nodular lesion in the left labia majora. Local excision was performed. Histological examination revealed an in situ malignant melanoma without any evidence of invasive disease. All suspicious lesions in the vulva region, even if there are no clinical symptoms, should be biopsied, and if an in-situ melanoma is identified, partial or total vulvectomy should be considered.

The effectiveness of misoprostol or dinoprostone in neonatal outcome after labour induction in post-term nulliparas.

OBJECTIVE: The object of this study was to investigate the efficacy of vaginal administration of misoprostol versus dinoprostone in neonatal outcome. MATERIALS AND METHODS: The first Group A included 77 pregnant women, who requested pregnancy termination one week after labour term and received vaginally misoprostol 50 µg, while the other 69 pregnant women in Group B were vaginally administrated three mg dinoprostone. According to the authors' protocol this procedure was repeated after six hours for a maximum of two times. RESULTS: The labour duration was longer in Group B (p = 0.000), while the APGAR score was better in Group A (p = 0.015). In Group A the labour modus was as follows: 86.9% normal vaginal labour, 3.8% vacuum extraction, and 9.3% cesarean section, while in Group B it was 82.83% normal vaginal labour, 3.07% vacuum extraction, and 14.1% cesarean section. CONCLUSION: Misoprostol has advantages according to neonatal outcome compared to administration of dinoprostone.

The riddle of Ballantyne's syndrome in the aspect of hydrops fetalis.

Ballantyne's syndrome, the combination of maternal generalized edema and fetal ascites, is rare and alarming in gestation. Early diagnosis might be useful in providing proper management of the fetus and aiming at an improved clinical result. The syndrome is an indication that HF is there, it has already started expanding the fetal torso and endangering the child-bearer's life. Despite the detailed investigation, no apparent cause for the emergence of the hydrops was identified.

Diagnostic approach and therapeutic management in early-stage endometrial cancer.

OBJECTIVE: The effectiveness of pelvic and para-aortic lymphadenectomy in the morbidity of patients affected by early-stage endometrial carcinoma (EC) is the subject of this study. STUDY DESIGN: Ninety-two cases with endometrial cancer that underwent para-aortic and pelvic lymphadenectomy, from June 1995 to June 2006, were studied and compared with 30 cases of patients with endometrial cancer without lymphadenectomy. RESULTS: According to the results, systematic pelvic and para-aortic lymphadenectomies improved disease-free and overall survival rates among the patients with endometrial cancer. The mean number of removed para-aortic lymph nodes was 19.01 +/- 5.88, whereas the mean number of removed iliac lymph nodes was 32.94 +/- 6.69. Forty-two and 31 metastatic iliac and para-aortic nodes were found, respectively. No surgery-related deaths and major intraoperative injuries occurred. The frequency and the type of postoperative complications were not affected by the performance of lymphadenectomy. The morbidity rate was 6.2%, similar to the group without lymphadenectomy (5.79%). No recurrence occurred in the group with lymphadenectomy, while in the other group the recurrence rate was 23.3%. CONCLUSIONS: Lymph nodes metastases can be observed in early stages of EC. Pelvic and para-aortic lymphadenectomies seems to provide profound information about the Stage of the disease and the patient's survival, identifying which patients are suitable for supplementary treatment, without significant clinical increase of morbidity.

Isolation of mesenchymal stem cells using the total length of umbilical cord for transplantation purposes.

BACKGROUND: Umbilical cord (UC) mesenchymal cells have the ability to differentiate into various cell types, which make them an easily obtainable source for therapeutic uses. Different approaches have been used to isolate mesenchymal stem cells (MSC). AIM: Here, we report a detailed enzymatic method where large number of cells can be efficiently isolated from the cord matrix and cryopreserved on the same day of arrival at the laboratory. METHODS/MATERIALS: Cells were successfully isolated from 12 samples, with a new procedure that uses the total length of the UC. MSC have been isolated using a detailed enzymatic method with collagenase and hyaluronidase followed by trypsin, without removing the vessels and without mincing the cord. Stem cells were measured with flow cytometry before cryopreservation and post-thaw. Cultured cells were assessed for MSC marker expression and adherence plasticity for three passages. Multilineage differentiation was performed. RESULTS: Nucleated cell yield was calculated at 0·95 × 10(6) /cm. MSC yield was calculated at 0·65 × 10(6) /cm of cord with flow cytometry while the mean length was 31 cm. Cultured cells expressed the mesenchymal markers CD29, CD90, CD105 and CD44. Mesenchymal marker expression remained intact over the three passages and post-thaw. Osteogenic and adipogenic differentiation was evaluated. CONCLUSIONS: Our findings provide a fast and efficient method for mesenchymal cell isolation from Wharton's jelly using the total length of the UC. This method resulted in a large number of cells while the cells retained their mesenchymal character after thawing. This method can be easily applied, along with UC blood, for UC banking.

Addition of adipose-derived stem cells in cord blood cultures stimulates their pluripotent differentiation.

INTRODUCTION: Adipose tissue is recognized as an important source of postnatal mesenchymal stem cells for generative medicine applications. Moreover, cord blood stem cells have been shown to contain pluripotent stem cells called unrestricted somatic stem cells (USSCs). However, this population is rare and cannot be generated from every cord blood sample. In this study, we have presented a new method of co-culture of adipose-derived stem cells (ADPCs) and cord blood stem cells that results in pluripotent differentiation. MATERIALS AND METHODS: ADPCs were obtained from a piece of adipose tissue after treatment with 0.075% collagenase, which was subsequently inactivated with DMEM/10% FBS. The cellular pellet of centrifugation was plated at 5-7 x 10(6) cells/mL in T25 culture flasks in a low-glycose DMEM with 30% FCS. Cord blood stem cells were obtained by centrifugation following double-processing in the presence of 2% HES 200/0.5 and plated at 5-7 x 10(6) cells/mL in the same medium. To investigate the crucial role of ADPCs in pluripotent cord blood differentiation, we added a ADPCS as (1 x 10(4) cells/mL) to the cord blood cultures and analyzed the contribution of ADPCs using a microscope as well as with flow cytometry. RESULTS: After only 3 days, adherent cells (USSC colonies) of fibroblastic morphology were detected in all co-cultured samples, whereas this was observed later or not at all in the non-co-cultured samples. The greater density of colonies in the co-coltured samples was another point. Hematopoietic CD45 cells were no longer detected after the first passage. Pluripotent stem cells were obtained from all co-cultured samples that contained stem cells positive for CD29, CD44, CD49e, CD90, CD105, CD51 Stro, and C-kit antibodies but negative for CD34, CD45, CD133, and glycophorin A. CONCLUSION: Addition of ADPCs was crucial to generate pluripotent-derived stem cells from cord blood samples. This double culture may be a useful tool for a universal allogeneic stem cell source for tissue repair or regeneration.

Cell recovery sufficient for adult transplantation by additional cord blood collection from placenta.

The amount of newborn blood that can be collected from a single cord donor is limited, but a significant amount remains in the placenta. We used a simplified perfusion method to collect this additional blood. Umbilical cord blood from 15 newborns was collected before placental delivery by umbilical vein puncture. After delivery, the placenta was placed on sterile gauze and 63 mL of citrate-phosphate-dextrose-adenine anticoagulant were injected into the umbilical vein that was then clamped near the placenta. The placenta was gently massaged, hung over a sterile vessel, and the umbilical cord cut sterilely near the embryonic surface. Additional blood was collected into the sterile vessel by pressuring a gauze bag around the placenta. We assessed the contribution of this second fraction to the total volume, total nucleated cell (NC), CD34+, hematopoietic progenitors cell, and colony forming unit count and bacterial contamination risk. The total collected volume was 127.3 mL (range 92-170) and the NC content was 1.6+/-0.73x10(9). The mean second fraction contribution from 15 units to the total nucleated and mononuclear cell content was 54+/-9.87% and 54+/-9.52%, respectively. The added percentage of CD34+ and hematopoietic progenitor cells was 54.3+/-10.35% and 46.7+/-11.5%, respectively, while the additional percentages of colony forming-granulocyte macrophage and colony forming-erythroid in the second fraction were 43.2+/-5.5% and 39.8+/-4.3%, respectively, indicating that the cells collected after placental perfusion (second fraction) had similar HPC content and in vitro hematopoietic potential. The method did not increase the risk of bacterial contamination.

A novel high-yield volume-reduction method for the cryopreservation of UC blood units.

BACKGROUND: For the application of umbilical cord blood (UCB) units as hematopoietic grafts, a dose of 3.7 x 10(7) nucleated cells (NC)/kg body weight is required. NC can be lost during volume-reduction processing and during thawing. A novel modification of the double-processing protocol with the aim of minimizing NC loss is described and evaluated. METHODS: One-hundred and fifty UCB were collected. The volume was reduced by a centrifugation step following double-processing in the presence of 2% HES 200/0.5. Pre- and post-processing cell counts and platelet parameters were measured with an automatic counter. The number of viable CD34+ hemopoietic stem cells was measured by flow cytometry. In 25 of the samples, colony-forming units (CFU) were also determined. The same samples were thawed 6 months after cryopreservation and re-evaluated. RESULTS: The volume was reduced to 6 +/- 1.5 mL. The recovery of NC, MNC, CD34+ hemopoietic stem cells, RBC depletion and CFU following double-processing was 93.6 +/- 3.2%, 95.8 +/- 2.2%, 98.4 +/- 1.5%, 96.8 +/- 1.1% and 107.1 +/- 6.1% (for 25 samples), respectively. The post-thaw recoveries of NC, MNC, CD34+ hemopoietic stem cells and CFU (for 25 samples) were 78.6 +/- 5.4%, 90.8 +/- 4.4%, 96.4 +/- 2.5%, 89.1 +/- 4.1%, respectively. No post-thaw cell aggregation was observed. A significant (P<0.05) post-thaw loss of platelets and signs of platelet activation was observed. DISCUSSION: The protocol uses non-expensive equipment and clinically approved materials and results in samples that can be used in patients with a mean weight of 32.7 kg.