Production and purification of higher molecular weight chondroitin by metabolically engineered Escherichia coli K4 strains.

The capsular polysaccharide obtained from Escherichia coli K4 is a glycosaminoglycan-like molecule, similar to chondroitin sulphate, that has established applications in the biomedical field. Recent efforts focused on the development of strategies to increase K4 polysaccharide fermentation titers up to technologically attractive levels, but an aspect that has not been investigated so far, is how changes in the molecular machinery that produces this biopolymer affect its molecular weight. In this work, we took advantage of recombinant E. coli K4 strains that overproduce capsular polysaccharide, to study whether the inferred pathway modifications also influenced the size of the produced polymer. Fed-batch fermentations were performed up to the 22 L scale, in potentially industrially applicable conditions, and a purification protocol that allows in particular the recovery of high molecular weight unsulphated chondroitin, was developed next. This approach allowed to determine the molecular weight of the purified polysaccharide, demonstrating that kfoF overexpression increased polymer size up to 133 kDa. Higher polysaccharide titers and size were also correlated to increased concentrations of UDP-GlcA and decreased concentrations of UDP-GalNAc during growth. These results are interesting also in view of novel potential applications of higher molecular weight chondroitin and chondroitin sulphate in the biomedical field.

Vascular endothelial growth factor (VEGF): can we use it as prognostic factor in endometrial cancer?

AIM: The aim of the study was to investigate if VEGF levels reflect the severity of endometrial cancer and the clinic relationship between microvasal density (MVD) and concentration of VEGF in tumor. METHODS: The study was conducted on 22 patients affected by endometrial cancer who were submitted to total abdominal radical hysterectomy plus bilateral salpingo-ophorectomy. VEGF (pg/mL) and MVD values were measured on histologic specimens of endometrial cancer obtained during the surgical treatment. The means and standard deviations of estimated values were calculated and a statistical comparison was effected by student t test for not coupled data. Pearson correlation test was used to analyze the eventual correlation among VEGF and MVD values in overall patients. RESULTS: We have documented that VEGF expression and MVD change according to FIGO stage, lympho-vascular infiltration and lymph node involvement. Pearson correlation test shows a good linear positive correlation in overall patients between VEGF and MVD values. CONCLUSIONS: Results obtained show a possible use of VEGF as prognostic factor in endometrial cancer. Confirmation of these data may permit both to identify high-risk patients, who must be treated with a more aggressive treatment, and to use an angiogenic therapy in endometrial cancer.

Double laparotomy wound recurrence of endometrial carcinoma.

INTRODUCTION: Abdominal scar recurrence of endometrial carcinoma after abdominal total hysterectomy is very rare. We report a case of a 65-year-old woman who had two recurrences in the abdominal incisional scar after total hysterectomy. CASE REPORT: A 65-year-old woman underwent total hysterectomy with bilateral salpingo-oophorectomy and pelvic lymphadenectomy because of well-differentiated endometrial adenocarcinoma (Stage IIB). Thus, the patient was treated by external beam radiotherapy. She developed two recurrences in the abdominal incisional scar two and three years after total hysterectomy, respectively. Surgery plus chemotherapy and surgery plus hormonal therapy were used for treatment of the first and second scar recurrence, respectively. CONCLUSIONS: It is a very intriguing and controversial biologic question how neoplastic cells can implant and grow in an abdominal scar without other concomitant metastases. We report a review of the literature and the possible mechanism of recurrences in laparotomy wounds.

Second-line with paclitaxel and carboplatin for recurrent disease following first paclitaxel and platinum compounds in ovarian carcinoma.

OBJECTIVE: The combination of paclitaxel and platinum compounds is considered the best first-line regimen for advanced ovarian carcinoma. The purpose of this study was to evaluate a paclitaxel and carboplatin combination in pretreated patients who recurred within 24 months after a complete clinical response with the same regimen used as first-line chemotherapy. METHODS: 18 patients were included in this study. Second-line chemotherapy consisted of paclitaxel, 175 mg/m2 as a 3-hour infusion, and carboplatin AUC 6 every 21 days. RESULTS: Among 15 evaluable patients, eight (53%) complete and five (34%) partial responses were observed, while two (13%) patients had stable disease (SD). The response rate was 67% among patients with measurable disease and 52% for evaluable disease. The median progression-free interval after second-line chemotherapy was 8.3 months. The median progression-free interval for patients with measurable disease was 8.6 months and for evaluable disease it was 7.9 months. Seven (46%) of 15 patients have developed recurrence after second-line chemotherapy with paclitaxel and carboplatin with a median time to recurrence of 9.8 months. CONCLUSION: Paclitaxel 175 mg/m2 and carboplatin AUC 6 as second-line chemotherapy in this sensitive population is effective in terms of response rate and progression-free interval.

Paclitaxel and carboplatin as outpatient therapy for stage III and IV epithelial ovarian cancer.

BACKGROUND: To determine the toxicity and the response rate of a three-hour paclitaxel infusion and carboplatin administered as outpatient treatment for stage III and IV epithelial ovarian cancer. METHODS: Forty-three patients with stage III/IV epithelial ovarian cancer underwent cytoreductive surgery and then received paclitaxel 175 mg/m2 over 3-hr infusion and carboplatin AUC5 every 21 days for six cycles. Elegible patients had adequate bone marrow, renal and hepatic function; G-CSF was recommended if white cell count fell under 3,000/mm3. RESULTS: No patients had hypersensivity reactions; 15 out of 43 patients (35%) required colony-stimulating factors, 39 patients (91%) had general alopecia, three patients (7%) had severe emesis, 20 patients (46%) had mild emesis, four patients (9%) had severe myalgias, eight patients (18%) had moderate myalgias, one patient (2%) had grade 3 neurotoxicity. Three patients experienced grade 3 thrombocytopenia (7%). At a median follow-up of 29 months, 32 of 43 patients are alive (74%). Median progression-free survival is 14 months. Median overall survival has not been reached. CONCLUSIONS: Three-hour infusion paclitaxel and carboplatin is an effective and safe outpatient therapy for epithelial ovarian cancer.

[Role of intestinal resection in primary cytoreduction of ovarian cancer]. / Il ruolo della resezione intestinale nella citoriduzione primaria del cancro dell'ovaio.

BACKGROUND: The aim of this study is to define the role of the intestinal removal for the therapy of ovarian cancer in advanced stadium. METHODS: We have examined 247 females with epithelial ovarian cancer in advanced stadium, that had intestinal removal. RESULTS: The survival in the females that had a very good intestinal removal is greatest than in the females that hadn't a very good intestinal removal.

[Cytoreductive surgery in patients with stage IV ovarian carcinoma]. / La chirurgia citoriduttiva nelle pazienti con carcinoma dell'ovaio allo stadio IV.

BACKGROUND: The aim of this study is to evaluate the efficacy of cytoriductive surgical in females with ovarian carcinoma in advanced stadium, and to define the role of this surgical for the survival of females with hepatic metastases. METHODS: This is a retrospective study. 164 females with ovarian cancer in IV stadium was examined. 64 females had hepatic metastasis. All patients had cytoriductive surgical. RESULTS: The survival in the patients without hepatic metastasis was 38 months if the cytoreduction was very good; it was 18.3 months if there is residual disease. The survival in the patients with hepatic metastasis was 50.1 months if the cytoreduction was very good; it was 27 months if there is residual disease. CONCLUSIONS: A very good surgical is very important for the survival of patients with ovarian cancer in advanced stadium. This is true also in the patients with hepatic metastases.

[Meigs' syndrome and "Meigs' pseudo-syndrome." Report of 2 cases]. / Sindrome di Meigs e "pseudo-sindrome di Meigs". Descrizione di due casi.

BACKGROUND: The aim of this study is to investigate similitudis and differences between the Meigs' syndrome and Meigs' pseudosyndrom. The Meigs' syndrome is an uncommon disease that is characterized by benign ovarian tumor, ascites and pleural effusion. The Meigs' pseudosyndrom is a serious disease that is characterized by malignant ovarian tumor, ascites, pleural effusion. METHODS: We have examined two cases: a case of Meigs' syndrome that is characterized by vomit, abdominal pain, ascites, height serum Ca 125 level; a case of Meigs' pseudosyndrom that is characterized by ovarian adenocarcinoma that is diagnosticated owing to ascites and pleural effusion. CONCLUSIONS: This study suggest that the surgical therapy have a very important role for the complete remission of the disease in the Meigs' syndrome and for the remission of ascites and pleural effusion in the Meigs' pseudosyndrom.

[HPV infection. Clinical features and treatment]. / Infezione da HPV. Aspetti clinici e trattamento.

The aspecific and exiguous symptoms and the lacking information are among the reasons of the diffusion of the vulvo-vaginal papillomatosis. We carried out the present study between 1995 and 1999 in the outpatient clinic of cervico-vaginal pathology of the Second University of Naples. 680 patients (aged between 18 and 56 years) underwent vulvoscopic and colposcopic examination. The did not show any relevant symptoms specific for HPV infection. Among the viral strains, HPV-16 and HPV 18 are able to induce a cervical cancer. To eliminate the pathology the primary prevention is necessary: it consist of both an adequate information about the micro-condilomatosis, the role of the activator agent, the modality of the infection, and the annual screening examinations such as pap-test and colposcopy. As first line treatment during secondary prevention, we utilize Roferon A, and perform diathermocoagulation according with the local diffusion and the degree of the disease (mild, moderate, severe). At the end of the therapy with Roferon A we observed that the infective focus was eliminated in about 60% of the cases and, only for moderate and severe micro-condilomatosis a diathermocoagulation was necessary.

[Second-look in ovarian cancer: laparoscopy or laparotomy?]. / Second-look per cancro dell'ovaio: laparoscopia o laparotomia?

BACKGROUND: The aim of the present study was to compare the laparoscopic second-look with laparotomic second-look as regards the consistency of diagnosis of residual tumoral disease after first step treatment in patients affected by ovarian cancer, and to evaluate the feasibility of the laparoscopic second-look. METHODS: Twenty-one patients affected by ovarian cancer underwent laparoscopic second-look followed by laparotomic second-look. Six months after the first surgical intervention all the patients showed no contraindications to laparoscopic second-look. All the surgeries were performed with the same procedure: after the introduction of the trocars the lysis of adherences was carried out, the whole abdominal cavity was explored, 18 abdominal-pelvic sites were examined, direct biopsies were performed and samples for the cyto- and histological analysis were obtained. RESULTS: Positive predictive value for laparoscopy was 100% (6 out of 6 cases), while negative predictive value was 84% (2 false negative cases out of 12). The complete abdominal-pelvic examination was possible in 95% of cases with laparotomy while in 41% of cases with laparoscopy, because of post-operative severe adherences. CONCLUSIONS: Laparoscopic second-look has a good consistency as regards the diagnosis of residual tumoral disease, but its feasibility is lower than laparotomy owing to the presence of severe adherences and the high risk of intra- and post-operative compliances.

Fetuses with cystic hygroma. A retrospective study.

BACKGROUND: The aim of this study is to analyse several theories of pathogenesis of cystic hygroma, its correlation with chromosomal abnormalities and the indicators of poor or good prognosis. EXPERIMENTAL DESIGN: This is a retrospective study that evaluates all cases of cystic hygroma seen during the four-year period from January 1994 to December 1997. SETTING: This study was performed in the center of prenatal diagnosis of institutional hospital in Caserta. PATIENTS: All cases of fetuses with cystic hygroma were examined in 2100 pregnant female who visited the ambulatory. INTERVENTIONS: The modality of diagnosis of this pathology and the presence of abnormal maternal serum levels of alpha-fetoprotein, human chorionic gonadotropin and unconjugated estriol were evaluated. Moreover, the presence of karyotype abnormalities or other non chromosomal abnormalities were also evaluated. Prognostic indicators such as the presence of septae seen by sonography were examined. RESULTS: Nine fetuses with cystic hygroma were diagnosed sonographically. Septae were identified in six cases. Chromosomal abnormalities were found in five cases. Two cases presented Turner's syndrome and one case Downs' syndrome. There were two cases with associated anomalies. The amniotic fluid alpha-fetoprotein (AFP) levels were high in all cases. CONCLUSIONS: Cystic hygroma is a malformation of the lymphatic system that is diagnosed by ultrasound very well from the first quarter of pregnancy. It is frequently associated with chromosomal and non chromosomal abnormalities. The presence of septae in it and amniotic fluid AFP levels are prognostic indicators.

[Obstetric conduct in IUGR]. / La condotta ostetrica nello IUGR.

Purpose of the study is to identify the correct attitude that the obstetrician must engage in the management of pregnancy and birth in case of IUGR. Different methods of diagnosis and therapy of IUGR and the formalities of assistance to the birth have been examined and compared. Accurate clinical examinations of the mother, the study of fetal kariotype and ultrasonography, are essential for the diagnosis of IUGR. The genetic study could be performed by collecting chorionic villi, amniocentesis, cordocentesis or placenta biopsy. Ultrasonography identifies the cases of IUGR, and distinguishes early IUGR from late IUGR. Color Doppler identifies the pathology of the flow in the umbilical artery, in the abdominal aorta and in the middle cerebral artery. After the 26th week, the follow-up of the fetus with IUGR is done with cardiotocography with or without acoustic stimulation or oxytocin. The amelioration of maternal conditions is obtained by avoiding the cigarette smoking, preferring to rest in bed and a balanced feeding; the hyperoxygenation doesn't find unanimous consent. The treatment off IUGR can consist of abdominal decompression, intra-abdominal infusion of amniotic liquid, or use of aspirin. The birth is carried out in the hospital, when the fetus has reached a sufficient maturity. The management of IUGR requires an accurate follow-up and an adequate antepartum therapy. The goal is a birth with less risk.

Influence of menstrual factors and dietary habits on menstrual pain in adolescence age.

OBJECTIVES: The aim of this study was to determine the frequency of the primary dysmenorrhea in adolescence age and investigate correlation between menstrual factors, dietary habits and this pathology. STUDY DESIGN: The sample was constituted from 356 students that were subjected to questionnaire, abdominal ultrasound, and in some cases, hormonal dosing. RESULTS: The frequency of the primary dysmenorrhea was 85%. Early menarche was related to an increase of its prevalence and its severity. A long and heavy menstrual flow was related to an increase of its severity. As far as dietary habits, it was noted that a higher consumption of fish, eggs, fruit and a lower consumption of wine is correlated with a lower frequency. CONCLUSION: Primary dysmenorrhea is very common in young women. The risk factors for this pathology are early menarche, long and heavy menstrual flow, and lower consumption of fish, eggs, and fruit.

[Climacteric and quality of life]. / Climaterio e qualità della vita.

BACKGROUND: Menopause is for women a critical period of their life, in which organic troubles and psychological problems cherish themselves. With particular reference to hot flushes and impaired sleep (the frequently found troubles), the benefits of physical exercise in women of this age have been evaluated. METHODS: A study based on a questionnaire has been carried out on 79 women of age between 51 and 58 (who practised regular physical activity, selected at sporting centers), and 481 women of age between 52 and 58, selected between patients who came to the University Hospital for reasons not ascribed to menopause. All the women selected in the study underwent to natural menopause, and didn't use hormonal therapy. The questionnaire evaluated symptomatology and its level of severity. RESULTS: In the first group 57% of the women didn't report symptoms, 30% light symptoms, 12% medium, 1% severe. In the second group 52% didn't report symptoms, 15% light symptoms, 22% medium, 11% severe. CONCLUSIONS: Besides verifying a bigger well-being among women who practice physical activity, the interesting result is that 52% of the second group didn't refer symptomatology. The advantages of a balanced diet, with particular attention to food with phytoestrogens, is discussed and the psychosocial problems of woman in menopause, covering a fundamental role to explain a great part of the symptomatology, are underlined. Finally, the guide lines for a right preparation to menopause are presented.

[Heparin-like factors in pregnancy. Report of a clinical case]. / Fattori heparin-like e gravidanza. Descrizione di un caso clinico.

Some authors have described the presence of circulating heparin-like anticoagulants; these substances are frequently associated with neoplastic pathology. Personal experience proves that this pathology of coagulation may be present also in pregnancy. The clinical case of a primipara, nullipara pregnant woman twenty-eight years aged is described; the case history showed that the patient presented circulating heparin-like substances. During this study, routine biochemical examinations were within normal limits as well as the coagulation tests. In the course of this experience, coagulation anomalies and hemorrhagic episodes in the intraoperative and postoperative period were not observed.

[Evaluation of bone loss in patient treated with Gn-Rh analogs]. / Valutazione della perdita di massa ossea in pazienti in trattamento con analoghi del Gn-RH.

BACKGROUND: The increasing use of analogs of Gn-RH during treatment of some benign gynaecological diseases, has induced the authors to investigate the principal collateral effects, fixing one's attention on the loss of bony mass. METHODS: This perspective research has considered 38 patients selected for two diseases "endometriosis" and "uterine fibromyomatosis". The therapy has been effected with triptorelin intramuscularly in a dose of 3.75 mg every 28 days for six months, in all six phials. RESULTS: After a half-yearly cycle of therapy, the loss of bony mass was valued about 3% medium. CONCLUSIONS: In the light of other studies too, it was decided to confirm the necessity of associating other medicines able to prevent the side effects caused by their analogs of Gn-RH.