Fetal and perinatal stem cells in cardiac regeneration: Moving forward to the paracrine era.

Cardiovascular disease (CD) is a major burden for Western society. Regenerative medicine has provided encouraging results, yet it has not addressed the focal defects causing CD and mainly related to the inefficient repair programme of the heart. In this scenario, stem cells have been broadly investigated and their paracrine effect proposed as a possible working strategy to boost endogenous mechanisms of repair and regeneration from within the cardiac tissue. The scientific community is now focusing on identifying the most effective stem cell secretome, as the whole of bioactive factors and extracellular vesicles secreted by stem cells and endowed with regenerative potential. Indeed, the adult stem cell-paracrine potential for cardiac regeneration have been widely analyzed with positive outcome. Nevertheless, low yield, invasive sampling and controversial self-renewal may limit adult stem cell application. On the contrary, fetal and perinatal stem cells, which can be easily isolated from leftover sample via prenatal screening during gestation or as clinical waste material after birth, can offer an ideal alternative. These broadly multipotent immature progenitors share features with both adult and embryonic stem cells, show high self-renewal, but they are not tumorigenic neither cause any ethical concern. While fetal and perinatal stem cells demonstrated to improve cardiac function when injected in the injured heart, the comprehensive characterization of their secretome for future applications is still at its infancy. In this review, we will discuss the paracrine potential of the fetal and perinatal stem cell secretome to provide cardiac repair and resurge the dormant mechanisms of cardiac regeneration for future therapy.

Sexual behaviour among Italian adolescents: knowledge and use of contraceptives.

BACKGROUND: Comprehensive data concerning the sexual behaviour of Italian adolescents are lacking; the planning of prevention programmes is therefore difficult. METHODS: The current study evaluates the knowledge of Italian adolescents about sexuality and their use of methods to prevent pregnancy and sexually transmitted infections. The age at first intercourse in the current sample was compared to that found in past samples to ascertain whether sex education was needed at an earlier age. The efficacy of one school-based sex education programme was analysed. RESULTS: Overall 630 students in three Italian cities were surveyed. Just over half the students were sexually active. Among these, the mean age at first intercourse was 15.6 years (SD +/- 1.3). The contraception most widely used was the condom although 'coitus interruptus', 'natural family planning' and 'no method' were also mentioned. The knowledge among students from Latina, after they had received sex education, was significantly better. CONCLUSION: Our data show a lower mean age at first intercourse than has been reported for earlier periods by other authors. School-based sexual education is effective in improving knowledge. Sexual health services for young people must be available and counselling services improved.

Heterogeneous nuclear ribonucleoprotein K: altered pattern of expression associated with diagnosis and prognosis of prostate cancer.

Using proteomic analysis of the nuclear matrix (NM), we found that heterogeneous nuclear ribonucleoprotein K (hnRNP K), a member of the hnRNP family with pleiotropic functions, was differentially expressed in prostate cancer (PCa) tissues. This study aimed to characterise the expression of hnRNP K and its subcellular localisation in PCa, utilising immunohistochemical and quantitative western blot techniques. Furthermore, the hnRNP K expression was studied in human PCa cell lines in order to determine its modulation by bicalutamide, the anti-androgen widely used in PCa therapy. Immunohistochemical staining of paraffin-embedded tissues showed that hnRNP K was overexpressed in PCa, where it was localised both in the cytoplasm and in the nucleus. Staining of non-tumour tissues showed exclusively nuclear localisation and a less intense or absent signal. Immunoblot analysis demonstrated that the hnRNP K level within the NM was higher in PCa compared with non-tumour tissues and closely correlated with Gleason score (P=0.008). Higher expression within the NM was significantly (P=0.032) associated with poor prognosis. In two-dimensional western blot analysis hnRNP K presented several isoforms; the one with pI 5.1 was the most differently expressed between non-tumour and PCa tissues. Preliminary results indicate that hnRNP K can be modulated in vitro by a non-steroidal anti-androgen. Taken together, our findings suggest that hnRNP K has potential implications at the diagnostic, prognostic and therapeutic levels in PCa.

Malignant melanoma metastasizing to the uterus in a patient with atypical postmenopause metrorrhagia. A case report.

UNLABELLED: The uterine metastases of melanoma are very rare. At the present time, only one case occurred in our department. CASE REPORT: a 54-year-old plurigravid woman showed a metrorrhagia of unknown origin. The patient underwent a diagnostic hysteroscopy and an endometrial biopsy, in order to investigate the symptomatic postmenopausal bleeding and exclude a neoplasia, such as the endometrial carcinoma. The patient was discharged with a diagnosis of uterine fibromatosis and called back to go through a complete laparoscopic hysterectomy and bilateral adnexectomy. During the operation, some metastases were found in the genital tract. An accurate physical examination allowed us to discover a cutaneous nevus, the excision and histology of which revealed its malignancy. The immunohistochemistry of the surgical sample was able to confirm the hypothesized relationship between the nevus and the metastases, thus leading to the diagnosis of malignant melanoma metastases, genital tract. It is important to make an accurate diagnostic passage to exclude tumoral pathology in patients with atypical uterine bleeding. Every uterine bleeding of the postmenopausal period (abnormal uterine bleeding, AUB) is considered atypical and it has to be early investigated, in order to exclude any endometrial cancer. The nature of the uterine bleedings can be ascribed to atrophy, dysfunctional matters (dysfunctional uterine bleeding, DBU), benign organic alterations, only in 7-10% of cases to endometrial cancer and more rarely to metastatic tumours, as well as this case of melanoma. Physicians should be aware of such unusual possibilities in order to look carefully for metastatic implants in adenomyomas.

Sequence specific peptidomimetic molecules inhibitors of a protein-protein interaction at the helix 1 level of c-Myc.

Our work is focused in the broad area of strategies and efforts to inhibit protein-protein interactions. The possible strategies in this field are definitely much more varied than in the case of ATP-pocket inhibitors. In our previous work (10), we reported that a retro-inverso (RI) form of Helix1 (H1) of c-Myc, linked to an RI-internalization sequence arising from the third alpha-helix of Antennapedia (Int) was endowed with an antiproliferative and proapoptotic activity toward the cancer cell lines MCF-7 and HCT-116. The activity apparently was dependent upon the presence of the Myc motif. In this work, by ala-scan mapping of the H1 portion of our molecules with D-aa, we found two amino acids necessary for antiproliferative activity: D-Lys in 4 and D-Arg in 5 (numbers refer to L-forms). In the natural hetero-dimer, these two side chains project to the outside of the four alpha-helix bundle. Moreover, we were able to obtain three peptides more active than the original lead. They strongly reduced cell proliferation and survival (RI-Int-VV-H1-E2A,S6A,F8A; RI-Int-VV-H1-S6A,F8A,R11A; RI-Int-VV-H1-S6A,F8A,Q13A): after 8 days at 10 muM total cell number was approximately 1% of the number of cells initially seeded. In these more potent molecules, the ablated side chains project to the inside in the corresponding natural four alpha-helix bundle. In the present work, we also investigated the behavior of our molecules at the biochemical level. Using both a circular dichroism (CD) and a fluorescence anisotropy approach, we noted that side chains projecting at the interior of the four alpha-helix bundle are needed for inducing the partial unfolding of Myc-H2, without an opening of the leucine zipper. Side chains projecting at the outside are not required for this biochemical effect. However, antiproliferative activity had the opposite requirements: side chains projecting at the outside of the bundle were essential, and, on the contrary, ablation of one side chain at a time projecting at the inside increased rather than decreased biological activity. We conclude that our active molecules probably interfere at the level of a protein-protein interaction between Myc-Max and a third protein of the transcription complex. Finally, CD and nuclear magnetic resonance (NMR) data, plus dynamic simulations, suggest a prevalent random coil conformation of the H1 portion of our molecules, at least in diluted solutions. The introduction of a kink (substitution with proline in positions 5 or 7) led to an important reduction of biological activity. We have also synthesized a longer peptido-mimetic molecule (RI-Int-H1-S6A,F8A-loop-H2) with the intent of obtaining a wider zone of interaction and a stronger interference at the level of the higher-order structure (enhanceosome). RI-Int-H1-S6A,F8A-loop-H2 was less active rather than more active in respect to RI-Int-VV-H1-S6A,F8A, apparently because it has a clear bent to form a beta-sheet (CD and NMR data).

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.

Women with a pelvic mass: indicators of malignancy.

OBJECTIVE: To evaluate the efficacy of five methods: pelvic examination (PE), transvaginal ultrasonography (US), Doppler ultrasonography, serum CA125 assay and serum CA72-4 assay, alone or associated, to predict malignancy in patients presenting a pelvic mass originating in the ovary. METHODS: 92 patients underwent a standard protocol for physical examination, CA125, CA72-4, transvaginal ultrasonography and Doppler ultrasonography. RESULTS: Eighteen women were dropped from the study because they had clearly benign masses; two women were dropped from the study because they had clearly malignant lesions. Twenty-two malignant (30%) and 50 benign (70%) pelvic tumors were found. When one method was considered alone the best sensitivity (SENS) was found in physical examination (90%) and the best specificity (SPEC) was found in CA72-4: 88%. If all indicators were positive, the SPEC was 100% but the SENS was 40%. Logistic regression analysis prediction of the character of the pelvic masses was correct in 86%. CONCLUSION: Some additional information to discriminate between malignant and benign pelvic masses can be obtained from the valuation of serum tumor markers, particularly CA72-4. Also Doppler ultrasonography appeared to be useful in the differential diagnosis of pelvic tumors. The prediction of the character of the pelvic masses calculated by a logistic model in which PE, US, CA 125, and CA72-4 are included is very good.

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.

Changes in the expression of cytokeratins and nuclear matrix proteins are correlated with the level of differentiation in human prostate cancer.

The nuclear matrix-intermediate filament complex (NM-IF) is a protein scaffold which spans the whole cell, and several lines of evidence suggest that this structural frame represents also a functional unit, which could be involved in the epigenetic control of cancer development. Here we report the characterization by high resolution two-dimensional gel electrophoresis and Western blot analysis of the NM-IF complex isolated from prostate cancer (PCa); tumor-associated proteins were identified by comparing the electrophoretic patterns with those of normal human prostate (NHP). Extensive changes in the expression of both the NM and IF proteins occur; they are, however, related in a different way to tumor progression. Poorly differentiated PCa (Gleason score 8-9) shows a strong down regulation of several constitutive cytokeratins (CKs 8, 18, and 19); their expression significantly (P < 0.05) decreases with respect to both NHP and benign prostatic hyperplasia (BPH) and, more interestingly, also with respect to moderately (Gleason score 6-7) and well (Gleason score 4-5) differentiated tumors. Moreover, we have identified a tumor-associated species which is present in all of the tumors examined, systematically absent in NHP and occurs only in a few samples of BPH; this polypeptide, of M(r) 48,000 and pI 6.0, represent a proteolytic fragment of CK8. At variance with these continuing alterations in the expression, the NM proteins undergo stepwise changes correlating with the level of differentiation. The development of less differentiated tumors is characterized by the appearance of several new proteins and by the decrease in the expression of others. Six proteins were found to be expressed with a frequency equal to one in poorly differentiated tumor, namely in all the samples of tumor examined, while in moderately and well differentiated tumors the frequency is less than one, and decreases with increasing the level of differentiation. When tumors of increasing Gleason score are compared with NHP a dramatic increase in the complexity of the protein patterns is observed, indicating that tumor dedifferentiation results in a considerable increase in the phenotypic diversity. These results suggest that tumor progression can be characterized using an appropriate subset of tumor-associated NM proteins.

[Changes of the pulsatile index in ovarian and uterine arteries of infertile women. Case-control study]. / Variazioni dell'indice di pulsatilità nelle arterie ovariche e uterine in donne infertili. Studio, caso, controllo.

BACKGROUND: This study was performed to evaluate the frequency of female pathology that cause infertility and to appraise the difference of pulsatility index in the ovarian and uterine arteries in infertile women and in a control group of healthy women. METHODS: In the period between January 1997 and March 1999 52 infertile couples were examined carrying out an accuracy anamnesis and objective examination. The dosing of FSH, LH, estrogens, progesterone, and an ultrasound examination for each woman was required. The pulsatility index in the ovarian and uterine vases was calculated with the color Doppler technique. RESULTS: A female pathology caused infertility in 24 couples, where only in nine couples the cause was masculine. In 14 couples there was a pathology in both the partners. In 5 couples any pathology was identified. The median of pulsatility index in healthy women was 1.8. Variations were observed in all the women with ovarian or uterine pathology unless in those with uterine malformations. An increase of the pulsatility index was observed also in 3 couples in which any cause of infertility was identified. CONCLUSIONS: Female pathology is a very common cause of infertility. The evaluation of the pulsatility index in ovarian and uterine arteries can be useful diagnosing the cause of infertility and in to explain an infertility due to unknown cause.

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.

[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.

Chromatin condensation is confined to the loop and involves an all-or-none structural change.

Using differential scanning calorimetry in combination with pulsed field gel electrophoresis, we relate here the changes in the thermal profile of rat liver nuclei induced by very mild digestion of chromatin by endogenous nuclease with the chain length distribution of the DNA fragments. The enthalpy of the endotherm at 106 degrees C, which reflects the denaturation of the heterochromatic domains, decreases dramatically after the induction of a very small number of double-strand breaks per chromosome; the thermal transition disappears when the loops have undergone on average one DNA chain scission event. Quantitative analysis of the experimental data shows that the loop behaves like a topologically isolated domain. Also discussed is the process of heterochromatin formation, which occurs according to an all-or-none mechanism. In the presence of spermine, a strong condensation agent, only the loops that have undergone one break are able to refold, in confirmation of the extremely cooperative nature of the transition. Furthermore, our results suggest a relationship between the states that give rise to the endotherms at 90 degrees C and 106 degrees C and the morphologies referred to as class II and class III in a previous physicochemical study of the folding of chromatin fragments (Widom, 1986. J. Mol. Biol. 190:411-424) and support the view that the overall process of condensation follows a sequential (two-step) pathway.

[Macrophage-colony stimulating factor and ovarian cancer]. / Macrophage-colony stimulating factor e carcinoma ovarico.

BACKGROUND: In this study the use of macrophage-colony stimulating factor (M-CSF) as tumor marker for ovarian cancer is evaluated. METHODS: Serum samples were obtained from 74 patients, 43 of these were affected by ovarian carcinoma and 31 by benign ovarian tumors. The M-CSF levels were assayed with an ELISA method and compared with those of 148 healthy women. CA 125 levels were also evaluated. RESULTS: In healthy women the M-CSF levels were 770.4 +/- 145.9 U/ml, the upper limit of normal level was considered 1056 U/ml. Serum M-CSF levels were significantly high in patients with ovarian cancer (1425.3 +/- 1007.1 U/ml; p < 0.001) and in 29 of the 43 patients exceeded the limit of 1056 U/ml. No differences were observed among the histologic types. There were no significant differences between patients with benign ovarian pathology and healthy women. No definite relationship was found with CA 125, but evaluating at the same time M-CSF and CA 125 positive results were found in 95.3% of cases. CONCLUSIONS: Therefore M-CSF can be considered a marker for ovarian cancer, and the assay of its serum levels can be particularly useful in association with those of CA 125.

[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.