Galectin-1 suppression delineates a new strategy to inhibit myeloma-induced angiogenesis and tumoral growth in vivo.

Galectin-1 (Gal-1) is involved in tumoral angiogenesis, hypoxia and metastases. Actually the Gal-1 expression profile in multiple myeloma (MM) patients and its pathophysiological role in MM-induced angiogenesis and tumoral growth are unknown. In this study, we found that Gal-1 expression by MM cells was upregulated in hypoxic conditions and that stable knockdown of hypoxia inducible factor-1α significantly downregulated its expression. Therefore, we performed Gal-1 inhibition using lentivirus transfection of shRNA anti-Gal-1 in human myeloma cell lines (HMCLs), and showed that its suppression modified transcriptional profiles in both hypoxic and normoxic conditions. Interestingly, Gal-1 inhibition in MM cells downregulated proangiogenic genes, including MMP9 and CCL2, and upregulated the antiangiogenic ones SEMA3A and CXCL10. Consistently, Gal-1 suppression in MM cells significantly decreased their proangiogenic properties in vitro. This was confirmed in vivo, in two different mouse models injected with HMCLs transfected with anti-Gal-1 shRNA or the control vector. Gal-1 suppression in both models significantly reduced tumor burden and microvascular density as compared with the control mice. Moreover, Gal-1 suppression induced smaller lytic lesions on X-ray in the intratibial model. Overall, our data indicate that Gal-1 is a new potential therapeutic target in MM blocking angiogenesis.

Nipple sparing mastectomy with immediate silicone implant reconstruction for malignant phyllodes tumor in a 19-year-old girl.

OBJECTIVE: Due to the rarity of MPT, the clinical records in the literature, collected along decades, lack to address a modern approach to breast reconstruction after mastectomy. CASE PRESENTATION: We report a case of a teen-aged female diagnosed to have a malignant phyllodes in her right breast. DISCUSSION: The surgical treatment of choice, taking in account the relation between the volume of the mass and the breast dimension, was considered to be a mastectomy. As the disease didn't involve the skin envelope a nipple-areolar-sparing gland removal allowed an immediate prosthetic reconstruction with a contralateral augmentation for symmetrization, so obtaining a satisfactory aesthetic outcome. CONCLUSIONS: At our knowledge we present for the first time this surgical approach that, in selected patients, can reach the oncologic radicality and an adequate cosmetic result too.

Thiophene-based compounds as fluorescent tags to study mesenchymal stem cell uptake and release of taxanes.

Human mesenchymal stem cells (hMSC) are multipotent cells that display the unique ability to home and engraft in tumor stroma. This remarkable tumor tropic property has generated a great deal of interest in many clinical settings. Recently, we showed that hMSC represent an excellent base for cell-mediated anticancer therapy since they are able to internalize paclitaxel (PTX) and to release it in an amount sufficient to inhibit tumor cell proliferation. In order to shed light on the dynamics of drug uptake and release, in the present paper we describe the synthesis of two novel thiophene-based fluorophore-paclitaxel conjugates, namely PTX-F32 and PTX-F35, as tools for in vitro drug tracking. We aimed to study the ability of these novel derivatives to be efficiently internalized by hMSC and, in a properly engineered coculture assay, to be released in the medium and taken up by tumor cells. In order to ensure better stability of the conjugates toward enzymatic hydrolysis, the selected oligothiophenes were connected to the taxol core at the C7 position through a carbamate linkage between PTX and the diamino linker. Antiproliferative experiments on both tumor cells and stromal cells clearly indicate that, in good correlation with the parent compound, cells are sensitive to nanomolar concentrations of the fluorescent conjugates. Moreover, in the coculture assay we were able to monitor, by fluorescence microscopy, PTX-F32 trafficking from hMSC toward glioblastoma U87 tumor cells. Our work paves the way for novel possibilities to perform extensive and high quality fluorescence-based analysis in order to better understand the cellular mechanisms involved in drug trafficking, such as microvescicle/exosome mediated release, in hMSC vehicle cells.

Analysis of the karyotype of expanded human adipose-derived stem cells for bone reconstruction of the maxillo-facial region.

Mesenchymal stem cells (MSC) and adipose-derived stem cells (ASC) were recently proposed for bone maxillofacial reconstruction in association with biomaterials. For this application MSC must be ex-vivo expanded in order to obtain, for a given volume of implanted biomaterial, a relevant number of bone forming cells. Previously conducted pre-clinical studies suggested that a concentration of 6 x 10(8) ASC associated with 900 mg of anorganic bovine bone (ABB) could be effective for human maxillary sinus floor elevation. A keystone issue to guarantee the quality and safety of Advanced Therapy Medicinal Products containing expanded MSC and ASC is their chromosome stability in culture: this topic has been widely investigated and conflicting results have been published. Abnormal karyotype of human ex-vivo expanded MSC and ASC was found by some authors, while, at the same time, several other studies showed the MSC and ASC karyotype to be normal. It is therefore important that all the results obtained on MSC and ASC karyotype analysis be published. Given this context, the aim of this manuscript, aim of this manuscript is to verify the karyotype stability of ASC in view of their applications in clinical trials. ASC obtained from the adipose tissue of 4 donors were expanded over extended culture time. Based on previous ASC expansions we hypothesized to be able to obtain 6 x 10(8) cells by passage 7. Karyotype analysis of 30 metaphases was planned to be investigated at passage 2, 7, and 15 in all the cultures. No abnormalities were found in the karyotype of two donors at all the passages tested, while a translocation was found in 2 metaphases of a donor at passage 7, but not at passage 15, and in the fourth donor in 5 metaphases a trisomy was found at passage 15. Chromosomal abnormalities were detected only after extended ASC expansion. Whether these anomalies can be related to risk for the patient's safety will have to be demonstrated by in-vivo studies.

Hypoxia-inducible factor (HIF)-1α suppression in myeloma cells blocks tumoral growth in vivo inhibiting angiogenesis and bone destruction.

Hypoxia-inducible transcription factor-1 (HIF-1α) is overexpressed in multiple myeloma (MM) cells within the hypoxic microenvironment. Herein, we explored the effect of persistent HIF-1α inhibition by a lentivirus short hairpin RNA pool on MM cell growth either in vitro or in vivo and on the transcriptional and pro-angiogenic profiles of MM cells. HIF-1α suppression did not have a significant impact on MM cell proliferation and survival in vitro although, increased the antiproliferative effect of lenalidomide. On the other hand, we found that HIF-1α inhibition in MM cells downregulates the pro-angiogenic genes VEGF, IL8, IL10, CCL2, CCL5 and MMP9. Pro-osteoclastogenic cytokines were also inhibited, such as IL-7 and CCL3/MIP-1α. The effect of HIF-1α inhibition was assessed in vivo in nonobese diabetic/severe combined immunodeficiency mice both in a subcutaneous and an intratibial MM model. HIF-1α inhibition caused a dramatic reduction in the weight and volume of the tumor burden in both mouse models. Moreover, a significant reduction of the number of vessels and vascular endothelial growth factors (VEGFs) immunostaining was observed. Finally, in the intratibial experiments, HIF-1α inhibition significantly blocked bone destruction. Overall, our data indicate that HIF-1α suppression in MM cells significantly blocks MM-induced angiogenesis and reduces MM tumor burden and bone destruction in vivo, supporting HIF-1α as a potential therapeutic target in MM.

Increased osteocyte death in multiple myeloma patients: role in myeloma-induced osteoclast formation.

The involvement of osteocytes in multiple myeloma (MM)-induced osteoclast (OCL) formation and bone lesions is still unknown. Osteocytes regulate bone remodelling at least partially, as a result of their cell death triggering OCL recruitment. In this study, we found that the number of viable osteocytes was significantly smaller in MM patients than in healthy controls, and negatively correlated with the number of OCLs. Moreover, the MM patients with bone lesions had a significantly smaller number of viable osteocytes than those without, partly because of increased apoptosis. These findings were further confirmed by ultrastructural in vitro analyses of human preosteocyte cells cocultured with MM cells, which showed that MM cells increased preosteocyte death and apoptosis. A micro-array analysis showed that MM cells affect the transcriptional profiles of preosteocytes by upregulating the production of osteoclastogenic cytokines such as interleukin (IL)-11, and increasing their pro-osteoclastogenic properties. Finally, the osteocyte expression of IL-11 was higher in the MM patients with than in those without bone lesions. Our data suggest that MM patients are characterized by a reduced number of viable osteocytes related to the presence of bone lesions, and that this is involved in MM-induced OCL formation.

Giant malignant phylloides tumor: case report.

The incidence of phylloides breast tumors is less than 1% in the population affected by breast cancers. The age at higher risk is between 35 and 45 years. These neoplasms are characterized by a proliferation of mesenchimal and epithelial cells. We present a rare case of giant malignant phylloides tumor (28 x 21 x 15 cm) with a complet substitution of the gland. The clinical presentation of phylloides tumors is heterogenous; the surgical treatment is a conservative one of the gland if the neoplastic lesion size is less than 5 cm with a free margin of 1 cm and a mastectomy if the diameter of lesion is more than 5 cm. Complementary therapies still remain controversial.

[Neural infiltration in benign gallbladder lesions. Description of a case]. / Infiltrazione neurale nelle lesioni benigne della colecisti. Descrizione di un caso.

Neural infiltration is considered a very useful marker of malignancy in the biliary tract. Aim of this study is to report a case of benign epithelium of the gallbladder with neural infiltration and to review the literature.

Pancreatic-polypeptide cell hyperplasia associated with pancreatic or duodenal gastrinomas.

An immunohistochemical investigation of pancreatic-polypeptide (PP) cells in the PP-rich region of the pancreas, of ventral embryological origin, was performed in three female patients affected by or previously operated on for functioning duodenal or pancreatic gastrinomas not associated with multiple endocrine neoplasia syndrome. A pronounced PP-cell hyperplasia showing histological patterns of endocrine cell dysplasia and focal adenomatosis as defined by Jaffe et al was found in all cases. Morphometric analysis showed that in these patients the fraction of ventral-type pancreatic lobules occupied by PP-immunoreactive cells was 14.77 +/- 5.73%, 8.94 +/- 2.92%, and 10.83 +/- 5.64%, respectively. These values were three to five times higher than the upper values found in controls (mean, 2.20%; range, 1.54 to 2.93%; P < .0001). PP-cell hyperplasia may contribute for the increased circulating levels of PP found in gastrinoma patients. In this regard, elevation of fasting blood PP was found in one of four determinations done in one patient, indicating that PP-cell hyperplasia may be responsible for, at least, transient PP hypersecretion. In one of our patients, PP-cell hyperplasia was found 15 years after normalization of gastrin levels by removal of a single pancreatic gastrinoma. This finding is against a trophic role for hypergastrinemia in the development of PP-cell hyperplasia. In one of two patients in whom the pancreatic regions of dorsal embryological origin (ie, body and tail of the gland) were examined, ventral-type, PP-rich islets were frequently encountered, a finding at variance with their exceptional detection in control cases. This finding suggests that PP cell hyperplasia of the PP-rich pancreatic region may be a feature of a more diffuse disorder of PP cell development in the pancreas of gastrinoma patients.

Mitral valve prolapse in a case of Marfan syndrome with congenital cardiac disease, chronic obstructive pulmonary disease and schizophrenia.

Mitral valve prolapse (MVP) is a common pathological finding in several inherited connective-tissue diseases among which the Marfan syndrome. In the literature several cases of Marfan syndrome characterized by schizophrenia, chronic obstructive pulmonary disease and severe mitral regurgitation due to a MVP have been reported. Our case of MVP was observed in a patient with Marfan syndrome with its characteristic musculoskeletal disorders associated with congenital cardiac disease, severe chronic obstructive pulmonary disease and schizophrenia. According to the latest genetic studies, Marfan syndrome seems to result from genetic mutations in an extracellular matrix glycoprotein, fibrillin. Authors believe that MVP represents only one of several pathological alterations which may be seen in the Marfan syndrome, as expression of a more generalized genetic disorder.

[Ag-NORs, nucleus-nucleolus-associated antigens (Ki67, PCNA, P-105 and p-120): prognostic markers of meningioma recurrence]. / Ag-NORs, antigeni nucleo-nucleolo associati (Ki67, PCNA, P-105 e p-120): markers di prognosi nella recidiva di meningioma.

The predictive significance of some nucleus-nucleolus associated markers, such as Ag-NOR, Ki67, PCNA, p-120, P-105, for the recurrency of meningiomas was investigated. A retrospective analysis was performed on a series of transitional meningiomas and of their recurrencies. Similar meningiomas but with no recidivism were used as controls. All cases were represented by women between the V and the VI decade of age. Besides, having all tumors presented with a cranial convexity localization total removal had been achieved. Recurrencies had taken place after 4.5 and 7.5 years in ten and two women, respectively. In all cases considered, the tumoral histotype did never present with structural elements in general suggestive of recurrence, such as high degree of cellularity, atypia, nuclear polymorphism, necrosis, appreciable mitotic index. On histological seriated 3 microns thick sections the silver staining technique of nucleolar organizer region-associated proteins (NORs) and the Ki67, PCNA, p-120, P-105 immunostaining ABC technique were applied. For each case 1000 tumoral cells were counted, with evaluation of the number of Ag-NOR dots and the percentage of Ki67, PCNA, p-120, P-105 positive cells. From the analysis, a mean value of Ag-NOR dots resulted of 6.44 +/- 0.65 in primitive meningiomas and of 6.53 +/- 0.88 in their respective recurrencies. In the control tumors the mean value of Ag-NOR dots resulted to be 3.53 +/- 0.55. Such difference between tumors that had repeated and controls that had not, was statistically significant (p < 0.001). For what concerns the expression of immunocytochemical relevant markers the percentage of positive cells, in primitive tumors (P), in recurrencies (R) and in the controls (C) was, namely, as follows: Ki67 (P: 14%, R: 12%, C: 6%); PCNA (P: 38%, R: 37%, C: 8%); and p-120 (P: 57%, R: 62%, C: 12%). The expression of P-105 was but occasional and without significance. From the data described, one can conclude that the evaluation of the whole of the markers considered in transitional meningiomas does predict recidivism.

[Stereotactic fine-needle aspiration biopsy (FNAB) of nonpalpable breast lesions: cyto-histological correlations]. / Agoaspirazione (FNAB) mediante centratura stereotassica di lesioni mammarie non palpabili: correlazioni cito-istologiche.

There are not complete consensus if mammographically guided FNABs of nonpalpable breast lesions can replace open biopsy. To elucidate this problem, we have reviewed the efficacy of stereotactic FNAB in a series of 502 cases of nonpalpable lesions observed in a period from 1990 to 1994, with a immediate cytologic control of adequacy of material. In 144 patients of the lesion is achieved with 4% sterile coal localisation and an open biopsy of the area performed. In each case cytologic findings, with a brief description of probable histotype of the lesion, are correlate with surgical pathologic diagnosis, with a comparison between two time period (1990-92; 1993-94). In our study particularly ductal and lobular hyperplasia (with or without atypia) restrict the efficiency of the FNAB. In fact the specificity varies from 83.3% to 90.3%, the sensitivity from 78.7% to 77.8%, positive predictive value from 86.0% to 90.3%, negative predictive value from 75.6% to 70% and diagnostic accuracy from 80.0 to 82.4%, considering ductal and lobular hyperplasias as true negatives or true suspects. Cytologically we have 2 false positive in the first period of time, 3 false suspects in the second period of time and 7 false negatives. However benign/malign biopsy ratio is well (0.97). Moreover histopathologic informations of cytological diagnosis is variable according the histotype of the lesion. In conclusion our study confirm that FNAB is an reliable method that avoid unnecessary biopsy but is insufficiently sensitive to replace open biopsy of nonpalpable breast lesions, also with an immediate cytological control of adequacy of aspirate.

[Our first experience with and considerations on the legal provisions of Law 194/78]. / Prime nostre esperienze e considerazioni sul disposto della legge 194/78.

PIP: During May 1978-November 1979, 2131 voluntary interruptions of pregnancy were carried out at the Obstetrical and Gynecological Clinic of the University of Naples. Most interventions done during the 1st months were done by dilatation and currettage; after that 99.65% were done by vacuum aspiration which seems to be safer within the 1st 12 weeks of pregnancy. General anesthesia was used in only 2.35% of cases and paracervical block in 97.65%. Hospitalization of patients was for the day only; 4.74% required a longer stay in the hospital. 50.87% of patients were in the age group 25-35; 35.48% were parity 1-2, 34.10% parity 3-4, and 13.94% parity 5 and over. Almost 80% of patients were married, and 64.80% of interventions were done between the 8th and 10th week. Only 8 patients were illiterate, and 79.02% had secondary school education; 73.81% were housewives, and 50.77% came from outside the city of Naples. Most partners of patients were employed as factory workers. There were 30 cases of hemorrhage and 9 cases of uterine perforation. All complications occurred in patients beyond the 10th week of pregnancy.^ieng

[Scope of the indications for abortion]. / Ampiezza delle indicazioni dell'aborto

PIP: Legalization of abortion in Italy generates never ending discussions. The problem should have been solved years ago with a national campaign for family planning, with the setting up of well organized family centers, and with contraception available and free to all. If it seems right and proper to perform abortion under certain circumstances, it does not seem proper to take into consideration socioeconomic conditions, and certainly not abortion on request; a new life must not be wasted because a woman does not feel like having a new child. Abortion, on the other hand, is certainly to be considered in case of danger for the mother, in case of fetal abnormalities, or when the pregnancy is result of incest or of rape. Abortion for psychological reasons is very valid if the reasons are real, evident, and have been thoroughly evaluated.^ieng