A Nicolini

Needle aspiration techniques in preoperative selection of patients with thyroid nodules: a long-term study.

PURPOSEnLong-term evaluation of the combination of two needle aspiration techniques (NAT) (fine-needle aspiration [FNA] and aspiration needle biopsy [ANB]) in performing an efficient preoperative selection of palpable thyroid nodules.nnnPATIENTS AND METHODSnEight years of extensive use of surgery for the detection of thyroid cancer was compared with 12 years of preoperative selection of by NAT.nnnRESULTSnA total of 1,140 operations were performed from 1972 to 1979, and 35 malignant nodules were discovered (3.1%). Five thousand four hundred three patients were examined by NAT from 1980 to 1992; 483 (9%) underwent surgery and 158 malignant nodules were excised. The number of malignant nodules identified by NAT was 166 (eight were not excised) (3.1% of the total population examined). The principal clinical and pathologic features were similar in both groups. ANB yielded a definite benign diagnosis in 88 patients with inadequate FNA findings, it correctly identified four malignant nodules diagnosed as benign by FNA, it showed a macrofollicular component in 115 nodules diagnosed by FNA as microfollicular nodules, and it significantly changed the predictive value of 79 suspicions FNA diagnoses.nnnCONCLUSIONnIntroduction of NAT reduced the number of operations for palpable thyroid nodules from 143 to 40 per year and increased from four to 13 the number of malignant nodules excised without any change in the overall incidence of malignant nodules. The combination of ANB to FNA significantly contributed to the high and efficient preoperative patient selection, principally by reducing the number of indeterminate or suspicious, as well as false-negative, preoperative FNA diagnoses.

Biomolecular markers of breast cancer.

Here, the structure, function, biological and pathological significance and clinical utility of the principal biomolecular markers of breast cancer is reviewed. Each marker was scored for clinical utility using a recently developed tumor marker utility grading system (TMUGS). Among the tissue markers, ERs and PRs are important prognostic/predictive factors and the only tissue markers routinely determined. ER cross-talks with other growth factors while co-regulatory factors enhance (co-activators) or decrease (co-repressors) its transcriptional activity. C-erbB-2 and Ki67/MIB-1 select for adjuvant chemotherapy a subgroup of lymph-node negative patients at a high risk of relapse. Monoclonal antibodies (trastuzumab, gefitinib, erlotinib and bevacizumab) targeting tissue markers and involved in tumor growth and metastasization (EGFR, C-erbB-2, VEGF) have been developed; they showed therapeutical single agent activity as well as potent synergy with chemotherapy agents in metastatic cancer. Among circulating markers, some are potentially useful in the early detection and monitoring of metastatic disease; nevertheless, none is routinely recommended. To suspect distant metastases, CEA-TPA-CA15.3 panel attained accuracy of about 90%. ECD HER2-neu, p53 and nucleophosmin antibodies seem suitable candidates for different associations. Preliminary observations suggest that an early detection with tumor markers and successive treatment of relapses significantly prolongs disease-free and overall survival in selected patients. In conclusion, biomolecular markers are improving understanding of biology and management of breast cancer.

Effect of L-lysine and L-arginine on primary osteoblast cultures from normal and osteopenic rats

A therapeutic role of amino acids L-lysine (Lys) and L-arginine (Arg) in osteoporosis and fracture healing was demonstrated previously by in vivo studies. In the present study, primary cultures of osteoblasts were used to investigate the effect of amino acids on gene expression (alkaline phosphatase activity, ALP; osteocalcin, OC; type I collagen), nitric oxide production (NO) and proliferation (MTT) of cells. Cells were isolated from the distal femurs of normal and osteopenic rats. Normal and osteopenic bone-derived cells were divided into four groups: control, Lys (0.587 mg/mL/d), Arg (0.625 mg/mL/d), and Lys + Arg (0.587 + 0.625 mg/mL/d). No evidence of differences between normal and osteopenic bone-derived cultures in basal conditions was observed. A significant (P = 0.002) increase of 10.4% in NO production was observed in normal bone-derived osteoblasts treated with Lys + Arg when compared to the control group at 7 days. At the same time, normal bone-derived osteoblasts treated with Arg and Lys + Arg showed significant increases in type I collagen synthesis of 25.3% and 28.4%, respectively, when compared to the control group. Osteopenic bone-derived osteoblasts showed significant (P = 0.002) increases of 27.6% in MTT and 28.7% in cell count at 48 hours when treated with Lys + Arg in comparison with the control group. At 7 days, NO production and type I collagen synthesis increased significantly (P< 0.005) both in osteopenic bone-derived osteoblasts treated with Arg (NO: 18.5%; type I collagen: 34.4%) and Lys + Arg (NO: 23.7%; type I collagen: 20.9%) compared to the control group. Finally, a significant (P = 0.025) decrease of 5.8% in OC level was observed in osteopenic bone-derived osteoblasts treated with Arg. Results suggest that the potential therapeutic effect of Lys and Arg on bone could be related, at least in part, to an improvement of NO production and type I collagen synthesis by osteoblasts both in normal and in osteopenic bone. In osteopenic bone-derived osteoblasts this synthetic phase is preceded by an initial increase of cell proliferation.

The role of tumour markers in predicting skeletal metastases in breast cancer patients with equivocal bone scintigraphy

SummaryBone scintigraphy (BS) is commonly performed in the staging and postoperative monitoring of breast cancer. Nevertheless, due to low specificity it often demonstrates hot spots with equivocal interpretation, which may be misleading in the management of these patients. The aim of this study was to assess the value of a serum tumour marker panel in selecting among the patients with equivocal BS those with bone metastases. Between January 1986 and December 1995, 297 breast cancer patients were followed-up after mastectomy with serial determinations of a CEA-TPA-CA15.3 tumour marker panel, BS and liver echography. The tumour marker panel was used to select patients with equivocal BS for examination of suspicious bone areas by further imaging techniques. Up to December 1995, 158 (53%) patients showed an equivocal BS and 47 patients developed bone metastases. In the 158 patients with equivocal BS, prolonged clinical and imaging follow-up over 45 months (mean; range 12–120) was used to ascertain the presence or absence of bone metastases. In these 158 patients the negative predictive value and positive predictive value of the tumour marker panel to predict bone metastases was 97% and 75% respectively. This study shows that in breast cancer patients the CEA-TPA-CA15.3 tumour marker panel has a high value in selecting those patients with bone metastases, or at high risk of developing clinically-evident bone metastases, among the large number of subjects with equivocal BS.

Comparative interspecies investigation on osteoblast cultures: data on cell viability and synthetic activity ☆

The aim of the present study was to evaluate and compare the most common parameters that characterize the expression of primary osteoblast cultures from different origin (human, rat, sheep), and of the human osteosarcoma cell line MG-63 before and after stimulation with vitamin 1,25(OH)(2)D(3). Cell viability was quite similar for primary osteoblast cultures (MTT: 1.64-2.11 OD); a significant (P < 0.005) difference was found between sheep osteoblasts and MG-63 (DeltaMTT: 0.52 +/- 0.20 OD). Osteocalcin synthesis ranged from 15.18 to 27.00 pg/ml in primary osteoblast cultures, while it was significantly (P < 0.01) lower in MG-63 (OC: 6.67 +/- 0.52 pg/ml) when compared with primary human osteoblasts. Alkaline phosphatase, C-terminal procollagen type I, and interleukin-6 were significantly (P < 0.005) lower in rat osteoblasts when compared with primary human osteoblasts, and similarly transforming growth factor-beta1 was significantly (P < 0.05) lower in rat and sheep osteoblasts when compared with primary human osteoblasts and MG-63. Nitric oxide synthesis did not show any significant difference either before or after vitamin 1,25(OH)(2)D(3) stimulation. In conclusion, the current findings confirm the presence of interspecies differences between the selected osteoblast lineages before and after stimulation with vitamin 1,25(OH)(2)D(3). Above all, the culture of sheep osteoblasts was seen to behave more similarly to that of primary human cells, mainly in terms of cell viability, osteocalcin, interleukin-6 and transforming growth factor-beta1 production.

Stem Cells: Their Role in Breast Cancer Development and Resistance to Treatment

About 20% of the total cells from primary breast tumors could generate palpable tumors in non-obese diabetic severe combined immunodeficient (NOD/SCID) immunocompromised mice. All the tumorigenic cells originate from a normal mammary stem cell. Human mammary stem cells are sensitive to oncogenic mutations and in mouse models they share similarities with breast cancer stem cells (BrCSCs). Tumorigenicity, invasion, progression and metastasization are further BrCSCs properties likely depending on their CD44+/CD24- phenotype. Local invasion and tumor metastasization seem to be facilitated by the epithelial to mesenchymal transition (EMT) program. This program may be reactivated from stable genetic alterations or through exposure of cancer cells to factors present in the surrounding micro-environment, or by an up-regulation of EMT-inducing transcription factors. One main explanation for resistance to treatment by cancer cells is that a rare subpopulation of cells in residual tumors with tumorigenic potential is intrinsically resistant to therapy. Consistent with this hypothesis, in human breast tumors, the subpopulation of tumor-initiating cancer cells with CD44(high)/CD24(low) cell surface-marker profile was found more resistant to cancer therapies (chemo, hormone and radiotherapy) than is the major population of more differentiated breast cancer cells. The reasons for CSC resistance to chemotherapy, hormone therapy and radiotherapy also have been examined and they opened new scenarios for cancer therapy.

Total parathyroidectomy without autotransplantation for the treatment of secondary hyperparathyroidism associated with chronic kidney disease: clinical and laboratory long-term follow-up.

Forty-six consecutive patients who underwent total parathyroidectomy (tPTX) for hyperparathyroidism associated with end-stage kidney disease (CKD5) in a University Hospital from 1990 to 1999 were included in a long-term observational study. Outcome parameters included symptoms (bone pain, pruritus and muscle weakness evaluated by visual analog scales [VAS]) and laboratory data (intact parathyroid hormone [iPTH], total calcium, and alkaline phosphatase) assessed before, shortly postoperatively and then at a later time point: 40 patients were on maintenance hemodialysis and six on conservative medical therapy. Forty-four patients had four glands removed, while only three glands were found in the remaining two. Perioperative complications consisted of acute symptomatic hypocalcemia in 10 (22%) patients and non-specific complaints in three (7%). No laryngeal nerve palsies occurred. After a median follow-up of eight years, 43 subjects were evaluated: 37 (86%) were cured, three (7%) had persistent and three (7%) recurrent disease. Eleven patients underwent successful renal transplantation and 23 died during the period of observation. iPTH decreased from a mean of 1084+/-505 pg/ml to 120+/-381 pg/ml (p < 0.0001). No subsequent bone fractures, persistent bone pain or disability were reported; this includes patients who later received a functioning renal graft. tPTX was able to correct hyperparathyroidism in most of the patients and was associated with a low long-term relapse rate. iPTH levels remained low in 17 cases without symptoms and no clinically significant side effects. The beneficial effects of tPTX occurred in the majority of patients while renal transplantation was performed in a minority of patients. tPTX should be considered a safe and successful procedure for the treatment of severe secondary hyperparathyroidism associated with chronic kidney disease.

Large needle aspiration biopsy for reducing the rate of inadequate cytology on fine needle aspiration specimens from palpable thyroid nodules

From 1980 to 1996, 1,907 consecutive euthyroid subjects with palpable thyroid nodules were examined by fine needle aspiration (FNA) cytology plus large needle aspiration biopsy (LNAB) histology. There were 1,630 (85%) women and 277 (14.5%) men aged from 17 to 80 years. A single nodule was palpated in 1,419 subjects (74.4%) while 488 (25.6%) showed multiple nodules. The nodule size ranged between 1 and 7.5 cm. The number of inadequate specimens at the first examination, FNA cytology of LNAB histology, were 261 (13%) or 398 (20.8%), respectively. LNAB performed on the 261 nodules with nondiagnostic cytology showed findings which were adequate for diagnosis in 130 (49.8%) and inadequate in 131 (50.2%). Among the 261 patients with inadequate initial cytological findings 61 were subjected to repeated FNA and 36 repeated LNAB. More than 60% of the nodules on which FNA was repeated achieved a cytological diagnosis; more than 80% of the nodules reinvestigated by LNAB were finally diagnosed by histology. The mean nodule size was larger in the group with inadequate result than in that with adequate FNA or LNAB result. Among the 261 patients with inadequate cytological finding at the first FNA 28 were operated on; 20 were in the group with adequate LNAB histological findings and eight in the group with an inadequate LNAB. Two papillary cancers, one per group, were found at postoperative histology. However, one was diagnosed by LNAB and one at the second FNA. The remaining 26 nodules were all found to be benign postoperatively. This study shows that the addition of LNAB to FNA leads to a histological diagnosis in 50% of the palpable thyroid nodules with inadequate cytology at the first FNA and that LNAB can be used even for those nodules which remain uncharacterized after repeat FNA.

Cell-mediated immunity in breast cancer patients.

In clinical practice, reports have been made on immunosuppression after surgical excision of primary tumor or at relapse. However, the relationship between undefined or overt metastases and the host immune system has not been sufficiently examined over a prolonged period. These aspects were investigated in 160 breast cancer patients followed up post-operatively with serial controls over a long period. One hundred and thirty-four cases (91 node negative, (N-), 43 node positive (N+)) were disease-free and 26 relapsed. In all patients, serum T cell populations, serum B lymphocytes and skin reaction of delayed hypersensitivity (SRDH) were serially determined for 39 +/- 12 months (m +/- SD). The reference values for these parameters were assessed as follows: T populations were evaluated in 24 healthy donors and SRDH in 95 healthy females. In non-relapsed patients, constant CD8+ T cell decrease and T4/T8 ratio increase were observed; the T4/T8 ratio was significantly higher (ranging from P < 0.05 - P < 0.001) than in the control group. The mean values of NK cells and B lymphocytes, the former parameter being highly significant (P < 0.001), were higher than in controls. In the 26 metastatic patients, the T4/T8 ratio from 20 months before to 30 months after the first sign of relapse decreased from 3.2 to about 1 (r = -0.256, P < 0.05) and from 30 to 92 months after relapse progressively increased to 2. Similarly, in the former subinterval a progressive decrease in the number of positive antigens and score was found (from 2.4 to 0 and from 10 to zero respectively). A significant inverse correlation between these two parameters and observation time occurred (P < 0.01 and P < 0.001 respectively). From 30 to 86 months after relapse, a progressive increase in the number of positive antigens and scores up to 2 and 12 were observed. A significant direct correlation (P < 0.05) was noted. In conclusion, these data indicate significant changes in T populations during the disease-free interval in breast cancer patients. The decrease in circulating CD8+ T cells is compatible with the hypothesis of CD8+ T cell localization at the site of the micrometastases. The increase in circulating B lymphocytes and NK cells suggests activation of aspecific humoral immunity and NK function. In addition, they show that progressive deficiency in cell-mediated immunity appears many months before and that recovery continues for a long time after overt metastatic disease.

Von Willebrand’s factor mediates the adherence of human tumoral cells to human endothelial cells and ticlopidine interferes with this effect

The aim of this study was to explore whether von Willebrands factor (vWF) plays a role in the adhesion of human colon tumor cells to human endothelial cells in our coculture system. Cell colony density was evaluated basally (endothelial plus colon tumor cells) and following the addition of: purified vWF, vWF plus vWF-blocking antibodies, antibodies against various integrins and adhesion molecules (alpha2 b integrin, beta1 integrin, beta3 integrin, intercellular adhesion molecule-I, intercellular adhesion molecule-II, vitronectin receptor CD61 CD51, laminin alpha6/beta4 receptor), and various drugs inhibiting the hemostatic system (ticlopidine, heparin, acetyl salicylic acid [ASA], defibrotide, indobuphen, dipyridamole, sulfinpyrazone). Furthermore, vWF concentration was measured in the supernatant fluid of the coculture system basally and following the addition of the above-listed drugs. Cell colony density (as measured by light absorption) increased by 33% following the addition of vWF and returned to a value similar to the basal level with antibodies against vWF, while it did not change significantly following the addition of antibodies against the other integrins or adhesion molecules tested. The same parameter was reduced by 35% following the addition of ticlopidine, while it showed a smaller or no change with the other drugs tested. Similarly, vWF concentration in the cell coculture supernatant showed the greatest reduction (from 0.22 to 0.11 mg/mL) following the addition of ticlopidine. These data suggest that vWF mediates the adherence of human tumor cells to human endothelial cells and that ticlopidine interferes with this effect.