Chromatin-associated CSF-1R binds to the promoter of proliferation-related genes in breast cancer cells.

The colony-stimulating factor-1 (CSF-1) and its receptor CSF-1R physiologically regulate the monocyte/macrophage system, trophoblast implantation and breast development. An abnormal CSF-1R expression has been documented in several human epithelial tumors, including breast carcinomas. We recently demonstrated that CSF-1/CSF-1R signaling drives proliferation of breast cancer cells via 'classical' receptor tyrosine kinase signaling, including activation of the extracellular signal-regulated kinase 1/2. In this paper, we show that CSF-1R can also localize within the nucleus of breast cancer cells, either cell lines or tissue specimens, irrespectively of their intrinsic molecular subtype. We found that the majority of nuclear CSF-1R is located in the chromatin-bound subcellular compartment. Chromatin immunoprecipitation revealed that CSF-1R, once in the nucleus, binds to the promoters of the proliferation-related genes CCND1, c-JUN and c-MYC. CSF-1R also binds the promoter of its ligand CSF-1 and positively regulates CSF-1 expression. The existence of such a receptor/ligand regulatory loop is a novel aspect of CSF-1R signaling. Moreover, our results provided the first evidence of a novel localization site of CSF-1R in breast cancer cells, suggesting that CSF-1R could act as a transcriptional regulator on proliferation-related genes.

AML1/ETO sensitizes via TRAIL acute myeloid leukemia cells to the pro-apoptotic effects of hypoxia.

We determined the effects of severe hypoxia (∼0.1% O2) on acute myeloid leukemia cells expressing the AML1/ETO oncogene. Incubation of Kasumi-1 cells in hypoxia induced growth arrest, apoptosis and reduction of AML1/ETO protein expression. The conditional expression of AML1/ETO in U937-A/E cells showed that hypoxia induces marked apoptosis in AML1/ETO-expressing cells only, pointing to AML1/ETO as a factor predisposing cells to hypoxia-induced apoptosis. In AML1/ETO-expressing cells, hypoxia enhanced TRAIL expression and its proapoptotic effects. AML1/ETO was found to bind TRAIL promoter and induce TRAIL transcription, although TRAIL expression was restrained by a concomitant relative transcription block. In hypoxia, such a TRAIL repression was removed and an increase of TRAIL expression was induced. Finally, blocking anti-TRAIL antibodies markedly reduced (Kasumi-1 cells) or completely inhibited (U937-A/E cells) hypoxia-induced apoptosis. Taken together, these results indicated that hypoxia induces apoptosis in AML1/ETO-expressing cells via a TRAIL/caspase 8-dependent autocrine loop and that TRAIL is a key regulator of hypoxia-induced apoptosis in these cells.

Selective anti-leukaemic activity of low-dose histone deacetylase inhibitor ITF2357 on AML1/ETO-positive cells.

We analysed the in vitro effects of a new hydroxamate derivative, ITF2357, on AML cells. ITF2357 potently induced histone acetylation. ITF2357 0.1 microM blocked proliferation and induced apoptosis in AML1/ETO-positive Kasumi-1 cells, while AML1/ETO-negative HL60, THP1 and NB4 cell lines were sensitive only to 1 microM ITF2357. Apoptosis was induced by 0.1 microM ITF2357 in AML1/ETO-positive primary blasts and U937-A/E cells induced to express AML1/ETO, but not in U937-A/E cells non-expressing AML1/ETO. In Kasumi-1 cells 0.1 microM ITF2357 induced AML1/ETO degradation through a caspase-dependent mechanism. ITF2357 0.1 microM also determined DNMT1 efflux from, and p300 influx to, the nucleus. Moreover, 0.1 microM ITF2357 determined local H4 acetylation and release of DNMT1, HDAC1 and AML1/ETO, paralleled by recruitment of p300 to the IL-3 gene promoter. ITF2357 treatment, however, did not induce re-expression of IL-3 gene. Accordingly, the methylation level of IL-3 promoter, as well as of several other genes, was unmodified. In conclusion, ITF2357 emerged as an anti-leukaemic agent very potent on AML cells, and on AML1/ETO-positive cells in particular. More relevantly, clearly emerged from our results that ITF2357 could be an ideal agent to treat AML subtypes presenting AML1/ETO fusion protein which determine HDAC involvement in leukaemogenesis.

Results of rebiopsy for suspected prostate cancer in symptomatic men with elevated PSA levels.

AIM OF THE STUDY: To develop indications for repeat biopsy in patients with suspected prostate cancer and first negative biopsy. MATERIALS AND METHODS: 148 consecutive patients, submitted to two or more biopsies for suspected prostate cancer, were extracted from our database on prostatic diseases. Patients were stratified according to the results of the last biopsy (benign or carcinoma) considering the results of the first and of the last biopsy when more than two biopsies had been performed. PSA velocity was calculated when the interval between PSA obtained before the initial and the final biopsy was at least 6 months; PSA velocities were annualized and absolute changes between the two groups were analyzed. RESULTS: Prostatic carcinoma was detected in 60 of the 148 patients (40.5%), including 19 of 41 (46.4%) with prostatic intraepithelial neoplasia (PIN) and 45 of 107 (42.1%) with normal tissue or prostatic epithelial atrophia on initial biopsy. 20% of patients (4 of 20) with low-grade PIN and 71.1% (15 of 21) with high-grade PIN had cancer at repeat biopsy. The mean PSA value of patients with carcinoma on the repeat biopsy was higher than that of patients without carcinoma (13.3 vs. 10.7 ng/ml). However, this difference was not statistically significant (p = 0.37). Mean PSA velocity increased for patients with a final diagnosis of carcinoma versus those without evidence of carcinoma (+0.3 vs. +1.4 ng/ml/year); this difference was statistically significant (p = 0.002). CONCLUSIONS: According to these results, patients with either PIN II-III, or high PSA and PIN I on initial biopsy, and/or with elevated PSA velocity (more than 1 ng/ml/year) should undergo repeat prostate needle biopsy, being at high risk of prostate carcinoma.

Procollagen type I carboxyterminal extension peptide in serum: a reliable marker of bone metastatic disease in newly diagnosed prostate cancer?

OBJECTIVE: This study evaluates the accuracy of type I procollagen, a bone matrix glycoprotein, and prostate-specific antigen (PSA) as markers for predicting the results of radionuclide bone scan in newly diagnosed, previously untreated patients with prostate cancer. METHODS: 74 patients underwent serum PSA and procollagen determination using specific antibodies. A staging radionuclide bone scan was then performed; patients with positive bone scan were submitted to x-rays of the suspicious zones. Then, we calculated sensitivity, specificity, positive predictive value, negative predictive value and overall accuracy of procollagen and PSA in the detection of bone metastases. RESULTS: Procollagen alone had 83.3% sensitivity, 96% specificity, 90.9% positive predictive value, 92.3% negative predictive value and 91.9% overall accuracy. PSA alone had 70.1% sensitivity, 86% specificity, 70.8% positive predictive value, 86% negative predictive value and 81.1% overall accuracy. CONCLUSIONS: According to our data, we no longer perform a staging radionuclide bone scan in patients with PSA < 20 ng/ml and normal procollagen level, diminishing the number of radionuclide bone scans and increasing the overall net savings for the health care system.

Serum levels of tumor-associated trypsin inhibitor (TATI) in benign and malignant gynecological diseases.

The behavior of tumor-associated trypsin inhibitor (TATI) as a marker for gynecological cancer was studied in a control population and in patients with different benign and malignant diseases. When a cut-off level of 21.4 micrograms/l was used the specificity was 100% in patients with benign diseases. The sensitivity in patients with malignant tumors was low for cervical and corpus cancer, 13% and 14%, respectively, whereas it was 33% in all the ovarian malignant tumors, reaching 60% in the mucinous type. There was a clear correlation between TATI level and stage.

[The renin-angiotensin-aldosterone system in liver cirrhosis]. / Il sistema renina-angiotensina-aldosterone nella cirrosi epatica.

The role of the RAA system in the genesis of ascites in liver cirrhosis patients is not yet perfectly clear. The present study was conducted on 176 cirrhosis patients in order to investigate RAA system function, to assess the changes taking place in the various stages of the disease and to correlate such changes with the various kidney function parameters. The patients were divided into 3 groups as follows: Group I: patients without ascites on admission and with no history of the condition; Group 2: patients with ascites of recent onset and/or response to diuretic treatment; Group 3: patients with ascites not responsive to diuretic treatment. In Group 1, 19 patients (38%) reveal a significant reduction in renin activity together with portal hypertension and increased hydrosaline retention. In Group 2 renin activity was reduced in 4 patients (6%), aldosterone activity in 3 (4%). Progressive deterioration in liver function parameters and progressive activation of the RAA system combined with reduced sodiuria content were found in over 50% of these patients. The presence or absence of portal hypertension in this group was not related to significant changes in diuresis or sodiuria. In Group 3 renin was activated in 54 patients (89%), aldosterone in 58 (95%) and there was also a distinct reduction in sodiuria (96% of patients) and chloruria (100%). A substantial increase was also noted in the incidence of low blood sodium (53%) while portal hypertension was found in 97% of patients. On the basis of those data it may be hypothesised that high pressure inside the liver creates the stimulus for primary sodium retention. The decrease in effective blood volume after vasodilation, accentuated by low blood albumin and splanchnic venous stagnation may the stimulate the sympathetic nervous system and RAA system. Hyperaldosteronism only becomes the dominant factor in renal imbalance when the cirrhosis reaches the resistant ascites phase.

[Changes in antidiuretic hormone (ADH) in liver cirrhosis with resistant ascites]. / Modificazioni dell'ormone antidiuretico (ADH) nella cirrosi epatica con ascite resistente.

The pathogenetic role of ADH in determining hyponatremia in patients with liver cirrhosis is still much debated. Osmotic stimuli are not able to inhibit secretion of ADH in refractory ascites and under such conditions the reduction in effective plasma volume has been put forward as the main cause. Twenty patients with liver cirrhosis and refractory ascites were studied before and during extraction-concentration-reinfusion (ECR) of ascitic fluid by means of Rhodiascit. ADH, renin, aldosterone, blood and urine osmolarity, plasma and urinary concentration of sodium, potassium, chlorine, and the clearance of free water were evaluated. All patients presented high renin values (15.4 +/- 11.7 ng/ml), aldosterone (341 +/- 172 ng/ml), ADH (6.3 +/- 5.2 pg/ml). During ECR, a significant drop was observed in renin (p less than 0.001), aldosterone (p less than 0.001) urinary osmolarity (p less than 0.001) and an equality significant increase in diuresis (p less than 0.001), natriuria (p less than 0.005), kaliuria (p less than 0.001) while ADH presented an irregular course: in 11 cases it remained unchanged, in 3 it fell and in 6 it presented a constant increase. To conclude, data suggest that the diminished filtrate reaching the distal tubule constitutes the greatest cause of the inability to dilute urine in many patients with cirrhosis and that ADH is a permissive rather than a primary factor.

[Ferritin, CEA and TPA as tumor markers in breast neoplasms]. / Ferritina-CEA-TPA come markers neoplastici nelle neoplasie mammarie.

In order to assess the sensitivity and specificity of Ferritin, CEA and TPA as neoplastic markers in breast carcinomas, 91 patients all classified according to the TNM-UICC system were studied in a cancer clinic. The results of the analyses indicate that ferritin is apparently only influenced by the presence of metastatic neoplasias and that greater sensitivity is obtained if all three markers are employed simultaneously.

[Advantages of preventing Rh isoimmunization]. / Korist od prevencije Rh imunizacije.

The authors analysed the frequency of Rh immunization from 1972 to 1983. The incidence of Rh-immunized women who after the birth of a Rh (D) positive child were not given anti-D immunoglobulin G and in subsequent pregnancies gave birth to a Rh (D) positive child was found to amount to 11.76%, while in women who were given anti-D immunoglobulin D this incidence was 0.77% (t = 5.98; p less than 0.05). Out of 29 Rh-immunized pregnant women, two developed Rh immunization in the course of the first pregnancy, three after the unsuccessful prevention of Rh immunization, and the rest after delivery or after delivery and abortion. Out of 29 Rh-immunized women, 27 (93.10%) were ABO-compatible and 2 (6.90%) ABO-incompatible with their child (p less than 0.05). In the first pregnancy the incidence of Rh immunization was 1.86 per 1000 deliveries in Rh negative pregnant women and 21.19 per 1000 deliveries in subsequent pregnancies (p less than 0.05). In the period observed there were 2.24 Rh immunizations per 1000 of all deliveries. From 1972 to 1977 there were 3.19 Rh immunizations per 1000 deliveries and from 1978 to 1983 only 1.43 (t = 2.08; p less than 0.05), which is a reduction by 55.17%. The perinatal mortality rate of children affected by Rh-hemolytic disease was 20%. In the last six years it has gone down by 60%, while the number of children with Rh-hemolytic diseases has been reduced by 50%.

[Study of blood ferritin in cancer patients and its possible use as an indicator of an unfavorable prognosis]. / Studio della ferritinemia in pazienti neoplastici e sua possibile utilizzazione come indice prognostico sfavorevole.

Ferritinaemia levels were measured in 97 neoplastic patients and compared with the levels found in a healthy control group, in order to discover whether ferritinaemia had any significance as a neoplastic marker. Higher levels were encountered in all neoplastic patients (P less than 0.005) than in the control group. Levels were particularly high in the patients with metastasised tumours (especially breast cancer: P less than 0.001). The highest ferritinaemia levels were found in terminal patients (P less than 0.001).

[Evaluation of blood ferritin in sideropenic anemia and its modifications in the course of divided-dose parenteral iron therapy]. / Valutazione della ferritinemia nell'anemia sideropenica e sue modificazioni in corso di terapia marziale parenterale refratta.

Changes in blood ferritin during divided dose parenteral iron therapy and the importance of ferritin evaluation in iron-deficiency anaemia were investigated in 20 women and 10 men with this diagnosis through withdrawals before and after treatment. In 6 subjects, blood ferritin values enabled the presence of iron deficiency to be ruled out, since they were high at the first control (in agreement with the histological examination of the marrow in the search for iron deposits). In sideropenic males, the difference between values at the time of diagnosis and those of normal controls was significant (p less than 0.001). The absence of this finding in the females may have been due to over-low values in the normal controls. Blood ferritin values during therapy gradually rose until its termination. The conclusion is drawn that at any rate in males the determination of blood ferritin can be a useful aid in the diagnosis of iron-deficiency anaemia, and in the demonstration of normal reserves after treatment.

[The importance of thyroid hormone determination in the course of acute and chronic liver diseases]. / Importanza del dosaggio degli ormoni tiroidei in corso di epatopatie acute e croniche.

The RIA values of thyroid hormones in the course of acute and chronic liver disease were studied to see whether they were related to the severity of the picture in a series of 50 healthy subjects and 133 with various hepatopathies: 26 with acute viral hepatitis, 18 with alcoholic liver disease, 16 with alcoholic cirrhosis without ascites and 33 with ascites, 14 non-alcoholic cirrhosis without ascites and 24 with ascites. A reduction in T3 proportional to the seriousness of the clinical and laboratory findings was noted in chronic forms, whereas both T3 and T4 were high in acute viral hepatitis. There was no difference in T3 values in alcoholic and non-alcoholic cirrhosis of similar gravity, showing that the fall in serum T3 is not a specific alcohol-induced lesion. T3 less than 25 ng/100 ml proved the best index in the prediction of mortality (chi 2 = 20,5; p less than 0,0005).