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We studied for 5 yr a homogeneous group of 83 patients randomly assigned to a levothyroxine (L-T4) suppressive therapy or to a control group to evaluate changes in nodule or thyroid size, appearance of new nodules, and correlations with clinical parameters. In the control group, mean nodule volume increased significantly after 5 yr (2.12 +/- 1.46 vs. 1.46 +/- 0.77 mL), whereas in the treatment group it decreased, although not significantly (1.45 +/- 1.17 mL vs. 1.53 +/- 0.61 mL). Baseline nodule volume was not different in the two groups, but a significant difference was observed at 5 yr. After 5 yr, sonograms detected 12 new nodules in the control group (28.5%) and 3 (7.5%) in the treatment group. Nodule shrinkage was more frequent in patients with complete TSH suppression, but no correlation was found with other parameters. A significant increase in thyroid size was observed in the control group. In conclusion, long term TSH suppression induced volume reduction in only a subgroup of thyroid nodules, but effectively prevented the appearance of new lesions and increases in nodule and thyroid volume. As the changes in the natural history of nodular goiter are related to prolonged TSH suppression, which can induce unfavorable side-effects, L-T4 suppressive therapy should be reserved for small nodules in younger patients.

In order to improve manufacturing processes and their automated integration, RoboTViRMo (Robot Teaching in Virtual Reality Mode) is aimed at designing and developing a user friendly system for the robot off-line programming in a virtual environment. The project, aiming at increasing the automation level in the small-medium enterprise plants by robot, proposes to develop tools for users, experts in the manufacturing processes, but who don't know programming languages for robotised systems. Using a Data Glove, or a 3D pointer, and Head Mounted Display (HMD), or stereoscopic glasses, it will be possible to program a robot for deburring operations without the knowledge of robot programming languages and of 3D graphical simulators, thanks to the use of the natural language performed by the user's hand movement and based on his experience. The system is composed by 4 main integrated environments: two are virtual environments, one is a 3D graphical simulator and one is a real deburring cell. The first environment allows the user to locate the burrs on the piece surface and to model, within a virtual environment and starting from the original 3D CAD model, the 3D model of the work piece. The second one allows the user to perform the virtual deburring on the piece: it could be done automatically, through a software for the deburring trajectory research (RATS project), or manually, through virtual operations of the user, that can select whether to furnish just few information (i.e. tool orientation, starting points, etc.) to the automatic system in order to decrease the time needed for the trajectory research or to perform the whole deburring process in the virtual environment. The third environment is a graphical 3D simulator that, on one hand, allows to value the designed tracks (automatic or manual ones) doing the feasibility analysis and the collision detection, and, on the other hand, realizes the down load on the real robot control for the physical deburring through a post-processor. The last environment is the physical one, where an anthropomorphic robot for the deburring operation onto sample pieces is modeled as described in the paper. The original and innovative approach is the application field, that is a complex manufacturing one that requires the presence of an expert of the job and, nowadays, a computer expert for technical support. RoboTViRMo helps the expert in utilizing the informative system without any middle-person and directly uses the expert experience in the simulation developing. The authors present the first prototype for deburring operations and the hardware and software configuration that allows to reach them.

SUMMARY. In chronic hepatitis C (HCV) infection, treatment with interferon is associated with a rather low rate of sustained response and many treated patients do not achieve significant benefit. Efforts have therefore been made to identify non-responders as early as possible to avoid unjustified costs and side-effects. We treated 106 cases of HCV with an algorithm based on the results of sequential alanine aminotransferase (ALT) and HCV RNA determinations, using an initial dose of 6 MU thrice weekly for 4 months, and modified the subsequent treatment according to the biochemical and virological profile. Thirty-three out of 48 patients (68.7%) who were HCV RNA negative with normal ALT at 4 months after initiation of treatment were sustained responders when treated for an additional 4-month period with a reduced 3 MU dose, while sustained response was achieved in 12.5% of HCV RNA positive patients treated with a higher dosage and for a more prolonged period of time. Our findings indicate that HCV RNA monitoring during interferon therapy may be useful in modifying of the treatment schedule for the individual patient.

Background Rheumatoid arthritis(RA) is a chronic autoimmune inflammatory disease. The inflammatory process causes joint destruction and disability. Patients have a shorter life expectancy and a relative risk of cardiovascular (CV) disease ranging from 1.5 to 4.0 with an increase in CV morbidity and mortality of 1.5 fold compared with the general population. The chronic inflammation seen in RA have its main effectors in factor alfa tumor necrosis (TNF) and interleukin 6 (IL-6) that, in addition to playing a key role in the initiation of joint damage and progression of atherosclerotic disease, are also potent inducers of the renin angiotensin system (RAS). The latter could contribute in these patients to the development of an early and accelerated CV damage. Objectives To evaluate the possible presence of hyperactivation of RAS and its association with the activity parameters of RA, with traditional risk factors and premature CV organ damage Methods there were evaluated 17 (F) patients (pts) with RA receiving stable therapy (3 months) with DMARDs or anti-TNF alpha, in the absence of previous CVevents and/or traditional risk factors. All pts underwent a visit with rheumatologic evaluation of patient history, activity of RA (DAS28), pain (VAS), quality of life (HAQ), patient GH. Blood samples were collected for the evaluation of: lipid profile, indices of insulin resistance (HOMA-IR), indices of activity of rheumatoid arthritis (autoantibodies, inflammatory markers) and CV risk according to risk charts based on the Framinghamstudy. The CV organ damage was assessed by imaging techniques such as Pulse Wave Velocity (PWV) and Augmentation Index (AIx), Intima-media thickness (IMT) and ankle-brachial index (ABI). From a sample of the patient’s blood leukocytes were separated and subsequently extracted RNA to perform Real time qPCR to verify the expression of Angiotensin II and AT1 and AT2 receptors. The mean disease duration was 9 ±7 years; the mean age (pts all female) was 55 ± 12 years. FR was positive in 49%, antiCC and ANA respectively in 64% and 59% Results data, though preliminary, showed a statistically significant correlation between the density of receptors for angiotensin II (AT1R and AT2R) and, respectively, the values of ABI(p Conclusions The results obtained so far make attractive and plausible the initial assumption and could, if confirmed in further continuation of the study, help to explain the paradoxical development of vascular disease in patients with RA and apparent low CV risk with important therapeutic implications Disclosure of Interest None Declared

AIMS: To determine cell proliferation in infiltrating breast carcinomas. METHODS: Using the MIB-1 monoclonal antibody, the proliferation index was measured in paraffin wax sections of 871 breast cancers. The MIB-1 proliferation index was compared with other markers of disease progression: size, lymph node status, histotype, oestrogen and progesterone receptor status, expression of p53 and Neu, and DNA ploidy. All parameters were measured using image analysis. In 347 tumours, the MIB-1 and Ki-67 proliferation indexes were compared. Follow up data were available for 170 cases (median 66.5 months). RESULTS: Of the tumours, 314 (36%) had a high proliferation index. The MIB-1 proliferation index was correlated directly with size, nodal status, overexpression of p53 and Neu, and the DNA index; and inversely with oestrogen and progesterone receptor status. The correlation between MIB-1 and Ki-67 proliferation indexes was statistically significant. In patients with pT1 tumours, a low proliferation index correlated with a longer relapse-free interval and overall survival; node negative patients with a low proliferation index had a longer overall survival. CONCLUSIONS: The MIB-1 proliferation index is a reliable, practical and useful method of measuring proliferative activity and is an important predictor of clinical behaviour.

Percutaneous ethanol injection (PEI) is an effective treatment for autonomously functioning thyroid nodules (AFTNs). The aims of this study were to assess: (1) the ultrasound (US) and colour-coded duplex sonography (CCDS) baseline and post-therapy patterns of AFTN, and (2) the contribution of CCDS in determining the efficacy and optimum duration of PEI. Forty AFTNs were evaluated by US and CCDS before, during and 6 and 12 months after PEI. Four grades of increasing vascularity were arbitrarily identified at CCDS (0-1-2-3). These data were compared with thyroid scintiscan and hormonal findings. All nodules showed a striking volume decrease. After treatment a hypoechoic pattern and blurred outlines on US were associated with a higher percentage volume reduction. Soon after PEI a reduction in the vascularity score was noticed in 29 of 40 of AFTNs. Pre- and post-treatment vascular score distributions were significantly different. It is concluded that a vascular score decrease to grade 0–1 at the end of PEI is an index of successful treatment.

Maldistribution of intrathecal local anesthetic has recently been implicated as a contributor to neurotoxic injury. In vitro modeling can be used to understand the distribution of anesthetic agents within the subarachnoid space. We describe an in vitro modeling technique that uses digital video image processing and its application to catheter injection of local anesthetic.A clear plastic model of the subarachnoid space, including a simulated spinal cord and cauda equina, was filled with lactated Ringer's solution. Phthalocyanine blue dye of known concentration was injected into the model through small-bore (28-G) and large-bore (18-G) catheters. Injections were performed at a variety of controlled rates and sacral catheter positions, and the propagation of dye throughout the model was recorded on videotape, digitized by computer, and converted to a two-dimensional image of dye concentration. A subset of data was compared with results obtained from spectrophotometric analysis.There was a strong correlation (r = 0.98) between data obtained with analysis by digital video image processing and those obtained spectrophotometrically. Catheter size, catheter angle, and injection rate significantly influenced the distribution and peak concentration of simulated anesthetic. No major differences in distribution or peak concentration were observed with the two types of 28-G catheters.The digital video image processing technique can be used to quantify anesthetic distribution rapidly within a model of the subarachnoid space without disturbing the distribution. The current results demonstrate a strong dependence of anesthetic distribution on catheter angle, catheter size, and injection rate. Comparisons between 28-G catheters suggest that the difference in reported incidence of cauda equina syndrome associated with different 28-G catheters cannot be explained on the basis of differences in anesthetic distribution.

GATA1, founding member of the GATA transcription factor family, is essential for cell maturation and differentiation within the erythroid and megakaryocyte (MK) lineages. Disruption of DNA- or protein-binding capacity of GATA1 causes severe hematopoietic dysfunction and plays a role in blood disorders such as thrombocytopenia, anemia or leukemia. GATA1 expression seems to be related to the MK commitment both in mice and in humans; indeed, similarly to the murine myeloid M1 cell line, in which the enforced expression of GATA1 induces the c-Mpl appearance and MK differentiation, transduction of human hematopoietic stem cells with a GATA1 highly expressing vector results in self-renewal block and in the exclusive generation of Meg-E lineages. More recently, a role for GATA1 also in myeloproliferative disorders (MPDs) was indicated by the “GATA1-low” mouse model which develop a disease closely resembling the human idiopathic myelofibrosis. Interestingly, patients affected by myelofibrosis was also shown to express decreased GATA1 levels by immunostaining of BM sections. In this study, we investigated by Real Time PCR the levels of GATA1 in a myeloproliferative disorders such as essential thrombocytemia (ET) tipically characterized by a neoplastic megakaryocitic proliferation. We have studied BM samples of 40 newly diagnosed patients (M:F ratio 1:1 - median age 53 years, range 18–84) affected by ET, as for the PVSG group criteria. These patients were selected from a cohort of 65 ET patients considering a similar erythroid/myeloid ratio at the FACS analysis to reduce a possible bias for the RT-PCR results due to the erythroid compartment interference. The median platelets count of the selected patients was 670,000/mL (range 493,000–1,400,000/mL), myelofibrotic index 0/1, and 18 out of 40 patients (45%) showed mild splenomegaly both at the physical examination and US scan (median spleen vol 550 ml - range 430–1400 mL). No chromosomal abnormalities were detected by cytogenetic analysis. JAK2 sequencing in 21/40 patients indicated that 9/21 patients (43%) were positive for the JAK2 V617F genomic mutation. At the end of observation time (median 18 mo.) no patients had evidence or signs of thrombotic or hemorrhagic complications. BM cells from six healthy donors were used as normal controls in the study. The relative GATA1 quantification was calculated in according to the DCt method with GAPDH as internal control. The results showed a significant increase of GATA1 expression in BM cells from ET patients (median DCt + 6,11 ; range −0,41/+18,11) compared to the controls (median DCt + 0,172 ; range −4,03/+1,7) (p < 0,003). Interestingly, the GATA1 overexpression is not a mere consequence of the proliferation and activation of MKs, indeed samples from three patients affected by idiopathic thrombocytopenic purpura, whose BM smears had the typical secondary megakaryocytic hyperplasia, showed GATA1 levels much lower than the ET patients (median DCt − 0,6 ; range − 3,21/−0,9). No significant difference in GATA1 level was found between patients harbouring a JAK mutation (median DCt +5,86 ; range 0,85/16,12) and those with wild type alleles (median DCt +4,75 ; range −0,41/10,21). In conclusion, our results suggest that GATA1 overexpression could be a trigger for MK neoplastic commitment and proliferation and, consequently, seems to have a central role for ET pathogenesis both in JAK2 mutated and in JAK2 WT patients.

EVI1 is a nuclear oncoprotein deregulated by recurring chromosomal abnormalities in MDS. The expression of this gene in MDS patients represents a very poor prognostic marker and is associated with erythroid and megakaryocytic defects. We have previously shown that the forced expression of EVI1 in mice results in a fatal disease with features characteristic of MDS including dyserythropoiesis, dysmegakaryopoiesis, and anemia. More recently we have shown that EVI1 directly interacts with GATA1 and disrupts GATA1−binding to DNA leading to deregulation of GATA1−dependent genes. Here we describe the effects of an EVI1 mutant unable to bind GATA1, on the regulation of GATA1 target genes and on the erythroid differentiation of murine bone marrow progenitors. The structure of two zinc finger motifs of this mutant, EVI1(1+6Mut), were destroyed by His to Ala and Cys to Ala changes. Semi−quantitative RT−PCR showed that most of the analysed genes were down−regulated in 32Dcl3 cells expressing EVI1 but not EVI1(1+6Mut). Bone marrow lineage negative cells infected with EVI1, EVI1(1+6Mut), or the empty vector as control, selected in G418, and plated in presence of Epo were utilized to determine the progenitors’ potential to differentiate in response to this cytokine. The EVI1−expressing cells were virtually unable to generate erythroid colonies after Epo stimulation and only scarce small colonies were observed. In contrast, the EVI1(1+6Mut)−cells produced about 65% of the colonies formed by the control cells. The cells were recovered and their morphology was analysed after Wright−Giemsa staining. All EVI1−positive erythroid cells showed an impaired differentiation that was arrested at the basophilic−erythroblast stage. These cells were bi− and tetra−nucleated with chromatin bridges, budding nuclei and nuclear−cytoplasmatic maturative asynchronizations. All of these features are known as dysplastic erythroid aspects in MDS patients. These dysplastic characteristics were less prominent in EVI1(1+6Mut)−positive cells and were observed only in a minority of cells rather than in the entire population. As expected, the control cells had the appearance of normal erythroblasts and none of the aberrant features were observed. These results parallel the morphology observation of peripheral blood smears obtained from EVI1−mice and EVI1(1+6Mut)−mice in that erythropoiesis defects such as aniso−poikilocytosis and orthochromatic erythroblasts were less striking and observed only in a limited number of erythroid cells in EVI1(1+6Mut)−mice. Based on these results, we propose that EVI1 blocks erythroid differentiation by direct interaction with GATA1 leading to impairment of GATA1 regulation. This differentiation block is significantly reduced in vitro and in vivo with the EVI1(1+6Mut) mutant, which re−establishes to a high degree the normal functions of GATA1 and diminishes the erythroid dysplastic aspects observed in EVI1 cells.

Blast localization in the brain occurring at diagnosis or at relapse in AL and in NHL patients is considered an adverse prognostic event. The management of meningeal or CNS disease includes intrathecal (IT) injection of cytotoxic drugs (methotrexate, cytarabine, steroids), cranial radiotherapy and high dose chemotherapy. IT therapy is currently carried out by weekly or biweekly drug administration, until clearance of leukemic cells from CSF. Liposomal cytarabine (Depocyte®), a newcomer in the strategy of lipid incapsulation of cytotoxic drugs, is a molecule registered exclusively for IT administration in lymphomatous meningeal diseases. Because of its long-lasting clearance profile, this drug can be administered every two weeks, thus resulting in better patient compliance. From September 2004 to April 2006, we treated 8 patients by IT Depocyte at the standard dose of 50 mg every two weeks, for a total of 25 doses. Three patients had acute leukemia: 2 ALL, 1 AML; 24, 54, and 55 year-old, respectively. At time of meningeal involvement, two patients were also in hematological relapse, while the other was in complete hematological remission. The patients had been previously treated by cytotoxic chemotherapy according to the current protocols; intrathecal prophylaxis with MTX and methylprednisolone had also been administered to ALL patients. A 46 year-old patient, had CML in blast crisis (CML-BP), with blast cells expressing both myeloid and lymphoid markers (CD 34+, CD19+, CD10+, CD24+, CD79a+, TdT+, CD33+, CD13+, CD11b+, CD7+). The blast crisis, involving simultaneously bone-marrow and CNS, had occurred during treatment with imatinib. Two elderly patients (> 60 year-old) suffering from NHL (mantle-cell and diffuse large-B-cell histotypes, respectively, had showed CNS localization (meningeal and meningeal + right orbital mass) during their disease progression. Two young patients, aged 28 and 39 yrs, suffered from primary cerebral (PCNS) lymphoma. 7/8 patients had blast cells detectable at CSF examination (mean, 857/μL; range, 23 – 2500/μL). AL patients received four doses of Depocyte as unique IT drug, and all showed early and complete clearance of blast cells from CNS. In the patient with CML-BP, CSF was not cleared after two doses of the drug, and the treatment was interrupted. As for NHL patients, the one with mantle cell lymphoma received the four scheduled Depocyte injections, and showed complete remission of the meningeal localization, while the other stopped the treatment after the 2nd dose because of CNS disease progression. Both patients with PCNS lymphoma received only two doses of Depocyte because of rapid neurological deterioration. No patient showed toxicity or adverse events due to the IT administration of liposomal cytarabine. In responding patients (4/8), CSF leukemic cell count fell rapidly after the first IT injection, and none of them had further meningeal involvement at hematological disease progression. Intrathecal therapy with Depocyte appears feasible, well tolerated and effective in patients with meningeal leukemic/lymphomatous disease, while it was ineffective, at least in our hands, in patients suffering from primary CNS lymphoma, for which the gold standard of treatment remains systemic high dose methotrexate and cytosine arabinoside followed by radiation therapy.

The molecular etiology of subgroups of essential thrombocythemia (ET), as well as some other myeloproliferative disorders (MPDs) has been recently better understood by the identification of an activating somatic point mutation (Valine 617 to Phenylalanine) in Jak2 detected in patients with MPDs. Approximately 23%–57% of ET patients harbor this mutation, which is thought to be one of the dominant factors contributing to the disease. However, the pathways disrupted in at least half of ET patients who have wild type Jak2 are not known. The fusion gene AML1/MDS1/EVI1 (AME), a product of the t(3;21)(q26;q22) translocation, is associated with several hematopoietic disorders, primarily chronic myelogenous leukemia and acute myeloid leukemia, and to a lesser extent with ET. We have investigated the role of AME by generating C57BL/6J mice that express this fusion oncogene in their bone marrow after bone marrow infection and transplantation. After a latency of 12 ± 1.8 months, all the reconstituted mice invariably developed an ET-like disease. The disease was fatal and the animals died of anemia probably caused by bleeding. Until 1–2 weeks before death, at which time the mice were severely cytopenic, the peripheral blood hemoglobin and leukocyte counts of the reconstituted AME mice were normal. The only striking difference was the platelet counts, which were consistently above the normal range (1456–3153 and 940–1608 K/ml for AME and control mice, respectively, p=0.00029). Platelets appeared dysplastic with anisocytosis, various degrees of degranulation, and giant size. The bone core biopsies of AME mice were especially remarkable for large and giant megakaryocytes with a tendency for clustering. Touch preparations of the biopsies showed progressive maturation in the granulocytic and erythroid precursor cell lines, a normal myeloid to erythroid ratio and no evidence of dyshematopoiesis. These finding coupled with markedly increased levels of TPO but normal concentrations of c-Mpl and Epo-R are concordant with the morphologic and laboratory features of patients with ET. The sequence analysis of exon 12 of Jak2 demonstrated no guanine-to-thymine transversion at nucleotide 1849 in all AME mice tested. These data are valuable because they provide a novel mouse model for understanding the deregulated pathways in ET patients with wild-type Jak2.

Essential thrombocytemia (ET) is a myeloproliferative disorder characterized by a primary increase in platelets count often associated with splenomegaly and/or hepatomegaly often developing/evolving in thrombosis. The t(3;21)(q26;q22) resulting in the AML1/MDS1/EVI1 (AME) fusion gene is associated with several hematological disorders often characterized by severe dysmegakaryopoiesis, suggesting that AME alters the megakaryocytic program. By BM infection and transplantation we generated two groups of mice that express either wild type AME or a point/internal-deletion mutant of AME (AME-D-C). This mutant lacks the eighth zinc finger motif and is unable to interact with the CtBP corepressor. Both groups of mice developed a fatal disease with features of human ET, however Kaplan-Meyer analysis showed a significant difference (p < 0.0032) in survival between the two groups with the AME-D-C-mice living considerably longer. The morphological analyses of the peripheral blood smears did not show significant differences between the AME- and AME-D-C-mice. However while the AME-mice were anemic, the AME-D-C-mice had normal counts. In both groups the platelet number was increased and atypical features like macro-platelets, agranular platelets and small aggregates were observed. Both groups of mice developed thrombocytosis with median platelet values consistently above the normal whereas the neutrophil counts were normal. It is possible that death resulted from bleeding complications maybe due to an intrinsic platelet dysfunction more evident in the AME population. Peripheral blood smears showed evidence of megakaryocytic fragments, large cytoplasm pieces containing pro-platelet, and nucleus scraps in both groups of mice. Seven out of 12 AME-mice and 8 out of 12 AME-D-C-mice showed slight neuthrophil dysplastic aspects such as hypersegmentation and open nuclei. The red blood cells were normocromic and normocytic in all mice however minimal anisocytosis was observed during the course of disease, with a variation in degree of polycromatophilia at the time of death. The bone core biopsies were normocellular in eight AME mice, three were hypocellular and only one was hypercellular. In contrast the cellularity in all the AME-D-C samples was normal. The most prominent aspects of the AME-mice were an increased number of BM megakaryocytes often grouped in clusters of four to six elements located in proximity of sinuses or in paratrabeculae and rare megakaryocytes that contained intact white cells in their cytoplasm. These two features previously described as endosteal dislocation and emperipolesis, often seen in patients with MPD or ET, were absent in AME-D-C-mice, in which only a minimum increase of megakaryocyte without atypical aspects was observed. No significant differences were found in spleen size, weight and histological features aside from a slight increase in fibrosis in AME-D-C-mice. These results suggest that a mutation in the eighth zinc finger motif and the abrogation of CtBP binding could attenuate the myeloproliferative phenotype caused by AME particularly in terms of histological features and overall survival.

Weed management systems for Norway spruce (Picea abies L.) Christmas trees were investigated under field conditions. Potential alternative methods to replace the use of herbicides included the application of three different soil covers: plastic film (100% recoverable), biodegradable film 40-μ thick, and biodegradable film 70-μ thick. On another group, weed control was carried out mechanically (control plants). Each treatment consisted of 27 plants with three replications. At the time of the first growing season, the percentage of dead and survived plants in treated plants and in the control plants did not differ significantly. Data were recorded on plant height and quality during the second year of growth. Control plants always showed height increases shorter than other plants. Application of these films significantly affected growth and plant quality. In plastic film-treated plants, the average height increases were significantly higher than those observed on control plants mechanically treated (19 vs. 12 cm, respectively), and did not differ from biodegradable film-treated plants (17 cm). Plants responded similarly to the biodegradable films of varying thickness. The results indicate that all three films have potential for use in Christmas tree production. However, the use of biodegradable films would be preferable because they do not need to be removed at the end of the rotation (about 6–7 years). Furthermore, this culture system would be an efficient way to achieve three objectives: 1) to enhance weed control, avoiding the application of herbicides by many Christmas tree producers; 2) to improve quality and plant commercial value; and 3) to obtain a shorter rotation.

The prognostic significance of the response to initial prednisone treatment in adult ALL has been recently emphasized. Prednisone response is usually defined on the basis of the peripheral leukemic blast count. The threshold value for the defintion as good or poor prednisone response is 1000 blasts/mmc on day 8 of prednisone pre-treatment. The drawback of this definition is the difficulty of classifying patients with less than 1000 blasts at diagnosis. In the LAL2000 GIMEMA study we therefore evaluated whether the blast reduction rate, which is not affected by the initial blast level, could be a factor with comparable prognostic value. The protocol design provided a 7-day (−6 to 0) pre-treatment phase with an escalating dose of prednisone up to 60 mg/sqm. On day 1 before starting the induction the response was assessed both according to the absolute blast count (< versus ≥ 1000/mmc) (criterion 1) and according to the blast reduction rate ≥ 75% (criterion 2) in the peripheral blood. The induction included high dose Daunorubicin; for patients in complete remission (CR) this was followed by consolidation with high dose ARA-C, chemo and radio prophylaxis of the central nervous system, and periodical reinduction over a three years maintenance period. Patients with adverse cytogenetic features [i.e. t(9;22), t(4;11), t(1;19)] who achieved a CR were treated according to the HAM protocol that included high dose ARA-C and Mitoxantrone followed by Imatinib for Ph+ ALL and by allogeneic or autologous hemopoietic stem cells transplantation for the others. Between September 2000 and December 2003 a total of 368 patients were evaluable for response to induction. The median age was 35 years (15–60) and median WBC count 15′109/L (0.3–872); 72 (20%) were T ALL and 121 (33%) had cytogenetic high risk features (104 (86%) Ph+, 4 (3%) t(4;11) and 13 (11%) t(1;19)). Eighty-seven percent of the patients were evaluable for response to steroid pre-treatment: ’responders’ were 75% according to criterion 1 (blast =/

The micropropagation of Cycas revoluta Thunb. via somatic embryogenesis was tested on immature and mature embryos using 2,4-dichlorophenoxyacetic acid (2,4-D) in combination with benzyladenine (BA) or kinetin. In addition, the effect of media manipulation on the induction of adventitious buds was studied to optimize culture parameters for the regeneration of plantlets via organogenesis from mature embryos. Schenk and Hildebrandt basal medium containing six different NO3-: NH4+ ratios (from 90:10 to 65:35) and BA at 9 μm was used. The development of induced buds occurred on the medium containing the identical NO3-: NH4+ composition without growth regulators. The effects of two IBA concentrations on the rooting of elongated shoots were evaluated. The treatment of immature embryos with 2.26 μm 2,4-D and 0.44 μm BA and that of mature embryos with 0.9 μm 2,4-D and 4.40 μm BA promoted growth of embryogenic tissue followed by embryo formation that failed to develop further. Bud induction was obtained through the whole range of NO3-: NH4+ ratios tested. Small decrease in these ratios affected subsequent differentiation and growth. Adventitious shoots obtained on 70% NO3- showed two-fold increase in height and diameter compared with those regenerated on 75% NO3-. Furthermore, 7% of the former produced at the shoot base adventitious shoots that have similar morphology to offset growing at the base of mature plant. The rooting percentage of shoots was low. Considering the difficulty in achieving somatic embryogenesis, these results suggested that among micropropagation techniques organogenesis can be applicable for production of Cycas, although rooting is still the limiting factor.

Cycasrevoluta Thunb., cultivated as an ornamental plant for indoor and outdoor use, is characterized by an extremely slow rate of growth. In spite of the occurrence in its coralloid roots of the nitrogen-fixing cyanobiont Nostoc, N fertilization is commonly used to accelerate Cycas growth. A greenhouse experiment was conducted to examine the effects of two forms of combined N on growth of Cycas plants and cyanobacterial nitrogenase activity, measured on intact plants. Cycas plants grown in pots were fertilized from June to September with nutrient solution containing macronutrients as P, K, Mg, and Hoagland's micronutrients. N (700 mg/plant) was supplied as KNO3, or applied as NH4NO3; control plants received nutrient solution without nitrogen. Treatments were applied monthly and repeated for three times. Each treatment consisted of 15 plants. The length of the new leaves was recorded during the growth cycle of the plants. The nitrogenase activity, based on acetylene reducing activity (ARA), was measured on the plants in situ in July and in October. N fertilization stimulated both the nitrogenase activity and the growth of the plants. In comparison with the control, average increases in ARA of more than 20% were observed in the treated plants. Nitrogenase activity was slightly better in the presence of NH4NO3 in July, whereas the values measured in October were about the same for two treatments. The two forms of nitrogen were the same also regarding the stimulus on growth: in N-treated plants the total length of the new leaves was more than double with respect to the control at the end of the growing season. Control plants grown without fertilizer N had a slow start to their growth cycles and were unable to recover and compensate later for the lack of transient N.

This study was aimed to characterize by Raman spectroscopy (excitation line 633 nm) different redox states of the mitochondrial cytochrome c oxidase. The results obtained from a systematic analysis carried out on the mitochondrial enzyme prepared under redox conditions, differently affecting the valence state of the metal prosthetic groups, and a comparison with homologous bacterial heme-copper oxidases, cytochrome c and pyridine hemo-chrome extract revealed a novel redox state marker specifically linked to the redox transition of heme a, peaking at 1645 cm-1, and tentatively assigned to the C=C and/or C=N streching mode of the imidazole ring of a proxymal histidine ligand. The possible involvment of this redox-linked conformational change in the catalytic activity of cytochrome oxidase is discussed.

The composition of ternary Si-B-O films some tens of nm thick, which grow during the thermal pre-deposition of boron in silicon, has been determined by X-ray microanalysis. The X-ray intensity data have been obtained by using an automated ARL-SEMQ microprobe operated at 20 kV. The experimental k ratios have been converted into concentrations using a Monte Carlo computer program based on the single-scattering approach. In the case of a film with an underlying substrate, which includes an element of the compound film, an independent determination of thickness is necessary. A general computer analysis on the effect of the substrate on electron trajectories and X-ray yields is performed; the possible sources of error are discussed. The composition of the films so obtained is in satisfactory agreement with the data previously deduced from Rutherford backscattering experiments. This result allows us to solve a discrepancy between the two analytical techniques found before in the case of substrate removal; a variation in chemical composition of the films during the removal of the substrate may be suggested.

Summary Cryptococcosis and aspergillosis in immunocompromised patients are extremely difficult clinical conditions to manage and treatment with available antifungal drugs often fails. Itraconazole, R-51211, Janssen Pharmaceutica, a new orally absorbed triazole, is a possible alternative drug which is potentially effective and nontoxic. Preliminary experience with 28 patients, eight with cryptococcosis and 20 with aspergillosis, is reported. Of these patients, 16 were immunocompromised (seven with the acquired immune-deficiency syndrome (AIDS), five heart transplant recipients and four with leukaemia or lymphoma). Overall, results of treatment were good (18 in remission, four markedly improved, four moderately improved and two failed). Prevention of relapses of cryptococcosis was obtained in all patients with AIDS on long-term itraconazole monotherapy (3 mg/kg). Treatment of invasive aspergillosis required a higher dosage (about 5 mg/kg) and prolonged administration. Besides its efficacy this antifungal agent allowed outpatient management.

TEM reveals (0001) stacking faults and periodic superstructures in cancrinite-related minerals, afghanite and franzinite, which are interpreted as aperiodic or periodic displacements of the hexagonal [(Si,AI)60~2] ring units. Franzinite also shows a superstructure when the structure is projected onto the xy plane; satellites occur in the hk0 diffraction patterns.

Mont Semiol (also called Mont Semiouse), Montbrison, Loire, France, is the only place where the zeolite offrettite is known to occur. Now, a new mineral, Mazzite, has been found in the same locality. It occurs in the form of tiny needles, and its properties closely match those of the synthetic “molecular sieve” Ω. The chemical formula is Na0.03K1.91Ca1.35Mg1.99[Al9.77Si26.54O72]·28.03 H2O. Lattice dimensions are a=18.392 A, c=7.646 A, with P 63/mmc as most probable space group; Dcalc.=2.108. Offrettite and mazzite occur in association with phillipsite, chabazite, calcite and siderite. The mineral has been so named in honour of Prof. Fiorenzo Mazzi.

A representative series of levyne samples shows a remarkably homogeneous chemical composition revealed by electron microprobe and thermogravimetric analyses (t.g.a.). The lattice parameters, accurately determined by X-ray powder diffractometry and refined by least squares treatment, have weak correlations with selected chemical parameters in accord with their very limited range of variation (average values with e.s.d., a = 1.335(3) nm, c = 2.289(12) nm). The fairly constant chemical and physical parameters of levyne are a distinguishing feature of this zeolite family and may testify to its narrow range of crystallization conditions. Levyne is often intimately associated with erionite and/or offretite, possibly in response to the changes of equilibrium conditions.

Melonite, merenskyite, and moncheite occur as minute grains (40 to less than 5 mu m) dispersed within the main sulfides (pyrrhotite, pentlandite, and chalcopyrite) in sulfide concentrations of the Ivrea-Verbano layered basic complex. Electron microprobe analyses indicate that melonite and merenskyite cover the whole range of Ni-Pd substitution, confirming the sofar inferred existence of a complete solid solution between these phases. Moncheite is much less abundant and always contains significant amounts of Pd and Ni. Substitution of Bi for Te is only occasional. Melonite is frequently found in paragenetic association with hessite, altaite, Bi tellurides, Ni-Co sulfarsenides, Au-Ag alloys (electrum), and irarsite.