In Vitro Apoptotic Effects of Farnesyltransferase blockade in Acute Myeloid Leukemia Cells.

Farnesyltransferase inhibitors (FTIs) are a class of oral anti-cancer drugs currently tested in phase I-II clinical trials for treatment of hematological malignancies. The in vitro effects of various FTIs (alpha-hydroxyfarnesylphosphonic acid, manumycin-A and SCH66336) were tested on CD34+ KG1a cell line and in primary acute myeloid leukemia (AML) cells from 64 patients. By cell viability and clonogeneic methylcellulose assays, FTIs showed a significant inhibitory activity in CD34+ KG1a and primary bone marrow (BM) leukemic cells from 56% of AML patients. FTIs also induced activation of caspase-3 and Fas-independent apoptosis, confirmed by the finding that inhibition of caspase-8 was not associated with the rescue of FTI-treated cells. We concluded that other cellular events induced by FTIs may trigger activation of caspase-3 and subsequent apoptosis, but the expression of proapoptotic molecules, as Bcl-2 and Bcl-XL, and antiapoptotic, as Bcl-X(s), were not modified by FTIs. By contrast, expression of inducible nitric oxide synthase (iNOS) was increased in FTI-treated AML cells. Our results suggest a very complex mechanism of action of FTIs that require more studies for a better clinical use of the drugs alone or in combination in the treatment of hematological malignancies.

Prolonged complete hematologic response in relapsed/refractory T-large granular lymphocyte leukemia after bendamustine treatment.

T-large granular lymphocyte leukemia (T-LGLL) is a chronic clonal proliferation of effector memory cytotoxic CD3+CD57+CD56- T cells and the current guidelines suggest immunosuppressive therapy as first-line therapy, but the treatment of refractory/relapsed patients is still challenging due to the lack of prospective studies. We describe a series of two refractory/relapsed T-LGLL patients successfully treated with bendamustine, a chemotherapeutic agent largely used for B-cell neoplasms, but poorly investigated for the treatment of T-cell diseases. Complete remission (CR) was achieved in 3 and 6 months, respectively, and maintained for at least 20 months. One patient relapsed after a 20-month CR, but she was responsive to bendamustine therapy again, obtaining a further prolonged CR. Bendamustine as single agent or in combination could be a feasible therapeutic option in refractory/relapsed T-LGLL, especially for elderly patients because of its safety profile.

OPSI threat in hematological patients.

Overwhelming post-splenectomy infection (OPSI) is a rare medical emergency, mainly caused by encapsulated bacteria, shortly progressing from a mild flu-like syndrome to a fulminant, potentially fatal, sepsis. The risk of OPSI is higher in children and in patients with underlying benign or malignant hematological disorders. We retrospectively assessed OPSI magnitude in a high risk cohort of 162 adult splenectomized patients with malignant (19%) and non malignant (81%) hematological diseases, over a 25-year period: 59 of them splenectomized after immunization against encapsulated bacteria, and 103, splenectomized in the previous 12-year study, receiving only life-long oral penicillin prophylaxis. The influence of splenectomy on the immune system, as well as the incidence, diagnosis, risk factors, preventive measures and management of OPSI are also outlined. OPSI occurred in 7 patients (4%) with a median age of 37 years at time interval from splenectomy ranging from 10 days to 12 years. All OPSIs occurred in non immunized patients, except one fatal Staphylococcus aureus -mediated OPSI in a patient adequately immunized before splenectomy. Our analysis further provides evidence that OPSI is a lifelong risk and that current immune prophylaxis significantly decreases OPSI development. Improvement in patients' education about long-term risk of OPSI and increased physician awareness to face a potentially lethal medical emergency, according to the current surviving sepsis guidelines, represent mandatory strategies for preventing and managing OPSI appropriately.

Accelerated bone mass senescence after hematopoietic stem cell transplantation.

Osteoporosis and avascular necrosis (AVN) are long-lasting and debilitating complications of hematopoietic stem cell transplantation (HSCT). We describe the magnitude of bone loss, AVN and impairment in osteogenic cell compartment following autologous (auto) and allogeneic (allo) HSCT, through the retrospective bone damage revaluation of 100 (50 auto- and 50 allo-HSCT) long-term survivors up to 15 years after transplant. Current treatment options for the management of these complications are also outlined. We found that auto- and allo-HSCT recipients show accelerated bone mineral loss and micro-architectural deterioration during the first years after transplant. Bone mass density (BMD) at the lumbar spine, but not at the femur neck, may improve in some patients after HSCT, suggesting more prolonged bone damage in cortical bone. Phalangeal BMD values remained low for even more years, suggesting persistent bone micro-architectural alterations after transplant. The incidence of AVN was higher in allo-HSCT recipients compared to auto-HSCT recipients. Steroid treatment length, but not its cumulative dose was associated with a higher incidence of bone loss. Allo-HSCT recipients affected by chronic graft versus host disease seem to be at greater risk of continuous bone loss and AVN development. Reduced BMD and higher incidence of AVN was partly related to a reduced regenerating capacity of the normal marrow osteogenic cell compartment. Our results suggest that all patients after auto-HSCT and allo-HSCT should be evaluated for their bone status and treated with anti-resorptive therapy as soon as abnormalities are detected.

HRCT in detection of pulmonary infections from nontuberculous mycobacteria: personal experience.

PURPOSE: The authors sought to assess the role of high-resolution computed tomography (HRCT) in the detection and follow-up of nontuberculous mycobacteria (NTM) pulmonary infection in immunocompetent patients and to identify the most common radiological patterns for diagnosis. MATERIALS AND METHODS: Plain chest radiographs and HRCT scans of 42 consecutive patients with NTM pulmonary infection (M/F 26/16; mean age 57, range 41-83) were retrospectively reviewed. Ten of these patients were followed up for 18 months after diagnosis. Small nodules (<10 mm), nodules 10- to 30-mm in diameter, lobar/segmental consolidation, cavitations, bronchiectasis and tree-in-bud pattern were analysed. RESULTS: Small nodules were more frequent than nodules 10- to 30-mm in diameter, and segmental consolidation was more frequent than lobar. Cavitations, tree-in-bud and bronchiectasis were more frequently located in the upper lobes. Four of the followed-up patients had cavitation of preexisting nodules, and five had progression of bronchiectasis. CONCLUSIONS: HRCT allows accurate detection and followup of the most frequent presentation patterns: diffuse small nodules, bronchiectasis, upper lobe segmental consolidation and cavitations. The appearance of new bronchiectasis and progression of old disease are due to pulmonary infection.

Surface antigens of human mesangial cells: impact of growth surface or IL-1alpha.

The interactions of mesangial cells (MC) with their environment are important events in glomerular physiology and pathology, yet a detailed characterization of the MC-surface antigens mediating these interactions is still lacking. In this study, a comparative phenotype analysis of primary human MC in culture using 191 monoclonal antibodies directed against 108 antigens was performed by flow-cytometry. The MC were grown on three different surfaces (human matrix, fibronectin, polystyrene) and cultured in the presence or absence of IL-1alpha. Seventy-one antibodies recognizing 35 different antigens (integrins: CD29, 49b, 49c, 49e, 51, 61; immunoglobulin gene family: CD54, 58, 90, 106, 146, 147, 166; growth factor receptors: CD105, 140b; apoptosis related: CD95; hemostatis related: CD141, 142; miscellaneous: CD44, 109, 138, 151, 157, 165, and 11 nonclustered antigens) reacted with mesangial cells. CD58, 109, 146, 147, 151, 157, 165, and 166 are reported for the first time to be present on human mesangial cells. In comparison to growth on polystyrene, CD44, 54, 95, 105, 109, 140b, 146, 147, 157, 165 and 166, were up-regulated on fibronectin, and CD44, 54, 90, 95, 105, 106, 109, 138, 140b, 141, 142, 146, 147, 151, 157, 165 and 166 were up-regulated on human matrix. The stimulation by IL-1alpha up-regulated CD44, 49e, 51, 54, 61, 106 on MC on polystyrene; CD49e, 51, 61, 106, 146, 165 on MC on fibronectin, and CD49e, 51, 54 on MC grown on human matrix. This analysis of surface antigen expression provides new information to enable a better understanding of the role of mesangial cells in glomerular pathophysiology.

Intrauterine insemination with frozen donor sperm. Pregnancy outcome in relation to age and ovarian stimulation regime.

BACKGROUND: The success of intrauterine insemination with donor semen (IUI-DI) is likely to be influenced by a number of variables, including age and ovarian stimulation (OS) regime. METHODS AND RESULTS: A retrospective analysis of 1056 treatment cycles in 261 women (212 single heterosexuals and 49 lesbians) was conducted to assess the influence of these two variables on pregnancy outcome during IUI-DI. The overall pregnancy rate was 10.6%, being 18.5% for women <35 years, 11.9% in women 35-40 years and 5.4% in women >40 years (P < 0.05). The cumulative pregnancy rate (CPR) after eight cycles was 0.86, 0.51 and 0.32 respectively (P < 0.05). A total of 445 inseminations were performed following spontaneous ovulation, 360 following OS with clomiphene citrate (CC) and 251 with hMG, the pregnancy rate per cycle being 13, 7.2 and 11.2% respectively. There was no statistically significant difference in the pregnancy rate per cycle, CPR or multiple pregnancy rate in the three treatment groups. CONCLUSIONS: These results indicate that the use of OS with either CC or hMG in women without ovulatory dysfunction does not improve the pregnancy rate during IUI-DI. The only factor associated with reduced effectiveness of fertility treatment was age, confirming that IUI is a poor treatment option for women >40 years of age.

The effects of atrial fibrillation on functional recovery in post-stroke patients.

PURPOSE: This study evaluates the negative effects of atrial fibrillation (AF) on after stroke recovery and its relationship with age and other concomitant diseases. METHOD: One hundred and ninety-nine consecutive inpatients, after their first stroke, were enrolled and entered a rehabilitation program lasting 60 days. The mean interval since the stroke was 19 days. The disability, both at admission and at discharge, was assessed by Functional Independence Measure (FIM). Atrial fibrillation was diagnosed at admission on the basis of electrocardiographic recordings. Gender, age, blood pressure and blood sugar level were also taken into account. RESULTS: Stroke patients suffering from AF, when compared to those lacking AF, have higher disability at admission as well as at discharge and had a lesser gain in functional recovery. No statistical difference of the AF occurrence was found between male and female patients. The negative correlation between age and functional gain was found only in AF patients. There was also a significant association between AF and hypertension, but not between AF and diabetes. At the beginning and at the end of the study, the median FIM scores of all the patients with AF were quite similar irrespective of manifesting hypertension, diabetes or neither of these two. CONCLUSIONS: The presence of AF has a negative prognostic value on post stroke outcome, particularly in the elderly patients. The pattern of recovery of stroke subjects having AF is not influenced by concomitant diseases like hypertension or diabetes.

Biocompatibility of a cuprophane charcoal-based detoxification device in cirrhotic patients with hepatic encephalopathy.

Extracorporeal detoxification has been proposed to treat patients with hepatic encephalopathy (HE) not responding to standard therapy. To investigate the biocompatibility of a cuprophane charcoal-based detoxification device, a prospective, randomized, controlled study was performed. Of 41 consecutive patients with cirrhosis and HE grade II or III who did not improve with conventional treatment, 20 patients (median age, 56 years; range, 33 to 71 years; 13 men) were randomly assigned to either ongoing conventional treatment or one additional 6-hour treatment with a sorbent suspension dialysis system. Main outcome parameters were physiological function and blood parameters of biocompatibility. In the 10 patients undergoing combined conventional and sorbent suspension dialysis treatment, blood pressure remained unchanged and body temperature and heart rate increased (P: < 0.01). Platelet count decreased (medians, from 75 to 26 g/L; P: < 0.001) and international normalized ratio increased after combined treatment (2.0 to 2.2; P: < 0.001). Three patients developed bleeding complications during treatment or shortly after. Treated patients showed increases in levels of plasma elastase (104 to 586 microg/L; P: = 0.001), tumor necrosis factor-alpha (5.4 to 7.5 pg/mL; P: = 0.04), and interleukin-6 (118 to 139 pg/mL; P: = 0.04), but not interferon-gamma and E-selectin. No changes were observed in the 10 patients treated conventionally. In conclusion, despite technical refinements compared with charcoal hemoperfusion, biocompatibility of sorbent suspension dialysis is still very limited. Clinical complications were apparently caused by blood-membrane interactions and disseminated intravascular coagulation. We suggest further developments in design and appropriate strategies of anticoagulation to improve the biocompatibility of artificial liver support.

Intrauterine donor insemination in single women and lesbian couples: a comparative study of pregnancy rates.

The outcome of intrauterine donor insemination (IUI-DI) with frozen spermatozoa was analysed retrospectively in 675 cycles in single women (n = 122; 536 cycles) and lesbian (n = 35; 139 cycles) couples. The lesbian patients were younger at the initiation of treatment (mean 34.5 years; range 26-44) than the single women (mean 38.5; range 29-47) (P = 0.005). Clinical pregnancy rate was 36% in single women and 57% in lesbians (P < 0.05), the cumulative pregnancy rate after six cycles being 47% and 70% respectively, although the outcome was similar when related to age. The miscarriage rate was higher (35%) in single women than in lesbians (15%; P < 0.05), the rate being independent of maternal age. There were no apparent differences seen between the two groups with respect to the possible effect of parity, duration of infertility, causes of infertility and type of treatment at initiation of treatment; the sole exception was that the age of lesbian women was statistically significantly younger than that of single women (P < 0.005). When corrected for age, the pregnancy rates and complications were lower and higher respectively in single women but these differences did not reach statistical significance. However, the disparity between the treatment outcomes of single women and lesbian patients of similar ages may also reflect the fact that single women are likely to have failed to conceive for a period of time prior to referral to a specialist centre for treatment.

Endothelial cell adhesion molecule and PMNL response to inflammatory stimuli and AGE-modified fibronectin.

BACKGROUND: Atherosclerotic vascular disease is the leading cause of death in patients with diabetes mellitus and end-stage renal disease. Advanced glycation end products (AGEs) are strongly suggested to be involved in the pathogenesis of atherosclerosis in these patients who also frequently experience infectious complications. We hypothesized that the interaction of AGEs and inflammatory mediators contributes to the up-regulation of endothelial cell activation. METHODS: We investigated the effect of advanced glycated fibronectin in the presence or absence of inflammatory stimuli on the endothelial cell surface and mRNA expression of cell adhesion molecules. Furthermore, the influence of advanced glycated fibronectin on the transendothelial migration pattern of polymorphonuclear cells was analyzed. RESULTS: Exposure to advanced glycated fibronectin together with inflammatory stimuli such as interleukin (IL)-1alpha, tumor necrosis factor-alpha (TNF-alpha) or lipopolysaccharide (LPS) led to a significant increase in the surface expression of the cell adhesion molecules E-selectin, ICAM-1, VCAM-1 and PECAM-1 on endothelial cells. Soluble AGEs in combination with advanced glycated fibronectin significantly enhanced the endothelial cell surface expression of ICAM-1, VCAM-1 and PECAM-1, whereas this was not the case for E-selectin. At the transcriptional level short-time exposure of endothelial cells to advanced glycated fibronectin and inflammatory mediators resulted in an increased expression of E-selectin, ICAM-1 and VCAM-1 mRNA levels, whereas PECAM-1 repeatedly showed a significant decrease of gene transcript levels. An increase of mRNA levels was also observed for E-selectin, ICAM-1, VCAM-1 and PECAM-1 following incubation with a combination of advanced glycated fibronectin and soluble advanced glycation end-products. Furthermore, polymorphonuclear cells responded with a sevenfold increase in transendothelial migration following exposure of endothelial cells to advanced glycated fibronectin and inflammatory mediators. CONCLUSIONS: These results suggest that the combination of matrix glycation and inflammation up-regulates the activation of the endothelial cell adhesion cascade, a mechanism that might contribute to the increased burden of atherosclerotic morbidity and mortality in patients suffering from diabetes mellitus or chronic renal failure.

Evaluation of a rapid protocol for the assessment of salt sensitivity against the blood pressure response to dietary sodium chloride restriction.

The "gold standard" for the assessment of salt sensitivity of hypertension is the blood pressure response to dietary NaCl restriction; nevertheless, for practical purposes, a more rapid test that would not depend on the patient's compliance to the dietary prescription would be very useful in clinical research and medical practice. The aim of this study was thus to evaluate the effectiveness and reliability of a rapid, easy-to-standardize protocol for the assessment of salt sensitivity against the blood pressure response to dietary salt restriction. A total of 108 hypertensive patients were screened for salt sensitivity by the modified protocol of Grim et al. Thereafter, nine patients identified by the test as salt sensitive and nine identified as salt resistant followed, for two consecutive periods of 1 week, a diet with normal (200 mmol/day) or low (50 mmol/day) NaCl content. Compliance to the diet was checked by repeated 24-h urine collections. The group as a whole experienced a significant fall in blood pressure during the low Na diet (mean pressure = 123 +/- 3 v 118 +/- 3 mm Hg; P < .05). However, whereas patients identified as salt sensitive by the Grim protocol had a marked and significant blood pressure decrease (systolic -12 mm Hg, diastolic -7 mm Hg), no change was observed in those classified as salt resistant (systolic -2 mm Hg, diastolic -2 mm Hg). A significant correlation between changes in urinary Na excretion and changes in blood pressure was found only in salt-sensitive hypertensive patients. In conclusion, the modified Grim protocol tested in this study was able to correctly predict a significant blood pressure response to dietary salt restriction in the majority of cases. A validation of this test in a larger patient population may be advisable.

NaCl sensitivity of essential hypertensive patients is related to insulin resistance.

OBJECTIVE: To evaluate insulin sensitivity of essential hypertensive patients with different salt sensitivities of blood pressure in the absence of confounding factors such as obesity, glucose intolerance and the inclusion both of normotensive and of hypertensive subjects that have affected most previous studies. PATIENTS: Ninety-nine patients with untreated mild or moderate essential hypertension, World Health Organization class I-II, participated in the study. METHODS: Salt sensitivity was estimated using the Weinberger protocol with minor modifications and the patients were classified into tertiles of salt sensitivity. RESULTS: Patients with high NaCl sensitivities were slightly older and had somewhat higher blood pressures than did subjects with low salt sensitivities. Plasma renin activity significantly decreased with increasing salt sensitivity. There were no differences among the three groups in terms of body mass index, fasting blood glucose and insulin plasma levels. There were no differences among the groups in the integrated glucose and insulin response to a standard oral-glucose tolerance test However, there was a significant difference in insulin sensitivity between two subgroups of the upper and lower tertile of salt sensitivity, the salt-sensitive hypertensives having a markedly lower utilization of glucose than did the salt-resistant ones, with a minor overlap (5.4 +/- 0.6 versus 7.4 +/- 0.3 mg/kg per min, P < 0.01). CONCLUSIONS: This study showed that essential hypertensive patients with high NaCl sensitivities were relatively insulin resistant compared with those with low NaCl sensitivities, independently of confounding factors such as age, obesity and glucose intolerance. Insulin resistance was not associated with overt hyperinsulinaemia among these patients.

[Ogilvie syndrome. Report of two cases]. / Sindrome di Ogilvie. Descrizione di due casi.

Ogilvie's syndrome or intestinal acute pseudo-obstruction is a rare form of intestinal obstruction, and is characterised by a dilatation of the distal colon and often of the tenue. The etiology of this syndrome is recognized as a block of sacral parasympathetic after abdominal surgery. We report two cases of post-cesarean section Ogilvie's syndrome verified at "Ospedale S. Maria del Popolo degli Incurabili", in March 1991 and June 1993 both resolved with a new surgery, after failure of the medical and the parendoscopic decompression treatment.

Antihypertensive and renal effects of acute and chronic therapy with a dihydropyridine Ca-antagonist in patients with different salt sensitivity.

We evaluated the effect of the dihydropyridine Ca-channel blocker nitrendipine on blood pressure (BP) and electrolyte urinary excretion after acute and chronic therapy in 33 patients with different NaCl sensitivity as assessed by a modification of the test of Grim and colleagues. Acute nitrendipine administration significantly reduced BP in the group as a whole, although the hypotensive effect was greater in patients with greater NaCl sensitivity; this difference was still evident after 1 month of chronic therapy. Furthermore, urinary sodium and calcium excretion significantly increased in the 3 h after nitrendipine administration during both acute and chronic therapy: these effects on electrolyte excretion were independent of the NaCl sensitivity of the subject.

Effect of pulsed ultrasound on the frequency of sce and on cell proliferation

Os efeitos de doses agudas e crônicas-ultra-som pulsado-foram estudados em linfócitos de três indivíduos do sexo masculino e três do sexo feminino, observando-se a frequência de trocas entre cromátides irmäs e o índice de proliferaçäo celular nas várias fases do ciclo celular. A diminuiçäo da frequência de células M2 em culturas de 48 horas (controle = 1.01;G1=0.48;S=0.29;1%<5%) tratadas com doses crônicas de ultra-som e a ausência total dessas células em culturas tratadas com doses agudas recomendam cautela no uso de ondas ultra-sônicas com finalidades terapêuticas. O tempo de cultura (42 e 72 horas) näo modificou a frequência de trocas entre cromátides irmäs tanto no grupo tratado pelo ultra-som como no grupo controle