A venetoclax and azacitidine bridge-to-transplant strategy for NPM1-mutated acute myeloid leukaemia in molecular failure.

NPM1-mutated acute myeloid leukaemia (NPM1mut AML) represents a mostly favourable/intermediate risk disease that benefits from allogeneic haematopoietic stem cell transplantation (HSCT) in case of measurable residual disease (MRD) relapse or persistence after induction chemotherapy. Although the negative prognostic role of pre-HSCT MRD is established, no recommendations are available for the management of peri-transplant molecular failure (MF). Based on the efficacy data of venetoclax (VEN)-based treatment in NPM1mut AML older patients, we retrospectively analysed the off-label combination of VEN plus azacitidine (AZA) as bridge-to-transplant strategy in 11 NPM1mut MRD-positive fit AML patients. Patients were in MRD-positive complete remission (CRMRDpos ) at the time of treatment: nine in molecular relapse and two in molecular persistence. After a median number of two cycles (range 1-4) of VEN-AZA, 9/11 (81.8%) achieved CRMRD -negative (CRMRDneg ). All 11 patients proceeded to HSCT. With a median follow-up from treatment start of 26 months, and a median post-HSCT follow-up of 19 months, 10/11 patients are alive (1 died from non-relapse mortality), and 9/10 patients are in MRDneg status. This patient series highlights the efficacy and safety of VEN-AZA to prevent overt relapse, achieve deep responses and preserve patient fitness before HSCT, in patients with NPM1mut AML in MF.

Simulating the selfing and migration of Luehea divaricata populations in the Pampa biome to investigate the conservation potential of their genetic resources.

Computer simulations are an important tool for developing conservation strategies for forest species. This study used simulations to investigate the genetic, ecological, and reproductive patterns that contribute to the genetic structure of the tree Luehea divaricata Mart. & Zucc. in five forest fragments in the Brazilian Pampa biome. Using the EASYPOP model, we determined the selfing and migration rates that would match the corresponding genetic structure of microsatellite marker data (based on observed and expected heterozygosity parameters). The simulated reproductive mode was mixed, with a high rate of outcrossing (rate = 0.7). This was consistent with a selfing-incompatible system in this species, which reduced, but did not prevent, selfing. The simulated migration rate was 0.02, which implied that the forest fragments were isolated by distance, and that the inbreeding coefficients were high. Based on Nei's gene diversity analysis, 94% of the genetic variability was distributed within the forest fragments, and only 6% of the genetic diversity was caused by differences between them. Furthermore, the minimum viable population and minimum viable area genetic conservation parameters (which determine conservation potential in the short and long term) suggested that only the Inhatinhum forest fragment had the short-term potential to maintain its genetic diversity. However, in the long term, none of the forest fragments proved to be sustainable, indicating that the populations will require intervention to prevent a decline in genetic variability. The creation of ecological corridors could be a useful solution to connect forest fragments and enhance gene flow between them.

Outcome of severe placental insufficiency with abnormal umbilical artery Doppler prior to fetal viability.

OBJECTIVE: To evaluate the outcome of pregnancies complicated by placental insufficiency and abnormal umbilical artery Doppler prior to viability. DESIGN: A retrospective cohort study. SETTING: Italy. POPULATION: Singleton pregnancies with fetal growth restriction and absence of end-diastolic velocities (AEDVs) in the umbilical arteries prior to 24 weeks. METHODS: A retrospective cohort study of singleton pregnancies with fetal growth restriction and AEDVs in the umbilical arteries prior to 24 weeks. MAIN OUTCOME MEASURES: Fetal growth restriction and AEDVs in the umbilical arteries prior to 24 weeks. RESULTS: Of 16 fetuses first seen at 20-23 weeks, only 12 survived and one of these developed cerebral palsy. Severe hypertensive disorders occurred in three mothers. In four women, the Doppler waveforms progressively improved and developed a normal pulsatility. These fetuses had a better outcome than those that had persistent alterations: they were delivered later (34 versus 28 weeks), had a larger birthweight (1598 versus 630 g) and developed fewer complications. CONCLUSIONS: Placental insufficiency with AEDV in the umbilical arteries prior to fetal viability is associated with a high probability of perinatal death and neonatal complications. However, progressive amelioration of Doppler indices occurs in a subset of women, and these fetuses have a much better outcome.

Occurrence of septic shock in a patient submitted to emergency cerclage following a negative amniocentesis: report of a case.

BACKGROUND: Second trimester emergency cerclage is an option for pregnant women presenting bulging fetal membranes. Despite a significant prolongation of pregnancy might be achieved, serious fetal and maternal events have been reported. Exclusion of infections through preprocedure amniocentesis has been proposed. METHODS: A 37-year-old woman, gravida 4 para 1, was admitted at 21 weeks of gestation to our University Hospital due to bulging fetal membranes. An amniocentesis was performed in order to exclude an actual amniotic infection. Our Microbiology Department found a negative amniotic culture for bacteria and Mycoplasma and a normal glucose and interleukin-6 level, so a cervical cerclage was performed. The patient was discharged home on oral erythromycin. RESULTS: After 48 h, the patient complained of hyperpyrexia, shivers and reduced fetal movements. Ultrasound at admission showed absent cardiac activity and after cerclage removal a non-viable fetus was delivered vaginally. Piperacillin and tazobactam were started, but the clinical course of the patient deteriorated and she developed a cold septic shock and was submitted to hysterectomy and transferred to the ICU of our hospital. CONCLUSION: This report heralds that even after negative amniocentesis, a life-threatening infection may not be excluded in women candidate for emergency cerclage due to bulging fetal membranes.

Metal-related artifacts in instrumented spine. Techniques for reducing artifacts in CT and MRI: state of the art.

The projectional nature of radiogram limits its amount of information about the instrumented spine. MRI and CT imaging can be more helpful, using cross-sectional view. However, the presence of metal-related artifacts at both conventional CT and MRI imaging can obscure relevant anatomy and disease. We reviewed the literature about overcoming artifacts from metallic orthopaedic implants at high-field strength MRI imaging and multi-detector CT. The evolution of multichannel CT has made available new techniques that can help minimizing the severe beam-hardening artifacts. The presence of artifacts at CT from metal hardware is related to image reconstruction algorithm (filter), tube current (in mA), X-ray kilovolt peak, pitch, hardware composition, geometry (shape), and location. MRI imaging has been used safely in patients with orthopaedic metallic implants because most of these implants do not have ferromagnetic properties and have been fixed into position. However, on MRI imaging metallic implants may produce geometric distortion, the so-called susceptibility artifact. In conclusion, although 140 kV and high milliamperage second exposures are recommended for imaging patients with hardware, caution should always be exercised, particularly in children, young adults, and patients undergoing multiple examinations. MRI artifacts can be minimized by positioning optimally and correctly the examined anatomy part with metallic implants in the magnet and by choosing fast spin-echo sequences, and in some cases also STIR sequences, with an anterior to posterior frequency-encoding direction and the smallest voxel size.

Advances in clinical NK cell studies: Donor selection, manufacturing and quality control.

Natural killer (NK) cells are increasingly used in clinical studies in order to treat patients with various malignancies. The following review summarizes platform lectures and 2013-2015 consortium meetings on manufacturing and clinical use of NK cells in Europe and United States. A broad overview of recent pre-clinical and clinical results in NK cell therapies is provided based on unstimulated, cytokine-activated, as well as genetically engineered NK cells using chimeric antigen receptors (CAR). Differences in donor selection, manufacturing and quality control of NK cells for cancer immunotherapies are described and basic recommendations are outlined for harmonization in future NK cell studies.

Efficient presentation of tumor idiotype to autologous T cells by CD83(+) dendritic cells derived from highly purified circulating CD14(+) monocytes in multiple myeloma patients.

To generate mature and fully functional CD83(+) dendritic cells derived from circulating CD14(+) cells highly purified from the leukapheresis products of multiple myeloma patients.CD14(+) monocytes were selected by high-gradient magnetic separation and differentiated to immature dendritic cells with granulocyte-macrophage colony-stimulating factor and interleukin-4 for 6-7 days and then induced to terminal maturation by the addition of tumor necrosis factor-alpha or stimulation with CD40 ligand. Dendritic cells were characterized by immunophenotyping, evaluation of soluble antigens uptake, cytokine secretion, capacity of stimulating allogeneic T cells, and ability of presenting nominal antigens, including tumor idiotype, to autologous T lymphocytes. Phenotypic analysis showed that 90% +/- 6% of cells recovered after granulocyte-macrophage colony-stimulating factor and interleukin-4 stimulation expressed all surface markers typical of immature dendritic cells and demonstrated a high capacity of uptaking soluble antigens as shown by the FITC-dextran assay. Subsequent exposure to maturation stimuli induced the downregulation of CD1a and upregulation of CD83, HLA-DR, costimulatory molecules and induced the secretion of large amounts of interleukin-12. Mature CD83(+) cells showed a diminished ability of antigen uptake whereas they proved to be potent stimulators of allogeneic T cells in a mixed lymphocyte reaction. Monocyte-derived dendritic cells, pulsed before the addition of maturation stimuli, were capable of presenting soluble proteins such as keyhole limpet hemocyanin and tetanus toxoid to autologous T cells for primary and secondary immune response, respectively. Conversely, pulsing of mature (CD83(+)) dendritic cells was less efficient for the induction of T-cell proliferation. More importantly, CD14(+) cells-derived dendritic cells stimulated autologous T-cell proliferation in response to a tumor antigen such as the patient-specific idiotype. Moreover, idiotype-pulsed dendritic cells induced the secretion of interleukin-2 and gamma-interferon by purified CD4(+) cells. T-cell activation was better achieved when Fab immunoglobulin fragments were used as compared with the whole protein. When dendritic cells derived from CD14(+) cells from healthy volunteers were analyzed, we did not find any difference with samples from myeloma patients as for cell yield, phenotypic profile, and functional characteristics. These studies demonstrate that mobilized purified CD14(+) cells represent the optimal source for the production of a homogeneous cell population of mature CD83(+) dendritic cells suitable for clinical trials in multiple myeloma.

Transforming growth factor beta3 inhibits chronic myelogenous leukemia hematopoiesis by inducing Fas-independent apoptosis.

OBJECTIVE: Transforming growth factor beta3 (TGF-beta3) is a potent suppressor of human hematopoietic progenitor cells. In this article, we compare the activity of TGF-beta3 on highly purified CD34+ cells and more immature CD34-DR(-) cells from chronic myelogenous leukemia (CML) patients in chronic phase and normal donors. MATERIALS AND METHODS: Primitive hematopoietic progenitors were stimulated in liquid cultures and clonogenic assays by early-acting growth factors such as stem cell factor (SCF) and interleukin 11 (IL-11) and the intermediate-late-acting stimulating factors IL-3, granulocyte-macrophage colony-stimulating factor, and erythropoietin. Molecular analysis of bcr/abl mRNA was performed on single CML colonies by nested reverse transcriptase polymerase chain reaction. Moreover, cell cycle analysis and assessment of apoptosis of normal and leukemic CD34+ cells were performed by propidium iodide (PI) alone and simultaneous staining with annexin V and PI, respectively. RESULTS: The colony-forming efficiency of CML CD34+ cells was generally inhibited by more than 90% regardless of whether the colony-stimulating factors were used alone or combined. When compared to normal CD34+ cells, leukemic cells were significantly more suppressed in 6 of 8 culture conditions. The inhibitory effect of TGF-beta3 on CD34+ cells was exerted within the first 24 hours of incubation as demonstrated by short-term preincubation followed by IL-3-and SCF-stimulated colony assays. Evaluation of bcr/abl transcript on residual CML colonies incubated with TGF-beta3 demonstrated a small subset of neoplastic CD34+ cells unresponsive to the inhibitory effect of the study cytokine. TGF-beta3 demonstrated a greater inhibitory activity on primitive CD34+DR cells than on more mature CD34+ cells. Again, CML CD34+DR(-) cells were significantly more inhibited by TGF-beta3 than their normal counterparts in 3 of 8 culture conditions. Kinetic analysis performed on CD34+ cells showed that TGF-beta induces cell cycle arrest in G(1) phase. However, this mechanism of action is shared by normal and leukemic cells. Conversely, TGF-beta3 preferentially triggered the programmed cell death of CML CD34-cells without increasing the proportion of leukemic cells coexpressing CD95 (Fas receptor), and this effect was not reversed by functional blockade of Fas receptor. Conclusion. We demonstrate that TGF-beta3 exerts a potent suppressive effect on CML cells that is partly mediated by Fas-independent apoptosis.

Mast cells in bronchiolitis obliterans organizing pneumonia. Mast cell hyperplasia and evidence for extracellular release of tryptase.

Idiopathic bronchiolitis obliterans organizing pneumonia (BOOP) is characterized by air space inflammation and fibrosis of unknown origin. The pathogenesis of the inflammatory reaction and fibrosis in fibrotic lung disorders remains unclear; however, recent attention has focused on the potential role of the mast cell in the genesis of fibrosis. To determine whether mast cells are implicated in the pathogenesis of BOOP, mast cells were identified in BAL fluid and in transbronchial lung biopsy specimens from 11 patients affected by BOOP and 17 control subjects. Mast cells and tryptase were significantly increased in BAL fluid of patients with BOOP (p = 0.001 and p = 0.03, respectively). In lung tissue of patients with BOOP, there was an increased number of mast cells per square millimeter of lung tissue with respect to control group (p = 0.001). Seventy-three percent of mast cells were found in the alveolar septa, 18% within alveoli often plunged in organizing pneumonia, 4% among alveolar lining cells, and 6% along blood vessels. No mast cells were located within alveoli in control subjects. Mast cell degranulation was evident in lung tissue specimens of patients with BOOP but not in those of control subjects (p = 0.01). This study shows the importance of mast cells and mast cell activation in the pathogenesis of BOOP.

Autologous transplantation of chemotherapy-purged PBSC collections from high-risk leukemia patients: a pilot study.

We have recently demonstrated that the combination of the alkylating agent nitrogen mustard (NM) and etoposide (VP-16) is capable of eliminating, ex vivo, leukemic cells contaminating PBSC collections and this is associated with a significant recovery of primitive and committed hematopoietic progenitor cells. Based on these data a pilot study on autologous transplantation of NM/VP-16 purged PBSC for high-risk leukemic patients was recently initiated. Twelve patients (seven females and five males) with a median age of 46 years (range 18-57) have been treated. Two patients had acute myeloblastic leukemia (AML) resistant to conventional induction treatment, four patients had secondary AML in I complete remission (CR), one patient was in II CR after failing a previous autologous BM transplantation, while two additional AML individuals were in I CR achieved after three or more cycles of induction treatment. Two patients with high-risk acute lymphoblastic leukemia (ALL) in I CR and one patient with mantle cell lymphoma and leukemic dissemination were also included. Eight patients showed karyotypic abnormalities associated with a poor clinical outcome. The mobilizing regimens included cytosine arabinoside and mitoxantrone with (n = 6) or without fludarabine (n = 3) followed by subcutaneous administration of G-CSF (5 microg/kg/day until the completion of PBSC collection) and G-CSF alone (n = 3) (15 microg/kg/day). A median of two aphereses (range 1-3) allowed the collection of 7.2 x 10(8) TNC/kg (range 3.4-11.5), 5 x 10(6) CD34+ cells/kg (range 2.1-15.3) and 9.2 x 10(4) CFU-GM/kg (0.3-236). PBSC were treated with a constant dose of 20 microg of VP-16/ml and a median individual-adjusted dose (survival < or = 5% of steady-state BM CFU-GM) of NM of 0.7 microg/ml (range 0.25-1.25). Eleven patients were reinfused after busulfan (16 mg/kg) and Cy (120 mg/kg) conditioning with a median residual dose of 0.3 x 10(4) CFU-GM/kg (0-11.5). The median time to neutrophil engraftment (>0.5 x 10(9)/l) for evaluable patients was 25 days (range 12-59); the median time to platelet transfusion independence (>20 and >50 x 10(9)/l) was 40 days (18-95) and 69 days (29-235), respectively. Hospital discharge occurred at a median of 25 days (18-58) after stem cell reinfusion. Four individuals are alive in CR (n = 3) or with residual nodal disease (n = 1 lymphoma patient) with a follow-up of 32, 26, 3 and 14 months, respectively. Seven patients died due to disease progression or relapse (n = 5) or extrahematological transplant toxicity (n = 2). Our data suggest that pharmacological purging of leukapheresis collections of leukemic patients at high-risk of relapse is feasible and ex vivo treated cells reconstitute autologous hematopoiesis.

Overwhelming post-splenectomy infection with Plesiomonas shigelloides in a patient cured of Hodgkin's disease. A case report.

The immune deficiencies of Hodgkin's disease persist to some degree even after the patients are clinically cured; these may be amplified by loss of splenic immunologic functions after staging laparotomy and splenectomy. The authors submit a case report wherein a bacterium of relatively low virulence, Plesiomonas shigelloides, was associated with a rapidly fulminant septicemia, disseminated intravascular coagulation, Waterhouse-Friderichsen syndrome, and death in a splenectomized patient free of Hodgkin's disease for approximately five years. This emphasizes the need for prolonged observation, rapid diagnosis, and aggressive intervention in immunocompromised patients, especially those supposedly cured of previous hematologic malignancy.

Dendritic cell differentiation from hematopoietic CD34+ progenitor cells.

Dendritic cells (DC) are the most powerful antigen presenting cells (APC) and play a pivotal role in initiating the immune response. In light of their unique properties, DC have been proposed as a tool to enhance immunity against infectious agents and in anticancer vaccine strategies. In the last few years, the development of DC has been extensively investigated. The present paper summarizes the most recent findings on the differentiation of myeloid DC from hematopoietic CD34+ progenitors and methods for DC generation in vitro. A better understanding of DC function has important implications for their use in clinical settings.

Two years' air mutagenesis monitoring in a northwestern rural area of Italy with an industrial plant.

The mutagenicity of organic extracts from inhalable airborne particles, collected in a northwestern rural area of Italy in which an industrial plant producing chemical intermediates is present, was assessed during the years 1989 and 1990. The Ames plate test with Salmonella strains TA98 and TA100 with and without metabolic activation was used. Eight sites in the first and three sites in the second year were monitored once and twice a month respectively. Results show that the mutagenicity of air particulate matter reaches maximum values in the cold months and is not dependent on plant activities. In addition, a correlation analysis between mutagenicity data and number of vehicles seems to indicate traffic emissions as the main source of mutagens.

Hypertension in a group of centenarians.

The aim of the present study was to evaluate the frequency of hypertension in a group of extremely old persons: 73 centenarians resident in Calabria were studied, aged 100-110 years, (mean age 102.95 +/- 2.0), 54 women (73.9%) and 19 men (26.1%), in the period January 1st and April 30th, 1994. All the centenarians were visited at their homes. Hypertensive centenarians were 14 (19.1%), women 12 (85.7%) and men 2 (14.3%). Even if blood pressure progressively increases with age, we have not found it in centenarians. The lesser frequency of hypertension in centenarians certainly depends on genetic and constitutional factors, but also their way of life and alimentary habits (decreased sodium intake, increase of calcium and potassium intake), as we have proven in a recent work, could be determinant. Moreover, the presence of hypertensive centenarians might mean that hypertension does not prevent to reach an age by far above the average, if it is the only risk factor.

A case of Paget's disease in an ultracentenarian subject with endoprosthesis of Moore type without cementation.

Paget's disease is an osteopathy the incidence of which increases with aging; in the 8th decade, it occurs in 11% of the population. Complications of this pathology often occur, and appear prevalently in form of basilar disease. We report a case of an ultracentenarian patient (aged 107) who suffered a cervical right femur fracture after falling in her own house at the age of 104 years. Radiological and laboratory findings revealed a mono-osseal Paget's disease of the right femur. Because of her good conditions, the patient underwent an operation of non-cemented endoprosthesis of Moore type, with a good post-operatory course and a physiokinetic therapeutic treatment. A kinesiologic evaluation was made, by measuring articular range of motion (ROM) through the neutral zero methods and evaluating the muscular strength: it showed a constantly progressive functional recovery of the lower limbs in both subjective and objective terms.

Aspects of sleep in centenarians.

Sleeping habits of 48 Calabrian centenarians (12 males, 36 females) were evaluated. Their average age was 102 +/- 1.87 years (range 100-107 years). Quantitative and qualitative aspects of sleep were recorded, such as the length of sleeping (hrs), the time of falling asleep (regular or variable) and the time of awakening (regular or variable). Moreover, we recorded and included in the global hours an eventual afternoon sleeping, during a 24-hour-period. The quality of sleeping was classified as: night sleeping, and morning sleeping; and for the subjects were grouped as short sleepers (subjects that fall asleep easily), and long sleepers (subjects with problems to fall asleep). We also considered if sleeping was uninterrupted or interrupted during the night and if the subjects followed particular habits to facilitate the sleeping (sleeping pills, alcohol, etc.). This study demonstrated that all the examined centenarians go to sleep early in the evening, have no problems in falling asleep, wake up early in the morning, take a nap in the afternoon and do not take pills before going to bed. Among the environmental factors, the quality, the quantity and the habits of sleeping might have great influence for longevity.

[Vaginal delivery following previous cesarean section]. / Il parto vaginale nelle donne precesarizzate.

BACKGROUND AND AIM: The increased use of cesarean section (CS) over the past 30 years has raised a problem which has been much debated in obstetric practice: is it always necessary to repeat CS in women who have previously undergone cesarean section? The aim of this study was to establish whether women previously undergoing CS can start trial labour? METHODS: The authors examined 195 pregnant women who had previously undergone 1 or 2 CS. Medical history, clinical examination and maternal and fetal monitoring techniques were used to select the women who could start trial labour.

[Estrogen and progestin treatment in postmenopause. Effects on the endometrium]. / Trattamento con estrogeni e progestinici in età post-menopausale. Effetti sull'endometrio.

BACKGROUND AND AIMS: It has been well demonstrated that menopausal disorders resulting from the lack of estrogens may be delayed and partly eliminated by appropriate hormone replacement therapy. The aim of this study was to demonstrate the absolute innocuity of the association of estrogen and progestin. METHODS: The authors used an association of conjugated equine estrogens and medroxi-progesterone acetate to treat menopausal syndrome in 80 women aged between 48 and 55 years old. The study lasted 2 years. RESULTS AND CONCLUSIONS: The results obtained showed the good tolerability and marked efficacy of the drugs used, as well as the absolute innocuity in relation to the target organs, namely the breast and endometrium. No cases of endometrial hyperplasia with atypical development or malignant evolution were reported during the study and there was a significant improvement in lipid metabolism, vasomotor or psychomotive disorders, as well as increased trophism of the genitourinary tract and skeleton.