Mutant p53 inhibits miRNA biogenesis by interfering with the microprocessor complex.

Downregulation of microRNAs (miRNAs) is commonly observed in cancers and promotes tumorigenesis suggesting that miRNAs may function as tumor suppressors. However, the mechanism through which miRNAs are regulated in cancer, and the connection between oncogenes and miRNA biogenesis remain poorly understood. The TP53 tumor-suppressor gene is mutated in half of human cancers resulting in an oncogene with gain-of-function activities. Here we demonstrate that mutant p53 (mutp53) oncoproteins modulate the biogenesis of a subset of miRNAs in cancer cells inhibiting their post-transcriptional maturation. Interestingly, among these miRNAs several are also downregulated in human tumors. By confocal, co-immunoprecipitation and RNA-chromatin immunoprecipitation experiments, we show that endogenous mutp53 binds and sequesters RNA helicases p72/82 from the microprocessor complex, interfering with Drosha-pri-miRNAs association. In agreement with this, the overexpression of p72 leads to an increase of mature miRNAs levels. Moreover, functional experiments demonstrate the oncosuppressive role of mutp53-dependent miRNAs (miR-517a, -519a, -218, -105). Our study highlights a previously undescribed mechanism by which mutp53 interferes with Drosha-p72/82 association leading, at least in part, to miRNA deregulation observed in cancer.

[Molecular biomarkers in workers and population exposed to inorganic arsenic: preliminary study in vitro]. / Biomarcatori molecolari in lavoratori professionalmente esposti ad arsenico inorganico e nella popolazione generale: studio preliminare in vitro.

The inorganic Arsenic (iAs) is a metalloid widely diffuse in all environmental matrices. The iAs has been classified as a Group 1 carcinogen by the International Agency for Research on Cancer. The microRNAs (miRNAs) are small non coding RNAs that negatively regulate the expression of hundreds of target genes in many key physiological and pathological cell processes, including stress response, differentiation, proliferation, apoptosis and cancer, miRNA expression profiles appear altered in most human cancers and it has been highlighted the potential of miRNA profiling in cancer diagnosis and prognosis. The present study evaluates the effect of iAs exposure on global miRNA expression in Jurkat cells. Treated cells show a reproducible increase in the expression levels of three miRNAs: miR-663, miR-222 and miR-638. This study supports the importance to proceeding in the investigation aimed to the possible application of some miRNAs as biomarkers in the environmental and occupational exposure to iAs.

Combined heart and kidney transplantation: long-term analysis of renal function and major adverse events at 20 years.

BACKGROUND: Combined heart-kidney transplantation (HKTx) is an accepted therapeutic option for patients with end-stage heart disease associated with severely impaired renal function. We report our long-term follow-up with this combined procedure. PATIENTS AND METHODS: Between April 1989 to November 2009, nine patients underwent combined simultaneous (HKTx) at our center. Seven patients were males (mean age 45.2 +/- 10.12 years); seven patients were on dialysis at the time of transplantation. RESULTS: Surgical procedures were uneventful in all patients. One patient died in the intensive care unit 41 days after transplantation. During long-term follow-up, three patients died: one due to infection and multiorgan failure 148 months after HKTx, one due to a lung neoplasm after 6 years, and one, a cerebral stroke at 34 months after transplantation. Only one patient experience renal allograft failure secondary to hypertension and cyclosporine nephrotoxicity at 10 years after HKTx with the need for renal replacement therapy. Last estimated glomerular filtration rates of all other patients was 61.3 +/- 17.4 mL/min. CONCLUSIONS: In selected patients, with coexisting end-stage cardiac and renal failure, combined HKTx with an allograft from the same donor proved to give satisfactory short- and long-term results, with a low incidence of both cardiac and renal allograft complications.

Percutaneous coronary interventions in cardiac allograft vasculopathy: a single-center experience.

OBJECTIVE: Cardiac allograft vasculopathy represents an accelerated form of obstructive coronary disease. It is the main cause of late death following heart transplantation. Percutaneous coronary intervention is considered a palliative procedure due to high restenosis rates. The aim of this study was to review our experience with percutaneous coronary interventions using stents in cardiac transplant recipients. METHODS: The present analysis included all primary adult heart transplanted patients who had been discharged from the hospital after transplantation, had a clinical follow-up of 12 months and underwent percutaneous coronary intervention (PCI). RESULTS: Seventy heart transplanted patients underwent percutaneous revascularization. Our analysis comprised 85 first-vessel procedures resulting in treatment of 135 lesions. The mean time from heart transplantation to first intervention was 9.3 +/- 4.8 years. Primary success was obtained in 96% lesions; at least 1 recurrent stenosis event occurred in 16 patients with primarily successful PCI. Lesions treated with drug-eluting stents experienced recurrent stenosis in 16% of cases. During a mean follow-up after PCI of 45.2 +/- 41.7 months, 27 deaths (19 cardiac) and 1 late re-transplantation occurred after PCI. CONCLUSION: In cardiac transplant recipients, percutaneous coronary intervention with stents can be performed safely with high rates of primary success. Restenosis rates were higher compared with coronary interventions in native coronary arteries. Drug-eluting stents seemed to favorably impact restenosis compared with bare-metal stents. The clinical benefit from percutaneous coronary intervention may be reduced due to disease progression in untreated coronary segments.

[Retrospective comparative analysis of antidementia medication persistence patterns in Spanish Alzheimer's disease patients treated with donepezil, rivastigmine, galantamine and memantine]. / Analisis comparativo de los patrones de persistencia de medicaciones antidemencia en una cohorte retrospectiva de pacientes con demencia de tipo Alzheimer tratados con donepecilo, rivastigmina, galantamina y memantina en Espana.

INTRODUCTION AND AIM: Dementia of Alzheimer type has become the most frequent type of dementia in our environment. Treatment persistence is a crucial factor to delay patient functional and cognitive impairment. The aim of the present study was to determine treatment persistence in usual care settings with four different antidementia drugs: donepezil, rivastigmine, galantamine and memantine in a cohort of patients with Alzheimer's dementia in Spain. PATIENTS AND METHODS: An Alzheimer type dementia retrospective cohort study was performed in 13 Primary Care Health Centers in Spain. The study included patients treated between January 2000 and March 2005. RESULTS: A total of 299 patients (44.8% female), mean age 77.9 years, were included: 101 donepezil (33.8.%), 105 rivastigmine (35.1%), 51 galantamine (17.1%) and 42 memantine (14.0%). Mean treatment duration was significantly different depending on therapy type, showing higher values for donepezil patients (mean: 83.3 weeks; 95% CI: 72.7-93.9) than for the other cholinesterase inhibitors: rivastigmine (mean: 76.6 weeks; 95% CI: 66.0-87.3), galantamine (mean: 65.8 weeks; 95% CI: 55.3-76.3) and memantine (60.9 weeks; 95% CI: 48.8-73.1), p = 0.049. Overall treatment persistence was significantly different between drugs, with again donepezil showing higher persistence (median time: 70.3 weeks; 95% CI: 49.8-90.7) than with the others drugs: rivastigmine (median time: 56.1 weeks; 95% CI: 36.1-76.2), galantamine (median time: 56.7 weeks; 95% CI: 41.1-72.3) and memantine (median time: 52.1 weeks; 95% CI: 35.2-69.1), log-rank = 10.16; p = 0.017. CONCLUSION: This study showed significative differences in the global treatment persistence among the considered drug-cholinesterase inhibitors, showing higher persistence resulting in patients treated with donepezil compared to those who received rivastigmine, galantamine or memantine.

Análisis comparativo de los patrones de persistencia de medicaciones antidemencia en una cohorte retrospectiva de pacientes con demencia de tipo Alzheimer tratados con donepecilo, rivastigmina, galantamina y memantina en España / Retrospective comparative analysis of antidementia medication persistence patterns in spanish alzheimer’s disease patients treated with donepezil, rivastigmine, galantamine and memantine

Introducción y objetivos. La demencia de tipo Alzheimeres la causa más frecuente de demencia en nuestro entorno. La persistenciadel tratamiento es un factor crucial para demorar el deteriorofuncional y cognitivo del paciente. El objetivo de este estudioha sido determinar la persistencia del tratamiento con cuatro medicacionesantidemencia (donepecilo, rivastigmina, galantamina ymemantina) en una cohorte de pacientes con enfermedad de Alzheimeren España. Pacientes y métodos. Se realizó un estudio retrospectivoen 13 centros de atención primaria en España. Los pacientesincluidos fueron tratados entre enero de 2000 y marzo de2005. Resultados. Se incluyeron un total de 299 pacientes (44,8%mujeres) con una edad media de 77,9 años: 101 se trataron condonepecilo (33,8%), 105 con rivastigmina (35,1%), 51 con galantamina(17,1%) y 42 con memantina (14,0%). La duración media deltratamiento fue significativamente diferente entre los anticolinesterásicos(p = 0,049), siendo superior en los pacientes tratados condonepecilo (media: 83,3 semanas; IC 95%: 72,7-93,9), que con elresto de agentes terapéuticos: rivastigmina (media: 76,6 semanas;IC 95%: 66,0-87,3), galantamina (media: 65,8 semanas; IC 95%:55,3-76,3) y memantina (media: 60,9 semanas; IC 95%: 48,8-73,1).La persistencia global del tratamiento fue significativamente diferenteentre los tratamientos (log-rank = 10,16; p = 0,017), siendosuperior con donepecilo (mediana: 70,3 semanas; IC 95%: 49,8-90,7) que con el resto de terapias: rivastigmina (mediana: 56,1 semanas;IC 95%: 36,1-76,2), galantamina (mediana: 56,7 semanas;IC 95%: 41,1-72,3) y memantina (mediana: 52,1 semanas; IC 95%:35,2-69,1). Conclusiones. El estudio ha mostrado diferencias significativasen la persistencia global de los distintos tratamientos antidemenciaconsiderados, siendo superior en aquellos pacientes tratadoscon donepecilo respecto a los tratados con rivastigmina,galantamina o memantina

Introduction and aim. Dementia of Alzheimer type has become the most frequent type of dementia in our environment.Treatment persistence is a crucial factor to delay patient functional and cognitive impairment. The aim of the present studywas to determine treatment persistence in usual care settings with four different antidementia drugs: donepezil, rivastigmine,galantamine and memantine in a cohort of patients with Alzheimer’s dementia in Spain. Patients and methods. An Alzheimertype dementia retrospective cohort study was performed in 13 Primary Care Health Centers in Spain. The study includedpatients treated between January 2000 and March 2005. Results. A total of 299 patients (44.8% female), mean age 77.9 years,were included: 101 donepezil (33.8.%), 105 rivastigmine (35.1%), 51 galantamine (17.1%) and 42 memantine (14.0%). Meantreatment duration was significantly different depending on therapy type, showing higher values for donepezil patients (mean:83.3 weeks; 95% CI: 72.7-93.9) than for the other cholinesterase inhibitors: rivastigmine (mean: 76.6 weeks; 95% CI: 66.0-87.3),galantamine (mean: 65.8 weeks; 95% CI: 55.3-76.3) and memantine (60.9 weeks; 95% CI: 48.8-73.1), p = 0.049. Overalltreatment persistence was significantly different between drugs, with again donepezil showing higher persistence (mediantime: 70.3 weeks; 95% CI: 49.8-90.7) than with the others drugs: rivastigmine (median time: 56.1 weeks; 95% CI: 36.1-76.2),galantamine (median time: 56.7 weeks; 95% CI: 41.1-72.3) and memantine (median time: 52.1 weeks; 95% CI: 35.2-69.1),log-rank = 10.16; p = 0.017. Conclusion. This study showed significative differences in the global treatment persistenceamong the considered drug-cholinesterase inhibitors, showing higher persistence resulting in patients treated with donepezilcompared to those who received rivastigmine, galantamine or memantine

Pharmacokinetics and metabolism in mice of IDN 5390 (13-(N-Boc-3-i-butylisoserinoyl)-C-7,8-seco-10-deacetylbaccatin III), a new oral c-seco-taxane derivative with antiangiogenic property effective on paclitaxel-resistant tumors.

IDN 5390 (13-(N-Boc-3-i-butylisoserinoyl)-C-7,8-seco-10-deacetylbaccatin III) is a new taxane, derived from 7,8-C-seco-10-deacetylbaccatin, selected for its ability to inhibit angiogenesis, mainly by acting on endothelial cell motility, and for its selective activity on class III beta-tubulin. In vivo, IDN 5390 shows activity against paclitaxel-sensitive and -resistant tumors when administered on a prolonged, continuous dosage schedule. We studied the pharmacokinetics and bioavailabilty of the drug in mice after single and repeated oral treatment. IDN 5390 was rapidly absorbed after oral administration, with good bioavailability (43%). After intravenous injection, it was extensively distributed in tissue, mainly the liver, kidney, and heart, with low but persistent levels in brain. The kinetics appear dose-dependent with a clearance of 2.6, 1.4, and 0.9 l/kg at, respectively, 60, 90, and 120 mg/kg, and a half-life 24, 36, and 54 min. After prolonged daily oral doses given for 2 weeks, we found that there was a decrease in drug availability; i.e., the area under the concentration-time curve value after p.o. daily administration on day 14 was 2-fold lower than that on day 1. Metabolism plays a major role in elimination of the drug, and at least 12 metabolites were identified in feces and urine. The percentage excreted as metabolites after an oral dose (42%) was higher than that after the i.v. dose (33%), suggesting a first-pass effect. Four metabolites were found in plasma at detectable levels; one of them, with restored taxane scaffold, is a species 3 times more potent than IDN 5390, possibly contributing to the observed anti-tumor activity.

[Stevens-Johnson syndrome, toxic epidermal necrolysis and phenytoin. Factors linked to a higher risk]. / Síndrome de Stevens-Johnson, necrólisis epidérmica tóxica y fenitoína. Factores asociados a un aumento del riesgo.

INTRODUCTION: Stevens-Johnson syndrome (SJS) and toxic epidermal necrolysis (TEN) are severe, although not very common, clinical skin pictures that are usually related to the use of medication. Several antiepileptic drugs, including phenytoin, have been linked to SJS/TEN. Some authors have described an increased risk for SJS/TEN when phenytoin is associated to radiotherapy, while others report the possibility of an increased risk when it is associated to corticoids. DEVELOPMENT: This work includes a review of the spontaneous reports of suspected cases of phenytoin-linked SJS/TEN recorded in the database of the Pharmacovigilance Department at Pfizer-España between October 2000 and December 2003. Nine cases compatible with SJS/TEN were found; four occurred in cancer patients that had received radiotherapy, three of whom were also treated with corticoids. DISCUSSION AND CONCLUSIONS: After reviewing the spontaneously reported cases in the database of the Pharmacovigilance Department at Pfizer-España as well as the cases in the literature, it can be concluded that when it comes to indicating a prophylactic antiepileptic treatment for cancer patients with cerebral metastasis, the clinician must take into account the existence of a greater risk of SJS/TEN if the patient is going to receive radiotherapy. If the patient already presents a history of skin rashes following administration of an antiepileptic drug, care must be taken in choosing another because phenytoin together with carbamazepine, phenobarbital and lamotrigine have all been linked to SJS/TEN. Cross-sensitivity of carbamazepine and barbiturates with phenytoin has been observed. Gabapentin and valproic acid could be considered as therapeutic options in such cases.

Síndrome de Stevens-Johnson, necrólisis epidérmica tóxica y fenitoína. Factores asociados a un aumento del riesgo / Stevens-Johnson syndrome, toxic epidermal necrolysis and phenytoin. Factors linked to a higher risk

Introducción. El síndrome de Stevens-Johnson (SSJ) y la necrólisis epidérmica tóxica (NET) son cuadros cutáneos graves, aunque poco frecuentes, que en la mayoría de los casos se relacionan con el consumo de fármacos. Varios antiepilépticos se han asociado a SSJ/NET, entre ellos la fenitoína. Algunos autores han descrito un aumento del riesgo de SSJ/NET cuando la fenitoína se asocia a radioterapia, y otros han informado de la posibilidad de un mayor riesgo cuando se asocia al empleo de corticoides. Desarrollo. En este trabajo se realiza una revisión de las notificaciones espontáneas de sospecha de SSJ/NET asociados a fenitoína registradas en la base de datos del Departamento de Farmacovigilancia de Pfizer-España, desde octubre de 2000 hasta diciembre de 2003.Se han recogido nueve casos compatibles con SSJ/NET; cuatro ocurrieron en pacientes oncológicos que habían recibido radioterapia y, además, tres de ellos se trataron también con corticoides.Discusión y conclusiones. Tras revisar los casos de notificación espontánea de la base de datos del Departamento de Farmacovigilancia de Pfizer-España, y tras revisar los casos de la literatura, se concluye que a la hora de indicar un tratamiento antiepiléptico profiláctico en pacientes oncológicos con metástasis cerebrales, debería valorarse la existencia de un mayor riesgo de presentar SSJ/NET si el paciente va a recibir radioterapia. Si el paciente ya presenta antecedentes de reacción cutánea con un antiepiléptico, la elección de otro debe realizarse con precaución, ya que tanto la fenitoína, como la carbamacepina, el fenobarbital y la lamotrigina se han asociado a SSJ/NET, y se ha observado sensibilidad cruzada de la carbamacepina y los barbitúricos con la fenitoína. La gabapentina y el ácido valproico podrían considerarse como opciones terapéuticas en estos casos (AU)

Introduction. Stevens-Johnson syndrome (SJS) and toxic epidermal necrolysis (TEN) are severe, although not very common, clinical skin pictures that are usually related to the use of medication. Several antiepileptic drugs, including phenytoin, have been linked to SJS/TEN. Some authors have described an increased risk for SJS/TEN when phenytoin is associated to radiotherapy, while others report the possibility of an increased risk when it is associated to corticoids. Development. This work includes a review of the spontaneous reports of suspected cases of phenytoin-linked SJS/TEN recorded in the database of the Pharmacovigilance Department at Pfizer-España between October 2000 and December 2003. Nine cases compatible with SJS/TEN were found; four occurred in cancer patients that had received radiotherapy, three of whom were also treated with corticoids. Discussion and conclusions. After reviewing the spontaneously reported cases in the database of the Pharmacovigilance Department at Pfizer-España as well as the cases in the literature, it can be concluded that when it comes to indicating a prophylactic antiepileptic treatment for cancer patients with cerebral metastasis, the clinician must take into account the existence of a greater risk of SJS/TEN if the patient is going to receive radiotherapy. If the patient already presents a history of skin rashes following administration of an antiepileptic drug, care must be taken in choosing another because phenytoin together with carbamazepine, phenobarbital and lamotrigine have all been linked to SJS/TEN. Cross-sensitivity of carbamazepine and barbiturates with phenytoin has been observed. Gabapentin and valproic acid could be considered as therapeutic options in such cases (AU)

Mechanical circulatory support in severe heart failure: single-center experience.

Ventricular assist devices (VADs) have become important therapeutic tools to treat patients with end-stage cardiac failure. VADs are an essential component of transplantation programs as they successfully bridge individuals who would otherwise die. Recently left ventricular VAD (LVAD) therapy has been proposed as alternative to heart transplantation (HTx) for patients who are not transplant candidates. Other indications have now expanded into areas such as postcardiotomy failure, acute myocarditis, and acute massive myocardial infarction. From 1988 to May 2003, 80 patients received left or biventricular mechanical circulatory support including 78 as a bridge to and two as an alternative to HT. All patients survived the operation. Mean duration of VAD support was 77 +/- 150 days. Fifty-one points (63.8%) underwent heart transplantation; 3 (3.8%) recovered and were weaned from VADs. Major bleeding episodes occurred in 11 patients (13.8%) and major neurologic events occurred in 8 (10%). Sixteen patients (20%) were discharged home while waiting for HTx. Twenty-two patients (27.5%) died on VAD. In conclusion, VAD therapy proved effective in bridging patients with end-stage heart failure to HTx. While on LVAD support patients who were assisted with implantable wearable devices could be discharged at home, improving their quality of life.

Impella recover 100 microaxial left ventricular assist device: the Niguarda experience.

BACKGROUND: The Impella Recover 100 (IR100) is an intravascular microaxial blood pump used to support blood circulation for a maximum of 7 days in cases of reduced left ventricular function, for example in postcardiotomy low output syndrome or in cardiogenic shock after acute myocardial infarction. MATERIALS AND METHODS: We supported five patients with the IR100. The mean age, cardiac index (CI), and ejection fraction (EF) of our population were 42 years, 1.83 L/min/m(2), and 20%, respectively. Two patients (group A) with ischemic dilated cardiomyopathy were bridged to heart transplant. Two patients (group B) with fulminan myocarditis and septic shock were bridged to recovery. One patient, with severe valvular cardiomyopathy who underwent aortic valve replacement and mitral valve annuloplasty, was supported to weaning from ECC. RESULTS: Mean support time was 9.8 +/- 2.3 days. Only one acute myocarditis patient died from a severe vasoplegic syndrome despite maximal inotropic and vasoactive support. Both group A patients were successfully transplanted. Among group B, the second patient resolved the septic status and was slowly weaned from the device and discharged home with moderate improvement of LV function (EF = 40%). Patient C was weaned from the IR100 and electively placed on the heart transplant recipient list. CONCLUSIONS: IR100 is a device that in our experience can be utilized for various indications for short-term support. In compromised patients where a traditional LVAD is contraindicated, the IR100 showed good results, for it is minimally invasive and does not need ECC or systemic anticoagulation.

The combination of yondelis and cisplatin is synergistic against human tumor xenografts.

Yondelis (trabectidin, ET-743) is a marine natural product that has shown activity both in preclinical systems and in human malignancies such as soft tissue sarcoma and ovarian cancers that are resistant to previous chemotherapies. Molecular pharmacological studies indicated that Yondelis interacts with DNA and DNA repair systems in a way that is different from Cisplatin (DDP). The current study was designed to investigate the effects of the combination of Yondelis and DDP in human cancer cell lines and in xenografts derived from different tumours. The in vitro studies performed in human TE-671 rhabdomyosarcoma, Igrov-1 and 1A9 human ovarian carcinoma cell lines showed additive effects or slight synergism. Several human tumour xenografts, such as TE-671 rhabdomyosarcoma, SK-N-DX neuroblastoma, FADU head and neck, LX-1 non-small cell lung cancer (NSCLC), H-187 melanoma and SKOV HOC 8 ovarian carcinoma, showed an antitumour effect for the combination that was greater than that of each drug when given as a single agent. No consistent changes in the activity were observed if Yondelis and DDP were given 1 h apart in sequence or simultaneously. An orthotopically transplanted human ovarian cancer HOC 8 growing in the peritoneal cavity of nude mice was used that is insensitive to Yondelis alone and only moderately sensitive to DDP alone. The combination of the two drugs produced a dramatic increase of survival lasting several months. In conclusion, the combination of Yondelis and DDP is synergistic in vivo (i.e. the antitumour effect is greater than that of each drug used as a single agent at the maximum tolerated dose (MTD)) in different human tumour xenografts. The two drugs can be combined at the MTD of each drug, thus indicating there are no overlapping toxicities. These results provide a rationale for testing the combination of Yondelis and DDP in the clinic.

Combined surgical approach to coexistent carotid and coronary artery disease: early and late results.

BACKGROUND: The management of patients with simultaneous coronary artery and carotid artery disease is still controversial. METHODS: A retrospective review of the records and follow-up data of 139 consecutive patients undergoing simultaneous coronary artery bypass graft and carotid endarterectomy from 1981 to 1999 was carried out. RESULTS: Early mortality was 2.1%, perioperative myocardial infarction and stroke rates were 2.8 and 1.4%, respectively. Survival at 7 years was 74.7+/-5.1% and event-free survival at 7 years was 67.9+/-5.6%. CONCLUSIONS: The combined surgical approach has proved to be effective and safe allowing the treatment of both diseases in a single operative procedure.

High-performance liquid chromatographic assay for the determination of the novel C-Seco-taxane derivative (IDN 5390) in mouse plasma.

A HPLC assay was developed to determine IDN 5390, a new paclitaxel analogue, in mouse plasma. The method involves solid-phase extraction from cyano cartridges (recovery >80%), HPLC separation on Symmetry C(18) (4.6 x 150 mm), on isocratic mobile phase of water-acetonitrile-acetic acid (49:50:1) and detection at 227 nm. Retention times of IDN 5390 and IDN 5517 (internal standard, I.S.) were 9.1 and 10.5 min, respectively. The assay was linear from 0.05 to 5 micro g/ml (r(2)>or=0.995), showed intra- and inter-day precision within 1.0 and 6.2%, and accuracy of 94.7-106.8%. LOQ was 0.050 micro g/ml. Using this method IDN 5390 pharmacokinetics was determined in mice.

High-performance liquid chromatography/tandem mass spectrometry for the quantitative analysis of a novel taxane derivative (BAY59-8862) in biological samples and characterisation of its metabolic profile in rat bile samples.

A sensitive, specific, accurate and reproducible high-performance liquid chromatography (HPLC) analytical method was developed and validated for the quantification of the novel oral taxane analogue BAY59-8862 in mouse plasma and tissue samples. A fully automated solid-phase extraction procedure was applied to the plasma after internal standard (IS) addition, with only 0.2 mL volume of the sample loaded on a CN-Sep-pak cartridge. In the case of the tissues a very simple acetonitrile extraction was used to recover BAY59-8862 and its internal standard from liver. The procedure for the quantification of BAY59-8862 and its IS (IDN5127) is based on high-performance liquid chromatography/ion spray-tandem mass spectrometry, operating in selected ion monitoring mode. The retention times of BAY and IS were 7.21 and 10.36 min, respectively. In both plasma and tissue specimens the assay was linear in the range 50-5000 ng/mL (ng/g). The overall precision and accuracy were assessed on three different days. The results for plasma were within 6.1% (precision) and between 99 and 112% (accuracy), and for the liver samples within 7.3% and between 104 and 118%, respectively. The LOD was 5 ng/mL and 20 ng/g in the plasma and liver, respectively. In addition, the biliary excretion of the compound in rats was studied. The study showed that an oxidative chemical reaction was the preferred metabolic pathway for biliary excretion, and two sets of mono- and dihydroxylated metabolites were detected by LC/ISP-MS/MS experiments. With this method, BAY59-8862 pharmacokinetic was determined in mice. The combined results demonstrate that the methodology can be considered a valid approach to conduct pharmacokinetic and metabolic studies during preclinical and clinical investigations.

First successful Italian clinical experience with DeBakey VAD.

The growing number of patients waiting for heart transplantation more than tripled between 1989 and 1998. Various non-pulsatile mechanical circulatory support devices have been developed as bridge to heart transplantation in recent years. We report the first successful Italian clinical experience with an axial-flow pump, DeBakey VAD, in a patient supported as bridge to transplantation for 55 days.

[Left monoventricular assistance with DeBakey VAD continuous flow pump: initial clinical Italian experience]. / Assistenza monoventricolare sinistra con pompa a flusso continuo DeBakey VAD: prima esperienza clinica italiana.

BACKGROUND: The shortage of heart donors causes a rise in mortality among candidates for cardiac transplantation and increases the waiting list. Consequently mechanical circulatory support for bridge to transplant is now a standard clinical procedure utilized in the most representative cardiac surgery centers. Recently, continuous-axial-flow pumps have been introduced in the clinical practice and have led to new perspectives. METHODS: Four patients suffering from end-stage heart failure were implanted with a DeBakey ventricular assist device (VAD) continuous-flow pump as a bridge to heart transplant. The DeBakey VAD is smaller than the pulsatile devices commonly employed, the pump is totally implantable and is connected to a small controller and two batteries by a transcutaneous drive line. RESULTS: One patient died of multiorgan failure during assistance; 3 patients were fully rehabilitated and were successfully transplanted after 55, 42 and 141 days respectively. In the early postoperative period the mean pump flow was 4.27 +/- 0.55 l/min, after 1 week of assistance the flow rose to 5.32 +/- 0.57 l/min and then progressively increased to 5.83 +/- 0.57 l/min. CONCLUSIONS: This experience demonstrated the possibility of continuous-flow left ventricular support with the DeBakey VAD for mid-term mechanical ventricular assistance. This pump presents new interesting aspects and opens new perspectives for the future of left ventricular mechanical assistance. Increasing experience will define the role of this device in the scenario of heart failure.

Tricuspid regurgitation secondary to mitral valve disease: tricuspid annulus function as guide to tricuspid valve repair.

METHODS: A prospective analysis was performed on 50 patients (pts) with rheumatic mitral disease and associate secondary tricuspid insufficiency who underwent mitral valve replacement from January 1995 to December 1998. Surgical indication to tricuspid annuloplasty was considered in patients with echocardiographic tricuspid annulus diameter > 21 mm/m2, regardless semiquantitative evaluation of tricuspid insufficiency. De Vega annuloplasty was performed in 33 out of 50 patients. RESULTS: Hospital mortality was 2.0% (CL 0.3-3.6). The follow up of the discharged patients ranged from 3 to 48 months (mean 25 +/- 15.9). Three late deaths occurred (6.1% CL 2.8-9.2). Forty-two patients out of the 46 followed up (91.3% CL 84.9-93.8) were in I or II NYHA class. In eight patients (16.3% of discharged patients) the obtained result has been considered as 'negative late results': persisting moderate (three cases) or moderate-severe (five cases) TrI, together with congestive heart failure requiring a furosemide intake of > 25 mg/day. No patients had severe TrI at follow up. The statistics analysis demonstrated the 'preoperative fraction shortening of the tricuspid annulus' (P = 0.038) as factor predictive of late negative result. The incidence of late negative result was 57.1% among patients with fractional shortening lower than 25% and 0% among those patients with fractional shortening greater than 25% (P = 0.0001). CONCLUSIONS: The choice to treat the tricuspid insufficiency according to indexed tricuspid annulus dimension (> 21 mm/m2) has been effective in terms of clinical efficacy and of late functional result. Fractional shortening of the tricuspid annulus, expression of right ventricular cardiomyopathy in patients with poorest prognosis, affects the postoperative evolution of tricuspid insufficiency.

[What treatment should be given for multiple sclerosis at the present time?]. / Qué tratamiento debemos emplear actualmente en la esclerosis múltiple?

OBJECTIVE: This communication aims to describe an approach suitable for the general neurologist or neurologist specialized in treating other disorders, to the current treatment of multiple sclerosis. DEVELOPMENT AND CONCLUSIONS: We discuss the treatments for recovery from the symptoms of an acute attack and those which modify the natural course of the illness (reduce the frequency and severity of attacks and/or prevent their progression). The attacks are treated with corticosteroids or ACTH. Both treatments have been shown on clinical trials to cause rapid improvement of the acute symptoms of an attack. In the progressive forms, the usefulness of high doses of corticosteroids has not been shown. Nor is there evidence that long term corticosteroid treatment, either daily or monthly, is of use in reducing the number of attacks or progression of the disease, although serious side-effects have been seen. At the moment, the interferons are the most popular treatment for multiple sclerosis. They have been the first drugs to modify the course of the disorder. Finally, we describe some of the most generally used treatments and some under investigation, although they are not widely used since it is still not clear exactly how they affect the course of the disease.