Do NPM1 and FLT3-ITD mutations modify prognosis in patients treated with non-intensive regimens?

FLT3-ITD and NPM1 mutations are key to defining the genetic risk profile of acute myeloid leukemia (AML). We aimed to assess the prognostic features of the FLT3-ITD and NPM1 mutations in old and/or unfit individuals with AML treated with non-intensive therapies in the era before azacitidine-venetoclax approbation. The results of various non-intensive regimens were also compared. We conducted a retrospective analysis that included patients treated with different non-intensive regimens, between 2007 and 2020 from PETHEMA AML registry. We compiled 707 patients with a median age of 74 years and median follow-up time of 37.7 months. FLT3-ITD patients (N = 98) showed a non-significant difference in overall survival (OS) compared to FLT3-ITD negative-patients (N = 608) (P = 0.17, median OS was 5 vs 7.3 months respectively). NPM1-mutated patients (N = 144) also showed a non-significant difference with NPM1 wild type (N = 519) patients (P = 0.25, median OS 7.2 vs 6.8 respectively). In the Cox regression analysis neither NPM1 nor FLT3-ITD nor age were significant prognostic variables for OS prediction. Abnormal karyotype and a high leukocyte count showed a statistically significant deleterious effect. Azacitidine also showed better survival compared to FLUGA (low dose cytarabine plus fludarabine). NPM1 and FLT3-ITD seem to lack prognostic value in older/unfit AML patients treated with non-intensive regimens other than azacitidine-venetoclax combination.

Identifying prognostic gene panels in acute myeloid leukemia.

INTRODUCTION: Acute myeloid leukemia (AML) is a heterogeneous disease currently including 12 entities defined by genetic findings with remarkable differences in prognosis and targeted therapies availability. Therefore, identification of genetic abnormalities by efficient techniques has become a necessary tool in routine clinical practice for AML patients. AREAS COVERED: In the present review, we will focus on our current knowledge of relevant prognosis gene mutations in AML, as recently updated by European Leukemia Net Leukemia risk classification. EXPERT OPINION: About 25% of newly diagnosed younger AML patients will be promptly classified as favorable prognosis by demonstrating the presence of NPM1 mutations or CBF rearrangements by qRTPCR, allowing for implementing molecular measurable residual disease-guided chemotherapy-based protocols. In fit AML patients, rapid detection of FLT3ITD is mandatory to associate midostaurin or quizartinib to treatment and assignment to intermediate prognosis. Conventional cytogenetics and FISH still have a role for detection adverse prognosis karyotypes and KMT2A, MECOM, or NUP98 gene rearrangements. Further genetic characterization is performed with NGS panels including favorable prognosis gene CEBPA bZIP and adverse prognosis genes, such as TP53 and myelodysplasia associated genes.

Awareness of obstetric warning signs in Ecuador: a cross-sectional study.

OBJECTIVE: In Ecuador, the reported maternal death rate was 45.71 per 100,000 live births in 2013. This may be partly due to a lack of maternal knowledge of obstetric warning signs during pregnancy, delivery and the post-partum period. This study sought to evaluate awareness of obstetric warning signs among pregnant women in relation to individual demographic and area-level socio-economic indicators. STUDY DESIGN: We conducted a cross-sectional analysis of data collected by Ecuador's Ministry of Health at the conclusion of a national maternal health campaign (2014-2015). A nationally representative sample of 3435 pregnant women from the nine administrative zones completed surveys regarding basic demographics and their awareness of obstetric warning signs. METHODS: We defined eight obstetrical warning signs according to the literature and Ecuadorian practice that could occur during pregnancy, delivery and the post-partum period (severe headache, strong abdominal ache, bleeding or presence of malodorous secretion, rupture of the amniotic sac, high fever, abnormal presentation of the baby, decrease in baby's movements and delayed labour). A woman was considered 'aware' if she recognised at least four of the eight warning signs and stated she would seek immediate healthcare at their presentation. For each administrative zone, four socio-economic indicators (poverty, illiteracy, unemployment and subemployment) were obtained from the National Institute of Statistics and Census. Correlates of awareness of the obstetric warning signs were evaluated using hierarchical logistic models clustered by the administrative zone. RESULTS: Nationally, 86.9% of women were 'aware' of obstetric warning signs. After adjustment for age, socio-economic indicators and clustering, indigenous participants were 59% less likely to be aware of obstetric warning signs than mestizos (odds ratio [OR] = 0.41, 95% confidence interval [CI] = 0.28-0.59). For every 1% increase in area poverty, participants had a 5% decreased likelihood of being aware of obstetric warning signs (OR = 0.95, 95% CI = 0.93-0.96), adjusting for age, ethnicity and other socio-economic indicators. The most effective source of campaign information about obstetric warning signs was personal communication with a healthcare professional, as opposed to mass media advertisements (OR = 1.90, 95% CI = 1.34-2.71). CONCLUSIONS: A majority of Ecuadorian pregnant and post-partum women are aware of obstetric warning signs. Indigenous ethnicity and area-level poverty are associated with a lack of awareness. Personal communication with a healthcare professional was the most effective source of information. These findings have implications for improvement of maternal awareness of warning signs.

Achromobacter xylosoxidans bacteremia: clinical and microbiological features in a 10-year case series.

OBJECTIVE: The treatment of Achromobacter xylosoxidans bacteremia is challenged by antimicrobial resistance and the paucity of data. We aimed at offering a contemporary description of this uncommon entity. METHODS: Retrospective case series of 13 episodes of A. xylosoxidans bacteremia diagnosed over a 10-year period (November 2007 to May 2017) in our tertiary care center. RESULTS: Solid organ cancer and heart failure were the most common comorbidities (4/13 [30.7%]). All but one episodes were hospital-acquired. Most patients had received previous antibiotic therapy (7/13 [53.8%]) and had a central venous catheter in place (6/13 [46.1%]). Primary and intravascular catheter were the most common sources (4/13 [30.7%] each). Meropenem was the agent with best in vitro activity (92.3% [12/13] of susceptible isolates). All-cause 30-day mortality (overall 23.1%) was higher in patients with primary bacteremia (50.0% vs. 11.1%; P-value=0.203) and prior chemotherapy (66.7% vs. 10.0%; P-value=0.108). CONCLUSIONS: Bacteremia due to A. xylosoxidans constitutes a serious infection among immunocompromised hosts. Carbapenem-based therapy may be appropriate in most cases.

Factores de riesgo para estancia hospitalaria prolongada después de colecistectomía laparoscópica electiva / Risk factors for prolonged hospital stay after elective laparoscopic cholecystectomy

Objetivo: Conocer los factores asociados a estancia hospitalaria prolongada en pacientes sometidos a colecistectomía laparoscópica electiva en población mexicana. Material y método: Estudio retrospectivo descriptivo en pacientes adultos sometidos a colecistectomía laparoscópica electiva. Se midieron las variables preoperatorias: edad, sexo, cirugía abdominal previa, ecografía vesicular; intraoperatorias: adherencias, perforación vesicular incidental, sangrado transquirúrgico, conversión quirúrgica, tiempo anestésico, tiempo quirúrgico, experiencia del cirujano, drenaje; postoperatorias: náusea o vómito, dolor, sangrado, reintervención. La relación entre cada factor perioperatorio y la estancia hospitalaria se analizó mediante la prueba t de Student para variables continuas y Chi cuadrado para variables categóricas. Resultados: Se incluyeron 384 pacientes, 334 mujeres (87 %) y 50 hombres (13 %), con edad media de 40,6 años. A 227 pacientes (59,1 %) se les dio el alta antes de las 24 horas y 157 (40,9 %) recibieron hospitalización de más de 24 horas. Se identificó la asociación de varios factores con estancia hospitalaria prolongada: edad > 65 años (p = 0,0034), pared vesicular > 3 mm (p = 0,0224), adherencias intraoperatorias (p < 0,0001), perforación vesicular incidental (p = 0,0001), tiempo quirúrgico mayor a 60 min (p < 0,00001), cirujano residente (p = 0,0093) y presencia de drenaje (p < 0,0001). Conclusiones: Los factores asociados a estancia prolongada en nuestra serie son edad, pared vesicular engrosada, adherencias, tiempo quirúrgico, perforación incidental y colocación de drenaje

Aim: To determine the main factors associated with longer hospital stay in patients undergoing laparoscopic cholecystectomy in the Mexican population. Material and methods: Retrospective descriptive study in adult patients undergoing elective laparoscopic cholecystectomy. The preoperative variables were age, sex, previous abdominal surgery, gallbladder ultrasound; intra-operative: adhesions, incidental gallbladder perforation, surgical bleeding, surgical conversion, anesthetic time, surgical time, surgeon experience, drainage; postoperative: nausea or vomiting, pain, bleeding, re-intervention. The relationship between each perioperative factor and the hospital stay was analyzed using Student’s t-test for continuous variables and chi-square for categorical variables. Results: We included 384 patients, 334 women (87 %) and 50 men (13 %), with an average age of 40.6 years. 227 patients (59.1 %) were discharged before 24 hours and 157 (40.9 %) received hospitalization of more than 24 hours. The association of several factors with prolonged hospital stay was identified: age > 65 years (p = 0.0034), gallbladder wall> 3 mm (p = 0.0224), intra-operative adhesions (p < 0.0001), incidental gallbladder perforation (p = 0.0001), surgical time greater than 60 min (p < 0.00001), resident surgeon (p = 0.0093), and presence of drainage (p < 0.0001). Conclusions: The factors associated with prolonged stay in our series are age, thickened gallbladder wall, adhesions, surgical time, incidental perforation and drainage placement

Lich-Gregoir technique and routine use of double J catheter as the best combination to avoid urinary complications in kidney transplantation.

INTRODUCTION: Urinary complications in kidney transplantation cause patient morbidity and can decrease graft survival. Most of the complications stem from the vesicoureteric anastomosis. Different techniques for ureteroneocystostomy (UNC) have been designed to avoid these complications. The routine use of a double J catheter after the anastomosis has the same purpose. Our aim was to show our experience and compare the use or non-use of a double J catheter and different techniques for ureteric reimplantation with the rate of urologic complications. MATERIAL AND METHODS: We conducted a retrospective, nonrandomized study of 1011 renal transplantations performed between July 1985 and April 2012. We recorded the surgical techniques for UNC, the use or non-use of a double J catheter, and urinary complications (ureteric fistulae and strictures). The first 700 kidney transplantations (group A) were performed using several UNC techniques (Taguchi, Leadbetter-Politano, and Lich-Gregoir) with a selective use of double J catheter according to the criteria of the surgeon. In the last 311 patients (group B), a surgical technique was established (Lich-Gregoir), as well as the universal use of double J catheters. RESULTS: Urinary fistula occurred in 7% of group A patients and 2% of group B patients (P = .0001). Ureteric stricture occurred in 5.3% of patients in group A and 3% of group B patients (P = .09). In our study, routine prophylactic stenting combined with the Lich-Gregoir UNC technique has decreased the incidence of postoperative fistulae. CONCLUSIONS: Individually, the use of double J catheters seems to lessen the incidence of fistulae, although statistically, the difference is not significant. Accurate knowledge of the complications rates, recommendations of guidelines, and the early diagnosis are essential to attaining reasonable results in kidney transplantation.

Síndrome de Felty: Presentación de un caso y revisión bibliográfica / Feltys syndrome: A case report and literature review

El síndrome de Felty se caracteriza por reunir la tríada compuesta por: artritis reumatoide, neutropenia y esplenomegalia. Es una enfermedad autoinmune poco frecuente, con compromiso sistémico, articular y extra articular. Se desarrolla en personas de mediana edad, con historia de artritis reumatoide crónica deformante. El diagnóstico es eminentemente clínico y el tratamiento está enfocado a disminuir el dolor articular, las altas tasas de infecciones y evitar las deformidades óseas. Presentamos una paciente de 69 años de edad, diagnosticada en nuestro hospital, motivo por el que realizamos revisión bibliográfica de la entidad.(AU)

Feltys syndrome has such as main feature the triad composed by: rheumatoid arthritis, neutropenia and splenomegaly. It is unusual autoimmune disease that compromises the nervous system as well as joint affectation and extra joint. This illness develops in middle aged subjects with arthritis rheumatoid chronic deform history. The diagnosis is clinical and the focus treatment is to diminish the articular pain, to reduce the infections high rates and to avoid the bony deformities. We report a clinical case of a patient who is 69 years-old, she was diagnosed in our hospital and we reviewed the bibliographic entity.(AU)

Síndrome de Felty: Presentación de un caso y revisión bibliográfica / Felty's syndrome: A case report and literature review

El síndrome de Felty se caracteriza por reunir la tríada compuesta por: artritis reumatoide, neutropenia y esplenomegalia. Es una enfermedad autoinmune poco frecuente, con compromiso sistémico, articular y extra articular. Se desarrolla en personas de mediana edad, con historia de artritis reumatoide crónica deformante. El diagnóstico es eminentemente clínico y el tratamiento está enfocado a disminuir el dolor articular, las altas tasas de infecciones y evitar las deformidades óseas. Presentamos una paciente de 69 años de edad, diagnosticada en nuestro hospital, motivo por el que realizamos revisión bibliográfica de la entidad.

Felty's syndrome has such as main feature the triad composed by: rheumatoid arthritis, neutropenia and splenomegaly. It is unusual autoimmune disease that compromises the nervous system as well as joint affectation and extra joint. This illness develops in middle aged subjects with arthritis rheumatoid chronic deform history. The diagnosis is clinical and the focus treatment is to diminish the articular pain, to reduce the infections high rates and to avoid the bony deformities. We report a clinical case of a patient who is 69 years-old, she was diagnosed in our hospital and we reviewed the bibliographic entity.

Comparative molecular analysis of therapy-related and de novo acute promyelocytic leukemia.

Therapy-related acute promyelocytic leukemia (t-APL) has been reported as a late complication of exposure to radiotherapy and/or chemotherapeutic agents targeting DNA topoisomerase II. We have analyzed in t-APL novel gene mutations recently associated with myeloid disorders. Unlike previous reports in acute myeloid leukemia (AML), our results showed neither IDHs nor TET2 mutations in t-APL. However we found an R882H mutation in the DNMT3A gene in a patient with t-APL suggesting a possible role of this alteration in the pathogenesis of t-APL.

Successful renal transplantation from a deceased donor with pesticide intoxication: a case report.

The number of individuals awaiting organ transplantation exceeds the number of organs. Patients who die from intoxication are rarely accepted as potential organ donors. Herein we have presented the results of kidney transplantations performed from a deceased 20-year-old female donor with suicidal ingestion of a pesticide (carbamate). The procured kidneys were successfully transplanted. Patients and grafts are doing well at 4 months following transplantation. There are few reports of successful transplantation of organs obtained from patients who die from various intoxications. Poisoned patients represent another pool of organ donors for transplantation services.

[Morbidity and risk factors of transrectal prostate biopsy]. / Morbilidad y factores de riesgo de la biopsia transrectal prostática.

OBJECTIVES: To evaluate the incidence of minor complications that rarely need treatment (haematuria, hematospermia and rectal bleeding) and major complications (fever and shock) in patients undergoing transrectal biopsy of the prostate and to identify risk factors. PATIENTS AND METHODS: We present an analytic and prospective study where we evaluated 146 patients subjected to transrectal biopsy of the prostate from December 2007 to September 2008. Complications rates and variables were analyzed. RESULTS: Eight patients (5,5%) suffered fever and seven (4,8%) of them were admitted. One of the patients (0,7%) suffered shock. The median of time between biopsy and fever was 3.5 days (1-10 days). Haematuria and hematospermia were more frequent in patients younger than 65 years (p<0.05) and fever and sepsis were more frequent in patients with prostate volume smaller than 55mL (p<0.05). CONCLUSIONS: The complications of the transrectal biopsy of the prostate are frequent, autolimited and they rarely suppose a health hazard for the patients. The most frequent are haematuria and hemospermia, specially in younger patients, whereas infectious complications are rarer and in our study are more frequent in patients with smaller prostates. We must take into account that the information to the patient is very important after a prostate biopsy, so we can avoid useless consultations (for example with haematuria) and it will enable to identify important signs like fever earlier.

Wnt signaling pathway is epigenetically regulated by methylation of Wnt antagonists in acute myeloid leukemia.

Activation of the Wnt signaling pathway has been implicated recently in the pathogenesis of leukemia. We studied the function of epigenetic regulation of the Wnt pathway and its prognostic relevance in acute myelogenous leukemia (AML). We used a methylation-specific polymerase chain reaction approach to analyze the promoter methylation status of a panel of Wnt antagonists including sFRP1, sFRP2, sFRP4, sFRP5, DKK1 and DKK3. Aberrant methylation of Wnt antagonists was detected in four AML cell lines and in up to 64% of AML marrow samples. Treatment of the cell lines with 5-aza-2'-deoxycytidine induced reexpression of methylated Wnt antagonists and inactivation of the Wnt pathway by downregulating the Wnt pathway genes cyclin D1, TCF1 and LEF1 and reducing nuclear localization of beta-catenin. In a subgroup of patients 60 years and younger with newly diagnosed AML and intermediate-risk cytogenetics, abnormal methylation of Wnt antagonists was associated with decreased 4-year relapse-free survival (28 vs 61%, respectively, P=0.03). Our results indicate a function of the epigenetic regulation of the Wnt pathway in predicting relapse in a subgroup of AML patients.

Implicación patogénica de dos nuevas mutaciones en cáncer de mama familiar / Pathogenic implication of two novel mutations in familial breast cancer

Se estima que entre el 5-10% de todos los casos de cáncerde mama (CM) son causados por mutaciones en los genessupresores de tumores BRCA1y BRCA2. El análisis mutacionalcompleto de estos genes ha permitido la identificación demutaciones patogénicas así como variantes de efecto desconocido(VED). Las VED se caracterizan porque su implicaciónpatogénica es confusa, y por lo tanto su importancia clínica esincierta, disminuyendo el valor de los resultados genéticos. Elobjetivo de este estudio ha sido caracterizar la significación clínicade dos nuevas variantes detectadas en un estudio previode 48 familias de la Comunidad Valenciana analizadas en elHospital Universitario La Fe de Valencia (España).Se han realizado análisis genéticos y bioinformáticos y se hanutilizado criterios bioquímicos para establecer si las VED sonpatogénicas. Estos análisis han determinado que la variantec.5025delT es una mutación deletérea y la variante c.8038C>Tafecta a secuencias consenso, requiriéndose estudios complementariospara determinar con precisión su significado

It is estimated that 5% to 10% of all breast cancer (BC)cases are caused by inherited mutations in the tumour suppressorgenes, BRCA1 and BRCA2. Whole gene mutationanalyses of these genes have led to the identification of pathogenicmutations and others variants known as genetic variantsof uncertain significance (USV). The pathogenicity of thesegenetic variants is unclear, and therefore their clinical relevanceis uncertain, diminishing the value of genetic test results. Theaim of the study has been to characterize the clinical significanceof two novel variants detected in a previous study of 48families from the Valencian Community analyzed in the UniversityHospital La Fe of Valencia (Spain). A range of genetic andbioinformatics analyses were performed and several biochemicalcriteria were used to establish whether the genetics variantsof study were pathogenic. These assays showed that thec.5025delT variant was a deleterious mutation and thec.8038C>T variant required of complementary studies to characterizeits meaning. This work highlights the importance ofstudying the USV in order to clarify their pathogenic effect thenthis information is essential for providing efficient counsellingfor BC families

Effect of aluminium and sulphate on anaerobic digestion of sludge from wastewater enhanced primary treatment.

The combined and individual effects of aluminium and sulphate at concentrations of 1,000 mg/l as Al(OH)3, and 150 mgSO4(2-)/L as K2SO4, respectively, on the anaerobic digestion of sludge from enhanced primary treatment (EPT) were evaluated in 1 L capacity semi continuous reactors. It was found that at 59 days, aluminium inhibits the specific methanogenic activity (SMA) of methanogenic and acetogenic bacteria resulting in a 50% to 72% decrease. Sulphate also inhibits (48% to 65%) the SMA of the same type of bacteria. Methanogenic and acetogenic bacteria were able to adapt, to a different extent, to the assayed concentrations of aluminium and sulphate. However, the combination of aluminium and sulphate resulted in a higher inhibition, especially of the hydrogenophilic methanogenic bacteria. Indeed, this effect remained during the time of the experiment, maintaining an inhibition of 44% at 114 days. Feeding with EPT sludge led to a bigger decrease in SMA of each bacterial group, with respect to the other treatments with time. It is concluded that the acidification of anaerobic reactors fed with EPT sludge is due, among other causes, to the concurrent presence of aluminium and sulphate.

Mechanical performance of acrylic bone cements containing different radiopacifying agents.

The effect that three different radiopacifying agents, two of them inorganic (BaSO4, ZrO2) and one organic (an iodine containing monomer, IHQM) have on the static and dynamic mechanical properties of acrylic bone cements was studied. Compressive and tensile strength, fracture toughness and fatigue crack propagation were evaluated. The effect of the inorganic fillers depends on their size and morphology. In relation to the radiolucent cement, the addition of zirconium dioxide improved significantly the tensile strength, the fracture toughness and the fatigue crack propagation resistance. In contrast, the addition of barium sulphate produced a decrease of the tensile strength, but did not affect the fracture toughness and improved the crack propagation resistance. When the iodine containing monomer was used, although the tensile strength and the fracture toughness increased, the fatigue crack propagation resistance remained as low as it was for the radiolucent cement.

Quantitative detection of AML1-ETO rearrangement by real-time RT-PCR using fluorescently labeled probes.

The persistence of the AML1-ETO rearrangement performed by reverse transcription polymerase chain reaction (RT-PCR) has been reported in acute myeloid leukemia (AML) patients in long-term complete remission (CR). This persistence, which is not associated with hematological relapse, limits the clinical use of qualitative RT-PCR. Here, we present a new quantitative real-time PCR method to detect AML1-ETO rearrangement using fluorescently labeled probes. Quantitative detection of AML1-ETO was performed in capillary tubes using two fluorescently labeled probes in the LightCycler equipment. The reliability of the method was checked in twenty-two bone marrow samples and one apheresis sample from eight patients with t(8;21) collected at diagnosis and during follow-up assessment. The regression coefficients obtained for standard curves of AML1-ETO and AML were all greater than 0.98. The sensitivity attained allowed the detection of rearrangements at a dilution of 10(-5) Kasumi-1 cDNA. The intra-assay coefficient of variation was 4% for AML1-ETO, and 7% for AML. The inter-assay coefficient of variation was 19% for AML1-ETO and 12% for AML. A log reduction from two to four in the AML1-ETO/AML ratio was evident after CR. The study of the method and first results obtained in patient samples support that quantitative real-time PCR with hybridization probes is a new reliable and sensitive method to monitor minimal residual disease in AML patients. Moreover, the fluorescent probes with the Light-Cycler technology offer the advantage of a rapid detection.

Standardized, unrelated donor cord blood transplantation in adults with hematologic malignancies.

The potential role of unrelated donor cord blood transplantation (UD-CBT) in adults remains unclear. This study reports the results of UD-CBT in 22 adults with hematologic malignancies following conditioning with thiotepa, busulfan, cyclophosphamide, and antithymocyte globulin in 21, with thiotepa, fludarabine, and antithymocyte globulin in 1, and graft-versus-host disease (GVHD) prophylaxis with cyclosporine and prednisone. Median age was 29 years (range, 18-46 years), and median weight was 69.5 kg (range, 41-85 kg). HLA match was 6 of 6 in 1 case, 5 of 6 in 13 cases, and 4 of 6 in 8 cases. Median number of nucleated cells infused was 1.71 x 10(7)/kg (range, 1.01 x 10(7)/kg to 4.96 x 10(7)/kg). All 20 patients surviving more than 30 days had myeloid engraftment, and only 1, who received the lowest cell dose, developed secondary graft failure. Median time to reach an absolute neutrophil count of at least 0.5 x 10(9)/L was 22 days (range, 13-52 days). Median time to platelets numbered at least 20 x 10(9)/L was 69 days (range, 49-153 days). Seven patients (32%) developed acute GVHD above grade II, and 9 of 10 patients at risk developed chronic GVHD, which became extensive in 4 patients. Twelve patients remained alive and disease-free 3 to 45 months after transplantation. Disease-free survival (DFS) at 1 year was 53%. Age strongly influenced DFS (P =.01). For patients aged 30 years or younger, the DFS at 1 year was 73%. These preliminary results suggest that UD-CBT should be considered a reasonable alternative in young adults with hematologic malignancy and no appropriate bone marrow donor.

Pretreatment characteristics and clinical outcome of acute promyelocytic leukaemia patients according to the PML-RAR alpha isoforms: a study of the PETHEMA group.

Of 167 newly diagnosed acute promyelocytic leukaemia patients, 83 patients were long (L)-form (50%), eight variable (V)-form (5%) and 76 short (S)-form (45%). The V-form and S-form groups presented a significantly higher percentage of patients with white blood cell counts > 10 x 10(9)/l (P < 0.05). The S-form cases displayed a significantly higher number of cases with M3v microgranular features (P = 0.005) and CD34 expression (P < 0.0001). There were no differences between the three isoforms in complete remission (CR) rate (overall CR 90%), but the 3-year disease-free survival was lower for V-form cases than it was for L- and S-form cases (62% vs. 94% and 89%, P = 0.056). We conclude that the V-form and S-form types are associated with some negative prognostic features at diagnosis. However, our data were only able to demonstrate an association with adverse prognosis in the V-form type and, moreover, as the number of cases was limited, needs to be confirmed in large, uniformly treated series.

Variability in the levels of PML-RAR alpha fusion transcripts detected by the laboratories participating in an external quality control program using several reverse transcription polymerase chain reaction protocols.

BACKGROUND AND OBJECTIVES: The detection of PML-RAR by reverse transcription (RT) polymerase chain reaction (PCR) in acute promyelocytic leukemia (APL) patients who are in hematologic remission influences therapeutic decision making in several trials. In the light of this, the Spanish group has recently designed an external quality assessment program (EQAP) of RT-PCR detection of PML-RAR, which includes a study of sensitivity of the participating laboratories. DESIGN AND METHODS: Eighteen laboratories were involved in the program. Ten laboratories followed the method of Biondi et al., 5 employed that of Borrow et al. and the 3 remaining used other protocols. The sensitivity was studied in five rounds of quality control. The first two shipments consisted of dilutions of NB4 RNA into non-APL RNA. The third round consisted of serial dilutions of the NB4 cell line into HL60 cells. The fourth and five rounds consisted of plasmid dilutions containing the bcr1 and bcr3 PML-RAR isoforms. RESULTS: The results showed that the distinct methods allow detection of the PML-RAR hybrid up to a dilution of 10(-4), and exceptionally, up to 10(-5). The laboratories following the method of Biondi et al. usually detected the 10(-3) dilution and less frequently the 10(-4) one, whereas those using other methods usually detected PML-RAR transcript in the 10(-4) dilution, and less commonly in the 10(-5) dilution. However, each of the PCR methods used by EQAP participating laboratories successfully detected at least 50 copies of PML-RAR alpha fusion transcript in plasmid dilution controls. INTERPRETATION AND CONCLUSIONS: The results point to heterogeneous sensitivity amongst participating laboratories. This may reflect differences in methodology, although variations in sample quality may also account for discrepant findings.

Unrelated donor cord blood transplantation in adults with chronic myelogenous leukemia: results in nine patients from a single institution.

The potential role of unrelated donor cord blood transplantation (UD-CBT) in adults is not well established. We report the results of UD-CBT in nine adult patients with chronic myeloid leukemia (CML). The median age was 27 years (range, 19-41 years), and the median weight was 62 kg (range, 45-78 kg). At transplant, six patients were in chronic phase (five in first, and one in second), two in blast crisis, and one in accelerated phase. Eight had received intensive chemotherapy, and three had undergone autologous peripheral blood hematopoietic stem cell transplantation. Four had received interferon with no cytogenetic response, and only three underwent UD-CBT within 1 year of diagnosis. After serological typing for class I antigens, and high-resolution DNA typing for DRB1, the degree of HLA match between patients and cord blood (CB) units was 4/6 in six cases and 5/6 in three cases. The median number of nucleated cells infused was 1.7 x 10(7)/kg (range, 1.2 to 4.9 x 10(7)/kg), and was above 2 x 10(7)/kg in only two cases. All patients received thiotepa, busulfan, cyclophosphamide and anti-thymocyte globulin as conditioning; cyclosporine and prednisone for graft-versus-host disease (GVHD) prophylaxis; and G-CSF from day +7 until engraftment. All seven evaluable cases engrafted. The median time to reach an absolute neutrophil count > or =0.5 x 10(9)/l and > or =1 x 10(9)/l was 22 days (range, 19-52 days) and 28 days (range, 23-64 days), respectively. In the four patients evaluable for platelet recovery time to levels of > or =20 x 10(9) platelets/l, > or =50 x 10(9) platelets/l, and > or =100 x 10(9) platelets/l, these ranged from 50 to 128 days, 60 to 139 days, and 105 to 167 days, respectively. Three patients developed acute GVHD above grade II, and three of the five patients at risk developed extensive chronic GVHD. Four patients, all transplanted in chronic phase, remain alive in molecular remission more than 18, 19, 24 and 42 months after transplantation. These preliminary results suggest that UD-CBT may be considered a reasonable alternative in adults with CML who lack an appropriate bone marrow donor.