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Most mealybug species in the Afrotropical Region have been described or redescribed adequately and their identities are now well established. The species are listed in the catalogue of world species by Ben-Dov (1994) and in ScaleNet, an online database of the scale insects (García-Morales et al. 2019). The genera found in South Africa, and their type species, were discussed by Millar (2002). However, two species remain that have not been discussed since they were described, and their identities remain obscure. Here we discuss these species.
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Hemípteros , AnimalesRESUMEN
Chronic myelomonocytic leukemia (CMML) is a complex clonal hematological disorder classified among myelodysplastic (MDS)/myeloproliferative neoplasms. Prognosis is poor and there is a lack of effective treatments. The hypomethylating agent decitabine has shown activity against MDS and elderly acute myeloid leukemia, but there is little data focusing specifically on its efficacy in CMML. In this prospective, phase 2 Italian study, CMML patients received intravenous decitabine 20 mg/m2 per day on Days 1-5 of a 28-day treatment cycle. Response was evaluated after four and six cycles; patients responding at the end of six cycles could continue treatment with decitabine. Forty-three patients were enrolled; >50% were high-risk according to four CMML-specific scoring systems. In the intent-to-treat population (n=42), the overall response rate after six cycles was 47.6%, with seven complete responses (16.6%), eight marrow responses (19%), one partial response (2.4%) and four hematological improvements (9.5%). After a median follow-up of 51.5 months (range: 44.4-57.2), median overall survival was 17 months, with responders having a significantly longer survival than non-responders (P=0.02). Grade 3/4 anemia, neutropenia and thrombocytopenia occurred in 28.6%, 50% and 38% of patients, respectively. Decitabine appears to be an effective and well-tolerated treatment for patients with high-risk CMML.
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Antimetabolitos Antineoplásicos/administración & dosificación , Decitabina/administración & dosificación , Leucemia Mielomonocítica Crónica/tratamiento farmacológico , Adulto , Anciano , Anciano de 80 o más Años , Femenino , Humanos , Italia , Leucemia Mieloide Aguda/tratamiento farmacológico , Leucemia Mieloide Aguda/patología , Leucemia Mielomonocítica Crónica/patología , Masculino , Persona de Mediana Edad , Pronóstico , Estudios Prospectivos , Resultado del TratamientoRESUMEN
Une nouvelle espèce de Dysmicoccus nuisible à la lavande en Provence (France) (Hemiptera, Sternorrhyncha, Pseudococcidae). Dysmicoccus lavandulae Germain, Matile-Ferrero & Williams n. sp. est décrite et illustrée. Ses séquences ADN sont présentées. L'espèce vit sur Lavandula x intermedia cultivée pour la production d'essence de lavande en Provence. La liste des espèces de pseudococcines vivant sur les lavandes spontanées en France est dressée. Le statut des 2 genres voisins Trionymus Berg et Dysmicoccus Ferris est discuté.
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Hemípteros , Lavandula/parasitología , Animales , Femenino , Francia , Hemípteros/anatomía & histología , Hemípteros/genética , Hemípteros/patogenicidadRESUMEN
The scope of this article is to show the Iveco activity in terms of comfort improvement in all its product Portfolio, focusing on innovation research and realization of tools to get better the life of the driver on commercial vehicles. Comfort related to the ergonomics, thermal, vibrational comfort and after-treatment system in order to improve the life of driver and passengers. It is to remember that Commercial vehicles have different use from a car. For example an heavy truck cabin is not only a place where to drive 8 hours a day, but it is at the same time, an office, a place where to eat, where to sleep and to have a rest. The effort in the last 10 years of Iveco is to improve the comfort of the life of the drivers, utilizing continuous research in standards and innovative systems in order to increase the security and life improvement, focusing also on worldwide legislation as a partner in European committees for health and safety.
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Automóviles , Comercio , Ergonomía , Invenciones , Diseño de Equipo , HumanosRESUMEN
We evaluated the analytical, work flow, and clinical performance of the Versant CT/GC DNA 1.0 assay (Versant CT/GC assay, where "CT" represents Chlamydia trachomatis and "GC" represents Neisseria gonorrhoeae). The assay simultaneously detects Chlamydia trachomatis and Neisseria gonorrhoeae in swab and first-catch urine (FCU) specimens. The limit of detection (LoD) was determined to be 342 copies/ml for C. trachomatis and 137 copies/ml for GC. The Versant CT/GC assay detected 15 C. trachomatis serovars and 46 GC strains. The Versant CT/GC assay demonstrated no cross-reactivity with 136 potentially cross-reacting organisms. Clinical concordance of the Versant CT/GC assay to the Aptima Combo 2 (AC2) assay from Gen-Probe was demonstrated using 1,129 patient specimens, including 589 urine and 540 swab specimens. Discrepant specimens were subjected to DNA sequencing to identify the presence of amplified targets and to identify false-positive and false-negative results. Overall percent agreement was greater than 98%. Positive and negative percent agreements for detection of C. trachomatis were 94.4% and 99.1%, respectively, in urine specimens and 95.8% and 99.8%, respectively, in swab specimens. Positive percent agreement for the detection of N. gonorrhoeae was 100% in both urine and swab specimens, and negative percent agreements were 99.6% and 99% in urine and swab specimens, respectively. In conclusion, the performance of the Versant CT/GC assay was comparable to that of the AC2 assay. The Versant CT/GC assay can be recommended for the detection of C. trachomatis and N. gonorrhoeae in swab and urine specimens of symptomatic and asymptomatic individuals.
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Técnicas Bacteriológicas/métodos , Infecciones por Chlamydia/diagnóstico , Chlamydia trachomatis/aislamiento & purificación , Gonorrea/diagnóstico , Técnicas de Diagnóstico Molecular/métodos , Neisseria gonorrhoeae/aislamiento & purificación , Juego de Reactivos para Diagnóstico , Infecciones por Chlamydia/microbiología , Chlamydia trachomatis/genética , Femenino , Genitales/microbiología , Gonorrea/microbiología , Humanos , Masculino , Neisseria gonorrhoeae/genética , Sensibilidad y Especificidad , Orina/microbiologíaRESUMEN
OBJECTIVES: We examined differences in demographic characteristics, HIV related risk behaviour, prevalence of sexually transmitted infections (STI), and HIV and other health concerns among women with and without a history of sex work. METHODS: A secondary analysis of a population based, cross sectional survey of young, low income women in northern California. RESULTS: Of the 2543 women interviewed, 8.9% reported a history of sex work. These women reported more lifetime male sexual partners, were more likely to use drugs before sex, and were more likely to have a history of having sex with partners at high risk for HIV (that is, men who have sex with men, inject drugs, or were known to be HIV positive). They were significantly more likely to have positive serology for syphilis, herpes simplex virus type 2 (HSV-2), and hepatitis C regardless of their personal injecting drug use history; however, they were no more likely to have HIV, chlamydia, gonorrhoea, hepatitis A or hepatitis B infection compared to women without a history of sex work. Women with a history of sex work were significantly more likely to have a history of sexual coercion and tobacco use. CONCLUSIONS: These data measure the population prevalence of sex work among low income women and associated STI. Women with a history of sex work have health concerns beyond STI and HIV treatment and prevention.
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Trabajo Sexual/estadística & datos numéricos , Enfermedades de Transmisión Sexual/epidemiología , Adolescente , Adulto , California/epidemiología , Estudios Transversales , Femenino , Infecciones por VIH/epidemiología , Infecciones por VIH/psicología , Humanos , Renta , Pobreza , Autorrevelación , Conducta Sexual , Parejas Sexuales , Enfermedades de Transmisión Sexual/psicología , Trastornos Relacionados con Sustancias/epidemiología , Sexo Inseguro/estadística & datos numéricosRESUMEN
The purpose of the study was to compare telomere length (TL) in peripheral blood progenitor cells (PBPC) collected after two tightly spaced high-dose (hd) chemotherapy courses. We assessed 37 previously untreated lymphoma patients undergoing a hd-chemotherapy program with autografting. They sequentially received hd-cyclophosphamide (CY) and hd-Ara-C, both followed by PBPC harvesting. Both post-CY and post-Ara-C harvests were assessed for TL by Southern blot analysis. In 12 patients, the assay was also performed on purified CD34+ cells. All patients displayed high PBPC mobilization following both hd-CY and hd-Ara-C. In all but one patient, TL was shorter in PBPC collected after Ara-C compared to CY: 7226bp (range: 4135-9852) vs 8282 bp (range 4895-14860) (P < 0.0001). This result was confirmed on CD34+ cells. Platelet recovery in patients receiving post-Ara-C PBPC was significantly slower compared to those receiving post-CY PBPC. In conclusion, (i) administration of tightly spaced hd-chemotherapy courses induces marked telomere shortening on harvested PBPC; (ii) engraftment kinetics seem slower, with delayed platelet recovery, in patients autografted with PBPC suffering marked TL erosion; (iii) long-term follow-up is required to verify whether PBPC with shortened telomeres display defective engraftment stability and/or risk of secondary leukemia; (iv) TL evaluation is advisable whenever new mobilization procedures are developed.
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Protocolos de Quimioterapia Combinada Antineoplásica/efectos adversos , Doxorrubicina/efectos adversos , Factor Estimulante de Colonias de Granulocitos/efectos adversos , Trasplante de Células Madre Hematopoyéticas , Células Madre Hematopoyéticas/patología , Linfoma no Hodgkin/tratamiento farmacológico , Linfoma no Hodgkin/patología , Prednisona/efectos adversos , Vincristina/efectos adversos , Adolescente , Adulto , Antibióticos Antineoplásicos/administración & dosificación , Antibióticos Antineoplásicos/efectos adversos , Antígenos CD34/metabolismo , Antimetabolitos Antineoplásicos/administración & dosificación , Antimetabolitos Antineoplásicos/efectos adversos , Antineoplásicos Alquilantes/administración & dosificación , Antineoplásicos Alquilantes/efectos adversos , Protocolos de Quimioterapia Combinada Antineoplásica/administración & dosificación , Terapia Combinada , Ciclofosfamida/administración & dosificación , Ciclofosfamida/efectos adversos , Citarabina/administración & dosificación , Citarabina/efectos adversos , Doxorrubicina/administración & dosificación , Femenino , Factor Estimulante de Colonias de Granulocitos/administración & dosificación , Movilización de Célula Madre Hematopoyética , Células Madre Hematopoyéticas/metabolismo , Humanos , Masculino , Persona de Mediana Edad , Recuento de Plaquetas , Valor Predictivo de las Pruebas , Prednisona/administración & dosificación , Telómero , Trasplante Autólogo , Vincristina/administración & dosificaciónRESUMEN
The greater sensitivity of nucleic acid amplification tests (NAATs) for Chlamydia trachomatis and Neisseria gonorrhoeae permits the use of urine and other noninvasive specimens, which can increase the reach and decrease the costs of public health screening programs aimed at controlling these infections. This study evaluated the performance of the APTIMA Combo 2 assay, a multiplex assay based on the transcription-mediated amplification reaction, for the simultaneous detection of both pathogens in endocervical swab and urine specimens from females. Combo 2 assay results were compared with patient infected status, which were available by using other commercial NAATs. Sensitivity and specificity for C. trachomatis were 94.2 and 97.6%, respectively, in swabs and 94.7 and 98.9%, respectively, in first-catch urine (FCU). Sensitivity and specificity for N. gonorrhoeae were 99.2 and 98.7%, respectively, in swabs and 91.3 and 99.3%, respectively, in FCU. The assay reliably detected both infections in coinfected patients. The Combo 2 assay can be recommended for use with endocervical swab and urine specimens from females, especially for screening tests for asymptomatic women in sexually transmitted disease surveillance programs. This Food and Drug Administration-cleared assay can be a useful tool in efforts to reduce the prevalence and incidence of C. trachomatis and N. gonorrhoeae infections in sexually active women and to prevent their costly and serious sequelae.
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Cuello del Útero/microbiología , Chlamydia trachomatis/aislamiento & purificación , Neisseria gonorrhoeae/aislamiento & purificación , Orina/microbiología , Femenino , Humanos , Juego de Reactivos para DiagnósticoRESUMEN
The performance of the Becton Dickinson BDProbe Tec ET System Chlamydia trachomatis and Neisseria gonorrhoeae Amplified DNA Assays (BD Biosciences, Sparks, Md.) was evaluated in a multicenter study. Specimens were collected from 2,109 men and women, with or without symptoms, attending sexually transmitted disease, family planning, and obstetrics and gynecology clinics. Both swab and urine samples were collected, and the results obtained from 4,131 specimens were compared to those from culture and the LCx nucleic acid amplification test (Abbott Industries, Abbott Park, Ill.). PCR and cytospin of the culture transport medium with chlamydia direct fluorescent antibody staining were used to adjudicate chlamydia culture-negative results. Sensitivity and specificity were calculated both with and without use of the amplification control (AC), with little apparent difference in the results. Without the AC result, sensitivity for C. trachomatis and N. gonorrhoeae were 92.8 and 96.6%, respectively, for cervical swabs and 80.5 and 84.9% for urine from women. C. trachomatis and N. gonorrhoeae sensitivities were 92.5 and 98.5%, respectively, for male urethral swabs and 93.1 and 97.9% for urine from men. This amplified DNA system for simultaneous detection of chlamydial and gonococcal infections demonstrated superior sensitivity compared to chlamydia culture and has performance characteristics comparable to those of other commercially available nucleic acid-based assays for these organisms.
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Infecciones por Chlamydia/microbiología , Chlamydia trachomatis/aislamiento & purificación , Gonorrea/microbiología , Neisseria gonorrhoeae/aislamiento & purificación , Técnicas de Amplificación de Ácido Nucleico/métodos , Juego de Reactivos para Diagnóstico , Cuello del Útero/microbiología , Chlamydia trachomatis/genética , Medios de Cultivo , ADN Bacteriano/análisis , Femenino , Humanos , Masculino , Neisseria gonorrhoeae/genética , Sensibilidad y Especificidad , Manejo de Especímenes , Uretra/microbiología , Orina/microbiologíaRESUMEN
Bcr/abl fusion gene, in experimental models, induces survival to growth factor deprivation and hypersensitivity to IL3. However, conflicting data were reported about chronic myeloid leukemia (CML) progenitors. We investigated the responsiveness of purified CML CFU-GM to GM-CSF/IL3 and their survival to growth factor deprivation. CFU-GM hypersensitivity to IL3 and/or GM-CSF was found in 3/11 CML cases only. CML CFU-GM survived well in stroma-free 'mass' culture (5 x 10(4) cells/ml) without cytokine addition, up to day 11, average recovery being around 95% in medium + 10% fetal bovine serum and 67-81% in serum-free medium. Conversely, normal progenitors declined steadily, particularly after extensive purification (18 +/- 10% recovery at the 7th day), and in serum-free medium (4 +/- 6% recovery). By contrast, normal and CML CFU-GM declined in a similar way in limiting dilution cultures (1-10 cells/50 microl). We also investigated the effects of retinoic acid and alpha-interferon on CFU-GM survival. Both all-trans- and 13-cis retinoic acid, particularly in combination with alpha-interferon, reduced CML CFU-GM recovery down to normal progenitors' values. In conclusion, hypersensitivity to CSFs is rare in CML, whereas resistance to growth factor deprivation has been confirmed in mass, but not in limiting, dilution cultures. Both stereoisomers of retinoic acid, at therapeutic concentrations and in combination with alpha-interferon, can overcome the survival advantage of CML progenitors.
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División Celular , Factor Estimulante de Colonias de Granulocitos y Macrófagos/farmacología , Interferón-alfa/farmacología , Interleucina-3/farmacología , Leucemia/patología , Retinoides/farmacología , División Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , HumanosRESUMEN
OBJECTIVE: Purging procedures are increasingly used to provide stem cell collections devoid of contaminating tumor cells. In follicle center lymphoma (FCL), most approaches eradicate polymerase chain reaction (PCR);-detectable disease in only a fraction of harvests undergoing ex vivo manipulation. In this study we evaluated whether there is a relationship between tumor burden of stem cell harvests and successful clearance of PCR-detectable disease following ex vivo manipulation. MATERIALS AND METHODS: To address this issue, we developed a real-time PCR approach for quantitative measurement of tumor contamination using the bcl-2 rearrangement. Real-time PCR was used to evaluate the relationship between tumor burden of stem-cell harvests and purging effectiveness in PCR(+) samples derived from 10 FCL patients. Ex vivo purging was performed using the MaxSep cell separator (Baxter Immunotherapy, Deerfield, IL, USA). RESULTS: Our real-time PCR method proved effective, sensitive, accurate, and reproducible. Four collections were successfully cleared of minimal residual disease (MRD) whereas six remained PCR(+). Real-time PCR showed that the four collections successfully cleared of MRD had a prepurging tumor burden significantly lower than those remaining PCR(+) (p = 0.04). CONCLUSION: This study provides the first evidence that evaluation of tumor burden in stem-cell harvests by real-time PCR can predict the effectiveness of therapeutic intervention in non-Hodgkin's lymphoma. Based on these findings, we foresee a more widespread use of this technique to evaluate the impact of different therapeutic approaches in FCL.
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Eliminación de Componentes Sanguíneos/métodos , Células Madre Hematopoyéticas/citología , Linfoma Folicular/sangre , Reacción en Cadena de la Polimerasa/métodos , Adolescente , Adulto , Secuencia de Bases , Reordenamiento Génico , Gliceraldehído-3-Fosfato Deshidrogenasas/genética , Trasplante de Células Madre Hematopoyéticas , Humanos , Linfoma Folicular/terapia , Persona de Mediana Edad , Datos de Secuencia Molecular , Neoplasia Residual/sangre , Proteínas Proto-Oncogénicas c-bcl-2/genética , Reproducibilidad de los Resultados , Sensibilidad y Especificidad , Translocación Genética , Trasplante AutólogoRESUMEN
In some studies the GM-CSF (granulocyte-macrophage colony-stimulating factor) increased the in vitro sensitivity of acute myeloid leukemia (AML) cells to cytosine arabinoside (Ara-C), however, in clinical trials no favorable effects were shown. We used a GM-CSF responsive AML cell line (AML 193) to test the effects of growth stimulation on in vitro efficacy of Ara-C and methotrexate (MTX). In 6 days continuous exposure, dose dependent Ara-C cytotoxicity was counteracted by GM-CSF. Conversely, MTX cytotoxicity was increased significantly. However, in a short term treatment (24 h, high doses) the GM-CSF increased both MTX and Ara-C cytotoxicity. These effects might depend on different drug regimens and cell features.
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Antimetabolitos Antineoplásicos/farmacología , Citarabina/farmacología , Factor Estimulante de Colonias de Granulocitos y Macrófagos/farmacología , Leucemia Mieloide Aguda/tratamiento farmacológico , Metotrexato/farmacología , Antimetabolitos Antineoplásicos/uso terapéutico , Muerte Celular/efectos de los fármacos , Citarabina/uso terapéutico , Relación Dosis-Respuesta a Droga , Resistencia a Antineoplásicos , Quimioterapia Combinada , Humanos , Leucemia Mieloide Aguda/patología , Metotrexato/uso terapéutico , Factores de Tiempo , Células Tumorales CultivadasRESUMEN
The feasibility and efficacy of a novel immunomagnetic ex vivo negative purging method was evaluated on peripheral blood progenitor cells (PBPC) from 13 non-Hodgkin's lymphoma patients (eight follicular, FL; three mantle cell, MCL; two FL with histologic transformation). A peculiar feature of the study was the collection of PBPC after prolonged tumor debulking. Our method included a stem cell enrichment phase followed by cell incubation with anti-B cell MoAbs (anti-CD19, CD20, CD22, CD23), addition of immunobeads, and then positive cell removal by passage on a Max-Sep (Baxter Immunotherapy) cell separator. Engraftment was rapid and stable. Hematological values were assessed 1 and 2 years after the autograft. Purging efficacy was molecularly assessed in a panel of 11 patients who showed persistence of PCR-detectable lymphoma cells on PBPC harvests despite intensified chemotherapeutic debulking. PCR-negativity was obtained in vitro and persisted in vivo after autograft in three FL patients; five more FL patients, whose purged PBPC were PCR+, converted to stable (3 patients) or fluctuating (two patients) PCR negativity after autograft. MCL patients never reached PCR negativity. Thus, ex vivo purging may have a role for FL patients harvesting PCR-positive PBPC after intensified chemotherapy. In contrast, the addition of ex vivo purging seems to be of little if any benefit for MCL patients.
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Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Trasplante de Células Madre Hematopoyéticas , Separación Inmunomagnética , Linfoma Folicular/terapia , Inducción de Remisión/métodos , Adulto , Relación Dosis-Respuesta a Droga , Estudios de Factibilidad , Femenino , Estudios de Seguimiento , Humanos , Masculino , Persona de Mediana Edad , Reacción en Cadena de la Polimerasa , Reproducibilidad de los Resultados , Factores de Tiempo , Trasplante AutólogoRESUMEN
Two recombinant human granulocyte colony-stimulating factor (rhG-CSF) isoforms were isolated from the medium conditioned by an engineered Chinese hamster ovary (CHO) cell line. The two rhG-CSFs were characterized and were found to differ in the carbohydrate structure attached to Thr-133. The glycoform, referred to as Peak 1, contains the O-linked glycan Neu5Ac(alpha 2-3)Gal(beta 1-3)GalNAc; the Peak 2 glycoform contains the O-linked glycan Neu5Ac(alpha 2-3)Gal(beta 1-3)[Neu5Ac(alpha 2-6)]GalNAc. The two glycoforms displayed a similar biological activity in cultures of a mouse 32D C13 cell line and human bone-marrow myelo-monocytic progenitor cells (CFU-GM). In the latter test both glycoforms displayed a higher activity than nonglycosylated rMet-hG-CSF from Escherichia coli. The pharmacokinetic profile and activity of the two rhG-CSF glycoforms and of a mixture of them (Pool) were investigated in mice treated with a single injection of rhG-CSF at the doses of 125 micrograms and 250 micrograms/kg, given via the intravenous (i.v.) and the subcutaneous (s.c.) route, respectively. The plasma concentration profiles obtained were similar for all three substances and did not show any relevant differences in absorption or elimination. The pharmacokinetic parameters indicate that the three substances have similar area under the curve (AUCs), volumes of distribution, and terminal half-life. Furthermore, our data indicate a high bioavailability of the two different glycoforms of rhG-CSF when given to mice via the s.c. route either singularly or as a mixture. Detectable levels of rhG-CSF persisted for more than 8 h in the i.v. and more than 24 h in the s.c. route of administration. All three substances induced early neutrophilia in mice. All rhG-CSF-treated mice developed a two-four-fold rise in neutrophil counts as early as 4 h after the intravenous and 2 h after the subcutaneous injection. Relatively high levels of neutrophils were maintained for at least 8 and 24 h after i.v. and s.c. administration, respectively.
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Factor Estimulante de Colonias de Granulocitos/aislamiento & purificación , Factor Estimulante de Colonias de Granulocitos/farmacología , Proteínas Recombinantes/aislamiento & purificación , Secuencia de Aminoácidos , Animales , Área Bajo la Curva , Disponibilidad Biológica , Células CHO , Cromatografía por Intercambio Iónico , Cricetinae , Relación Dosis-Respuesta a Droga , Electroforesis en Gel de Poliacrilamida , Factor Estimulante de Colonias de Granulocitos/química , Semivida , Inyecciones Intravenosas , Inyecciones Subcutáneas , Focalización Isoeléctrica , Recuento de Leucocitos/efectos de los fármacos , Masculino , Ratones , Ratones Endogámicos C57BL , Datos de Secuencia Molecular , Neutrófilos/efectos de los fármacos , Mapeo Peptídico , Isoformas de Proteínas/química , Isoformas de Proteínas/aislamiento & purificación , Isoformas de Proteínas/farmacología , Proteínas Recombinantes/química , Proteínas Recombinantes/farmacología , Análisis de SecuenciaRESUMEN
BACKGROUND AND OBJECTIVE: All culture systems exploring the early (pre-CFU) hematopoietic compartment are generally complex, time-consuming and unsuitable for routine application. The aim of our study was to develop a stroma-free culture system to quantify early bone marrow (BM) myeloid progenitor cells. DESIGN AND METHODS: Low density, progenitor cell enriched BM cells underwent a double cytotoxic treatment with CD38 and CD33 monoclonal antibodies + rabbit complement, which depleted 99% of CFU-GM and BFU-E. Then they were cultured, both in agar and in limiting-dilution liquid culture, in the presence of 5637 cell line supernatant (containing GM-CSF, G-CSF and interleukin 1 ), stem cell factor (SCF) and interleukin 3 (IL3). RESULTS: The largest number (median 14.9 on 1x10(5) cells) and size (>50,000 cells) of myelomonocytic cell clones from CD33Eth /CD38Eth progenitors was reached after 3-4 weeks of liquid culture. SCF, but not IL3, was essential for that growth. The frequency of CD33-/ CD38- progenitors grown in liquid culture was approximately three times greater than the LTC-IC frequency in the same cell suspension. An average 93% of CD33-/CD38- progenitors displayed HLA-DR antigens and 43% generated secondary CFU-GM. In the BM of 9/10 patients, previously exposed to chemotherapy, CD33-/CD38- progenitor frequency was quite low (median 0.9 on 1x10(5) cells), in spite of normal cellularity and morphology and sustained disease remission. INTERPRETATION AND CONCLUSIONS: CD33-/CD38- progenitors can be grown and quantified in a stroma-free culture system in a relatively short time. The test can reveal long-lasting, subclinical BM damage induced by chemotherapy and could also be valuable for estimating the amount of early myeloid progenitors for transplantation purposes.
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Antígenos CD/análisis , Antígenos de Diferenciación Mielomonocítica/análisis , Antígenos de Diferenciación/análisis , Células Madre Hematopoyéticas/efectos de los fármacos , NAD+ Nucleosidasa/análisis , Factor de Células Madre/farmacología , ADP-Ribosil Ciclasa , ADP-Ribosil Ciclasa 1 , División Celular , Supervivencia Celular/efectos de los fármacos , Células Madre Hematopoyéticas/inmunología , Humanos , Glicoproteínas de Membrana , Lectina 3 Similar a Ig de Unión al Ácido SiálicoRESUMEN
Several studies indicate that a number of signal-transducing molecules involved in the proliferation, differentiation, and functional activation of normal hemopoietic cells may be constitutively activated in primary leukemic cells and play a role in the outcome or in the progression of these neoplastic disorders. In this study we show that the product of the proto-oncogene c-Cbl, whose function is still unknown, is constitutively tyrosine phosphorylated not only in cells from chronic myelogenous leukemias (CMLs) in the blast phase, but also in cells from acute myeloblastic leukemias (AMLs), Ph-negative acute T-lymphoblastic leukemias (T-ALLs), and Ph-negative pre-B lymphoblastic leukemias (pre-B ALL). Moreover, in acute leukemia cells, c-Cbl was not stably complexed with the tyrosine-phosphorylated adaptor protein CrkL. The analysis of Grb2/c-Cbl interaction demonstrated that, in both acute leukemia and CML blasts, c-Cbl was stably complexed with the N-terminal Src homology (SH) 3 domain of Grb2 and, in blasts from ALL patients, with the Grb2 SH2 domain. The analysis of c-Cbl subcellular distribution showed that in all cases of leukemia tested, as well as in growth factor-stimulated M-07e cells, c-Cbl was present in the cytosolic, in the membrane, and in the detergent-insoluble fractions. Finally, in polymorphonuclear neutrophils (PMNs) from CML patients, c-Cbl was found stably associated with the detergent-insoluble fraction, whereas in PMNs from normal donors, it was detected only in the cytosolic fraction. Our findings that c-Cbl is constitutively tyrosine phosphorylated and associated with the detergent-insoluble fraction in AML and ALL blasts and in PMNs from CML patients suggest that this event represents a common step in the neoplastic transformation of both myeloid and lymphoid progenitor cells.
Asunto(s)
Proteínas Adaptadoras Transductoras de Señales , Leucemia/metabolismo , Fosfotirosina/metabolismo , Proteínas Proto-Oncogénicas/análisis , Proteínas Proto-Oncogénicas/metabolismo , Fracciones Subcelulares/química , Ubiquitina-Proteína Ligasas , Western Blotting , Proteína Adaptadora GRB2 , Humanos , Técnicas de Inmunoadsorción , Leucemia/patología , Leucemia Mielógena Crónica BCR-ABL Positiva/metabolismo , Leucemia Mielógena Crónica BCR-ABL Positiva/patología , Leucemia Mieloide Aguda/metabolismo , Leucemia Mieloide Aguda/patología , Leucemia-Linfoma de Células T del Adulto/metabolismo , Leucemia-Linfoma de Células T del Adulto/patología , Proteínas Nucleares/análisis , Proteínas Nucleares/metabolismo , Fosforilación , Leucemia-Linfoma Linfoblástico de Células Precursoras B/metabolismo , Leucemia-Linfoma Linfoblástico de Células Precursoras B/patología , Leucemia-Linfoma Linfoblástico de Células Precursoras/metabolismo , Proteínas/metabolismo , Proto-Oncogenes Mas , Proteínas Proto-Oncogénicas c-cbl , Dominios Homologos srcRESUMEN
The Gen-Probe AMPLIFIED Chlamydia Trachomatis Assay (AMP CT) uses transcription-mediated amplification and hybridization protection assay procedures to qualitatively detect Chlamydia trachomatis rRNA in urine, endocervical swab, and urethral specimens. The performance of the AMP CT was compared to that of cell culture for endocervical swab and urine specimens from women and urethral and urine specimens from men. Analysis of specimens with discrepant results was performed by a combination of reculture, direct fluorescent-antibody (DFA) staining of specimen sediment, and amplification which targeted a different chlamydial rRNA. A total of 800 urine samples were tested by the AMP CT (607 from women and 193 from men), and 7. 1% were positive for C. trachomatis, with a sensitivity of 91.2% and a specificity of 99.6% upon discrepant analysis. A total of 926 swab specimens were tested by culture and AMP CT (717 endocervical swab specimens and 209 urethral swab specimens from men), and 7.7% were positive for C. trachomatis, with a sensitivity and specificity of 100% upon discrepant analysis. The AMP CT is a sensitive and specific nucleic acid hybridization assay for the detection of C. trachomatis in endocervical swab specimens from women, urethral swab specimens from men, and urine specimens from men and women.
Asunto(s)
Infecciones por Chlamydia/diagnóstico , Infecciones por Chlamydia/microbiología , Chlamydia trachomatis/genética , Chlamydia trachomatis/aislamiento & purificación , Técnicas de Amplificación de Ácido Nucleico , Instituciones de Atención Ambulatoria , Técnicas Bacteriológicas/estadística & datos numéricos , California , Cuello del Útero/microbiología , Estudios de Evaluación como Asunto , Femenino , Humanos , Masculino , Sensibilidad y Especificidad , Uretra/microbiología , Orina/microbiologíaRESUMEN
Peripheral blood leukocytes are becoming the preferred source of hematopoietic progenitor/stem cells for autologous transplantation. However, in vitro purging procedures are complex and expensive when applied to peripheral blood progenitor cells harvests. This is mainly due to the large quantities of nucleated cells present in leukapheresis collections. Aiming to reduce total cellularity without significant loss of CD34+ cells, we developed an in vitro cell separation procedure based on ficoll/metrizoate gradient used at a final density of 1.067 g/ml. To obtain this density, standard Lympho-prep (1.077 g/ml) was diluted with normal saline solution (NaCl 9 g/l). Twenty-six leukapheresis collections (median cellularity 21.1 x 10(9), range 2.8-60) from 14 patients with non-Hodgkin's lymphoma, multiple myeloma or plasma cell leukemia were processed (median two leukaphereses per patient). Mean (+/- s.d.) recovery of total nucleated cells, CD34+ cells and CFU-GM was 20.9 +/- 10%, 74.7 +/- 22% and 70.5 +/- 19%, respectively. Cumulative per patient progenitor cell recovery was always above 50%, and as high as 80% in 10/14 patients, while total cellularity was reduced to a median 21.5% (10-33%) of pre-separation values. Contaminating neoplastic cells, identified by immunofluorescence in five collections, were reduced by 1-2 logs. The results indicate that our density gradient separation is an effective method to reduce total cellularity prior to immunological purging, without significant loss of progenitor cells.
Asunto(s)
Centrifugación por Gradiente de Densidad/métodos , Trasplante de Células Madre Hematopoyéticas , Leucaféresis/métodos , Antígenos CD34/metabolismo , Purgación de la Médula Ósea , Ensayo de Unidades Formadoras de Colonias , Ficoll , Células Madre Hematopoyéticas/inmunología , Células Madre Hematopoyéticas/patología , Humanos , Técnicas In Vitro , Leucemia de Células Plasmáticas/terapia , Recuento de Leucocitos , Linfoma no Hodgkin/terapia , Ácido Metrizoico , Mieloma Múltiple/terapia , Trasplante AutólogoRESUMEN
Many laboratories use a commercial enzyme immunoassay (EIA) with verification testing to diagnose Chlamydia trachomatis infections in an effort to contain costs. This study was designed to compare the performance and cost-effectiveness of direct fluorescent-antibody assay (DFA), commercial PCR, and ligase chain reaction (LCR) for the verification of EIA results. Cervical specimens were screened by EIA. DFA, PCR, and LCR were compared as verification tests for EIA-reactive specimens and negative greyzone (NGZ) specimens at 50% below the cutoff value. These samples were also tested by in-house PCR, which was used in the analysis of verification results. A total of 477 (7%) of 6,571 samples were reactive or within the NGZ. EIA results with verification by DFA testing (EIA/DFA results) agreed with 93% of the true results compared with 97% for EIA/PCR results for one set of 242 samples; there was 97% agreement with true results for EIA/DFA results versus 95% for EIA/LCR results for another set of 235 samples. Ten samples were false positive by LCR. Time and costs were equivalent for EIA with the DFA, PCR, or LCR as the verification test but were two- to threefold greater for PCR or LCR alone than for EIA with verification. Since it is important to balance cost containment with public health objectives, DFA, PCR, and LCR as EIA verification tests for cervical samples offer acceptable sensitivities and specificities at reasonable cost for low- to moderate-risk populations and therefore can be extended to a broader spectrum of at-risk populations.
Asunto(s)
Infecciones por Chlamydia/diagnóstico , Chlamydia trachomatis , Reacción en Cadena de la Polimerasa , Enfermedades del Cuello del Útero/diagnóstico , Adolescente , Adulto , Análisis Costo-Beneficio , Femenino , Técnica del Anticuerpo Fluorescente Directa , Humanos , Técnicas para InmunoenzimasRESUMEN
BACKGROUND: The granulocyte-colony stimulating factor (G-CSF) and the interleukin 6 (IL-6) are part of a large family of cytokines that regulate the production and the functional activity of hemopoietic cells. Recent studies have shown that IL-6 and G-CSF share structure homology and have partially overlapping functions. OBJECTIVES: Our research gives some information for planning the production of new artificial cytokines in order to establish whether we could obtain molecules that had two functions or an inhibitory function. STUDY DESIGN: Fourteen different chimeric molecules (called Harlequin molecules) of human IL-6 and G-CSF have been produced exchanging the sequence encoded by the 2nd to the 5th exon of the genes of the two cytokines. RESULTS: In order to test their biological activity we performed different assays: proliferation of murine B9-cells and immunoglobulin production from human Epstein-Barr virus (EBV)-transformed B-cell lines for IL-6; induction of granulocytic differentiation of the murine 32DC13(G) cell line and normal bone marrow progenitor cells for G-CSF. Some chimeric molecules maintain the activity of either IL-6 and G-CSF and at least one (Harlequin 11) has both biological activities. One chimeric protein has no biological activity but competes, presumably at the receptor level, for the activity of the intact cytokine. CONCLUSION: These studies can provide important information on the structure/function of the two cytokines. The bifunctional Harlequin molecule 11 could be a potential candidate as therapeutic agent.