RESUMEN
New diagnostics are needed to improve clinicians' ability to detect tuberculosis (TB) disease in key populations such as children and persons living with HIV and to rapidly detect drug resistance. Circulating cell-free DNA (ccfDNA) in plasma is a diagnostic target in new obstetric and oncologic applications, but its utility for diagnosing TB is not known. Here we show that Mycobacterium tuberculosis complex DNA can be detected in plasma of persons with sputum smear-positive TB, even in the absence of mycobacteremia. Among 40 participants with bacteriologically-confirmed smear-positive TB disease who had plasma tested by quantitative PCR (qPCR), 18/40 (45%) had a positive result on at least one triplicate reaction. Our results suggest that plasma DNA may be a useful target for improving clinicians' ability to diagnose TB. We anticipate these findings to be the starting point for optimized methods of TB ccfDNA testing and sequence-based diagnostic applications such as molecular detection of drug resistance.
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Ácidos Nucleicos Libres de Células/genética , ADN Bacteriano/sangre , Mycobacterium tuberculosis/genética , Tuberculosis/diagnóstico , Adulto , Femenino , Humanos , Masculino , Reacción en Cadena en Tiempo Real de la Polimerasa , Sensibilidad y Especificidad , Esputo/microbiología , Tuberculosis/sangreRESUMEN
BACKGROUND: Distant metastasis accounts for 90% of deaths from colorectal cancer (CRC). Genomic heterogeneity has been reported in various solid malignancies, but remains largely under-explored in metastatic CRC tumors, especially in primary to metastatic tumor evolution. PATIENTS AND METHODS: We conducted high-depth whole-exome sequencing in multiple regions of matched primary and metastatic CRC tumors. Using a total of 28 tumor, normal, and lymph node tissues, we analyzed inter- and intra-individual heterogeneity, inferred the tumor subclonal architectures, and depicted the subclonal evolutionary routes from primary to metastatic tumors. RESULTS: CRC has significant inter-individual but relatively limited intra-individual heterogeneity. Genomic landscapes were more similar within primary, metastatic, or lymph node tumors than across these types. Metastatic tumors exhibited less intratumor heterogeneity than primary tumors, indicating that single-region sequencing may be adequate to identify important metastasis mutations to guide treatment. Remarkably, all metastatic tumors inherited multiple genetically distinct subclones from primary tumors, supporting a possible polyclonal seeding mechanism for metastasis. Analysis of one patient with the trio samples of primary, metastatic, and lymph node tumors supported a mechanism of synchronous parallel dissemination from the primary to metastatic tumors that was not mediated through lymph nodes. CONCLUSIONS: In CRC, metastatic tumors have different but less heterogeneous genomic landscapes than primary tumors. It is possible that CRC metastasis is, at least partly, mediated through a polyclonal seeding mechanism. These findings demonstrated the rationale and feasibility for identifying and targeting primary tumor-derived metastasis-potent subclones for the prediction, prevention, and treatment of CRC metastasis.
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Neoplasias Colorrectales/patología , Secuenciación del Exoma , Heterogeneidad Genética , Metástasis de la Neoplasia/genética , Neoplasias Colorrectales/genética , Humanos , Mutación , Siembra NeoplásicaRESUMEN
OBJECTIVE: The current study examined the effects of partial and total sleep deprivation on emotional reactivity. METHODS: Twenty-eight partially sleep-deprived participants and 31 totally sleep-deprived participants rated their valence and arousal responses to positive and negative pictures across four testing sessions during the day following partial sleep deprivation or during the night under total sleep deprivation. RESULTS: The results suggest that valence and arousal ratings decreased under both sleep deprivation conditions. In addition, partial and total sleep deprivation had a greater negative effect on positive events than negative events. CONCLUSION: These results suggest that sleep-deprived persons are more likely to respond less to positive events than negative events. One explanation for the current findings is that negative events could elicit more attentive behavior and thus stable responding under sleep deprivation conditions. As such, sleep deprivation could impact reactivity to emotional stimuli through automated attentional and self-regulatory processes.
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Nivel de Alerta/fisiología , Emociones/fisiología , Privación de Sueño/fisiopatología , Atención/fisiología , Femenino , Humanos , Masculino , Sueño/fisiología , Privación de Sueño/psicología , Adulto JovenRESUMEN
PURPOSE: (1) To determine the safety of the epidermal growth factor receptor (EGFR) antibody cetuximab with concurrent gemcitabine and abdominal radiation in the treatment of patients with locally advanced adenocarcinoma of the pancreas. (2) To evaluate the feasibility of pancreatic cancer cell epithelial-mesenchymal transition (EMT) molecular profiling as a potential predictor of response to anti-EGFR treatment. METHODS: Patients with non-metastatic, locally advanced pancreatic cancer were treated in this dose escalation study with gemcitabine (0-300 mg/m(2)/week) given concurrently with cetuximab (400 mg/m(2) loading dose, 250 mg/m(2) weekly maintenance dose) and abdominal irradiation (50.4 Gy). Expression of E-cadherin and vimentin was assessed by immunohistochemistry in diagnostic endoscopic ultrasound fine-needle aspiration (EUS-FNA) specimens. RESULTS: Sixteen patients were enrolled in 4 treatment cohorts with escalating doses of gemcitabine. Incidence of grade 1-2 adverse events was 96%, and incidence of 3-4 adverse events was 9%. There were no treatment-related mortalities. Two patients who exhibited favorable treatment response underwent surgical exploration and were intraoperatively confirmed to have unresectable tumors. Median overall survival was 10.5 months. Pancreatic cancer cell expression of E-cadherin and vimentin was successfully determined in EUS-FNA specimens from 4 patients. CONCLUSIONS: Cetuximab can be safely administered with abdominal radiation and concurrent gemcitabine (up to 300 mg/m(2)/week) in patients with locally advanced adenocarcinoma of the pancreas. This combined therapy modality exhibited limited activity. Diagnostic EUS-FNA specimens could be analyzed for molecular markers of EMT in a minority of patients with pancreatic cancer.
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Adenocarcinoma/terapia , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Receptores ErbB/antagonistas & inhibidores , Neoplasias Pancreáticas/terapia , Adenocarcinoma/patología , Adulto , Anciano , Anciano de 80 o más Años , Anticuerpos Monoclonales/administración & dosificación , Anticuerpos Monoclonales Humanizados , Antineoplásicos/uso terapéutico , Protocolos de Quimioterapia Combinada Antineoplásica/administración & dosificación , Biopsia con Aguja Fina , Cadherinas/genética , Cetuximab , Estudios de Cohortes , Terapia Combinada , Desoxicitidina/administración & dosificación , Desoxicitidina/análogos & derivados , Relación Dosis-Respuesta a Droga , Transición Epitelial-Mesenquimal , Estudios de Factibilidad , Femenino , Perfilación de la Expresión Génica , Regulación Neoplásica de la Expresión Génica , Humanos , Masculino , Persona de Mediana Edad , Neoplasias Pancreáticas/patología , Tasa de Supervivencia , Vimentina/genética , GemcitabinaRESUMEN
Despite the development of nomograms designed to evaluate the prognosis of a patient with prostate cancer (CaP), the information has been limited to prostate-specific antigen (PSA), clinical stage, Gleason score, and tumor volume estimates. To improve our ability to predict prognosis, information regarding the molecular properties of CaP is needed. Hyaluronic acid (HA) is a glycosaminoglycan that promotes tumor metastasis. Hyaluronidase (HAase) is an enzyme that degrades HA into angiogenic fragments. We recently showed that in CaP tissues, whereas HA is localized mostly in the tumor-associated stroma, HYAL1 type HAase is exclusively localized in CaP cells (Lokeshwar et al. J. Biol. Chem., 276: 11922-11932, 2001). We evaluated the prognostic potential of HA and HYAL1 in CaP by immunohistochemistry. Archival CaP specimens were obtained from patients who underwent radical retropubic prostatectomy for clinically localized CaP. Group 1 (n = 25) included patients who showed biochemical recurrence (PSA >0.4 ng/ml; mean recurrence: 21.3 months). Group 2 included patients with no clinical or biochemical recurrence (n = 45; mean follow-up: 80.9 months). For HA and HYAL1 staining, a biotinylated HA-binding protein and an anti-HYAL1 antibody were used. The staining was evaluated on the basis of intensity (0 to 3+) and as dense or sparse (for HA staining only) and then grouped as low grade and high grade. In CaP specimens, HYAL1 was exclusively expressed in tumor cells. Although the stroma was stained positive for HA, 40% of tumor cells also expressed HA. HA, HYAL1, and combined HA-HYAL1 staining predicted progression with 96%, 84%, and 88% sensitivity, 55.5%, 80%, and 84.4% specificity, and 70%, 81.4%, and 85.7% accuracy, respectively. In the univariate analysis, preoperative PSA, Gleason sum, stage, margin, seminal vesicle, extra-prostatic extension (EPE), HA, HYAL1, and HA-HYAL1 were significant in predicting progression (P < 0.05). However, in the multiple logistic regression analysis, only EPE [odds ratio (OR) = 33.483; P = 0.002), HYAL1 (OR = 12.42; P = 0.009), HA-HYAL1 (OR = 18.048; P = 0.0033), and margin (OR = 26.948; P = 0.006)] were significant. Thus, in this 5-year follow-up study, HYAL1, together with EPE and margin, was found to be an independent prognostic indicator.
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Biomarcadores de Tumor/metabolismo , Ácido Hialurónico/metabolismo , Hialuronoglucosaminidasa/metabolismo , Neoplasias de la Próstata/metabolismo , Adulto , Anciano , Progresión de la Enfermedad , Humanos , Inmunohistoquímica , Modelos Logísticos , Masculino , Persona de Mediana Edad , Pronóstico , Neoplasias de la Próstata/enzimología , Neoplasias de la Próstata/cirugíaRESUMEN
BACKGROUND: One of the goals of a noninvasive test for bladder carcinoma screening would be to reduce surveillance cystoscopies among patients with a history of bladder carcinoma. In addition, an accurate bladder carcinoma marker could be used to screen a high-risk population. The authors examined the efficacy of the hyaluronic acid-hyaluronidase (HA-HAase) and BTA-Stat tests to detect and predict bladder carcinoma recurrence and tested their specificity for bladder carcinoma screening. METHODS: Over a four year period, the authors prospectively collected 225 urine specimens from 70 bladder carcinoma patients and analyzed them by the HA-HAase test. Tumors were identified during 178 visits, and in 47 specimens there was no evidence of disease (NED). Twenty six of these 70 patients were randomly selected to have the BTA-Stat test (111 surveillance visits). In a separate study, 401 former Department of Energy (DOE) workers, who are likely to be at a higher risk for bladder carcinoma, were screened by the HA-HAase and BTA-Stat urine tests. RESULTS: The HA-HAase test had an approximately 91.0% sensitivity, 70% specificity, 87% accuracy, 92% positive predictive value (PPV), and 67% negative predictive value (NPV) in the 70 bladder carcinoma patients. There were 14 false-positives; however, 6 of these had recurred in approximately 5 months. Only 4 out of 33 NED cases recurred in that time period (chi-square = 5.43; degrees of freedom [DF] = 1; P = 0.0198). Thus, a false-positive HA-HAase test carried a significant risk of recurrence within five months (relative risk [RR] = 3.5; odds ratio [OR] = 5.44). In a direct comparison, the HA-HAase and BTA-Stat had 94% and 61% sensitivity, 63% and 74% specificity, 87% and 64% accuracy, 89% and 88% PPV, and 77% and 38% NPV, respectively. While 6 of the 10 false-positive on the HA-HAase test recurred in 5 months (chi-square = 9.6; DF = 1; P = 0.004), only 1 of the 7 false-positives on the BTA-Stat test recurred in that time period (chi-square = 0.096; DF = 1; P = 0.756). The RR and OR for the HA-HAase test were 10.2 and 24, and for the BTA-Stat, 1.4 and 1.5, respectively. In the DOE worker screening study, the HA-HAase and BTA-Stat had 14% (56 out of 401) and 16.7% (67 out of 401) positive rates, respectively. Sixty three percent of the positives on the BTA-Stat test, but only 25% of the positives on the HA-HAase test, had benign urologic conditions. None of the biomarker positive cases with clinical follow-up (n = 29) had evidence of bladder carcinoma. CONCLUSIONS: The HA-HAase test is efficient and superior to the BTA-Stat for detecting and predicting bladder carcinoma recurrence. Noninvasive tests with low false positive rates could be used for bladder carcinoma screening in high-risk populations (e.g., those with occupational exposure to carcinogens or smokers).
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Biomarcadores de Tumor/orina , Factor H de Complemento/orina , Ácido Hialurónico/orina , Hialuronoglucosaminidasa/orina , Neoplasias de la Vejiga Urinaria/diagnóstico , Humanos , Recurrencia Local de Neoplasia , Neoplasias de la Vejiga Urinaria/orinaRESUMEN
In the causative agent of syphilis, Treponema pallidum, the gene encoding 3-phosphoglycerate mutase, gpm, is part of a six-gene operon (tro operon) that is regulated by the Mn-dependent repressor TroR. Since substrate-level phosphorylation via the Embden-Meyerhof pathway is the principal way to generate ATP in T. pallidum and Gpm is a key enzyme in this pathway, Mn could exert a regulatory effect on central metabolism in this bacterium. To study this, T. pallidum gpm was cloned, Gpm was purified from Escherichia coli, and antiserum against the recombinant protein was raised. Immunoblots indicated that Gpm was expressed in freshly extracted infective T. pallidum. Enzyme assays indicated that Gpm did not require Mn(2+) while 2,3-diphosphoglycerate (DPG) was required for maximum activity. Consistent with these observations, Mn did not copurify with Gpm. The purified Gpm was stable for more than 4 h at 25 degrees C, retained only 50% activity after incubation for 20 min at 34 degrees C or 10 min at 37 degrees C, and was completely inactive after 10 min at 42 degrees C. The temperature effect was attenuated when 1 mM DPG was added to the assay mixture. The recombinant Gpm from pSLB2 complemented E. coli strain PL225 (gpm) and restored growth on minimal glucose medium in a temperature-dependent manner. Increasing the temperature of cultures of E. coli PL225 harboring pSLB2 from 34 to 42 degrees C resulted in a 7- to 11-h period in which no growth occurred (compared to wild-type E. coli). These data suggest that biochemical properties of Gpm could be one contributing factor to the heat sensitivity of T. pallidum.
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Operón , Fosfoglicerato Mutasa/genética , Fosfoglicerato Mutasa/metabolismo , Treponema pallidum/enzimología , Treponema pallidum/genética , Adenosina Trifosfato/metabolismo , Animales , Cromosomas Bacterianos/genética , Clonación Molecular , Medios de Cultivo , Escherichia coli/enzimología , Escherichia coli/crecimiento & desarrollo , Geobacillus stearothermophilus/enzimología , Glucosa/metabolismo , Concentración de Iones de Hidrógeno , Cinética , Manganeso/farmacología , Músculo Esquelético/enzimología , Fosfoglicerato Mutasa/aislamiento & purificación , Conejos , Proteínas Recombinantes/aislamiento & purificación , Proteínas Recombinantes/metabolismo , Lugares Marcados de Secuencia , Temperatura , Treponema pallidum/crecimiento & desarrolloRESUMEN
The angiogenic response of a progressing malignancy is characterized by a shift in the balance of stimulatory and inhibiting factors of angiogenesis. Recognition of the regulated steps in tumor angiogenesis provides unique targets for developing anti-tumor therapy. Vitaxin is a humanized monoclonal antibody, which has specificity for the integrin alpha v beta 3 (vitronectin receptor). This antibody can impair the vascular response of endothelial cell growth factors in vitro and inhibit tumor cell mediated angiogenesis in pre-clinical animal models. Patients with metastatic cancer who failed standard therapy received intravenous doses of 10, 50 or 200 mg in cohorts of three patients. The unlabeled dose of Vitaxin was infused on days 0 and 21 of a treatment cycle. All patients received a pre-therapy imaging dose of 1 mg of Tc-99m Vitaxin with gamma camera imaging studies. There was no significant toxicity noted in these three dose levels. There were no objective anti-tumor responses. Three patients received two cycles of therapy and had stable disease at day 85 when taken off study. Radioimaging of tumor vasculature was unsuccessful although one patient with alpha v beta 3 positive melanoma had imaging of tumor sites. There was no immune response to Vitaxin in any patient. Patients receiving 10 mg doses of Vitaxin had poor plasma recovery of injected doses and brief circulation in plasma. Doses of 50 and 200 mg had plasma recovery that better approximated the predicted levels in plasma and circulation half-lives of approximately 7 days. This data suggests that an every three-week schedule of Vitaxin at doses of 200 mg (2.5-3.5 mg/kg) can maintain circulating levels of antibody with little or no toxicity. Future studies will be challenged to define anti-tumor activity in malignancy or appropriate surrogates of anti-tumor effect and explore escalating doses and alternate schedules of administration.
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Anticuerpos Monoclonales/administración & dosificación , Neoplasias/tratamiento farmacológico , Receptores de Vitronectina/inmunología , Adolescente , Anticuerpos Monoclonales/efectos adversos , Anticuerpos Monoclonales/farmacocinética , Anticuerpos Monoclonales Humanizados , Relación Dosis-Respuesta Inmunológica , Femenino , Humanos , Masculino , Neoplasias/diagnóstico por imagen , Neoplasias/metabolismo , Compuestos de Organotecnecio , Proyectos Piloto , CintigrafíaRESUMEN
A fundamental tenet of microbial pathogenesis is that bacterial pathogens must overcome host iron limitation to establish a successful infection. Surprisingly, the Lyme disease pathogen Borrelia burgdorferi has bypassed this host defense by eliminating the need for iron. B. burgdorferi grew normally and did not alter gene expression in the presence of iron chelators. Furthermore, typical bacterial iron-containing proteins were not detected in cell lysates, nor were the genes encoding such proteins identified in the genome sequence. The intracellular concentration of iron in B. burgdorferi was estimated to be less than 10 atoms per cell, well below a physiologically relevant concentration.
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Grupo Borrelia Burgdorferi/fisiología , Hierro/metabolismo , Proteínas Bacterianas/biosíntesis , Proteínas Bacterianas/genética , Grupo Borrelia Burgdorferi/genética , Grupo Borrelia Burgdorferi/crecimiento & desarrollo , Grupo Borrelia Burgdorferi/patogenicidad , Calcio/metabolismo , Carbonil Cianuro m-Clorofenil Hidrazona/farmacología , Quelantes/farmacología , Medios de Cultivo , Expresión Génica , Genoma Bacteriano , Quelantes del Hierro/farmacología , Magnesio/metabolismo , Manganeso/metabolismo , Poliestirenos/farmacología , Polivinilos/farmacología , Desacopladores/farmacología , Zinc/metabolismoRESUMEN
Primary signet ring cell carcinoma of the urinary bladder and colon are rare disease entities that are aggressive, difficult to manage, and portend a poor prognosis. We present a case report of a 25-year-old man born with an imperforate anus who developed signet ring cell carcinoma of the pulled-through sigmoid colon that mimicked a primary invasive bladder tumor. Despite radical surgery and adjuvant radiation, the patient died of his disease 7 months after surgery.
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Adenocarcinoma Mucinoso/patología , Ano Imperforado/cirugía , Carcinoma de Células en Anillo de Sello/patología , Neoplasias del Colon Sigmoide/patología , Neoplasias de la Vejiga Urinaria/patología , Adulto , Carcinoma de Células en Anillo de Sello/etiología , Colon Sigmoide/cirugía , Diagnóstico Diferencial , Resultado Fatal , Humanos , Masculino , Neoplasias del Colon Sigmoide/etiologíaRESUMEN
OBJECTIVE: To assess the clinical behaviour of clinically localized prostate cancer in elderly patients monitored until progression, and the impact of clinical variables, i.e. clinical stage, Gleason score, the dynamics of prostate specific antigen (PSA) and age, on the natural history of the disease. PATIENTS AND METHODS: Between February 1991 and January 1998, 54 patients (mean age 76.4 years, median 77 at the time of diagnosis) with clinically localized prostate cancer who elected for watchful waiting were identified. They were monitored regularly and treatment deferred until progression. Progression was defined as local stage progression (as assessed on a digital rectal examination), biochemical progression or metastasis. All patients who progressed were offered either radiation therapy or hormonal treatment. Each clinical variable was assessed by univariate and multivariate analysis to predict disease progression. The mean follow-up was 47 months. RESULTS: Of the 54 patients, 28 (52%) progressed; 10 had biochemical, 11 local and four biochemical and local progression, and three developed metastasis. All the patients who progressed elected to receive hormonal treatment. The mean time to progression was 35 months. Gleason score ( 6), age ( 75 years) and serum PSA level ( 10 ng/mL) were statistically significant predictors of disease progression (P = 0.04, < 0.001 and 0.02, respectively). The clinical stage at the time of diagnosis had a borderline effect on disease progression (P = 0.06). On multivariate analysis, Gleason score and PSA level were statistically significant predictors of disease progression. CONCLUSION: These results suggest that the treatment of prostate cancer should not be deferred in patients aged > 75 years with a good performance status when the biopsy has a Gleason score >/= 6 and the serum PSA level is >/= 10 ng/mL.
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Selección de Paciente , Neoplasias de la Próstata/cirugía , Factores de Edad , Anciano , Anciano de 80 o más Años , Distribución de Chi-Cuadrado , Progresión de la Enfermedad , Estudios de Seguimiento , Humanos , Masculino , Estadificación de Neoplasias , Antígeno Prostático Específico/sangre , Neoplasias de la Próstata/sangre , Neoplasias de la Próstata/patologíaRESUMEN
The NCCN Colorectal Cancer Guidelines panel believes that a multidisciplinary approach is necessary for the management of the patient with colorectal cancer. The panel endorses the concept that treatment of patients in a clinical trial has priority over standard or accepted therapy. The recommended surgical procedure for resectable colon cancer is an en bloc resection; laparoscopic surgery should be done only in the context of a clinical trial. For patients with stage III disease, 5-FU-based adjuvant therapy is recommended. A patient who has metastatic disease in the liver or lung should be considered for surgical resection if he or she is a candidate for surgery and if surgery can extend survival. Surgery should be followed by adjuvant chemotherapy. The panel advocates a conservative post-treatment surveillance program for colon and rectal carcinoma patients. Serial CEA determinations are appropriate if the patient is a candidate for aggressive surgical resection, should recurrence be detected. Abdominal and pelvic CT scans should be utilized only when there are clinical indications of possible recurrence. Patients whose disease progresses during 5-FU-based therapy should be treated with irinotecan or encouraged to participate in a phase I or phase II clinical trial.
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Neoplasias Colorrectales , Neoplasias Colorrectales/diagnóstico , Neoplasias Colorrectales/terapia , Humanos , Ganglios Linfáticos/patología , Recurrencia Local de Neoplasia , Estadificación de Neoplasias , Estados UnidosRESUMEN
Genome sequence analysis of Treponema pallidum, the causative agent of syphilis, suggests that this bacterium has a limited iron requirement with few, if any, proteins that require iron. Instead, T. pallidum may use manganese-dependent enzymes for metabolic pathways. This strategy apparently alleviates the necessity of T. pallidum to acquire iron from the host, thus overcoming iron limitation, which is a primary host defense. Interestingly, a putative metal-dependent regulatory protein, TroR, which has homology with the diphtheria toxin regulatory protein, DtxR, from Corynebacterium diphtheriae was identified from T. pallidum. We describe here the characterization of TroR, a regulatory protein. Mobility-shift DNA binding and DNase I footprint assays indicated that purified TroR bound to a 22-nt region of dyad symmetry that overlaps the -10 region of the promoter of the tro operon, which contains the genes for a putative metal transport system, the glycolytic enzyme phosphoglycerate mutase, and TroR. Unlike other metal-dependent regulatory proteins like diphtheria toxin regulatory protein and the ferric ion uptake regulator, Fur, which can be activated by divalent metals such as Fe(2+), Mn(2+), Co(2+), Ni(2+), and Zn(2+), TroR is activated only by Mn(2+). The TroR-Mn(2+) complex binds its target sequence and blocks transcription of the troPO/lacZ fusion, suggesting that TroR acts as a metal-dependent repressor in vivo. In addition, TroR exists as a dimer in both its inactive (metal free) and active states as indicated by chemical crosslinking experiments. Based on these data, we propose that TroR represents a unique regulatory system for controlling gene expression in T. pallidum in response to Mn(2+).
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Proteínas Bacterianas , Manganeso/metabolismo , Proteínas Represoras/química , Proteínas Represoras/genética , Treponema pallidum/química , Secuencia de Bases , Huella de ADN , Regulación Bacteriana de la Expresión Génica , Manganeso/farmacología , Modelos Genéticos , Datos de Secuencia Molecular , Regiones Operadoras Genéticas , Regiones Promotoras Genéticas , Unión Proteica , Proteínas Recombinantes/aislamiento & purificación , Homología de Secuencia de Ácido Nucleico , Transcripción Genética , Treponema pallidum/patogenicidadRESUMEN
MDX-H210 is a chemically, cross-linked, half-humanized bispecific antibody composed of F(ab') fragment from monoclonal antibody (mAb) H22 that binds to the high-affinity receptor Fc gamma RI and F(ab') of mAb 520C9 that recognizes the erbB-2 (HER2/neu) oncoprotein. In a previous trial, the murine bispecific, MDX-210 at a dose of 7 mg/m2, was well tolerated and activated monocytes and macrophages in vivo in doses as low as 0.35 mg/m2. In our multidose trial, granulocyte-macrophage colony-stimulating factor, which increases and activates potential effector cells, was given on days 1-4 at 250 micrograms/m2 s.c. and MDX-H210 was given on day 4 weekly for 4 consecutive weeks. Thirteen patients were treated at dose levels of 1, 3.5, 7, 10, 15, and 20 mg/m2 without dose-limiting toxicity. Fever, chills, and rigors occurred during and up to 2 h postinfusion and correlated with the time to peak levels of tumor necrosis factor-alpha (median 88.2 pg/ml; range 15.6-887 pg/ml) and interleukin-6 (median 371 pg/ml; range 175-2,149 pg/ml). By the fourth consecutive week of treatment the side effects and cytokine levels decreased significantly. Human antibispecific antibody (HABA) levels were increased by 200- to 500-fold above pretreatment levels in 5 of 11 evaluable patients after 3 weeks of treatment. The monocyte and granulocyte population increased on days 4 and 11 (median 44%; range 18-68% and 42%; 19-71%), respectively, for monocytes and (60%; 43-75% and 74%; 54-82%) on days 4 and 11 for granulocytes. There was a significant decrease in the monocyte populations immediately after MDX-H210 administration (median decrease 73%; range 42-94%) and (52%; 12-72%) on days 4 and 11, respectively. Ten patients completed 4 weeks of treatment. One patient had a 48% reduction in an index lesions and six patients had stable disease at the time of evaluation. Three patients progressed before the fourth week. The therapy was generally well tolerated with toxicity, primarily, limited to the days of treatment.
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Adenocarcinoma/tratamiento farmacológico , Anticuerpos Biespecíficos/administración & dosificación , Anticuerpos Monoclonales/administración & dosificación , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Factor Estimulante de Colonias de Granulocitos y Macrófagos/administración & dosificación , Receptor ErbB-2/efectos de los fármacos , Adenocarcinoma/inmunología , Adolescente , Adulto , Anciano , Anticuerpos Biespecíficos/efectos adversos , Anticuerpos Monoclonales/sangre , Anticuerpos Monoclonales Humanizados , Formación de Anticuerpos/efectos de los fármacos , Citocinas/efectos de los fármacos , Citocinas/metabolismo , Supervivencia sin Enfermedad , Relación Dosis-Respuesta a Droga , Esquema de Medicación , Femenino , Estudios de Seguimiento , Factor Estimulante de Colonias de Granulocitos y Macrófagos/efectos adversos , Humanos , Inyecciones Subcutáneas , Recuento de Leucocitos/efectos de los fármacos , Masculino , Persona de Mediana Edad , Proyectos Piloto , Receptor ErbB-2/sangre , Índice de Severidad de la Enfermedad , Tasa de Supervivencia , Resultado del TratamientoRESUMEN
We have cloned a 3.6-kb genomic DNA fragment from Pseudomonas aeruginosa harboring the rpoA, rplQ, katA, and bfrA genes. These loci are predicted to encode, respectively, (i) the alpha subunit of RNA polymerase; (ii) the L17 ribosomal protein; (iii) the major catalase, KatA; and (iv) one of two iron storage proteins called bacterioferritin A (BfrA; cytochrome b1 or b557). Our goal was to determine the contributions of KatA and BfrA to the resistance of P. aeruginosa to hydrogen peroxide (H2O2). When provided on a multicopy plasmid, the P. aeruginosa katA gene complemented a catalase-deficient strain of Escherichia coli. The katA gene was found to contain two translational start codons encoding a heteromultimer of approximately 160 to 170 kDa and having an apparent Km for H2O2 of 44.7 mM. Isogenic katA and bfrA mutants were hypersusceptible to H2O2, while a katA bfrA double mutant demonstrated the greatest sensitivity. The katA and katA bfrA mutants possessed no detectable catalase activity. Interestingly, a bfrA mutant expressed only approximately 47% the KatA activity of wild-type organisms, despite possessing wild-type katA transcription and translation. Plasmids harboring bfrA genes encoding BfrA altered at critical amino acids essential for ferroxidase activity could not restore wild-type catalase activity in the bfrA mutant. RNase protection assays revealed that katA and bfrA are on different transcripts, the levels of which are increased by both iron and H2O2. Mass spectrometry analysis of whole cells revealed no significant difference in total cellular iron levels in the bfrA, katA, and katA bfrA mutants relative to wild-type bacteria. Our results suggest that P. aeruginosa BfrA may be required as one source of iron for the heme prosthetic group of KatA and thus for protection against H2O2.
Asunto(s)
Proteínas Bacterianas , Catalasa/metabolismo , Grupo Citocromo b/genética , Grupo Citocromo b/metabolismo , Ferritinas/genética , Ferritinas/metabolismo , Peróxido de Hidrógeno/farmacología , Pseudomonas aeruginosa/genética , Pseudomonas aeruginosa/metabolismo , Secuencia de Aminoácidos , Catalasa/genética , Grupo Citocromo b/química , Farmacorresistencia Microbiana , Escherichia coli/efectos de los fármacos , Escherichia coli/genética , Escherichia coli/metabolismo , Evolución Molecular , Ferritinas/química , Prueba de Complementación Genética , Genotipo , Datos de Secuencia Molecular , Filogenia , Plásmidos , Pseudomonas aeruginosa/efectos de los fármacos , Proteínas Recombinantes de Fusión/metabolismo , Mapeo Restrictivo , Alineación de Secuencia , Homología de Secuencia de AminoácidoRESUMEN
The present project is a step by step description of the creation of a computerized surveillance system using historical information derived from automated expert system acquisition. Since historical information is in many cases not sufficient for establishing an individual's medical diagnosis, the accuracy of surveillance is measured against the "gold standard" diagnosis provided by a panel of physicians. It was possible to survey within acceptable limits of accuracy in the conditions of the project. The results reveal a high level of sensitivity by computer surveillance as well as an accurate ability of electronic tracking of disease incidence over a period of time. However, further investigation into the accuracy of electronic surveillance and selection of symptoms used to define a disease should be studied. The feasibility of employing electronic historical medical information to survey disease has potential in providing real-time epidemiological data.
Asunto(s)
Sistemas Especialistas , Sistemas de Registros Médicos Computarizados , Vigilancia de la Población/métodos , Enfermedades Respiratorias/epidemiología , Estudios de Factibilidad , Humanos , Anamnesis , Atención Primaria de Salud , Enfermedades Respiratorias/diagnósticoRESUMEN
OBJECTIVE: To study the relationship between the genetic degree of Cherokee ancestry, the apolipoprotein E *E4 (APOE*E4) allele type, and the development of Alzheimer disease (AD) in individuals from the Cherokee Nation who reside in northeastern Oklahoma. SETTING: Alzheimer disease center satellite clinic and university departments of neurology, psychiatry, and academic computing. DESIGN: Standardized dementia evaluations based on criteria from the National Institute of Neurological and Communicative Disorders and Stroke and the Alzheimer's Disease and Related Disorders Association were performed on 26 patients aged 65 years or older to establish a diagnosis of AD. Twenty-six control subjects were recruited and similarly assessed. The APOE allele type determinations were obtained on all patients and control subjects. Appropriate statistical analyses were used to compare the genetic degree of Cherokee ancestry, the APOE allele type, and the development of AD. RESULTS: The data indicated that as the genetic degree of Cherokee Indian ancestry increased, the representation of AD decreased. The 9 patients with AD with a greater than 50% genetic degree of Cherokee ancestry constituted 35% of the group with AD. The 17 remaining patients with AD who were less than 50% Cherokee constituted 65% of the group with AD. In contrast, 17 (65%) of the control subjects were more than 50% Cherokee; only 9 (35%) were less than 50% Cherokee. These percentages of AD were not changed by the *E4 allele. This inverse relationship between the genetic degree of Cherokee ancestry and AD, independent of the APOE*E4 allele status, diminished with increasing age, suggesting an age-related protective effect of being Cherokee. For a decrease of 10% in Cherokee ancestry, the odds of developing AD are estimated to be 9.00 times greater at age 65 years but only 1.34 times greater at age 80 years. CONCLUSIONS: A greater genetic degree of Cherokee ancestry reduces the risk of developing AD and, thus, seems protective. This protective genetic factor is independent of APOE allele type and diminishes with age.
Asunto(s)
Enfermedad de Alzheimer/genética , Indígenas Norteamericanos/genética , Anciano , Anciano de 80 o más Años , Envejecimiento/fisiología , Alelos , Apolipoproteína E4 , Apolipoproteínas E/genética , Femenino , Humanos , Masculino , Análisis Multivariante , Factores de RiesgoRESUMEN
Microfibrillar collagen is a bovine collagen material that promotes hemostasis. When mixed with contrast material it makes a fine slurry that is easily injected through small catheters. Experience with preoperative embolization of head and neck neoplasms in six patients indicates that microfibrillar collagen slurry is a highly effective, easy to use embolic material for occlusion of highly vascular neoplasms.
Asunto(s)
Colágeno/uso terapéutico , Embolización Terapéutica , Neoplasias de Cabeza y Cuello/cirugía , Hemostáticos/uso terapéutico , Adolescente , Adulto , Anciano , Angiografía , Arterias Carótidas/diagnóstico por imagen , Tumor del Cuerpo Carotídeo/cirugía , Medios de Contraste , Femenino , Neoplasias de Cabeza y Cuello/irrigación sanguínea , Neoplasias de Cabeza y Cuello/terapia , Humanos , Masculino , Meningioma/cirugía , Persona de Mediana Edad , Neoplasias Nasofaríngeas/cirugía , Cuidados PreoperatoriosRESUMEN
Vascular changes, in the absence of respiratory complications, were observed in monkeys and dogs after infusion of 20% ethanol solutions.