RESUMEN
BACKGROUNDS: The Ann Arbor system is typically used for the staging of Non-Hodgkin's lymphomas. This classification was nevertheless originally developed in the 1970s for Hodgkin's lymphoma, a disease usually confined to the lymph nodes with less frequent dissemination to extralymphatic organs/tissues and extremely rare primary extranodal involvement. Non-Hodgkin's lymphomas, however, are more often associated with extralymphatic involvement and primary extranodal lymphomas are relatively common (approximately 1/3 of cases). Therefore, the value of the Ann Arbor staging system appears to be limited in these cases. An analysis of data from centres participating within the Czech Lymphoma Study Group showed that staging of Non-Hodgkin's lymphomas with extranodal involvement is not uniform. DESIGN: At the end of 2009, a draft for a Non-Hodgkin's lymphomas staging system was put forward for use by the lymphoma register of the Czech Lymphoma Study Group with special regard paid to the involvement of extralymphatic organs/tissues. This draft was further refined following comments from members of the Czech Lymphoma Study Group committee and the final form was accepted at the meeting of the Czech Lymphoma Study Group committee in January 2010. RESULTS: A consensus was reached at the meeting of the Czech Lymphoma Study Group committee regarding the staging of various combinations of nodal and extranodal involvement. For the purpose of suitable staging and appropriate treatment intensity, extranodal organs were divided into "major"--liver, lungs, bones, mesothelium (pleura, peritoneum, pericardium) and soft tissues. All other organs were defined as "minor". CONCLUSION: The Ann Arbor staging system is suitable for the staging of Non-Hodgkin's lymphomas with lymph node/lymphatic tissue involvement. As regards the extralymphatic spread of the disease or primary extranodal lymphomas, this classification should rather be adapted to practical needs. The validity of the updated classification system will be assessed in both prospective and retrospective Czech Lymphoma Study Group studies.
Asunto(s)
Linfoma no Hodgkin/patología , Humanos , Linfoma no Hodgkin/clasificación , Estadificación de NeoplasiasRESUMEN
BACKGROUND: Clostridium difficile associated diarrhoea / disease (CDAD) is an inflammatory disease of the colon. It affects patients who have been exposed to wide-spectrum antibiotics and long-term in-patient care, with immunosuppression. The most difficult form of this disease is manifested as pseudomembranous enterocolitis, it runs fulminantly in significantevents. In recent years there has been an increase in the incidence of this disease worldwide. Several serious epidemics caused by virulent strains of Clostridium difficile have been discovered in Western Europe, North America and Asia. PATIENTS AND METHODS: We observed an increased occurrence of this disease at our clinic during 2004-2007. A group of 36 patients with CDAD was analyzed in the article. Patients with different severity courses were identified in the group--from slightly running post-antibiotic diarrhoea to serious pancolitis with the manifestation of sepsis and MODS (multiple organ dysfunction syndrome). MATERIALS AND METHODS: It is a retrospective analysis of a patients' group with CDAD. RESULTS AND CONCLUSIONS: According to our experience, in the group of oncological patients, post-antibiotic clostridia diarrhoea often develops in a very complicated and protracted way. It also affects relatively young patients. Protein malnutrition and febrile neutropenia have a significant occurrence during its genesis. A higher risk of CDAD is found in the group of patients with malignant lymphomas and colorectal malignancy. 20% of our patients did not have any previous ATB exposure, so we can express the theory of oncology therapy as a predisposition factor of CDAD. The CDAD relevance in oncological patients cannot be evaluated according to leukocytosis (a significant part of febrile neutropenia in our group).The disease could require a combined causal therapy and intensive supporting treatment.There is a higher risk of heavy MODS illness course in the group of oncological patients. The article also deals with the case report of one complicated case.
Asunto(s)
Clostridioides difficile , Enterocolitis Seudomembranosa/complicaciones , Neoplasias/complicaciones , Infecciones Oportunistas/complicaciones , Adulto , Enterocolitis Seudomembranosa/microbiología , Femenino , Humanos , Infecciones Oportunistas/microbiologíaRESUMEN
BACKGROUND: Evaluation of practical value of monitoring t(14:18) in peripheral blood in follicular lymphoma. METHODS AND RESULTS: t(14;18) was tested in 115 follicular lymphoma patients by methods: FISH, nested and multiplex PCR of blood, bone marrow and lymph node specimens. We tested the patients with rearrangement MBR quantitatively by real-time PCR. Testing intervals of t(14;18) in peripheral blood were 1 month during treatment, 2-3 months during the first year after the end of treatment, then every 4 to 6 months. Patients were clinically examined in the same intervals and regular restaging was done by CT/PET. Each patient was evaluatee separately. Total detection of t(14;18) was 97% regardless tissue and methods of detection, FISH was superior to PCR (95% vs. 72%). The higher number of copies were observed in lymph nodes in comparison to bone marrow (p = 0.036) and peripheral blood (p = 0.016); 46/115 (40%) patients were positive for MBR, we followed up behaviour of t(14;18) in peripheral blood in 33 of them in long intervals (>6 months, med. 33 months). Molecular and clinical courses correlated in 20/33 (61%) patients, 7/33 (21%) clinically relapsed in lasting molecular remission. We found very short interval to clinical relaps in 7 cases of molecular relapses (0-5 months, median 3 months). We could not define "threshold quantity" of clinically important molecular relaps. Lasting molecular remission was associated with clinical in about 60% cases; lasting molecular activity corresponded with clinical relaps in 86% patients. CONCLUSIONS: t(14;18) is highly associated with follicular lymphoma. In practice, monitoring of t(14;18) is feasible only in part of patients. Even if there is some correlation of clinical and molecular course, monitoring of t(14;18) in blood bears only limited prognostic value for the concrete patient. The treatment of patient can not be accomplished on the basis of these results only.
Asunto(s)
Cromosomas Humanos Par 14/genética , Cromosomas Humanos Par 18/genética , Linfoma Folicular/genética , Translocación Genética , Adulto , Anciano , Anciano de 80 o más Años , Femenino , Humanos , Hibridación Fluorescente in Situ , Linfoma Folicular/tratamiento farmacológico , Masculino , Persona de Mediana Edad , Reacción en Cadena de la Polimerasa , Recurrencia , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
BACKGROUND: One of the perspective therapeutic possibilities in follicular lymphomas (FL) is the fludarabine-based regimen FND (fludarabine, mitoxantron, dexamethason). However serious signs of myelotoxicity and significant immunosuppression with appearance of the opportunistic infections were described after the fludarabine treatment. METHODS AND RESULTS: Follicular lymphoma patients with advanced disease grade I-II were treated with FND (6-8 cycles). The immunotoxicity was evaluated by measuring of immunoglobuline levels (IgA, IgG, IgM) and that of lymphocytes subpopulations (CD3+, CD4+, CD8+, CD20+, CD56+) in peripheral blood. The myelotoxicity was evaluated by cultures of progenitor cells (CFC and LTC-IC). Totally 34 patients (median age 55.5 years) were evaluated, the overall response was 72% (CR 61%, PR 11%, progression 28%). 73% patients of 11 after 6-8 FND show persisting CR (27% relapsed) with median follow-up about 15 months. The dominating toxicity was myelotoxicity. The leucopenia grade III-IV occurred in about 30% cycles. Because of toxicity it was necessary to reduce doses of FND in 10% cycles and this treatment had to be finished ahead of schedule in 29% patients. The significant immunotoxicity was found only in the decrease of whole lymphocyte population (p < 0.05) and of IgG level (p < 0.05). The decrease of lymphocyte subpopulations did not reach any statistical significance. The long-term myelotoxicity caused the decrease of LTC-IC that had a clinical correlation with the very difficult mobilization of PBSC. CONCLUSIONS: FND is efficient in treatment of follicular lymphoma. However myelotoxicity seems to be limiting. Myelotoxicity doesn't allow administering scheduled dose of FND in substantial amount of patients, long-term myelotoxicity complicates PBSC-mobilization. Lymphotoxicity is apparent, but seems not to be clinically important.
Asunto(s)
Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Linfoma Folicular/tratamiento farmacológico , Adulto , Anciano , Protocolos de Quimioterapia Combinada Antineoplásica/administración & dosificación , Protocolos de Quimioterapia Combinada Antineoplásica/efectos adversos , Médula Ósea/efectos de los fármacos , Dexametasona/administración & dosificación , Dexametasona/efectos adversos , Femenino , Humanos , Inmunoglobulinas/efectos de los fármacos , Linfocitos/efectos de los fármacos , Masculino , Persona de Mediana Edad , Mitoxantrona/administración & dosificación , Mitoxantrona/efectos adversos , Vidarabina/administración & dosificación , Vidarabina/efectos adversos , Vidarabina/análogos & derivadosRESUMEN
Kinesin motors move on microtubules by a mechanism that involves a large, ATP-triggered conformational change in which a mechanical element called the neck linker docks onto the catalytic core, making contacts with the core throughout its length. Here, we investigate the thermodynamic properties of this conformational change using electron paramagnetic resonance (EPR) spectroscopy. We placed spin probes at several locations on the human kinesin neck linker and recorded EPR spectra in the presence of microtubules and either 5'-adenylylimidodiphosphate (AMPPNP) or ADP at temperatures of 4-30 degrees C. The free-energy change (DeltaG) associated with AMPPNP-induced docking of the neck linker onto the catalytic core is favorable but small, about 3 kJ/mol. In contrast, the favorable enthalpy change (DeltaH) and unfavorable entropy change (TDeltaS) are quite large, about 50 kJ/mol. A mutation in the neck linker, V331A/N332A, results in an unfavorable DeltaG for AMPPNP-induced zipping of the neck linker onto the core and causes motility defects. These results suggest that the kinesin neck linker folds onto the core from a more unstructured state, thereby paying a large entropic cost and gaining a large amount of enthalpy.
Asunto(s)
Dominio Catalítico , Espectroscopía de Resonancia por Spin del Electrón/métodos , Cinesinas/química , Microtúbulos/química , Proteínas Motoras Moleculares/química , Movimiento (Física) , Sitios de Unión , Transferencia de Energía , Humanos , Cinesinas/genética , Mutación , Unión Proteica , Conformación Proteica , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Marcadores de Spin/síntesis química , Estrés Mecánico , TemperaturaRESUMEN
Genetically-encoded affinity tags constitute an important strategy for purifying proteins. Here, we have designed a novel affinity matrix based on the his-arsenical fluorescein dye FlAsH, which specifically recognizes short alpha-helical peptides containing the sequence CCXXCC (Griffin BA, Adams SR, Tsien RY, 1998, Science 281:269-272). We find that kinesin tagged with this cysteine-containing helix binds specifically to FlAsH resin and can be eluted in a fully active form. This affinity tag has several advantages over polyhistidine, the only small affinity tag in common use. The protein obtained with this single chromatographic step from crude Escherichia coli lysates is purer than that obtained with nickel affinity chromatography of 6xHis tagged kinesin. Moreover, unlike nickel affinity chromatography, which requires high concentrations of imidazole or pH changes for elution, protein bound to the FlAsH column can be completely eluted by dithiothreitol. Because of these mild elution conditions, FlAsH affinity chromatography is ideal for recovering fully active protein and for the purification of intact protein complexes.
Asunto(s)
Arsénico , Cromatografía de Afinidad/métodos , Fluoresceínas , Colorantes Fluorescentes , Compuestos Organometálicos , Proteínas/aislamiento & purificación , Marcadores de Afinidad , Secuencia de Aminoácidos , Escherichia coli/genética , Fluoresceínas/síntesis química , Colorantes Fluorescentes/síntesis química , Cinesinas/química , Cinesinas/genética , Cinesinas/aislamiento & purificación , Datos de Secuencia Molecular , Compuestos Organometálicos/síntesis química , Proteínas/química , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/aislamiento & purificaciónRESUMEN
A series of ATP analogs, in which moieties of various sizes have been added to the gamma-phosphorus of ATP, bind to the active site of myosin and to the actomyosin complex in myofibrils and in chemically skinned fibers. The affinity of the analogs for the active site shows only a slight dependence on the size of the added moiety. Addition of even our smallest group (CH3) reduced the binding affinity of ATPgamma-CH3 for S1 to 40 microM, a factor of 10(5) less than observed for ATP. Computer molecular docking of ATP-gammaCH3 into the myosin-ADP.BeF3 crystal structure of Dictyostelium discoideum indicates no steric interference to prevent binding. This suggests that the maintenance of charge at the gamma-phosphate is crucial for tight nucleotide binding. Addition of larger groups, (1) an EPR probe (ATP-gammaSL) or (2) ADP (i.e., P1, P5-diadenosine pentaphosphate, AP5A), reduced the affinity by only approximately a factor of 10 over that of ATP-gammaCH3. In the crystal structure of S1 complexed with nucleotides, the phosphates are buried within a protein structure called "the phosphate tube". Both the bulk of the modifying groups and the lack of dependence on the size of the group are incompatible with threading of the phosphates down the Pi-tube, showing that the tube must open. Similar domain movements have been found in other proteins including members of the G-protein superfamily, a family that has structural homologies to myosin.
Asunto(s)
Adenosina Trifosfatasas/metabolismo , Adenosina Trifosfato/metabolismo , Miosinas/metabolismo , Animales , Unión Competitiva , Reactivos de Enlaces Cruzados , Colorantes Fluorescentes/metabolismo , Fluorometría , Cinética , Maleimidas/farmacología , Modelos Moleculares , Fibras Musculares Esqueléticas/metabolismo , Músculo Esquelético/metabolismo , Unión Proteica/genética , Conejos , Especificidad por Sustrato , Compuestos de Sulfhidrilo , TriptófanoRESUMEN
We have synthesized 2'-deoxy-2'-iodoadenosine-5'-triphosphate (2'-IATP), a heavy-atom analog of adenosine-5'-triphosphate. This compound was made for X-ray structural studies to target the nucleotide site of ATP binding proteins. It was diffused successfully into crystals of the microtubule-based motor proteins ncd (non-claret disjunctional protein from Drosophila melanogaster) and kinesin. With ncd, the nucleotide binding site was 70% occupied and the crystals were able to diffract X-rays to 2.5 A. The iodo-analog provided a useful isomorphous derivative with overall phasing power 1.89 in the range of 25.0-2.5 A. With kinesin, 2'-IATP co-crystallized with the protein. The crystals diffracted to at least 2.8 A with a phasing power of 1.73 in the range of 20.0-5.0 A. The analog was also found to be a substrate for all of the enzymes tested, including creatine kinase, pyruvate kinase, hexokinase, and myosin, with values of Km and Vmax that were within a factor of 10 of those for ATP. The analog supported muscle contraction, relaxing fibers, and producing active tension with values not statistically different from those obtained with ATP. These results all suggest that this analog should be useful for providing a heavy-atom derivative for crystals of enzymes that bind ATP.
Asunto(s)
Adenosina Trifosfatasas/química , Adenosina Trifosfato/análogos & derivados , Proteínas de Drosophila , Cinesinas/química , Proteínas de Microtúbulos/química , Nucleótidos/metabolismo , Proteínas/química , Adenosina Trifosfatasas/metabolismo , Adenosina Trifosfato/síntesis química , Adenosina Trifosfato/metabolismo , Sitios de Unión , Creatina Quinasa/metabolismo , Cristalización , Cristalografía por Rayos X , Hexoquinasa/metabolismo , Cinesinas/metabolismo , Proteínas de Microtúbulos/metabolismo , Estructura Molecular , Miosinas/metabolismo , Proteínas/metabolismo , Piruvato Quinasa/metabolismo , Relación Estructura-Actividad , Especificidad por SustratoRESUMEN
In a series of 2038 paraproteinemic sera, 9 IgD paraproteins (0.44%) were found. Clinical diagnosis of all the 9 was multiple myeloma. One patient had an IgD-lambda plasmacytoma which developed into plasmocellular leukemia. Our series of IgD paraproteins consisted of 6 females and 3 males with a mean age of 58.5 years. The mean concentrations of IgD paraproteins were 6.7 +/- 4.26 g/l and the mean proteinuria was 5.1 +/- 4.76 g/24 h. Eight IgD paraproteins had light chains lambda and one had light chains kappa.
Asunto(s)
Inmunoglobulina D/sangre , Mieloma Múltiple/inmunología , Paraproteinemias/diagnóstico , Adulto , Factores de Edad , Anciano , Anciano de 80 o más Años , Femenino , Humanos , Inmunoglobulina D/orina , Cadenas Ligeras de Inmunoglobulina , Cadenas kappa de Inmunoglobulina , Cadenas lambda de Inmunoglobulina , Inmunoglobulinas/sangre , Masculino , Persona de Mediana Edad , Mieloma Múltiple/complicaciones , Paraproteinemias/etiología , Paraproteinemias/inmunología , Factores Sexuales , Microglobulina beta-2/análisisRESUMEN
A 23-year-old woman was noted to have a hypoesthetic patch over the right mandible. A biopsy specimen revealed alopecia mucinosa. Anesthesia in the involved area has been reported rarely in the literature.