Advances and controversies in the classification and grading of pituitary tumors.

BACKGROUND: Pituitary tumors are common lesions, and they represent the second most frequent primary brain tumor. Their classification has undergone several changes over time. The World Health Organization conducts periodic expert review/consensus meetings and publishes the results as recommendations for changes in classification, based on advances in molecular and genetic advances. This paper summarizes the results of the 2017 WHO Classification, which recommends several important changes. PURPOSE: This paper provides a review of the major changes and issues leading to an understanding of the basis for a new pituitary tumor classification. They include the rejection and modification of prior conceptual and pathological characteristics of these neoplasms. There is also considerable concern related to invasive and recurrent pituitary tumors which follow a less benign course than the typical pituitary adenoma. METHODS: A review of the outcome data for the previously designated "atypical" pituitary tumor category revealed that the former criteria were not adequate to support their ability to predict with accuracy the clinical course of a given tumor. A similar review was accomplished regarding the role of the p53 tumor suppressor mutation. Again, there was no reliable contribution of p53 status to tumor aggressiveness. Other changes have occurred regarding the cytogenetic lineage of the various subtypes of pituitary adenoma. The transcription factors Pit-1, SF-1, and TPit play a major role in determining tumor subtypes and have become part of the classification criteria. RESULTS: These advances now help provide the background for more reliable and consistent classification of pituitary adenomas. Further definition of aggressive characteristics such as cavernous sinus and dural invasion remain to be considered in the quest to make more accurate prognostic projections based on histopathological analysis. CONCLUSIONS: The 2017 WHO Classification of Pituitary Tumors provides a more solid basis for accurate and reliable prognostic assessment of these lesions. Further progress undoubtedly will be made as the recommendations of this update are incorporated in to routine use.

Proteinograma sérico de cães sadios e com linfoma obtido por eletroforese em gel de poliacrilamida (SDS-PAGE) / Serum protein concentrations in healthy dogs and dogs with lymphoma by means of sodium dodecil sulphate-polyacrylamide gel electrophoresis (SDS-PAGE)

Foram avaliadas amostras de soro sanguíneo de 10 cães sadios e de 12 com linfoma, utilizando-se a eletroforese em gel de poliacrilamida contendo dodecil sulfato de sódio. Houve diferença entre as médias dos teores de proteína total de cães sadios, 7,68g/dL±0,46 e de cães com linfoma, 7,93g/dL±2,49. As concentrações de IgA e IgG não foram diferentes entre os grupos. Os teores das proteínas de pesos moleculares 142000, 110000, 52000, 49000, 24000 e 18000 dáltons foram mais elevados em cães com linfoma. Os cães com linfoma apresentaram concentrações mais elevadas de ceruloplasmina, 43,95mg/dL±18,19, e haptoglobina, 554mg/dL±449,51, e menores de albumina, 2908mg/dL±476,67, em comparação aos cães sadios (ceruloplasmina: 3,42mg/dL±7,44; haptoglobina: 94,54mg/dL±59,50 e albumina: 4207mg/dL±206,18). Conclui-se que concentrações séricas mais elevadas de ceruloplasmina e haptoglobina e menores de albumina podem estar associadas ao linfoma em cães.

Blood serum samples of ten healthy dogs and 12 dogs with lymphoma were evaluated by means of sodium dodecil sulphate-polyacrylamide gel electrophoresis. There was difference in total protein concentrations among healthy dogs, 7.68g/dL±0.46 and dogs with lymphoma, 7.93g/dL±2.49 values of immunoglobulins A and G presented no difference between groups. Serum proteins with molecular weights 142,000; 110,000; 52,000; 49,000; 24,000; and 18,000 Daltons presented increased concentrations in dogs with lymphoma. Dogs with lymphoma presented increased ceruloplasmin (43.95mg/dL±18.19) and haptoglobin (554mg/dL±449.51) values and lesser albumin concentration (2,908mg/dL±476.67) when compared to healthy dogs (ceruloplasmin: 3.42mg/dL±7.44; haptoglobin: 94.54mg/dL±59.50, and albumin: 4,207mg/dL±206.18). In conclusion, increased ceruloplasmin and haptoglobin and lesser albumin serum concentrations are possibly related to lymphoma in dogs.

Associação do methimazole e do ondansetron à quimioterapia com cisplatina em cães submetidos a quatro diferentes protocolos de fluidoterapia / Association of methimazole and ondansetron to cisplatin chemotherapy in dogs submitted to four different fluid therapy protocols

Utilizaram-se 12 cães, machos, distribuídos em quatro grupos (G) experimentais, selecionados de acordo com o tempo de fluidoterapia com solução fisiológica 0,9 por cento: G1 (sem fluidoterapia), G2 (uma hora de fluidoterapia antes da cisplatina), G3 (uma hora de fluidoterapia antes da cisplatina e uma hora após) e G4 (duas horas de fluidoterapia antes da cisplatina e uma após). Todos os animais receberam a cisplatina (70mg/m²), pela via intravenosa, sendo os ciclos de quimioterapia realizados em intervalos de três semanas, num total de três ciclos. O ondansetron (0,4mg/kg) foi administrado pela via intravenosa, a cada oito horas, no dia da quimioterapia e, a seguir, a cada 12 horas, por dois dias. O methimazole (40mg/kg) foi pela via oral, 30 minutos antes da cisplatina e quatro horas após. Avaliaram-se os parâmetros hematológicos, bioquímicos, urinários e dosagem de tiroxina e triiodotironina a cada sete dias até o término do experimento. Esse protocolo foi eficaz e seguro em cães que permaneceram sob fluidoterapia durante duas a três horas. Os animais que não receberam fluidoterapia e os que ficaram somente uma hora sob infusão intravenosa de solução fisiológica apresentaram alterações que resultaram em não-recomendação do protocolo.

Twelve male dogs were divided in four experimental groups (G), which were selected according to the time of 0.9 percent saline fluid therapy: G1 (without saline therapy), G2 (an hour of saline therapy before receiving cisplatin), G3 (an hour of saline therapy before receiving cisplatin and one hour after) and G4 (two hours of saline therapy before receiving cisplatin and one hour after). All the animals received 70mg/m² cisplatin, via intravenous, and the chemotherapy cycles were accomplished in intervals of three weeks, in a total of three cycles. Ondansetron (0.4mg/kg) was administered via intravenous, every 8 hours, on the day of the chemotherapy and, from that, every 12 hours for two days. Methimazole (40mg/kg) was orally administered 30 minutes before the cisplatin injection and four hours after. The hematological, biochemical and urinary parameters and dosages of tiroxin and triiodotironine were evaluated every seven days until the end of the experiment. This protocol was effective and safe in the animals that stayed for a period of two or three hours receiving saline therapy, because they did not present clinical and laboratory alterations of the use of chemotherapy. The animals that did not receive saline therapy and those that received infusion of physiologic solution via IV only during one hour presented alterations that made unfeasible the use of this protocol.

Un caso de leucemia aguda linfocítica (ALL), tipo B con un inusual rearreglo cromosómico / A case of acute lymphocytic leukemia (ALL), B with an unusual type chromosomal rearrangement

Presentamos los hallazgos cromosómicos de una mujer de cuatro años de edad con trombocitopenia. El cariotipo demostró un 1(7) q(10) como una posible deleción en 11q23 y un cuestionable rearreglo en 9p. Los estudios por FISH de ambas interfase del núcleo y metafase de la célula, usando la fase de reposo MLL y caracterización de la prueba instrumental en el gen MLL, el cual fue encriptado por análisis citogenético convencional. Específicamente, el patrón FISH fue consistente con una inserción de la región 5' del gen MLL dentro de un cromosoma 4 hacia la banda q21, mas estrechamente una variante 1(4;11) (q 21;g23). Este caso ejemplifica la importancia del FISH y su consiguiente caracterización de casos precursores B-cell all, sin algún significado pronóstico de anormalidad cromosómica.

We present the chromosome findings in a 4-year-old female with thrombocytopenia. The karyotype showed an i(7)(q10) as well as a possible deletion on 11q23 and a questionable rearrangement on 9p. FISH studies on both interphase nuclei and metaphase cells using the MLL break apart rearrangement probe were instrumental in the characterization of an MLL gene rearrangement , which was cryptic by conventional cytogenetic analysis. Specifically, the FISH pattern was consistent with an insertion of the 5' region of the MLL gene into one chromosome 4 at band q21, most likely a variant t(4;11)(q21;q23). This case exemplifies the importance of FISH in the further characterization of precursor B-cell ALL cases without any apparent prognostically significant chromosome abnormalities.

Cryptic ins(4;11)(q21;q23q23) detected by fluorescence in situ hybridization: a variant of t(4;11)(q21;q23) in an infant with a precursor B-cell acute lymphoblastic leukemia report of a second case.

We report the chromosomal findings in a 4-year-old female with precursor B-cell acute lymphoblastic leukemia (ALL). The diagnostic karyotype showed an isochromosome 7q, i(7)(q10), as well as questionable rearrangements on 9p and 11q. Fluorescence in situ hybridization (FISH) studies on both interphase and metaphase cells using the MLL "break-apart" and the centromeric chromosome 4 probes were instrumental in the characterization of an MLL gene rearrangement, which was cryptic by conventional cytogenetic analysis. Specifically, the FISH pattern was consistent with an insertion of the 5' region of the MLL gene into chromosome 4 at band q21, most likely a variant t(4;11)(q21;q23). This is the second case of FISH detection of an ins(4;11) in ALL. Our case exemplifies the importance of FISH in the further characterization of precursor B-cell ALL cases without any apparent prognostically significant chromosomal abnormalities.

Enzymatic treatments on paraffin blocks for DNA flow cytometry.

The Hedley method for DNA ploidy analysis on paraffin-embedded tissue allows retrospective studies of large numbers of common and rare tumors for which treatment, progression, and outcome are known. However, the technique is cumbersome and has many variables, only some of which can be controlled at the time of laboratory analysis. We performed DNA ploidy analyses on two blocks from two islet cell tumors and on five blocks from two colon carcinomas. Sections of 50-microns thickness were deparaffinized in xylene, rehydrated in graded alcohols and in distilled water, and disaggregated with various enzymatic treatments: 0.05% pepsin (30 and 90 min), 0.5% pepsin (30 and 90 min), 0.05% protease (60 min), and 0.1% protease (60 min). The cell suspensions obtained were filtered, washed in PBS, and visually evaluated in a hemocytometer. Nuclei were treated with RNAse (0.1%) and stained with 50 micrograms/ml propidium iodide. Results were evaluated with the following criteria: (a) recovery of DNA aneuploid and/or G2M cells (cell-cycle analysis and visual evaluation); (b) coefficient of variation of the major peak (DNA diploid or DNA aneuploid depending on the case); (c) amount of debris (background events and visual evaluation); (d) mean channel for the G0G1 peak; (e) event rate; and (f) G2M/G0G1 ratio. The best results were observed with 0.05% protease when there was tissue necrosis and hence cell fragility, with 0.1% protease when there was significant tissue fibrosis, and with 0.05% pepsin (90 min) when there were intact cellular specimens without fibrous entrapment. The original procedure using 0.5% pepsin for 30 min produced less cell recovery, and histogram quality similar to or worse than these modifications in all cases studied.(ABSTRACT TRUNCATED AT 250 WORDS)

Evaluation of a rapid screening immunoassay for antibodies to Toxoplasma gondii.

A total of 349 human serum samples were examined for anti-Toxoplasma antibodies by the Murex Single Use Diagnostic System (SUDS) qualitative screening test, indirect hemagglutination assay (IHA), and indirect immunofluorescence assay (IFA). Concordant results with SUDS and IHA were obtained for 91.9% of serum samples; 8.9% were SUDS+ and IHA-; none were SUDS- and IHA+. Comparison of the SUDS assay with IFA showed a concordance of 95.3%, with a sensitivity of 97.5% and a specificity of 94.7%. Moreover, the positive and negative predictive values were 84.9 and 99.2%, respectively, when results of the SUDS assay and IFA were compared. The SUDS assay is a rapid, simple test requiring no instrumentation and can be performed on 50 microliters of serum, features which make this an excellent screening test for detecting anti-Toxoplasma antibodies, particularly in outpatient settings.

Transplacental immunization of the human fetus to tetanus by immunization of the mother.

Experimental studies in rats showed that immunization of the pregnant female led to the transplacental immunization of her fetuses. The possibility that this also occurred in humans was explored by immunizing 42 pregnant women with tetanus toxoid (2.5 or 5 Lf) in the fifth and eighth months of pregnancy and comparing the immune responses of their offspring with the responses of the offspring of 25 unimmunized mothers. Only the offspring of the immunized mothers were sensitized to tetanus. IgM antitetanus antibodies were in their blood before immunization with diphtheria, pertussis, tetanus vaccine (DPT), they had a more rapid (P less than 0.01) response to DPT immunization, and they were still highly sensitized (P less than 0.01) to tetanus 13 mo after birth. In addition, pregnancy had no immunosuppressive effect (P less than 0.05) on the responses of the mothers to tetanus toxoid. Thus, transplacental immunization occurs in humans; it enhances the response of the offspring to subsequent immunization, and it could be used to circumvent the necessity for immunization in early neonatal life.

Radioimmunoassay of IgM and IgG antitetanus toxoid antibody.

A radioimmunoassay was developed to quantitate antitetanus toxoid antibody in whole plasma which can detect 0.75 ng IgG antibody (0.75 micrograms/ml plasma) and 1.60 ng IgM antibody (0.40 micrograms/ml plasma). The IgG antibody was measured by direct binding to a tetanus toxoid-Sepharose immunoadsorbent. The IgM antibody was measured by inhibition of binding by soluble tetanus toxoid because of the relatively high and erratic nonspecific reaction between IgM and the immunoadsorbent. The immunoglobulin standards were [131I]IgG or [131I]IgM coupled to Sepharose, respectively, and the amount of antibody activity in the 125I-labeled antiglobulin reagents could be calculated from the amount bound to the immunoadsorbent and the specific activity of the appropriate immunoglobulin standard. The assay is sensitive, reproducible and suitable for use with large numbers of samples: it should find wide applicability in both clinical and experimental settings.