Identification of the novel HLA-DQB1*03:03:02:04 allele in a Spanish individual.

HLA-DQB1*03:03:02:04 and DQB1*03:03:02:02 alleles differ by a single point mutation in intron 2.

Different patterns of A*80:01:01:01 allele generation based on exon or intron sequences.

Generation of the HLA-A*80:01:01:01 allele has been analysed using its complete sequence. Direct comparison of the sequences and phylogenetic trees using the human leukocyte antigen (HLA)-A representative alleles and the major histocompatibility complex (MHC)-A sequences of non-human primates has been made. Results based on exon sequences confirm previously published, but considering only the sequences of the introns, two distinct regions can be differentiated. The first one comprises from the 5' untranslated region region to the first part of intron 3 sequence (shared with A2 family), and the second one includes the sequence from the end of intron 3 to intron 7 (shared with A1/A3/A11/A36/A30 family). Each of them clusters with Gorilla and Chimpanzee MHC-A sequences, respectively, suggesting an origin coming from a common ancestor to Gorilla and Chimpanzee.

Promoter sequences confirm the three different evolutionary lineages described for HLA-G.

HLA-G alleles follow a different pattern of polymorphism generation that those of the HLA classical I alleles. However, this polymorphism maintenance could have an evolutionary specific pathways based on non coding regions as introns, 14 bp deletion/insertion (exon 8) or promoter regions. For this reason, a systematic sequencing study of HLA-G promoter region was done in 36 individuals with a total of 15 different alleles. From the 12 sequences obtained, 7 were new sequences and not previously described. Results show that the sequences have three different patterns of evolution confirming the results obtained in the rest of the sequence regions (exons, introns and 3'UTR) where three different lineages were established. Only one of these lineages includes the non-human primate promoter sequences suggesting the possibility of this lineage could come directly from non-human primates while the other two could be generated after the speciation. More non-human primates MHC-G promoter sequences must be obtained to confirm this hypothesis. Expression and functional assays could be done considering the differences obtained in the promoter regions involving the HLA-G function (mRNA expression, isoforms).

Genomic predictors of response to doxorubicin versus docetaxel in primary breast cancer.

UNLABELLED: Taxanes and anthracyclines improve the outcome of early breast cancer, although the benefit is limited to a small proportion of patients and are toxic. We prospectively looked for predictors of response to these drugs. EXPERIMENTAL DESIGN: Four cycles of doxorubicin (75 mg/m²) or docetaxel (100 mg/m²) were compared as presurgical chemotherapy for breast cancer. Biomarkers were determined by immunohistochemistry and fluorescent in situ hybridization using prechemotherapy core biopsies. Tumors were also classified into one of the molecular intrinsic subtypes using an immunohistochemical panel of five biomarkers and genomic profiles. Single genes and intrinsic subtypes were correlated with response to doxorubicin versus docetaxel. Among the 204 evaluable patients, significant predictors of sensitivity in multivariate analysis were low topo2a expression and ER-negative status for doxorubicin and small tumor size and ER-negative status for docetaxel. Predictors of resistance in multivariate analysis were triple-negative status (ER/PgR/HER2 negative by IHC/FISH) for doxorubicin, and high TNM stage for docetaxel. Triple-negative tumors were associated with topo2a overexpression more than the other subtypes. In 94 patients with gene expression profiles, docetaxel was superior to doxorubicin in the basal-like subtype (good pathological response rate - PCR + class I of 56 vs. 0%; P = 0.034); no significant differences were observed in the other subtypes when comparing these two drugs. Low topo2a expression and ER-negative status were predictors of response to doxorubicin, while small tumor size and ER-negative status predicted response to docetaxel. Docetaxel was superior to doxorubicin in triple-negative/basal-like tumors, while no significant differences were seen in the remaining intrinsic subtypes.

The novel HLA-G*01:03:01:02 allele differs from G*01:03:01:01 by a possible inversion event in intron 3.

HLA-G*01:03:01:02 and G*01:03:01:01 differ by a two nucleotide changes in intron 3.

The HLA-B*83:01 allele is generated by a gene conversion event including whole of exon 2 and partial introns 1 and 2 between B*44 and B*56 alleles.

Several studies have indicated the gene conversion as the most important mechanism about the MHC polymorphism generation when intron sequences are studied. The data obtained confirm that the B*83:01 allele is generated by gene conversion event including exon 2 and partial intron 1 and 2 between B*44 and B*56 alleles.

¿Es necesaria la técnica combinada para la detección del ganglio centinela en el cáncer de mama? / No disponible

Objetivos: Determinar si la punción con colorante mejora los resultados de la biopsia del ganglio centinela. Material y métodos: Ciento cincuenta casos de carcinoma invasor de la mama fueron sometidos a biopsia del ganglio centinela mediante técnica combinada, para determinar si la punción con colorante es rentable. Se realizó inyección subareolar indérmica de una dosis de 0,4 mCi de radioisótopico el día previo a la cirugía. Se practicó gammagrafía en todos los casos. La inyección intraparenquimatosa de 4 cc. de colorante se realizó 20 minutos antes de la cirugía, y se siguió de masaje mamario. Resultados: La tasa de migración fue 92,3% para el radioisótopo y 75% para el colorante (p = 0,01). La media de ganglios resecados fue mayor para la técnica con colorante: 2,6 vs. 1,2 (p = 0,02). No se observaron diferencias en la tasa de falsos negativos (0,2 vs. 0,4) ni en el valor predictivo negativo. La precisión diagnóstica fue mayor para el radioisótopo (90,3% vs. 75% (p = 0,001). El mismo resultado se obtuvo para el porcentaje de éxito técnico (92,3 vs. 75% (0,001)). Conclusiones: La punción con colorante no añade información a la realizada con radioisótopo. A pesar de que la técnica se debe adaptar a las necesidades del centro y a las habilidades del cirujano, una vez superada la curva de validación la técnica radioisotópica por sí misma aporta suficiente información, con menor morbilidad y coste(AU)

Objetives: To determine whether blue dye enhances sentinel node biopsy detection. Material and methods: One hundred fifty hundred consecutive cases of breast cancer were submitted to sentinel node biopsy by combined technique in order to analyze if vital blue was cost-efective. Radioisotope dose was 0,4 mCi of Tc, subareolar intradermic inyection, the day before surgery. Gammagraphy was performed in every case. Colorant was methylene blue, 4 cc administered by intraparenchimatous inyection in upper-outer quadrant 20 minutes previous to surgery, followed by breast massagge. Results: Migration rate was 92.3% for radioisotope and 75% for colorant (p = 0.01). Mean number of nodes excised was higher for colorant: 2,6 vs. 1,2 (p = 0.02). False negative rate showed no difference (0.2 vs. 0.4) nor did negative predictive value. Accuracy to staging (True neg+ true pos./total) was higher for technecium (90.3% vs. 75% (p = 0.001) and so happenned with percentage of technical success (total- no migration): 92.3% vs. 75% (0.001). Conclusions: Colorant did no add any information concerning axillary status. More nodes had to be excised, and bigger incisions were made to achieve direct visualization. Accuracy to stage the lesion and percentage of technical success were higher with radioisotope after the learning curve is achieved and blue injection can be spared(AU)

A new allele, HLA-G*010120, is generated by a recombination event between HLA-G*01010101/02 and HLA-G*01010201.

Recombination between HLA-G*01010101/02 and HLA-G*01010201 generates HLA-G*010120.

The novel HLA-G*01010302 allele differs from G*01010301 by a single nucleotide change in intron 5.

HLA-G*01010301 and G*01010302 differ by a single point mutation in intron 5.

A new allele, HLA-G*01010106, with changes in intron 2.

Three nucleotide changes and one insertion in intron 2 upon human leucocyte antigen (HLA)-G*01010105 generate HLA-G*01010106.

Brain metastasis and carcinomatous meningitis from vulvar squamous cell carcinoma: case report.

BACKGROUND: Brain metastasis and carcinomatous meningitis from gynecological tumors are an uncommon event, usually related to choriocarcinoma, ovarian and cervical cancer. CASE: A 74-year-old woman was diagnosed with locally advanced vulvar squamous carcinoma. Initial therapy consisted of multiagent chemotherapy and vulvar, pelvis and groin irradiation. The patient subsequently developed widely spread metastatic disease including brain and meningeal metastases. CONCLUSION: The rising incidence of central nervous system metastasis in the last two decades is probably associated with treatment-related improvement in life expectancy. To our knowledge, this is the first case reported of brain metastases and meningeal carcinomatosis associated with vulvar squamous cell carcinoma.

P53 overexpression predicts endometrial carcinoma recurrence better than HER-2/neu overexpression.

OBJECTIVE: to investigate the prognostic value of p53 and HER-2/neu overexpression in endometrial cancer. STUDY DESIGN: p53 and HER-2/neu immunostaining was performed in 114 paraffin-embedded specimens of endometrial cancer diagnosed and treated between 1990 and 1997. Nuclear p53 and membrane HER-2/neu immunostaining were used. RESULTS: p53 and HER-2/neu overexpression was observed in 17 cases (14.9%) and in 19 cases (16.7%), respectively. In univariate analysis p53 (P<0.001) and HER-2/neu (P=0.018) overexpression had a positive correlation with a high risk of recurrence. In multivariate analysis, age (P<0.001), FIGO stage (P<0.001), differentiation (P=0.013), non-endometrioid subtypes (P<0.001) and p53 overexpression (P<0.001), but not HER-2/neu overexpression, were independent prognostic indicators of recurrence. Simultaneous p53 and HER-2/neu overexpression made worse the prognostic (P<0.001). CONCLUSIONS: p53 overexpression was an independent predictor of recurrent disease in endometrial cancer. HER-2/neu overexpression had a more limited effect but enhance the effect of p53.

A comparison of epithelial membrane antigen overexpression in benign and malignant endometrium.

OBJECTIVE: The aim of this study was to analyze the value of epithelial membrane antigen (EMA) overexpression in benign and malignant endometrium and its prognostic significance. METHODS: EMA immunostaining was performed in 178 paraffin-embedded specimens including 105 endometrial cancers, 40 endometrial hyperplasias, and 33 benign endometriums. EMA immunostaining was correlated with traditional prognostic factors and progression-free survival in endometrial cancer specimens. RESULTS: EMA overexpression was observed more frequently in adenocarcinomas (60%) than in hyperplasias (15%) or benign endometrium (9.1%). EMA overexpression was observed in two patients with endometrial hyperplasia who progressed to carcinoma. In adenocarcinomas, EMA overexpression had a positive correlation with nonendometrioid subtypes (P = 0.012). In multivariate analysis, FIGO stage (P = 0.025) and EMA overexpression (P = 0.017) were independent prognostic factors for progression-free survival. CONCLUSIONS: EMA overexpression appears to be a marker of malignant transformation in the endometrium and it is an independent predictor of recurrent disease in endometrial cancer.

Prognostic value of DNA quantification in early epithelial ovarian carcinoma.

OBJECTIVE: To evaluate the prognostic value of flow cytometric DNA quantification and immunohistochemical expression of c-erbB-2 and p53, and traditional clinicopathologic variables in stages I-II invasive epithelial ovarian cancer. METHODS: We retrospectively reviewed 77 cases of stages I-II ovarian cancer after comprehensive surgical staging. We recorded anthropometric data (age, menopausal status, weight loss, Karnofsky index) and pathologic variables (tumor size, bilaterality, capsular status, ascites, peritoneal cytology, histologic type, and grade). In 72 cases representative paraffin-embedded samples were available for DNA quantification and immunohistochemical evaluation of c-erbB-2 and p53 overexpression. Most women (87%) had received cisplatin-based adjuvant chemotherapy. RESULTS: The median follow-up was 90 months (range 50-148 months). The 6-year overall disease-free survival rate was 70% (95% confidence interval [CI] 60%, 81%), and overall global survival was 77% (95% CI 67%, 87%). Multivariable analysis using Cox stepwise regression identified DNA content (odds ratio [OR] 12.3; P <.001) and stage (OR 1.4, P =.09) as independent poor prognosis factors for relapse, and DNA content (OR 9.8, P <.001) as the main independent factor for survival. In stepwise discriminant analysis the combination of DNA content and stage provided a correct prediction of relapse in 78% of women. CONCLUSION: Flow cytometric DNA quantification was the main independent prognostic factor of relapse and survival in these women with stages I-II epithelial ovarian cancer.

Tissue quantification of CA 125 in epithelial ovarian cancer.

The objectives of this study were the determination of CA 125 in the cytosol of healthy and carcinomatous ovarian tissue by immunoanalysis, analysis of its correlation with the biological characteristics of ovarian carcinoma, determination of serum CA 125 levels, and study of the prognostic value of the marker in cytosol. The levels of the marker depend not only on the tumor's production rate, so its determination in tissue can indicate more accurately if the tumor is a producer of the marker and establish its value for the prognosis of the disease. Determination of CA 125 in tissue was performed by immunoanalysis in 50 ovarian epithelial cancer samples, 13 benign pathology samples and 32 healthy ovary samples. The presurgical serum level of the marker was also obtained. The correlation between the CA 125 level in the cytosol and the different biological characteristics of the ovarian carcinoma, the serum levels of the marker and survival were analyzed. The CA 125 level proved to be higher in malignant tissue (p < 0.0001). There was a significant association between the tissue marker and histological type (high CA 125 was associated with serous and endometrioid tumors) and between the marker and survival. No relation with stage was found. There was a correlation between the CA 125 level in the cytosol and serum both variables being dependent, with a correlation coefficient of 0.44. This good correlation speaks in favor of the usefulness of CA 125 determination in serum in the follow-up of ovarian cancer. Tumors having high tissue expression of CA 125 were found to have a double relative risk of death, independently of tumor stage.

Treatment of advanced ovarian cancer with cisplatin, adriamycin and cyclophosphamide (PAC).

Forty two ovarian cancer patients with residual disease after the first laparotomy were treated with the combination of cisplatin (80 mg/m2 day 1), adriamycin (50 mg/m2 i.v. day 2) and cyclophosphamide (500 mg/m2 i.v. day 2) (PAC). Forty women were considered evaluable for analysis, with an overall response rate (partial, plus complete responses) of 62.5%. Twelve patients (30%) obtained a complete response (histologically confirmed after second look surgery in 6 cases, surgical complete response, residual tumor completely resected in the second look-in 5 cases and maintained complete clinical remission without second look confirmation in 1 case). Main side effects were nausea and vomiting (90%), leukopenia (70%), mucositis (45%), and anemia (37%). Seventeen percent of the patients were free of disease at 60 months, after a median follow-up of 48 months. The prognostic factors that showed significant influence on survival were the Karnofski index (90-100 vs 80 or less), stage of the disease (II + III vs IV) and the volume of residual tumor after the first surgical procedure (less than or equal to 2 cms vs greater than 2 cms). Patients who achieved a complete remission have not reached the median 5 years survival, which was 10 months for the remaining patients. These results confirm the activity of PAC in ovarian cancer, mainly in those patients with residual tumor of less than 2 cms and good performance status.