Description of the novel HLA-DQB1*02:02:01:02 allele in a Spanish individual.

HLA-DQB1*02:02:01:02 has an intron sequences coming from a combination of DQB1*02:01:01 and DQB1*02:02:01:01.

Description of the novel HLA-DQB1*02:02:01:02 allele in a Spanish individual.

HLA-DQB1*02:02:01:02 has an intron sequences coming from a combination of DQB1*02:01:01 and DQB1*02:02:01:01.

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).

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.

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.

Characterisation of human outbreaks of brucellosis and sporadic cases by the use of hyper-variable octameric oligonucleotide fingerprint (HOOF) variable number tandem repeats.

Hyper-variable octameric oligonucleotide fingerprints (HOOFs) enable typing of Brucella spp. by targeting the 8-bp tandem repeat in eight loci that vary in number (variable number tandem repeats; VNTRs). Brucella is one of the most important zoonotic pathogens, because of its public health and economic consequences. To assess the role of HOOFs as epidemiological markers for Brucella melitensis, which is the main species involved in human brucellosis in Spain, 87 sporadic and outbreak isolates were investigated; these originated from broad or more restricted geographical locations, including unrelated (n = 42), semi-related (n = 19) and closely related (n = 26) groups of isolates. Distinct HOOFs were detected in the entire (n = 74), unrelated (n = 42), semi-related (n = 19) and closely related (n = 13) groups. Seven of the eight VNTR markers investigated identified multiple alleles in the four groups of isolates. Using the composite data for eight VNTRs, a diversity value of 0.98 was calculated for the entire population, taking into account single- and double-locus variants. A high correlation (R = 0.98) between the maximum copy number and the number of alleles was observed. The most polymorphic markers were VNTR-1, VNTR-4, VNTR-5 and VNTR-7 (D > OR = 0.8). Characterisation of B. melitensis isolates by HOOFs enabled the recognition of related human cases and the exchange of molecular epidemiological information concerning a spreading clone, thus improving brucellosis surveillance.

Central nervous system metastases of a pulmonary epitheloid haemangioendothelioma.

The case of a 55-yr-old male with a right pleural effusion and multiple bilateral nodules is reported. A diagnostic thoracothomy was necessary to obtain a definitive histological diagnosis. During the postoperative course, the subject's neurological condition deteriorated and multiple cerebral mass lesions were discovered. The pathological analysis of both lung and cerebral tumours revealed an atypical endothelial cell proliferation; vascular immunohistochemical markers, such as factor VIII and CD34, were strongly positive. His general condition remained poor and the patient died 18 months after the initial diagnosis. The final diagnosis was pulmonary epitheloid haemangioendothelioma with synchronous central nervous system dissemination, the first time the authors believe that association has been reported. Little is known of the prognosis and treatment of these tumours, due to their rarity. Negative prognostic factors appear to be the presence of symptoms, pleural effusion or multifocal presentations. Treatment should include surgical resection if possible; chemotherapy appears to have little effect. Watchful waiting is an acceptable option, especially in asymptomatic patients.

[Epidemiologic features of tetanus in adults form Yucatan, Mexico]. / Características epidemiológicas del tétanos en adultos en Yucatán, México.

OBJECTIVE: To describe epidemiologic features from tetanus in adult patients (TIAP) treated at hospital general O'Horan in Merida, Yucatan, Mexico and compare them with another Mexican series analyzed 25 years ago. MATERIAL AND METHODS: From 1985 to 1999, 121 TIAP cases aged 13 years or older were identified from which 112 were analyzed. Diagnosis of TIAP was made just in a clinical basis. To analyze information inferencial statistics were used. RESULTS: People affected by tetanus averaged 43 +/- 21, 95% CI 39 to 47. A 3:1 male to female ratio was documented. Eighty two (73%) patients come from the rural area; 43 (38%) did work as peasants. Sixty two cases (55%) were diagnosed during the Fall and Winter seasons. In 91 patients (81%) no anti-tetanus vaccination was documented. In 89 cases (79%) incubation period averaged 5.4 +/- 4 days, 95% CI 5 to 6. According to this 89 cases (79%) with incubation period < 10 days were graded as severe tetanus and 23 (21%) with incubation period > or = 10 days were graded as non-severe tetanus. Tetanus-prone wounds were documented in 95 (85%) cases, 59 (62%) of which (62%) were localized in the lower extremities. Final outcome dichotomized either as death patient (group one) or surviving patient (group two) was documented in 103 cases of whom 67 (65%) were in group one and 36 (35%) were in group two. By comparing them, differences were seen in mean age (P = 0.004, 95% CI 3.9 to 19.8), age categories (< 50 vs. > or = 50) (chi 2 P = 0.001, 95% CI 0.6 to 0.60), severity of tetanus (Fisher exact test P = 0.0009, 95% CI 2 to 53) and mean hospitalization time (mean difference 14.8, P = 0.0001, 95% CI 11 to 18) but not in sex (chi 2 0.69, P = 0.40). CONCLUSIONS: In the State of Yucatan, Peninsula de Yucatan, Mexico, TIAP is still an endemic process with high mortality rate specially among young productive people. Secondary preventive measures as routinely tetanus toxoid booster vaccination are still not enough, at least in adulthood.

The antigenic structure of Mycobacterium tuberculosis examined by immunoblot and ELISA. Influence of the age of the culture and of the obtaining method on the composition of the antigenic extracts.

In this study we have examined by immunoblot (IB) and enzyme-linked immunosorbent assay (ELISA) the humoral immune response in pulmonary tuberculosis. As a previous step, in an attempt to obtain the optimal antigen preparation for these studies, the influence of the culture age and of the obtention method on the composition of the extracts was analyzed. The highest number of antigenic bands was found in culture filtrates of 6 and 8 weeks; at these times two thick bands of 65 and 63 kilodaltons (kDa) were identified. These bands were absent from younger and older cultures. When analyzing the source of antigens, we found that culture filtrates contained more antigenic bands than sonic extracts. In view of these findings, culture filtrates of 6 weeks of age were used as test antigens. With 19 tuberculous sera a total of 16 antigenic bands were observed by IB. The response was very heterogeneous with respect to the intensity of the detected reactions and the number of reacting bands. The most frequently recognized bands were those of 31, 32, 38, 58 and 94 kDa. By ELISA with 49 tuberculous sera and with 48 control sera, a specificity of 0.98 and a sensitivity of 0.70 were obtained.

A 38-kD Mycobacterium tuberculosis antigen associated with infection. Its isolation and serologic evaluation.

To identify antigens that could be specifically associated with tuberculosis infection, the antibody response to Mycobacterium tuberculosis antigens of patients with pulmonary tuberculosis and of healthy individuals were compared by immunoblot. In healthy individuals, serum antibodies were found in the majority of cases. Bands of 60 and 32-31 kilodaltons (kD) were the antigens more frequently recognized by antibodies of normal sera (55.8 and 64.7%, respectively). In patients with pulmonary tuberculosis, the number and intensity of the developed antigen bands were much higher than in normal individuals. Antigens reacting preferentially with tuberculosis sera were also identified. Furthermore, a unique disease-associated protein antigen of 38 kD was found to react with 57% of patients' sera but with none of the controls. This antigen was isolated by elution from nitrocellulose membranes and tested as an ELISA reagent in the serodiagnosis of pulmonary tuberculosis. A specificity of 0.96 and sensitivity of 0.68 were obtained.

Complejo principal de histocompatibilidad y complotipo en sarcoidosis. / Major histocompatibility system and complotype in sarcoidosis

La sarcoidosis es una enfermedad multisistemica que se acompana de profundas modificaciones inmunologicas cuyos mecanismos intimos no se han esclarecido. En este articulo se informa un estudio del complejo principal de histocompatibilidad y del complotipo en un paciente y tres familiares sanos, en un intento por esclarecer algunos factores geneticos que pudieran participar en esta enfermedad. Se encontro en el paciente presencia de una recombinacion de genoma en el brazo corto del cromosoma 6 que abarcaba desde los genes del complotipo hasta el de glioxalasa