Next generation sequencing characterizes the extent of HLA diversity in an Argentinian registry population.

Next generation DNA sequencing is used to determine the HLA-A, -B, -C, -DRB1, and -DQB1 assignments of 1472 unrelated volunteers for the unrelated donor registry in Argentina. The analysis characterized all HLA exons and introns for class I alleles; at least exons 2, 3 for HLA-DRB1; and exons 2 to 6 for HLA-DQB1. Of the distinct alleles present, there are 330 class I and 98 class II. The majority (~98%) of the cumulative allele frequency at each locus is contributed by alleles that appear at a frequency of at least 1 in 1000. Fourteen (18.2%) of the 77 novel class I and II alleles carry nonsynonymous variation within their exons; 52 (75.4%) class I novel alleles carry only single, apparently random, nucleotide variation within their introns/untranslated regions. Alleles encoding protein variation not usually detected by typing focused only on the exons encoding the antigen recognition domain are 1.0% of the class I assignments and 7.3% of the class II assignments (predominantly DQB1*02:02:01, DQB1*03:19:01, and DRB1*14:54:01). Updates to the common and well documented list of alleles include 10 alleles previously thought to be uncommon but that are found at least 30 times. Five locus haplotypes estimated using the expectation-maximization algorithm as present 3 or more times total 187. While the known HLA diversity continues to increase, the conservation of known allele sequences is remarkable. Overall, the HLA diversity observed in the Argentinian population reflects its European and Native American ancestry.

Identification of TEM-135 ß-lactamase in Neisseria gonorrhoeae strains carrying African and Toronto plasmids in Argentina.

One hundred forty-three penicillinase-producing Neisseria gonorrhoeae (PPNG) isolates obtained in Argentina from 2008 and 2012 were examined to detect blaTEM-135 genes and to investigate plasmid profiles and multiantigen sequence types. Forty-two PPNG isolates were found to carry TEM-135, and two contained a new TEM derivative characterized as TEM-220. The blaTEM-135 allele was carried by the Toronto/Rio and African plasmids. Molecular epidemiology revealed that two blaTEM-135 isolates were related to previously described isolates from Thailand and China, indicating a common evolutionary origin.

HLA antigenic and haplotype frequencies estimated in hematopoietic progenitor cell donors from Argentina.

BACKGROUND: Hematopoietic progenitor cell transplantation is considered a standard-of-care treatment for defined hematological and non-hematological conditions affecting bone marrow-derived cells. METHODS: Patients and potential donors are HLA typed for their HLA-A, -B, -C, -DRB1, and -DQB1 alleles. The best allogeneic donor is one for which each allele matches the patient at HLA-A, -B, -C, and -DRB1 (8/8). For patients with no related donor, the transplant physician will start a search for unrelated donors. The search is performed through a local registry and often includes the search for donors worldwide. The Argentinean HPC Donors Registry was established in 2003. Our National HPC Donor Registry has already typed more than 31,000 donors for HLA-A, -B, and -DR. RESULTS: We present the analysis of HLA frequencies and haplotypes estimates for the subset of our donor database that is additionally typed for HLA-C. We analyzed HLA data from 2657 donors. Antigen and haplotype frequencies were estimated through the use of expectation maximization. CONCLUSIONS: Our analysis showed for the first time the antigenic HLA frequency distribution from HPC donors in Argentina. Knowing haplotype frequencies in our population will help us to select potential donors for high-resolution typing for the patients.

Síndrome Poliglandular Autoinmune Tipo II: Posible Asociación con HLA DRB1*-DQB1* / Possible association of Type II Autoimmune Polyendrocrine Syndrome with HLA DRB1*-DQB1*

Los síndromes poliendocrinos autoinmunes (APS) asocian enfermedades endocrinas autoinmunes con otros desórdenes autoinmunes no endocrinos. El APS tipo II se caracteriza por compromiso primario suprarrenal, tiroideo y/o DM tipo I. Presentamos un paciente masculino de 46 años que fue internado por astenia, adinamia, hiporexia, severa disminución de peso, mareos y vómitos. Antecedente de obesidad y diabetes diagnosticada 3 años antes. Presentaba hipotensión arterial, hiperpigmentación de mucosas y pliegues, anemia, hiponatremia e hipoglucemias frecuentes a pesar de la disminución de la dosis de insulina. Se diagnosticó insuficiencia suprarrenal, concomitantemente con hipotiroidismo y diabetes tipo 1, todas de origen autoinmune, iniciándose reemplazo hormonal. Se encontró una posible asociación del HLA DRB1*-DQB1* en los estudios genéticos. Conclusiones: Nuestro paciente presenta el HLA DQB1*0302 descripto en el APSII, pero el HLA DRB1 *08 encontrado no está descripto en este síndrome ni en ningún otro desorden autoinmune. En pacientes con Diabetes tipo 1 que disminuyan el requerimiento insulínico, habría que descartar insuficiencia suprarrenal, un componente del APS II, como factor etiológico, a pesar de su baja prevalencia.

Autoimmune polyendocrine syndromes (APS) are the association of autoimmune endocrine diseases with other non-endocrine autoimmune disorders. Type II APS is defined by occurrence of Addison´s disease with thyroid autoimmune disease and/or type 1 diabetes mellitus. We present a 46-year-old male patient who was hospitalized because of asthenia, adynamia, hyporexia, severe loss of weight, dizziness and vomiting. Diabetes mellitus had been diagnosed 3 years earlier when he was obese. He presented arterial hypotension, anemia, darkening of the skin and oral mucosa, hyponatremia and frequent hypoglycemia although his insulin dose was decreased. Adrenal insufficiency was diagnosed together with hypothyroidism and type 1 diabetes, all of them of autoimmune origin. Hormonal replacement treatment was initiated. Genetic studies were performed and a new polymorphism was found. Conclusions: HLA DRB1 *08 found in our patient has not been described in APS II or in any other autoimmune disorders. He also has HLA DQB1*0302 described in previous reports related to APS II. In type 1 diabetic patients whose insulin requirement decreases, it would be advisable to rule out adrenal insufficiency, a component of APS II, as an etiologic factor in spite of its low prevalence. In diabetic obese patients (mainly young) who lose weight without a defined cause, type 1 diabetes mellitus should be excluded.

Asociación de polimorfismos de simple nucleótido (SNP) de CD28 (IVS3+17T/C) Y CTLA-4 (+49A/G) con autoinmunidad tiroidea (AIT) / Association of single nucleotide polymorphism (SND) of CD28 (IVS3+17+K) and CTLA-4 (+49 A/G) with thyroid autoimmunity AIT)

La enfermedad tiroidea autoinmune es la patología autoinmune más prevalente y afecta hasta el 5% de la población general. Su desarrollo está dado por la interacción entre susceptibilidad genética y otros factores. Una característica es la temprana producción de anticuerpos antiperoxidasa tiroidea (aTPO) que a menudo predicen el desarrollo clínico de la enfermedad. La susceptibilidad genética para AIT es generada por genes del HLA y por otros candidatos del cromosoma 2q33. Esta región contiene los genes CTLA-4 y CD 28, y sus polimorfismos estarían asociados. Objetivo: Analizar y comparar la distribución de los polimorfismos de simple nucleótido (SNP) de CD28 (IVS3+17 T/C) y CTLA-4 (+49 A/G) en pacientes con aTPO >10 IU/ml (AIT) comparados con un grupo control aTPO ≤ 10 IU/ml sin AIT. Sujetos y métodos: Estudiamos 69 pacientes y 36 sujetos considerados controles. Para determinar aTPO se utilizó IMMULITE 1000 y muestra sérica. Para el estudio de los SNP se extrajo ADN de sangre periférica. La amplificación de los genes se realizó por PCR. Las diferencias entre grupos fueron comparadas usando el test de Chi Cuadrado. Resultados: Observamos diferencia significativa en el genotipo CD28 C/T entre AIT y controles (p=0.026). Analizando los genotipos de los polimorfismos CTLA-4 no observamos diferencia significativa entre AIT y controles. Del análisis de asociación de genotipos CD28 C/T y CTLA-4 A/A o A/G, observamos diferencia significativa comparando AIT vs. controles (p=0,013). Conclusión: Encontramos una posible asociación significativa del genotipo CD28 C/T en individuos con AIT, y estos portadores tendrían un riesgo tres veces mayor de adquirir AIT. La combinación de los genotipos CD28 C/T y CTLA-4 A/A o A/G incrementaría cuatro veces el riesgo de adquirir AIT. Estos resultados permitirían llevar a cabo un diagnóstico precoz, con la adecuada caracterización de una posible enfermedad tiroidea autoinmune en pacientes con AIT.

The autoimmune thyroid disease is the most prevalent autoimmune affection and affects until 5% of the general population; its development is given by the interaction between genetic susceptibility and other factors. One particularity is the early production of thyroid autoantibodies against thyroid peroxidase (aTPO) which often predicts the clinical development of the disease. The genetic susceptibility for the thyroid autoimmunity (AIT) is generated by genes of the HLA and by other genes candidates of the chromosome 2q33. This region contains the genes: CTLA-4 and CD 28. Several polymorphisms of both would be associated according to previous studies. Objective: To analyze and to compare the simple nucleotide polymorphism distribution (SNP) of CD28 (IVS3+17 T/C) and CTLA-4 (+ 49 A/G) in patients with aTPO> 10 IU/ml (AIT) compared to a control group aTPO ≤ 10 IU/ml with no AIT. Subjects and Methods: We have studied 69 patients with AIT and 36 control subjects. Serum aTPO were measured by using chemiluminescence immunoassay (IMMULITE1000, Siemens). Genomic DNA was prepared from peripheral white blood cells. The amplification of the genes was carry out by polymerase chain reaction (PCR). Statistical analyses : the differences between groups were made using the chisquare test. P less than 0.05 was considered statistically significant. Results: There was a significant difference of genotype CD 28 C/T in patients with AIT compared with controls (p=0.026). The genotypes of CTLA-4 was analyzed and there was no significant difference between AIT and controls. Analysis of genotypes association CD 28 C/T and CTLA-4 A/A or A/G, revealed significant difference comparing AIT versus controls (p= 0.013). Conclusions: We found a possible association of genotype CD 28 C/T in individuals with AIT, since carriers of genotype C/T would have a risk three times higher to acquire AIT. The combination of genotypes CD 28 C/T and CTLA-4 A/A or A/G would increase the risk of acquiring AIT four times. These results could be useful in order to make a premature diagnosis, with adequate characterization of a possible autoimmune thyroid disease in patients with AIT.

Ulnar variance and scaphoid fracture.

Between 1997 and 2006, radiographs of 66 scaphoid fractures were retrospectively reviewed to evaluate ulnar variance. Twenty-one (31.8%) patients had an 'ulna neutral' wrist, six (9.1%) had an 'ulna plus' and 39 (59.1%) had an 'ulna minus' wrist. The mean ulnar variance was -1.3 (SD 1.8) mm (range -5.5, 2.5). We observed a significant difference in the distribution of ulnar variance (P < 0.00001) and in the proportion of cases with ulna minus (OR = 5.0; 95% CI: 2.7, 9.3) compared to previous publications.

[Resistance surveillance of Neisseria gonorrhoeae in a hospital in Santa Fe province, Argentina: 1997-2004]. / Vigilancia de la resistencia de Neisseria gonorrhoeae en un hospital de la provincia de Santa Fe, Argentina: 1997-2004.

Resistance phenotypes characterized by minimum inhibitory concentration, disk diffusion and beta-lactamase production were determined in 434 isolates from patients attending the Sexually Transmitted Disease Service at Dr. José Maria Cullen Hospital in Santa Fe, Argentina. Susceptibility tests to penicillin, tetracycline, ciprofloxacin, espectinomycin, azithromycin and ceftriaxone were performed. Pulsed-field gel electrophoresis was conducted made to on three ciprofloxacin-resistant isolates. Epidemiologically speaking, three interesting events should be highlighted: during 1997, plasmid-mediated high level tetracycline-resistant strains were observed (33.3%); from 2002 to 2004 a significant increase of plasmid-mediated penicillin-resistant strains was registered (9.7% to 34.8%); and in the year 2000 the first two quinolone-resistant strains emerged in the province. In our hospital, the first azithromycin-resistant isolate emerged in 2004. We therefore emphasize the importance of the Clinical Microbiology Laboratory in order to provide information for the empiric treatment of this infection.

Vigilancia de la resistencia de Neisseria gonorrhoeae en un hospital de la provincia de Santa Fe, Argentina: 1997-2004 / Resistance surveillance of Neisseria gonorrhoeae in a hospital in Santa Fe province, Argentina: 1997-2004

Se determinaron los fenotipos de resistencia caracterizados por la concentración inhibitoria mínima, la difusión con discos y la producción de β-lactamasa de 434 aislamientos de Neisseria gonorrhoeae obtenidos de pacientes atendidos en el Servicio de Enfermedades de Transmisión Sexual del Hospital Dr. José María Cullen, Santa Fe, Argentina. Se realizaron pruebas de sensibilidad a los siguientes antimicrobianos: penicilina, tetraciclina, ciprofloxacina, espectinomicina, azitromicina y ceftriaxona. A tres aislamientos resistentes a ciprofloxacina se les realizó electroforesis de campo pulsado. Se destacaron tres situaciones epidemiológicas de interés: en el año 1997, alta incidencia de aislamientos con resistencia plasmídica a tetraciclina (33,3%); en el período 2002-2004, un aumento significativo de la resistencia plasmídica a penicilina (9,7% a 34,8%); y en el año 2000, la emergencia de dos de los tres primeros aislamientos con resistencia a quinolonas del país. El primer aislamiento resistente a azitromicina en nuestro hospital emerge en el 2004. Este trabajo jerarquiza el rol del Laboratorio de Microbiología Clínica en la orientación del tratamiento empírico de la gonorrea.

Resistance phenotypes characterized by minimum inhibitory concentration, disk diffusion and β-lactamase production were determined in 434 isolates from patients attending the Sexually Transmitted Disease Service at Dr. José María Cullen Hospital in Santa Fe, Argentina. Susceptibility tests to penicillin, tetracycline, ciprofloxacin, espectinomycin, azithromycin and ceftriaxone were performed. Pulsed-field gel electrophoresis was conducted made to on three ciprofloxacin-resistant isolates. Epidemiologically speaking, three interesting events should be highlighted: during 1997, plasmid-mediated high level tetracycline-resistant strains were observed (33.3%); from 2002 to 2004 a significant increase of plasmid-mediated penicillin-resistant strains was registered (9.7% to 34.8%); and in the year 2000 the first two quinolone-resistant strains emerged in the province. In our hospital, the first azithromycin-resistant isolate emerged in 2004. We therefore emphasize the importance of the Clinical Microbiology Laboratory in order to provide information for the empiric treatment of this infection.

Análisis de la asociación de HLA DRB1-DQB1 y autoanticuerpos en familias con Síndrome Poliendocrino Autoinmune (APS) / An analysis of the relationship between HLA DRB1-DQB1 and autoantibodies, in families with autoinmune polyendocrine syndrome (APS)

El APS es la asociación de enfermedades endocrinas autoinmunes, con otros desórdenes autoinmunes no endocrinos, denominados componentes mayores y menores. Este síndrome se clasificó en 4 tipos. Las alteraciones de la respuesta inmune provocan fallas regulatorias de la misma; y polimorfismos de HLA, entre otros, sumado a factores adquiridos o permanentes, representan gatillos disparadores de la autoinmunidad. Nuestro objetivo fue buscar la asociación de HLA-DRB1*-DQB1* en individuos pertenecientes a dos familias, con diagnóstico en uno o más de ellos de APS, o con enfermedades autoinmunes aisladas. Determinar los anticuerpos séricos: a21-OH, aGAD y aTPO y observar la asociación con el haplotipo HLA. Estudiamos padres e hijos de dos familias, dos integrantes padecían APS tipo 2 y 3; y otros con enfermedades autoinmunes. Buscamos HLA-DRB1*-DQB1* y cuantificamos a21-OH, aTPO y aGAD. Los pacientes con APS 2 y 3 presentaron el HLA-DRB1*0301-DQB1*0201. De los individuos estudiados, 5/9 tenían este haplotipo HLA y al menos un autoanticuerpo positivo. Hallamos el factor genético en 2/3 de los integrantes con enfermedades autoinmunes correspondientes a componentes mayores. La relación observada, entre APS y HLA-DRB1*0301-DQB1*0201, aumenta la posibilidad de identificar personas en riesgo de contraer afecciones autoinmunes en grupos familiares, en los cuales algún integrante padece APS.

The APS is the association of autoimmune endocrine diseases, with other non-endocrine autoimmune disorders, named mayor and minor components. This syndrome was classified in 4 types. The alterations of the immune response cause regulatory faults; and HLA polymorphisms, among others; taken in conjunction with acquired or permanent factors, these represent triggers of autoimmunity. Our objective was to find out the association of HLA-DRB1*-DQB1* in individuals belonging to two families, with diagnosis in at least one of them APS, or with isolated autoimmune diseases. To determine serum antibodies: a21-OH, aGAD and aTPO and to observe the association with HLA haplotype. We have studied parents and offspring of two families, two members who suffered APS type 2 and 3, and others with autoimmune diseases. We have looked for HLA-DRB1*-DQB1* and quantified a21-OH, aTPO and aGAD. Patients with APS 2 and 3 showed HLA-DRB1*0301-DQB1*0201. Among the population we have studied, 5/9 had this HLA haplotype and at least one positive auto antibody. We have found the genetic factor in 2/3 of the members with autoimmune diseases corresponding to greater components. The observed relation between APS and HLA-DRB1*0301-DQB1*0201, increases the possibility of identifying people at risk of catching autoimmune affections in familiar groups in which at least one member suffers APS.

An unusual transmission of Neisseria meningitidis: neonatal conjunctivitis acquired at delivery from the mother's endocervical infection.

BACKGROUND: Primary meningococcal conjunctivitis is assumed to be due to the direct inoculation of Neisseria meningitidis into the conjunctival sac from an exogenous source. According to a literature review, no case of neonatal conjunctivitis infection acquired at delivery from maternal endocervicitis has been published. GOAL: To report a case of meningococcal neonatal conjunctivitis acquired at delivery because of the mother's endocervical infection and cross-transmission of the strain with her partner. STUDY DESIGN: Strains were characterized by bacteriologic and serologic methods including grouping (agglutination), typing, and subtyping (enzyme-linked immunoabsorbent assay). Molecular analysis was done by pulsed-field gel electrophoresis. RESULTS: The three strains (newborn infant, mother, partner) were of the same antigenic formula (C:NT:P1.NT) and exhibited similar NheI and SpeI pulsed-field gel electrophoresis patterns. CONCLUSION: The identical phenotypic and genomic analysis of strains is the evidence for N meningitidis transmission at delivery from a maternal endocervical infection to the newborn infant and cross transmission between sexual partners.

Dedo en gatillo: frecuencia y presentación en el Hospital Eugenio Espejo entre enero de 1994 y marzo de 1999

El dedo en gatillo es una tenosinovitis estenosante que compromete el funcionamiento de la mano al afectar los tendones flexores, poco conocida por el Médico general. Con el objetivo de determinar su frecuencia y presentación clínica en nuestro medio se revisaron las historias clínicas de los pacientes intervenidos quirúrgicamente por el servicio de Ortopedia y Traumatología del Hospital Eugenio Espejo. De 438 cirugías de mano el 9.8 por ciento obedecieron a un diagnóstico de dedo en gatillo. Los grupos etareos mas afectados fueron los comprometidos entre 11 a 20 años y de 41 a 50 años con predominio del sexo femenino (p mayor a 01). El dígito mas comprometido fue el pulgar (67,4 por ciento) y su etiología más frecuente fue de tipo adquirido...

Characterization of an outbreak of tetM-containing Neisseria gonorrhoeae in Argentina.

Phenotypic and molecular characterization of an outbreak of 9 Neisseria gonorrhoeae (NG) isolates exhibiting high-level plasmid mediated resistance to penicillin and tetracycline (PP-TRNG) that took place in Tandil, Argentina between February and April 1995. Comparison with the patterns of the 3 PP-TRNG strains previously isolated were made. We determined the following markers for each strain: antimicrobial susceptibility, serogroup, auxotype, plasmid profile, presence of tetM determinant and restriction pattern of the tetM-containing plasmid. Antimicrobial tests values were: tetracycline disk diameter 12-14 mm, minimum inhibitory concentration (MIC) 32 micrograms/ml; penicillin disk diameter 6 mm, MIC 32 micrograms/ml and sensitive by both methods to spectinomycin, cefuroxime, ceftriaxone and ciprofloxacin. All isolates were of the same serogroup (WI). Ten of the strains, including the 9 from Tandil outbreak, were arginine-requiring, while the other 2 were methionine and arginine-requiring. All of them demonstrate the same plasmid profile (2.6, 3.2, 25.2 MDa). They were positive for the tetM determinant and the restriction analysis identified it is a Dutch-type plasmid. In spite of the temporal and geographical dispersion, PP-TRNG strains in Argentina seem to be highly homogeneous in terms of antimicrobial susceptibility, serogroup, plasmid profiles and even auxotype.

PIP: This study determines the phenotypic and molecular characterization of an outbreak of 9 Neisseria gonorrhea isolates exhibiting high-level plasmid-mediated resistance to penicillin and tetracycline (PP-TRNG) that took place in Tandil, Argentina. Subjects included patients attending Tandil's Sanatamarina Hospital. Individuals infected with sexually transmitted disease were examined using a standardized questionnaire. Results show that all isolates produced B-lactamase, were penicillin resistant (disk zone diameter 6 mm, MIC 32 mcg/ml) and high-level tetracycline resistant (disk zone diameter 12-14 mm, MIC 32 mcg/ml). Moreover, all strains were fully sensitive to cefuroxime, ceftriaxone, ciprofloxacin, and spectinomycin; while autotype and serogroup results showed that all the strains were arginine-requiring and of serogroup WI. In spite of the temporal and geographical dispersion, PP-TRNG strains in Argentina seem to be highly homogeneous in terms of antimicrobial susceptibility, serogroup, plasmid profiles, and even auxotype.

Primer informe en la República Argentina de resistencia a penicilina en Neisseria gonorrhoeae mediada por el plásmido de 3,2 MDal (africano) / First report in Argentine of resistance to penicillin in Neisseria gonorrhoeae madiated by the 3,2 MDal African plasmid

En la República Argentina, Buenos Aires (INM), detectamos la primer cepa productora de ß-lactamasa en 1980, incrementándose la prevalencia de estas cepas resistentes de 1,9 por ciento (1980-1984) a 30 por ciento (1984-1988) y a 37 por ciento (1989-1991). El objetivo de este trabajo es la caracterización de los plásmidos responsables de esta resistencia, en un estudio retrospectivo que abarca de 1985 a 1991. Se estudiaron 28 cepas de Neisseria gonorrhoeae productoras de penicilinasa (PPNG), enviadas para su estudio o aisladas en nuestro Servicio de exudados genitales. El 100 por ciento de las cepas presentó el plásmido críptico de 2,6 MDal. El plásmido de 3,2 MDal se encontró en 13 cepas (46,4 por ciento) y en el 92,3 por ciento de ellas acompañado por el de 24,4 MDal. En 15 cepas (53,60 por ciento) se detectó la presencia del plásmido de 4,4 MDal, sólo en 9 de ellas (60 por ciento) junto con el plásmido conjugativo. Este es el primer informe sobre la presencia del plásmido de 3,2 MDal (africano) en las cepas PPNG aisladas en la Argentina. Nuestro primer hallazgo de este plásmido data de 1988

Dedo en gatillo: frecuencia y presentación en el Hospital Eugenio Espejo de 1994 a 1999

El dedo en gatillo es una tenosinovitis estenosante que compromete el funcionamiento de la mano al afectar los tendones flexores poco conocida por el médico general. Con el objetivo de conocer su frecuencia y presentación clínica en nuestro medio se revisaron las historias clínicas de los pacientes intervenidos quirúrgicamente de 1994 a 1999 por el Servicio de Ortopedia y Traumatología del Hospital Eugenio Espejo. De 438 cirugías de mano el 9.8 por ciento obedecieron a un diagnóstico de dedo en gatillo. Los grupos etareos más afectados fueron los comprendidos entre 11 a 20 años y de 41 a 50 años con predominio del sexo femenino (p<0.01). El dígito más comprometido fue el pulgar (67.4 por ciento) y su etiología más frecuente fue de tipo adquirido (Nódulo de Notta 62.8 por ciento)...