In-vivo efficacy of alcohol-based hand rubs against noroviruses: a novel standardized European test method simulating practical conditions.

BACKGROUND: Non-enveloped viruses are particularly resistant to disinfectants, so it is necessary to use disinfectants with proven virucidal activity in order to prevent and control the spread of viral infections. However, a test such as EN 1500, which uses an internal standard as the reference treatment for determining the bactericidal efficacy of hand rubs, is still lacking. This study aimed to establish a European standard for testing the in-vivo efficacy of hand rubs against non-enveloped viruses. METHODS: The concentration and mode of application of ethanol as the reference were determined, and compared with the efficacies of two commonly used hand rubs. The hands of volunteers were contaminated with murine norovirus strain S99. RESULTS: 70% wt/wt ethanol (2 x 3 mL in 2 x 30 s) was used as the internal reference treatment. The commercial ethanol-based hand rub was able to reduce the titre of murine norovirus significantly in 30 s, whereas a hand rub based on ethanol and propan-2-ol was significantly less effective compared with the reference. CONCLUSION: This study established a possible standard for testing the in-vivo efficacy of hand rubs against non-enveloped viruses using murine norovirus, a low contamination volume technique and ethanol as the internal reference. These findings need to be confirmed in European ring trials.

[Epidemiological aspects of gastrointestinal infections]. / Epidemiologische Aspekte gastrointestinaler Infektionen.

BACKGROUND AND OBJECTIVES: The aim of this study was to investigate seasonal patterns and age-associated trends of the main bacterial, viral, and parasitic enteric pathogens in Southwest Germany. PATIENTS AND METHODS: From January 2002 through December 2008 a total of 99,057 patients were tested for Norovirus, Rotavirus, bacterial pathogens, Cryptosporidium parvum (C. parvum), and Giardia lamblia (G. lamblia). RESULTS: All these pathogens were detected throughout the whole year. But there were distinctive seasonal patterns of activity of the following pathogens being detected: norovirus was detected mainly from September through April. The highest rotovirus activity was observed from December through June. But bacterial pathogens und C. parvum were found mainly from June to November. The percentage of positive results during the months with the highest activity was 10 - 49% for norovirus, 25% - 41% for rotavirus, 14 - 18% for bacterial infection and 3 - 4 % for C. parvum. G. lamblia and adenovirus were found throughout the year in 7 - 15% and 3 - 10% of samples, respectively. Moreover, the detection rate of different pathogens depended on patient age. In infants younger than one year, rotavirus, norovirus and adenovirus were most frequently isolated pathogenes. Stool samples from kindergarden- and school-age children were positive largely for bacterial pathogens such as Salmonella and Campylobacter particularly in late summer or early autum. In patients older than 60 years, norovirus, rotavirus, and toxin producing Clostridium difficile strains were the most common pathogens. CONCLUSIONS: In view of the age and season related frequency of detection of enteric pathogens, a step-by-step diagnosis of gastrointestinal tract infections is recommended. Considering that most pathogens are detected sporadically over the whole year, the analysis of negative samples should be appropriately expanded. The knowledge of seasonal occurrence can also be applied to improve the application of hygienic measures.

Deficiencia somatotrófica en niños: características clínicas y bioquímicas según la etiología en 75 casos / Somatotrophic deficiency in children: report of 75 cases

Somatotrophic deficiency (SDMT) can be due to a deficiency of growth hormone releasing hormone(GHRH), growth hormone (GH) or insulin like growth factor I (IGF-I). Although its clinical features have been thoroughly described, the diagnosis is still controversial. Now there is an effective treatment with GH or IGF-I for these patients. AIM: To analyze the main clinical, etiological and laboratory characteristics of 75 SD patients (44 males), aged 9.4 + 4.5 years, with severe growth retardation. The diagnosis was confirmed by the lack of response to two GH stimulation tests (Clonidine, Glugagon or Insulin) and low levels of IGF-I or insulin-like growth factor binding protein- 3 (IGFBP-3). RESULTS: In 34 patients (46 percent), the cause of DSMT was considered idiopathic (DSMT-I), in 31 (41 percent) there was an organic cause (DSMT-O), most commonly caused by malformations or pituitary tumors and in 10 (13 percent), it was genetic (DSMT-G) (three patients with Laron's Syndrome, five with mutations of GH gene and 2 with probable mutations of Prop-1 and Pit-1 genes). IGF-1 levels, were significantly lower in DSMT-O and DSMT-G thanin DSMT-I (21.2 +/- 46.1, 23.4 +/-30.3 ng/mL and 50.2 +/- 48.3 ng/mL, respectively). The lowest height score corresponded to DSMT-G, compared to DSMT-O and DSMT (­5.7 +/- 0.9, -4.0 +/- 1.6 and ­4.3 +/- 1.2 DS, respectively) CONCLUSIONS: The high percentage of organic and genetic etiologies in our patients can be due to the systematic search of these diseases. DSMT-G (Laron, mutations in GH and Pit-1 genes) had the most severe growth retardation.

Kaposi's sarcoma-associated herpesvirus serology in Europe and Uganda: multicentre study with multiple and novel assays.

A multicentre study was undertaken to define novel assays with increased inter-assay concordance, sensitivity, specificity and predictive value for serological diagnosis of human herpesvirus type 8 (HHV-8) infection. A total of 562 sera from European and Ugandan human immunodeficiency virus (HIV)-infected or uninfected individuals with or without Kaposi's sarcoma (KS) and blood donors were examined under code by 18 different assays in seven European laboratories. Sera from KS patients and all non-KS sera found positive by at least 70%, 80%, or 90% of the assays were considered "true positive." The validity of the assays was then evaluated by univariate logistic regression analysis. Two immunofluorescence assays (IFA) for detection of antibodies against HHV-8 lytic (Rlyt) or latent (LLANA) antigens and two enzyme-linked-immunosorbent assays (ELISA) (M2, EK8.1) for detection of antibodies against HHV-8 structural proteins were found to be highly concordant, specific, and sensitive, with odds ratios that indicated a high predictive value. When used together, the two IFA (Rlyt-LLANA) showed the best combination of sensitivity (89.1%) and specificity (94.9%). The performance of these assays indicate that they may be used for the clinical management of individuals at risk of developing HHV-8 associated tumours such as allograft recipients.

Use of recombinant glycoprotein antigens gB and gH for diagnosis of primary human cytomegalovirus infection during pregnancy.

The major risk factor for intrauterine transmission of human cytomegalovirus (HCMV) is a primary infection during pregnancy. The neutralizing antibody response appeared after an average of 13 weeks after seroconversion and therefore the absence of neutralizing titers in HCMV IgG positive pregnant women is a reliable marker for primary infection. Determination of neutralizing antibody, however, is time-consuming and labor-intensive. For this reason an immunoblot assay for detection of neutralizing antibodies was developed based on the use of recombinant antigens representing neutralizing epitopes of glycoproteins (gp) gB (gpUL55) and gH (gpUL75) of HCMV. In this study, 93.6% of sera of pregnant women with prior infection recognized the gp-specific epitopes corresponding to a nonresponder rate of 6.4% relative to the neutralizing antibody. In primary infection the gp-response in general coincided with the appearance of neutralizing antibody. Intriguingly, lack of HCMV gB-specific antibodies was correlated with a lower risk of intrauterine fetal infection (P < 0.05).

Near final height in pubertal growth hormone (GH)-deficient patients treated with GH alone or in combination with luteinizing hormone-releasing hormone analog: results of a prospective, randomized trial.

To study the effects of delaying puberty in GH-deficient (GHD) children, we studied 21 GHD (9 boys, 14 girls), treatment-naive, pubertal patients in a prospective, randomized trial. Their chronological age was 14.3 +/- 1.6 yr, and their bone age was 11.3 +/- 1.1 yr (mean +/- SD) at the beginning of the study. Four patients who developed hypogonadotropic hypogonadism were subsequently excluded from the study. Patients were randomly assigned to receive GH + LH-releasing hormone analog (LHRH-A) (n = 7), or GH alone (n = 10). GH and LHRH-A treatment started simultaneously in each patient. GH (Nutropin) was administered at a dose of 0.1 U/kg x day sc, until patients reached a bone age (BA) of 14 yr in girls and 16 yr in boys, and LHRH-A (Lupron depot) was administered at a dose of 300 microg/ kg every 28 days in during 3 yr. We defined GH deficiency as patients with a growth velocity less than 4 cm/yr, BA delay more than 1 yr in relationship to chronological age, GH response to two stimulation tests less than 7 microg/L, associated with low serum insulin-like growth factor I and insulin-like growth factor binding protein 3 levels. Statistical analysis was performed by ANOVA or Kruskall Wallis when variances were not homogeneous. We observed a significant decrease in the rate of BA maturation in the group treated with GH+LHRH-A (1.5 +/- 0.2 yr) compared with the group treated with GH alone (4.2 +/-0.5 yr) during the 3 years of LHRH-A therapy (P < 0.05). This delay in BA maturation produced a significant gain in final height in the group treated with GH+LHRH-A, which reached - 1.3 +/- 0.5 SD score compared with -2.7 +/- 0.3 SD score (P < 0.05) in the group treated with GH alone. These results indicate that delaying puberty with LHRH-A in GHD children during treatment with GH increases final height.

The proteasome regulator PA28alpha/beta can enhance antigen presentation without affecting 20S proteasome subunit composition.

PA28alpha/beta is a regulatory complex of the 20S proteasome which consists of two IFN-gamma inducible subunits. Both subunits, alpha and beta, contribute equally to the formation of hexa- or heptameric rings which can associate with the 20S proteasome. Previously, we have shown that overexpression of the PA28alpha subunit enhanced the MHC class I-restricted presentation of two viral epitopes and that purified PA28alpha/beta accelerated T cell epitope generation by the 20S proteasome in vitro, indicating a role for PA28alpha/beta in antigen presentation. This conclusion was recently confirmed in PA28beta gene targeted mice which were severely deficient in MHC class I-restricted antigen presentation. These mice displayed a defect in the assembly of immunoproteasomes, suggesting that a lack of the proteasome subunits LMP2, LMP7, and MECL-1 may account for the deficiency in antigen presentation. In this study we investigated whether the effect of PA28alpha/beta on antigen presentation is dependent on a change of proteasome subunit composition. We have analyzed the assembly and subunit composition of proteasomes in fibroblast transfectants overexpressing both, alpha and beta subunits of PA28. In these transfectants we found a marked enhancement in the presentation of the immunodominant H-2Ld-restricted pp89 epitope of murine cytomegalovirus, although the 20S proteasome composition was the same as in recipient cells. We, therefore, conclude that PA28alpha/beta can enhance antigen processing independently of changes in 20S proteasome subunit composition or assembly.

High-throughput microarray-based enzyme-linked immunosorbent assay (ELISA).

A new generation biochip is described as capable of supporting high-throughput (HT), multiplexed enzyme-linked immunosorbent assays (ELISAs). These biochips consist of an optically flat, glass plate containing 96 wells formed by an enclosing hydrophobic Teflon mask. The footprint dimensions of each well and the plate precisely match those of a standard microplate. Each well contains four identical 36-element arrays (144 elements per well) comprising 8 different antigens and a marker protein. Arrays are formed by a custom, continuous flow, capillary-based print head attached to a precise, high-speed, X-Y-Z robot. The array printing capacity of a single robot exceeds 20,000 arrays per day. Arrays are quantitatively imaged using a custom, high-resolution, scanning charge-coupled device (CCD) detector with an imaging throughout of 96 arrays every 30 s. Using this new process, arrayed antigens were individually and collectively detected using standard ELISA techniques. Experiments demonstrate that specific multiplex detection of protein antigens arrayed on a glass substrate is feasible. Because of the open microarray architecture, the 96-well microarray format is compatible with automated robotic systems and supports a low-cost, highly parallel assay format. Future applications of this new high-throughput screening (HTS) format include direct cellular protein expression profiling, multiplexed assays for detection of infectious agents and cancer diagnostics.

Gps mutations in Chilean patients harboring growth hormone-secreting pituitary tumors.

Hypersecretion of GH is usually caused by a pituitary adenoma and about 40% of these tumors exhibit missense gsp mutations in Arg201 or Gln227 of the Gs, gene. We studied 20 pituitary tumors obtained from patients with GH hypersecretion. One tumor was resected from an 11 year-old boy with a 3 year history of accelerated growth, associated with increased concentrations of serum GH and IGF-I, which were not suppressed by glucose administration. The remaining 19 tumors were obtained from adult acromegalic patients, who had elevated baseline serum GH levels that did not show evidence of suppression after administration of glucose. The gsp mutations were studied by enzymatic digestion of the amplified PCR fragment of exon 8 (Arg201) and exon 9 (Gln227) with the enzymes NlaIII and NgoAIV, respectively. The tumors obtained from the boy and from nine of the 19 patients with acromegaly exhibited the gsp mutation R201H. None of the tumors had the Gln227 mutation. The gsp positive patients tended to be older, had smaller tumors, and had preoperative basal serum GH levels which were significantly lower (21 +/- 6 vs 56 +/- 16 microg/l, p<0.05) than the gsp negative patients. In this study, we documented the presence of a gsp mutation in Arg201 in a boy with gigantism and in approximately half of 19 Chilean adult patients with acromegaly, similar to other populations.

Inactivation of a defined active site in the mouse 20S proteasome complex enhances major histocompatibility complex class I antigen presentation of a murine cytomegalovirus protein.

Proteasomes generate peptides bound by major histocompatibility complex (MHC) class I molecules. Avoiding proteasome inhibitors, which in most cases do not distinguish between individual active sites within the cell, we used a molecular genetic approach that allowed for the first time the in vivo analysis of defined proteasomal active sites with regard to their significance for antigen processing. Functional elimination of the delta/low molecular weight protein (LMP) 2 sites by substitution with a mutated inactive LMP2 T1A subunit results in reduced cell surface expression of the MHC class I H-2Ld and H-2Dd molecules. Surface levels of H-2Ld and H-2Dd molecules were restored by external loading with peptides. However, as a result of the active site mutation, MHC class I presentation of a 9-mer peptide derived from a protein of murine cytomegalovirus was enhanced about three- to fivefold. Our experiments provide evidence that the delta/LMP2 active site elimination limits the processing and presentation of several peptides, but may be, nonetheless, beneficial for the generation and presentation of others.

Effects of luteinizing hormone-releasing hormone analog-induced pubertal delay in growth hormone (GH)-deficient children treated with GH: preliminary results.

To study the effect of delaying epiphyseal fusion on the growth of GH-deficient children, we studied 14 pubertal, treatment naive, GH-deficient patients (6 girls and 8 boys) in a prospective, randomized, placebo-controlled trial. Chronological age was 14.5 +/- 0.5 yr, and bone age was 11.6 +/- 0.3 yr (mean +/- SEM) at the beginning of the study. Patients were assigned randomly to receive GH and LH-releasing hormone (LHRH) analog (n = 8) or GH and placebo (n = 6) during 3 yr, with planned continuation of GH treatment until epiphyseal fusion. Patients were measured with a stadiometer and had serum LHRH tests, serum testosterone (boys), serum estradiol (girls), and bone age performed every 6 months. Patients treated with GH and LHRH analog showed a clear suppression of their pituitary-gonadal axis and a marked delay in bone age progression. We observed a greater gain in height prediction in these patients than in the patients treated with GH and placebo after 3 yr of treatment (mean +/- SEM, 14.0 +/- 1.6 vs. 8.0 +/- 2.4 cm; P < 0.05). These preliminary findings suggest that delaying epiphyseal fusion with LHRH analog in pubertal GH-deficient children treated with GH increases height prediction and may increase final height compared to treatment with GH alone.

A single residue exchange within a viral CTL epitope alters proteasome-mediated degradation resulting in lack of antigen presentation.

CTL epitope (KSPWFTTL) encoded by AKV/MCF type of murine leukemia virus (MuLV) differs from the sequence in Friend/Moloney/Rauscher (FMR) type in one residue (RSPWFTTL). CTL experiments indicated defective processing of the FMR peptide in tumor cells. Proteasome-mediated digestion of AKV/MCF-type 26-mer peptides resulted in the early generation and higher levels of epitope-containing fragments than digestion of FMR-type peptides, explained by prominent cleavage next to R in the FMR sequence. The fragments were identified as 10- and 11-mer peptides and were efficiently translocated by TAP. The naturally presented AKV/MCF peptide is the 8-mer, indicating ER peptide trimming. In conclusion, a single residue exchange can cause CTL epitope destruction by specific proteasomal cleavage.

A role for the proteasome regulator PA28alpha in antigen presentation.

Cytotoxic T cells recognize viral proteins as peptide fragments which are produced in the cytosol and transported on major histocompatibility complex (MHC) class I proteins to the cell surface. Viral peptides that meet the stringent binding characteristics of class I proteins are generated by the 20S proteasome. The interferon (IFN)-gamma-inducible activator of the 20S proteasome, PA28, strongly influences the proteasomal cleavage pattern in vitro. This led us to investigate whether changes in cellular levels of PA28 affect the efficiency of viral antigen processing. A mouse fibroblast line expressing the murine cytomegalovirus pp89 protein was transfected with either the human or murine gene encoding the PA28alpha subunit, which is sufficient to activate the peptide-hydrolysing activity of the 20S proteasome in vitro. Here we report that enhanced expression of PA28alpha at a level similar to that obtained after IFN-gamma induction resulted in a marked enhancement of recognition by pp89-specific cytotoxic T cells; the presentation of influenza nucleoprotein was also significantly improved. These results demonstrate a fundamental in vivo function for PA28alpha in antigen processing.

Hepatitis crónica activa en un caso de hipertiroidismo severo / Active chronic hepatitis in a severe hyperthiroidism case

Se describe una adolescente que sufría de hipertiroidismo severo de larga evolución, con pobre respuesta a medicamentos antitiroideos y dos episodios de ictericia previos, que ingresó con un hipertiroidismo clínicamente activo a pesar de estar recibiendo propiltiouracilo, ictericia intensa y hepatomegalia. El estudio y manejo de su hepatitis fue complejo, dado que las posibilidades de disfunción hepática en el contexto de un hipertiroidismo, podrían obedecer a toxicidad por drogas, insuficiencia cardíaca, hipertiroidismo por sí mismo o hepatitis autoinmune. Se manejó con suspensión del propiltiouracilo, régimen hipercalórico, propanolol y dos dosis de I131, con lo cual se logró controlar el hipertiroidismo. El estudio de laboratorio e histopatológico fue compatible con hepatitis crónica activa que se manejó con corticoides orales, coincidiendo con lo cual la paciente está asintomática, si bien la biopsia hepática muestra aún signos de actividad

The cleavage preference of the proteasome governs the yield of antigenic peptides.

Proteasomes degrade endogenous proteins in the cytosol. The potential contribution of the proteasome to the effect of flanking sequences on the presentation of an antigenic epitope presented by the major histocompatibility complex class I allele Ld was studied. Peptides generated in cells from minigenes coding for peptides of 17- and 19-amino acid length were compared with the in vitro 20S proteasome degradation products of the respective synthetic peptides. The quality of generated peptides was independent of ubiquitination. In vivo and in vitro processing products were indistinguishable with respect to peptide size and abundance. Altering the neighboring sequence substantially improved the yield of the final antigenic nonapeptide by 20S proteasome cleavage. These results suggest that, in addition to the presence of major histocompatibility complex class I allelic motifs, the cleavage preference of the proteasome can define the antigenic potential of a protein.

Gigantismo pituitario / Pituitary gigantism

Se describe un niño afectado por gigantismo hipofisiario. Las manifestaciones clínicas de su enfermedad comenzaron a los 8 años de edad, con aceleración del crecimiento, cefalea y un año después, amaurosis derecha que motivó su ingreso al hospital. Su talla era >p95 para edad, facies tosca, manos pies grandes, amaurosis derecha y cuadrantopsia izquierda. La concentración sérica de hormona de crecimiento era anormalmente alta y no supresible con sobrecarga oral de glucosa, la somatomedina C estaba aumentada y sufría déficit de hormonas tiroídea y cortisol. En la tomografía axial computadorizada se registró un tumor hipofisiario con extensión supraselar. Los padres rechazaron cualquier tratamiento. Reingresó un año después por hipertensión endocraneana, amaurosis bilateral y aumento del tamaño del tumor en la resonancia nuclear magnética. Se realizó resección trasesfenoidal del tumor hipofisiario, cuya histología correspondía a macroadenoma con necrosis y hemorragia y calcificaciones. Su evolución postoperatoria fue satisfactoria, manteniéndose en tratamiento de sustitución con hormona tiroidea y suprarrenal. Un año después de operado el niño está asintomático, las concentraciones séricas de hormona de crecimiento y somatomedina C son normales y no hay signos radiológicos de recidiva del tumor

Talla baja por resistencia a la hormona de crecimiento / Low height due to growth hormone resistance

El retraso de crecimiento de un niño por alteraciones en la hormona de crecimiento puede producirse por déficit en su producción o por falta de respuesta a su efecto. Ambos grupos de niños tienen características clínicas bastante similares. La hormona de crecimiento ejerce su efecto biológico a través de receptores tisulares, y si éstos están alterados se producirá un cuadro de resistencia periférica a dicha hormona. Presentamos un niño que consultó a los 14 años y 7 meses de edad por retraso severo del crecimiento, con talla de 95 cm, obesidad centrípeta, manos y pies pequeños, micropene y facies infantil, características sugerentes de un déficit de hormona de crecimiento. El estudio de laboratorio mostró concentraciones plasmáticas elevadas de hormona de crecimiento elevados y disminución de las de somatomedina-C. El paciente fue tratado con hormona de crecimiento biosintética, sin respuesta bioquímica ni clínica, lo que sugiere resistencia periférica a la acción de la somatotropina

Inhibition of human cytomegalovirus maturation by brefeldin A.

Brefeldin A (BFA) was found to interfere with specific events of human cytomegalovirus (HCMV) maturation in human fibroblasts. Ultrastructural as well as biochemical studies suggested that short-term exposure of infected cultures to BFA during the late infectious cycle primarily prevented Golgi-dependent processes, e.g. envelopment of naked cytoplasmic nucleocapsids in the trans-Golgi network (TGN) and normal processing of glycoprotein B. In contrast, the nuclear phase of viral morphogenesis, e.g. transport budding at the nuclear envelope, was not impaired. These observations were compatible with the interpretation that HCMV morphogenesis may involve sequential budding events at the nuclear envelope and at cisternae of the TGN. BFA treatment during the early infectious cycle efficiently inhibited HCMV-DNA synthesis and thus late viral functions, preventing production of viral progeny. Cytotoxicity was excluded as a cause for these findings.

Respuesta al tratamiento con hormona de crecimiento humana (hGH) biosintética en pacientes con deficiencia aislada de GH de diferentes tipos / Response to biosynthetic human growth hormone in patients with isolated tipe I-A and idiopatic growth hormone deficiency

Se estudió la respuesta al tratamiento con hormona de crecimiento (HC) biosintética en cuatro niños con deficiencia aislada de hGH tipo I-A, y se la comparó con la de cuatro con deficiencia de HC de tipo idiopático sometidos a igual tratamiento. La edad de los pacientes fue de 5 a 16 años, 5 hombres y 3 mujeres. Todos los pacientes crecían 3 cm o menos al año. Todos tenían edad ósea retrasada, que variaba entre 1 año 9 meses y 11 años. Los pacientes con deficiencia aislada de GH tipo I-A presentaba retardo de crecimiento severo y facie peculiar, 3 de ellos con antecedentes familiares de un hermano con idéntico compromiso clínico, los que no se incluyeron en el estudio. Todos los pacientes se trataron con HC biosintética (Genotropin Kabi) subcutánea, en dosis de 0,5 U* Kg * semana durante un período aproximado de un año. La respuesta fue buena en los 4 pacientes con déficit de hGH idiopático, creciendo aproximadamente 11 cm el primer año de tratamiento. De los 4 pacientes con aparente deficiencia aislada GH de tipo I-A dos mostraron una buena respuesta, similar a la de los pacientes idiopáticos y los 2 restantes sólo crecieron bien los primeros 3 meses, disminuyendo luego su velocidad de crecimiento a la que tenían antes del tratamiento. Se concluye, como está descrito, que los pacientes con deficiencia idiopática responde bien a la administración de HC exógena; por el contrario, los pacientes con déficit de HC de tipo I-A presentan una respuesta variable a este tratamiento

Hirsutismo en niñas adolescentes: estudio clínico y de laboratorio / Hirsutism in adolescent girls

Se estudiaron prospectivamente 14 niñas con hirsutismo, entre los 12,5 y 16 años de edad, normotensas, de peso corporal normal, sin tumores, afecciones ováricas primarias o hiperplasia suprarrenal clásica. El hirsutismo se calificó según Ferriman. Se hicieron determinaciones basales de 17 ketosteroides, 17 hidroxicorticoides, pregnanetriol y dehidroespiandrosterona en orina de 24 horas, así como de LH, FSH, testosterona, cortisol (F), 17-hidroxiprogesterona (17-OHP), dehidroepiandrosterona-sulfato (DHEA-S) en plasma, mediciones de las tres últimas 60 min después de inyectar por vía endovenosa 0,25 mg de corticotrofina de acción rápida y ultrasonografía ovárica. De acuerdo al cuadro clínico y de laboratorio dos de las pacientes eran probables portadoras heterozigotas del gen de deficiencia de 21 hidroxilasa, nueve sin trastornos menstruales tenían DHEA-S basal elevada (promedio 8.289, márgenes 4.750 a 14.800 ng/ml) como único hallazgo de laboratorio y su hirsutismo era, por lo tanto, de etiología probablemente suprarrenal, estando aún en estudio para el diagnóstico definitivo. Finalmente, las otras tres pacientes tienen hirsutismo idiopático. Las dos niñas probablemente portadoras heterozigotas del gen de deficiencia de la 21-OH y cinco de las que tenían la DHEA-S elevada, fueron tratadas con dexametasona, con normalización de la DHEA-S, mejoría del aspecto ecográfico ovárico y del hirsutismo