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Mucocutaneous lesions are the most frequent symptoms of Behçet’s disease (BD). Recently, new therapies are being used to treat refractory cases, but the effect of these treatments on mucocutaneous manifestations has been scarcely reported. Our objective was to describe the mucocutaneous response to the different therapies used to treat BD in routine clinical practice. We retrospectively reviewed the clinical records of all patients diagnosed with BD seen at our institution between January 2010 and January 2022. Patients with BD without mucocutaneous manifestations were excluded. We included 109 patients diagnosed with BD: 51 males (46.8%) and 58 females (53.2%). The mean age at diagnosis was 31.58 years (standard deviation (SD) 12.110) and the mean time of disease evolution was 14.94 years (SD 11.094). Oral ulcers were the most frequent symptom present in 100% of patients, followed by genital ulcers (GU) in 76.1% of patients. Twenty-four patients (22%) had severe mucocutaneous symptoms (>12 lesions/year) before treatment. We found that among patients with GU there was a higher prevalence of episodes of posterior uveitis and venous thrombosis ( p = 0.011 and p = 0.045 , respectively). In our series, we observed a lower complete cutaneous response to colchicine in patients with GU, pathergy or severe mucocutaneous symptoms ( p < 0.05 ). Regarding the choice of a TNF-α inhibitor, we observed a lower prevalence of complete cutaneous response to adalimumab among patients with GU (53.3% complete response in patients with GU vs. 100% in patients without GU, p = 0.022 ), whereas no differences were found between clinical characteristics in the response to infliximab.

Resumo Em busca de melhorias na criopreservação do sêmen, substâncias naturais são comumente estudadas com o objetivo de melhorar a qualidade do sêmen. O objetivo deste estudo foi avaliar o efeito da adição de sucos de laranja, abacaxi e beterraba em diferentes concentrações e combinações ao diluidor de criopreservação de sêmen ovino. Foram utilizados cinco ejaculados de cinco carneiros adultos. O pool de sêmen foi diluído em diluente à base de gema de ovo e misturado com os seguintes 15 tratamentos (na concentração final de 400x10⁶ sptz/ml): laranja 10% (O10) e 15% (O15); abacaxi 10% (P10) e 15% (P15); beterraba 10% (B10) e 15% (B15); abacaxi + laranja 10% (PO10) e 15% (PO15); abacaxi + beterraba 10% (PB10) e 15% (PB15); beterraba + laranja 10% (BO10) e 15% (BO15); abacaxi + beterraba + laranja 10% (PBO10) e 15% (PBO15); e o grupo controle (CON). Pós-descongelação em palhetas de 0,25 ml a análise da qualidade do sêmen criopreservado foi realizada pelo CASA e citometria de fluxo. A análise de variância foi realizada e as médias comparadas pelo teste SNK. O teste de correlação de Pearson também foi realizado. Nenhum efeito foi observado na adição de sucos ao diluidor de sêmen antes da criopreservação. Após o descongelamento, embora estatisticamente semelhante ao grupo controle, a motilidade total do grupo B10 atingiu padrões aceitáveis de motilidade total. Além disso, o grupo B10 apresentou os maiores valores (p

Searching for improvements in semen cryopreservation, natural substances are commonly studied focusing to improve the sperm quality. The aim of this study were evaluated the effect of adding orange, pineapple, and beet juices in different concentrations and combinations to the ram semen cryopreservation extender. Five ejaculates from five adult rams were used. The semen pool was diluted in egg yolk-based extender and mixed with the following 15 treatments (at a final concentration of 400.106 sptz/mL): orange 10% (O10) and 15% (O15); pineapple 10% (P10) and 15% (P15); beet 10% (B10) and 15% (B15); pineapple + orange 10% (PO10) and 15% (PO15); pineapple + beet 10% (PB10) and 15% (PB15); beet + orange 10% (BO10) and 15% (BO15); pineapple + beet + orange 10% (PBO10) and 15% (PBO15); and the control group (CON). Post-thaw in 0.25 mL straws semen quality analysis of cryopreserved semen was performed by CASA and flow cytometry. Analysis of variance (PROC GLM) was carried out and the averages were compared using the SNK test. Pearson's correlation test was also performed. No effect was noted in the addition of juices to the semen extender prior to cryopreservation. Post-thawed, although, statistically similar to the control group, the total motility of the B10 group reached acceptable standards of total motility. In addition, B10 group showed the highest values (p

Background Our aim was to study the association between case rates and reductions in urban mobility in state capitals of Colombia. Methods We designed an ecological time-series study to correlate the Colombian incidence rate with reductions in mobility trends of retail stores. Results The meta-analysis of β coefficients describing the association between case rates and reductions in mobility trends of retail stores resulted in a mean estimate of 0.0637 (95% confidence interval 0.027 to 0.101; p Conclusions We recommend continuing to consider mobility restrictions when the number of cases starts to climb in each local jurisdiction.

Transgenic 'Duncan' grapefruit (Citrus paradisi Macf.) and 'Valencia' sweet orange (Citrus sinensis [L.] Osbeck) plants ectopically expressing C. sinensis (cv. Washington navel orange) APETALA1 (CsAP1) or LEAFY (CsLFY) genes under control of the Arabidopsis thaliana stress-inducible promoter AtRD29A flowered under non-inductive (warm temperature, well-watered) greenhouse conditions, whereas their wild-type (WT) counterparts did not. The transgenic plants that flowered exhibited no altered morphological features, except the lack of thorns characteristic of juvenile WT plants. The most precocious T0 line, 'Duncan' grapefruit (Dun134-3) expressing the CsAP1 gene, flowered and fruited when it was 4.5 years old and the T1 siblings from this line flowered and fruited when they were just over 18 months old. In contrast, T1 seedlings from three lines of 'Duncan' grapefruit expressing the CsLFY gene flowered within 3 months after germination, but were unable to support fruit development. Transcript levels of corresponding transgenes in leaves were not correlated with earliness of flowering. To further study the activity of AtRD29A, leaves from three 'Carrizo' citrange (C. sinensis × Poncirus trifoliata) rootstock seedlings transformed with the green fluorescent protein (GFP) gene under regulation of the AtRD29A promoter were subjected to drought stress or well-watered conditions. Expression of GFP was not stress-dependent, consistent with the observation of flowering of CsAP1 and CsLFY transgenic plants under non-inductive conditions. Taken together, the results suggest that AtRD29A is constitutively expressed in a citrus background. Despite the loss of control over flowering time, transgenic citrus lines ectopically expressing C. sinensis AP1 or LFY genes under control of the A. thaliana RD29A promoter exhibit precocious flowering, fruit development and viable transgenic seed formation. These transformed lines can be useful tools to reduce the time between generations to accelerate breeding.

The objective was to evaluate the performance, carcass and meat quality of 40 lambs classified by RFI (residual feed intake) and RIG (residual intake and gain). Dry matter intake (DMI) was recorded to calculate the RFI and RIG, classified as efficient, moderately or inefficient. After the confinement period, they were slaughtered and the carcass and meat quality were determined. The efficient animals had DMI scores of 0.700 RFI and 0.400 kg/d RIG, lower than the inefficient ones with similar weight gain. The RFI efficient animals showed greater shear force, without effect in the RIG classification. In general, the variables analyzed were not influenced by RFI or RIG. Efficiency measures do not affect the carcass and meat quality of sheep, but they do have the advantage of identifying animals with lower feed consumption, making the system more efficient. However, as the dataset is limited to fully assess the effects, this manuscript can be used as preliminary results for future studies.

To investigate the prevalence, associated factors, and effects of primary overt renal disease on morbidity in patients with primary Sjögren's syndrome (pSS).All patients in the Sjögrenser (registry of adult pSS patients of the Spanish Society of Rheumatology) cohort were retrospectively investigated for the presence of clinically significant renal involvement directly related to pSS activity.Of the 437 patients investigated, 39 (9%) presented overt renal involvement during follow-up. Severe renal disease necessitating kidney biopsy was relatively rare (2%). Renal involvement may complicate pSS at any time during the disease course and is associated with severe disease (indicated by higher scores of involvement, activity, and damage), systemic multiorgan involvement, and a higher frequency of lymphoma. Multivariate analysis showed that older age (odds ratio [OR] 1.03, 95% confidence interval [CI] 1.00-1.07), higher European League Against Rheumatism Sjögren's Syndrome Disease Activity Index scores (OR 1.1, CI 1.03-1.18), serum anti-La/SSB positivity (OR 6.65, CI 1.41-31.372), and non-vasculitic cutaneous involvement (OR 5.47, 1.03-29.02) were independently associated with this complication. Chronic renal failure developed in 23 of 39 patients (59%); only 1 of them progressed to end-stage renal disease necessitating renal replacement therapy. Patients with overt renal disease showed higher Sjögren's syndrome disease damage index scores, higher rates of hospitalisation due to disease activity and higher rates of clinically relevant comorbidities.Overt renal involvement in pSS is not uncommon. Although it usually shows a favourable prognosis, is associated with significant morbidity.

Objectives To investigate the prevalence, associated factors, and effects of primary renal disease on morbidity in patients with primary Sjögren’s syndrome (pSS).Methods All patients in the SJÖGRENSER (registry of adult SSp patients of the Spanish Society of Rheumatology) cohort were retrospectively investigated for the presence of clinically significant renal involvement directly related to pSS activity.Results Of the 437 patients investigated, 39 (9%) presented overt renal involvement during follow-up. Severe renal disease necessitating kidney biopsy was relatively rare (23%). Renal involvement may complicate pSS at any time during the disease course and is associated with severe disease (indicated by higher scores of involvement, activity, and damage), systemic multiorgan involvement, and a higher frequency of lymphoma. Multivariate analysis showed that older age (odds ratio [OR] 1.03, 95% confidence interval [CI] 1.00–1.07), higher European League Against Rheumatism Sjogren’s Syndrome Disease Activity Index scores (OR 1.1, CI 1.03–1.18), serum anti-La/SSB positivity (OR 6.65, CI 1.41–31.372), and non-vasculitic cutaneous involvement (OR 5.47, 1.03–29.02) were independently associated with this complication. Chronic renal failure developed in 23 of 39 patients (59%); only 1 of them progressed to end-stage renal disease necessitating renal replacement therapy. Patients with renal disease showed higher Sjögren’s syndrome disease damage index scores, higher rates of hospitalization due to disease activity and higher rates of clinically relevant comorbidities.Conclusion Renal involvement is an uncommon complication in pSS that was observed in 9% of patients. Although categorized as a non-negligible comorbidity, this condition shows a favorable prognosis.

En el cáncer de endometrio, el subtipo endometrioide, las variables clínicas e histológicas son las más importantes para el pronóstico de la enfermedad, entre estas se encuentran: el estadio según la Federación Internacional de Ginecología y Obstetricia (FIGO), el grado del tumor, la profundidad de la invasión miometrial, el compromiso del espacio linfático-vascular y/o de los ganglios linfáticos. Existen hasta el momento varias investigaciones que evalúan diferentes variables biológicas en muestras de suero y de tejido tumoral para detectar biomarcadores capaces de predecir un resultado clínico. En el presente artículo se revisa la evidencia sobre la relevancia en el campo diagnóstico, de pronóstico y seguimiento de marcadores moleculares como el p53, PTEN, PIK3, mTOR, B-cateninas, inestabilidad de microsatélites, la expresión del factor de crecimiento endotelial vascular, aneuploidía del DNA entre otros y de marcadores bioquímicos como CA125, CA15-3, YKL-40, VEGF, HE-4 que ayuden al médico a una mejor estratificación de los pacientes en categorías con diferentes riesgos de recurrencias y brindarles un mejor tratamiento adyuvante.

Bioconversion of the aromatase inhibitor formestane (4-hydroxyandrost-4-ene-3,17-dione) (1) by the fungus Rhizopus oryzae ATCC 11145 resulted in a new minor metabolite 3,5α-dihydroxyandrost-2-ene-4,17-dione (2) and the known 4β,5α-dihydroxyandrostane-4,17-dione (3) as the major product. The structural elucidation and bioactivities of these metabolites are reported herein. Molecular modeling studies of the interactions between these metabolites and the aromatase protein indicated that acidic (D309), basic (R115), polar (T310), aromatic (F134, F221, and W224), and non-polar (I133, I305, A306, V369, V370, L372, V373, M374, and L477) amino acid residues contribute important interactions with the steroidal substrates. These combined experimental and theoretical studies provide fresh insights for the further development of more potent aromatase inhibitors.

Dentro de los eventos tromboembólicos considerados están: la trombosis venosa profunda y el embolismo pulmonar, principales complicaciones que se pueden presentar durante y después de una cirugía ginecológica, por vía abierta o laparoscópica. Existe incertidumbre sobre el riesgo atribuido de tromboembolismo venoso después de procedimientos laparoscópicos, siendo controvertida la recomendación de tromboprofilaxis en este campo. El American College of Chest Physicians y el American Congress of Obstetricians and Gynecologist sugieren la clasificación de riesgo en 4 categorías: bajo, medio, alto y muy alto de las pacientes programadas para cirugía ginecológica. El riesgo de evento tromboembólico va a depender del tipo de procedimiento, tiempo quirúrgico, edad y la presencia de otros factores de riesgo. Hasta tener mayor evidencia, las pacientes llevadas a cirugía laparoscópica deberán ser estratificadas por categoría de riesgo y proveer profilaxis similar a pacientes llevadas a laparotomía. En mujeres de alto riesgo (mayores de 60 años, historia de trombosis venosa profunda, embolismo pulmonar o cáncer) se pueden beneficiar de la profilaxis dual y de prolongar su profilaxis hasta por 28 días.

In an effort to generate new steroidal aromatase inhibitors, formestane (4-hydroxyandrost-4-ene-3,17-dione) (1) was biotransformed by Rhizopus oryzae to yield the known 4β,5α-dihydroxyandrostane-3,17-dione as the major product (5) and bioconverted by Beauveria bassiana to afford the known reduced 4,17β-dihydroxyandrost-4-en-3-one (6) and 3α,17β-dihydroxy-5β-androstan-4-one (7) and the new 4,11α,17β-trihydroxyandrost-4-en-3-one (8). All the metabolites showed more potent activities than their parent congener in the aromatase and MCF-7 breast cancer assays. The bioactivities and structural elucidation of these metabolites as well as the semisynthesis of formestane (1) from testosterone (2) are reported herein.

Objective: To identify characteristics of early arthritis units, that may be associated with better referral eficiency. Methods: A national survey of the 36 early arthritis units (EAU) in Spanish Rheumatology Units in 2004 (SERAP project). Survey collected information about general practitioners (GP) educational program to improve knowledge and practical skills of early arthritis, networking and feed-back system and referral efficiency. EAU were classified in two groups according to 25 and 50% of inappropriate referral process, respectively.

Adult-onset Still's disease (AOSD) is frequently refractory to standard therapy. Tocilizumab (TCZ) has demonstrated efficacy in single cases and in small series of patients with AOSD. The aim of this multicenter study was to assess the efficacy of TCZ in patients with AOSD refractory to conventional treatment.This was a retrospective open-label study of TCZ treatment in 34 patients with AOSD who had experienced an inadequate response to corticosteroids and at least 1 standard synthetic immunosuppressive drug and also, in many cases, biologic agents.The mean ± SD age of the patients (8 men and 26 women) was 38.7 ± 16.1 years. The median duration of AOSD before TCZ was initiated was 4.2 years (interquartile range [IQR] 1-9 years). The initial dosages of intravenous TCZ were 8 mg/kg every 4 weeks in 22 patients, 4 mg/kg every 4 weeks in 2 patients, and 8 mg/kg every 2 weeks in 10 patients. TCZ treatment resulted in rapid and maintained improvement in both clinical and laboratory parameters. After 1 year of TCZ therapy, the incidence of joint manifestations had decreased from 97.1% at baseline to 32.4%, the incidence of both cutaneous manifestations and fever had decreased from 58.8% to 5.9%, and the incidence of lymphadenopathy had decreased from 29.4% to 0%. A dramatic reduction in laboratory markers of inflammation, including the C-reactive protein level, the erythrocyte sedimentation rate, and the ferritin level, was achieved. The median dosage of prednisone was also reduced, from 13.8 mg/day (IQR 5-45) at the initiation of TCZ to 2.5 mg/day (IQR 0-30) at 12 months. After a median followup of 19 months (IQR 12-31 months), only 2 patients required permanent discontinuation of TCZ therapy because of severe infections.TCZ treatment was associated with rapid and maintained clinical and laboratory improvement in patients with AOSD refractory to standard treatment. However, joint manifestations seem to be more refractory to treatment compared with systemic manifestations.

Objective.We evaluated whether a single-nucleotide polymorphism (SNP) of theTRAF6gene previously associated with systemic lupus erythematosus and rheumatoid arthritis may be a common risk factor for systemic sclerosis (SSc) and giant cell arteritis (GCA).Methods.A total of 1185 patients with SSc, 479 patients with biopsy-proven GCA, and 1442 unrelated healthy controls of white Spanish origin were genotyped for the rs540386 variant using a specifically designed TaqMan©allele discrimination assay.Results.No significant associations of this SNP with global SSc or GCA were found. This was also the case when the potential associations of theTRAF6polymorphism with the main clinical phenotypes of the 2 diseases (e.g., limited cutaneous and diffuse cutaneous SSc, or presence of polymyalgia rheumatica and visual ischemic manifestations in GCA) were assessed.Conclusion.Our data do not support a role of the rs540386TRAF6variant as a key component of the genetic network underlying SSc and GCA.

Identifying early predictors of response to biological agents is important for both the individual patient and health economics. The aim here was to identify clinical variables that are easily assessed in clinical practice which are associated with a major response to rituximab (moderate to good EULAR response, according to DAS28 values) in patients with active rheumatoid arthritis and inadequate response to anti-TNF agents or traditional DMARDs.Rituximab (2x1g, two weeks apart) was administered to 108 patients in four different Spanish hospitals. The primary efficacy endpoint was the percentage of patients who achieved a major response at six months. Potential predictors of a major response were identified using multivariate binary logistic regression models.At six months of treatment 75.9% of patients achieved a major response (24% good and 52% moderate). Comparing the clinical features at baseline between patients who did or did not achieve a major response, significant differences were found in rheumatoid factor (RF) and anti-CCP positivity, as well as in the number of failed anti-TNF agents prior to rituximab. While rituximab delivers clinical benefit in seronegative patients, the presence of RF and/or anti-CCP consistently enriches clinical responses. The multivariate analysis showed that the best model for predicting a major EULAR response to rituximab was comprised of the following two variables: the anti-CCP antibody positivity (p=0.045) and the number of previous anti-TNF agents used (p=0.028). Using a cut-off level for CCP of 300 U/ml we found that patients with an anti-CCP titre300 U/ml were 3-4 times more likely to achieve a major EULAR response [odds ratio (OR): 3.38; 95% CI: 1.025-11.17]. By contrast, those patients who had failed to respond to 2 or more anti-TNF agents had a 72.5% lower probability of achieving a moderate to good EULAR response (OR: 0.275; 95% CI: 0.087-0.871) than did patients who had only failed to respond to one such agent.A lower number of previously-failed TNF blockers and high anti-CCP titre can help select the best candidates for RTX therapy in patients with RA.

Hepatitis A vaccination is justified in areas with an intermediate endemicity of the infection.To estimate the epidemiological impact of hepatitis A infection in Colombia.Epidemiological indicators of hepatitis A infection prevalence by age, morbidity by age, and lethality by age were estimated from a literature search. These measures were projected on a hypothetical cohort of children followed from birth until 15 years of age. The number of cases of infection, jaundice, hepatic failure, hospitalizations and deaths were estimated.From birth to adolescence, a cohort of 872 923 urban children in Colombia would generate between 312,331 and 598,591 infections, between 13,586 and 25,960 hospitalizations, between 213 and 407 hepatic failures and between 107 and 204 deaths.The impact of hepatitis A infection in Colombia, is important. Introducing hepatitis A vaccination would reduce a substantial number of severe hepatitis A cases.

To assess the influence of the interleukin (IL)2-IL21 rs6822844 G/T polymorphism in the susceptibility to biopsy-proven giant cell arteritis (GCA) and in the clinical spectrum of manifestations of this vasculitis.Two hundred and seventy-two biopsy-proven GCA patients were included in this study. DNA from patients and matched controls (n=791) was obtained from peripheral blood. Samples were genotyped for the rs6822844 polymorphism using a predesigned TaqMan allele discrimination assay and by polymerase chain reaction amplification.No significant differences in the allele and genotype frequencies between biopsy-proven GCA patients and controls were observed. However, the stratification of GCA patients disclosed some differences according to gender and ischemic manifestations of the disease. In this regard, the frequency of the minor allele T was increased in males (14.8%) compared to females (8.4%) (odds ratio-OR:1.89 (95% confidence interval-CI: 1.09-3.28); p=0.02; Bonferroni adjustment p=0.12). Also, minor allele T frequency was increased in GCA patients with severe ischemic complications (12.8%) compared to those without severe ischemic complications (7.7%) (OR:1.72 (95% CI: 0.97-3.05); p=0.05; Bonferroni adjustment p=0.30), and specifically in patients with jaw claudication (13.7% versus 8.2% in those without jaw claudication; OR:1.76 (95% CI: 1.02-3.04); p=0.04; Bonferroni adjustment p=0.24).IL2-IL21 rs6822844 polymorphism does not appear to be a genetic risk factor for susceptibility to biopsy-proven GCA. However, this gene polymorphism may contribute to the different phenotypic expression of this vasculitis, in particular in the development of ischemic complications of the disease.

Resume Prevotella bivia est une bacterie anaerobie Gram negative qui fait classiquement partie de la famille des Bacteroides. Elle a ete rapportee dans des infections obstetricales et gynecologiques. A ce jour, il n'y a eu qu'un seul cas publie d'arthrite septique due a ce germe. Les auteurs rapportent le premier cas d'arthrite septique a ce germe chez un patient atteint d'une polyarthrite rhumatoide grave et ancienne traitee par de faibles doses de corticoides. La polyarthrite rhumatoide est une cause favorisante classique d'arthrite septique. Ses causes sont multifactorielles (immunosuppression induite par la maladie et ses traitements, protheses…). Cependant, les arthrites septiques dues a des germes anaerobies sont rarement signalees. En general, leur aspect clinique est tres variable : debut insidieux, apyrexie, absence de signes inflammatoires ou systemiques sont souvent observes. L'infection articulaire succede generalement a une dissemination hematogene. Les bacteries du groupe Bacteroides fragilis sont celles qui sont le plus souvent isolees. Un diagnostic precoce et un traitement rapide, comportant un drainage et un debridement de l'articulation atteinte associes a un traitement antibiotique specifique avec du metronidazole sont indispensables pour obtenir un resultat favorable.

Objective. To estimate the incidence of early arthritis (EA) and of RA in adults (>16 yrs) in Spain. Methods. Primary care physicians were instructed in the detection of new cases using a checklist. All cases were evaluated at EA units (EAUs) within 15 days of detection. ACR criteria for the classification of RA were assessed every 6 months thereafter. Results. In an area covering 4 342 378 inhabitants over the age of 16 yrs, 2467 patients were referred to the EAU, of whom 1063 fulfilled EA criteria (43.1%). After 6 months, 362 patients fulfilled RA criteria. The estimated annual incidence of EA was 25/100 000 population (95% CI: 23, 26). The annual incidence of RA was 8.3 cases/100 000 (95% CI: 7.5, 9.2): 11.3/100 000 in women (95% CI: 10.0, 12.8), and 5.2/100 000 in men (95% CI: 4.3, 6.3). The incidence of RA increased with age in both sexes. At the 6 months’ assessment, 187 (51.7%) of the patients with RA were RF positive. The presentation of RA was mainly polyarticular (n ¼ 268; 74.0%). There were 701/1063 patients with EA who did not fulfil RA criteria by 6 months after the first rheumatologist visit. If all cases of undifferentiated arthritis (n ¼ 118; 17%) became RA, the incidence would be in the range of 10 cases/100 000 population. Conclusions. RA incidence in Spain is in the lower range of published data. The incidence of EA is about three times that of RA.

Although there is relevant information on frequency of osteoporosis in women with rheumatoid arthritis (RA), data about male patients are limited. We evaluated the frequency of osteoporosis in a group of Spanish men with RA followed in a university hospital.From the database of our bone densitometry unit, we searched for men with RA evaluated between January 1991 and December 2004 and identified 187 patients, 156 of whom were older than 50 years. Previously recorded demographic, disease, and treatment-related variables were collected. Bone mineral density (BMD) was measured by dual x-ray absorptiometry (DEXA). Osteoporosis was defined according to the criteria of the World Health Organization (WHO), recommended for postmenopausal Caucasian women, as a T scoreor= -2.5 SD, and the International Society for Clinical Densitometry (ISCD), which indicates the same diagnostic threshold, but only in men over age 50 years.BMD was lower than in the general population in both lumbar spine [Z score -0.41 +/- 1.00 (95% CI -0.55 to -0.26)] and femoral neck [Z score -0.46 +/- 0.95 (95% CI -0.60 to -0.31)]. When the WHO threshold for postmenopausal women was applied, frequency of osteoporosis was 13% at lumbar spine, 12% at the femoral neck, and 21% in at least one of the evaluated sites. When ISCD criteria were applied, the frequency of osteoporosis was 13%, 14%, and 23%, respectively.Frequency of osteoporosis in men is considerably lower than that traditionally established in women with RA, independent of the diagnostic criteria applied.

En este estudio se estandarizó una metodología para la caracterización molecular de cepas nativas de Bacillus thuringiensis, la cual se basó en la amplificación de los genes cry mediante la Reacción en Cadena de la Polimerasa (PCR). Se utilizaron cuatro mezclas de oligonucleótidos: dos Generales (I y II), los cuales reconocen genes de las familias cry1, cry2, cry3, cry4 y cry1Ia, y dos Específicos (A y B), que identifican los genes de la familia cry1 (cry1Aa, cry1Ab, cry1Ac, cry1Ba, cry1Ca y cry1Da). La calidad y concentración del ADN bacteriano influyó sobre la especificidad y concentración de los productos obtenidos mediante la amplificación de los genes cry. La calidad del ADN purificado a través del método rápido reportado por M. He et al. (Nucleic Ac. Res. 18:1660, 1990) permitió una amplificación eficiente. Para realizar la PCR en condiciones óptimas, se utilizó una mezcla de reacción con un volumen final de 20 µl, la cual contenía 0,4 µM de cada oligonucleótido, 1X PCR buffer (50 mM KCL, wmM Tris-HCl, pH 8,3 y 3,0 mM MgCl 2), 200 µM de cada dNTP y 1U de Taq-ADN-polimerasa, además de 10-100 y 300- 500 ng de ADN bacterial para las mezclas de los oligonucleótidos Generales y Específicos, respectivamente. El programa de amplificación incluyó 30 ciclos de desnaturalización a 94°C, hibridación a 53°C y síntesis a 72°C durante 20 segundos cada uno. La metodología estandarizada se puede utilizar rutinariamente para amplificar los genes cry procedentes de aislamientos nativos de B. thuringiensis, lo cual permite clasificarlos y seleccionarlos de una manera rápida y precisa de acuerdo con su actividad biológica y potencial biotecnológico; adicionalmente, como paso previo de los ensayos de toxicidad contra diversas especies de insectos plaga de interés agrícola.Utilization of Polymerase Chain Reaction for Characterization of Native Isolates of Bacillus thuringiensisA methodology based on Polymerase Chain Reaction (PCR) techniques was standardized for the molecular characterization of cry genes in Bacillus thuringiensis. Four oligonucleotides mixes (primers) were used: two General (I and II) -which recognize the genes families cry1, cry2, cry3, cry4 and cry1Ia-, and two Specific (A and B) which recognize the genes family cry1 (cry1Aa, crylAb, cry1Ac, cry1Ba, cry1Ca and cry1Da). The quality of bacterial DNA influenced the amplification product specificity and concentration. The quality of the DNA purified by M. He et al. fast method (Nucleic Ac. Res. 18:1660, 1990) allowed an efficient amplification. The optimum conditions for PCR were achieved using a mixture reaction with a final volume of 20 µL which contained 0.4 µM of each primer, 1X PCR buffer (50 mM KCL, 10mM Tris-HCI, pH 8.3 and 3.0 mM MgCI,), 200 µM dNTP's, 1 U Taq-DNA-polymerase, 10-100 ng of bacterial DNA for the General mixtures and 300-500 ng of bacterial DNA for the Specific mixtures. The amplification program included 30 cycles as follows: denaturation at 94°C, annealing at 53°C and synthesis at 72°C during 20 seconds each one. The standardized methodology could be used routinely in the cry genes amplification of native isolates of B. thuringiensis for the classification, rapid and precise selection according to the potential biological activity as a previous step to toxicity trials against diverse insect pests of agriculture interest.

Genes containing the cauliflower mosaic virus 35S promoter fused to open reading frames coding for tomato proteinase inhibitor I, tomato inhibitor II, and potato inhibitor II were expressed in transgenic tobacco plants. Inhibitor I and II proteins were identified by immunoblotting and quantified by immunoradial diffusion. Both inhibitors exhibited the molecular weights found for the native proteins in their natural environments. Extracts of leaves from transformed plants contained inhibitory activities against trypsin and chymotrypsin that reflected the levels of inhibitor I or II protein present. The results demonstrate that in tobacco leaves the introns of both inhibitor I and inhibitor II genes were excised correctly and that pre and prepro inhibitor I and II proteins were correctly processed. Growth of Manduca sexta larvae (tobacco hornworms) feeding on leaves of transgenic plants containing inhibitor II, a powerful inhibitor of both trypsin and chymotrypsin, was significantly retarded, compared to growth of larvae fed untransformed leaves. Levels of inhibitor II protein as low as 50 micrograms/g of tissue moderately affected larval growth, whereas levels above 100 micrograms/g severely reduced growth. The presence of tomato inhibitor I protein, a potent inhibitor of chymotrypsin but a weak inhibitor of trypsin, in transgenic tobacco leaves had little effect on the growth of the larvae. These experiments indicated that trypsin inhibitory activity, but not chymotrypsin inhibitory activity, was mainly responsible for the inhibition of larval growth.