The importance of factorial design on the optimization of biosensor performance: immobilization of glucose oxidase as a case study.

Conventionally, the optimization of glucose biosensors is achieved by varying the concentrations of the individual reagents used to immobilize the enzyme. In this work, the effect and interaction between glucose oxidase enzyme (GOx), ferrocene methanol (Fc), and multi-walled carbon nanotubes (MWCNTs) at different concentrations were investigated by a design of experiments (DoE). For this analysis, a factorial design with three factors and two levels each was used with the software RStudio for statistical analysis. The data were obtained by electrochemical experiments on the immobilization of GOx-Fc/MWCNT at different concentrations. The results showed that the factorial DoE method was confirmed by the non-normality of the residuals and the outliers of the experiment. When examining the effects of the variables, analyzing the half-normal distribution and the effects and contrasts for GOx-Fc/MWCNT, the factors that showed the greatest influence on the electrochemical response were GOx, MWCNT, Fc, and MWCNT:Fc, and there is a high correlation between the factors GOx, MWCNT, Fc, and MWCNT:Fc, as shown by the analysis of homoscedasticity and multicollinearity. With these statistical analyses and experimental designs, it was possible to find the optimal conditions for different factors: 10 mM mL-1 GOx, 2 mg mL-1 Fc, and 15 mg mL-1 MWCNT show a greater amperometric response in the glucose oxidation. This work contributes to advancing enzyme immobilization strategies for glucose biosensor applications. Systematic investigation of DoE leads to optimized immobilization for GOx, enables better performance as a glucose biosensor, and allows the prediction of some outcomes.

Global Situation of Bioremediation of Leachate-Contaminated Soils by Treatment with Microorganisms: A Systematic Review.

This systematic review presents the current state of research in the last five years on contaminants in soils, especially in leachates from solid waste landfills, with emphasis on biological remediation. In this work, the pollutants that can be treated by microorganisms and the results obtained worldwide were studied. All the data obtained were compiled, integrated, and analyzed by soil type, pollutant type, bacterial type, and the countries where these studies were carried out. This review provides reliable data on the contamination of soils worldwide, especially soils contaminated by leachate from municipal landfills. The extent of contamination, treatment objectives, site characteristics, cost, type of microorganisms to be used, and time must be considered when selecting a viable remediation strategy. The results of this study can help develop innovative and applicable methods for evaluating the overall contamination of soil with different contaminants and soil types. These findings can help develop innovative, applicable, and economically feasible methods for the sustainable management of contaminated soils, whether from landfill leachate or other soil types, to reduce or eliminate risk to the environment and human health, and to achieve greater greenery and functionality on the planet.

In vitro ovicidal activity of Brongniartia montalvoana against small ruminant gastrointestinal nematodes.

Several plants of the Fabaceae family have been assessed regarding their high nutritional value and anthelmintic properties. The ovicidal effect of the hydroalcoholic extract (Bm-HAE) and subfractions from the aerial parts of Brongniartia montalvoana (Fabaceae) against a mixed strain of gastrointestinal nematodes (GIN) (Haemonchus spp., Trichostrongylus spp. and Oesophagostomum spp.) resistant to albendazole sulfoxide, ivermectin and levamisole was evaluated by the egg hatch test (EHT). The Bm-HAE was subjected to liquid-liquid chemical separation with ethyl acetate giving two fractions, an aqueous (Bm-Aq) and an organic (Bm-EtOAct). The purification of the bioactive fraction (Bm-EtOAct) through chromatographic separation resulted in four bioactive subfractions (BmR6, BmR7, BmR8 and BmR10). The treatments were designed as follows: Bm-HAE at 800, 1,500, 3,000 and 6,000 µg/mL, and Bm-Aq, Bm-EtOAct and subfractions (BmR6, BmR7, BmR8 and BmR10) at 100, 200, 400 and 800 µg/mL. Two properly negative controls (distilled water and 2% methanol) and thiabendazole (100 µg/mL) as a positive control were used for each bioassay. The chemical identification of the extract, fractions and subfractions was performed through chromatographic processes like open column chromatography, thin-layer chromatography (TLC) and high-performance liquid chromatography (HPLC-PDA). Additionally, the GIN eggs exposed to the bioactive compounds were observed through confocal laser scanning microscopy (CLSM). The Bm-HAE showed 99.5% egg hatching inhibition (EHI) at 6,000 µg/mL with a lethal concentration (LC50) of 1110 µg/mL. The Bm-EtOAc fraction displayed 99.1% EHI at 800 µg/mL with LC50 = 180 µg/mL. The ovicidal activity of the four subfractions was similar at 800 µg/mL: BmR6 (92% EHI); BmR7 (100% EHI); BmR8 (97.8%); and BmR10 (99.1%). The HPLC-PDA analysis of the bioactive subfractions allowed identification of p-coumaric acid, ferulic acid and coumarin derivatives as major compounds. The CLSM analysis allowed observation of morphological alterations in unhatched larvae caused by bioactive compounds present in the Bm-EtOAc and BmR10. In addition, the flavonoids eriodyctiol, luteolin and cynaroside were described for the first time for B. montalvoana.

Differences in moral reasoning among medical graduates, graduates with other degrees, and nonprofessional adults.

BACKGROUND: Reasoning and moral action are necessary to resolve day-to-day moral conflicts, and there are certain professions where a greater moral character is expected, e.g., medicine. Thus, it is desirable that medical students develop skills in this field. Some studies have evaluated the level of moral reasoning among medical students; however, there are no comparative studies involving other types of populations. Therefore, the objective of this study was to compare the moral reasoning among medical graduates with that of a group of young graduates with other degrees and of a group of nonprofessional adults. METHODS: An exploratory cross-sectional study was conducted. Pediatric residents and pediatric subspecialty residents at a pediatric hospital were invited to participate, forming the group of "medical graduates". A group of young people from a social program and students with a master's degree in a science from the same pediatric hospital were also invited to participate, comprising the group of "graduates with other degrees". Finally, a group of beneficiaries of a family clinic was invited to participate, which we categorized as "nonprofessionals". To evaluate the differences in moral reasoning between these 3 groups, we applied the Defining Issues Test (DIT), a moral reasoning questionnaire designed by James Rest using Kohlberg's theory of moral development. RESULTS: The moral reasoning of 237 subjects-88 from the "medical graduates" group, 82 from the "graduates with other degrees" group and 67 from the "nonprofessionals" group- was evaluated. We found differences in the profiles of moral development of the groups. The profile of the "nonprofessionals" showed a very high predominance of subjects at the preconventional level, 70%, but only 4.5% at the postconventional level. Among the "medical graduates", we observed 37.5% at the preconventional level and 34% at the postconventional level (X2 p < 0.001); this group had the highest percentage in this category. This large difference could be because the differences in the ages and socioeducational levels of nonprofessionals are much wider than those among medical graduates. However, significant differences were also found when the profiles of medical graduates were compared with those of graduates with other degrees, since the latter demonstrated 56% at the preconventional level and 18% at the postconventional level (X2 test, p = 0.02). CONCLUSIONS: Significant differences were found in moral reasoning among the groups that we evaluated. Among the group of medical graduates, there was a higher percentage of subjects at the postconventional level than among the group of graduates with other degrees and a much higher percentage than among the group of nonprofessionals. Our conclusions give the first evidence that studying medicine seems to influence the development of moral reasoning in its students. Therefore, we consider it relevant to develop educational strategies where the student is involved in simulated but realistic decision-making situations, where there are moral dilemmas to resolve from their early years of training.

In vitro ovicidal activity of Baccharis conferta Kunth against Haemonchus contortus.

The indiscriminate use of chemical drugs to deworm livestock tends to trigger an anthelmintic resistance problem. In this context, the use of plant extracts rich in secondary metabolites could be an alternative method for the control of gastrointestinal nematodes. Baccharis conferta Kunth is a native plant species from Mexico that is widely used by several ethnic groups as forage for farm animals and medicinally to treat gastrointestinal diseases such as acute stomach ache, dysentery, diarrhoea, vomiting, indigestion, colic, intestinal spasms, urinary problems, and cramps. The aim of the present study was to isolate and characterise the ovicidal constituents of B. conferta and to determine a possible mode of action against Haemonchus contortus. The ovicidal activity was determined using the egg hatching inhibition test (EHI) to assess the methanol extract obtained from B. conferta foliage. The dry extract was partitioned (water/ethyl acetate) to obtain an ethyl acetate (BcEtOAc-F) and aqueous fraction. BcEtOAc-F showed an ovicidal activity of 72.32% EHI at 1 mg/mL. The chromatographic fractionation of BcEtOAc-F resulted in three active sub-fractions with higher ovicidal activity: BcC1R4 (99.15% EHI at 1.0 mg/mL); BcC1R5 (92.51% EHI at 0.75 mg/mL); and BcC1R8 (96.8% EHI at 3.0 mg/mL). Chemical analysis of the BcC1R4 fraction allowed the identification of the major active compound, isokaempferide (1, 98.06% EHI at 1 mg/mL). While, 4,5-di-O-acid caffeoylquinic (3; 96.8% EHI at 3 mg/mL) and an inactive flavone (vicenin-2, 2) were identified as the main compounds in BcC1R8. Chemical characterisation of the isolated compounds was performed via spectroscopic (NMR) and spectrometric (UPLC-MS) analyses. Additionally, the environmental and confocal scanning microscopy analyses revealed that isokaempferide was able to cross the eggshell layer without breaking it and attach itself to the embryo, causing its death. The flavonol, isokaempferide, and the hydroxycinamic acid, 4,5-di-O-caffeoylquinic, displayed powerful ovicidal effects, proving to be a potential alternative for the development of a phytodrug for the control of haemonchosis.

Análisis estadístico de los espectros de frecuencia de las regiones reguladoras del ENCODE / Statistical analysis of the Fourier Spectra of the ENCODE regulatory regions

RESUMEN En la actualidad, nuevas bases de datos genómicos (secuencias de ADN) son puestas al alcance del dominio público para su análisis. La bioinformática ha desarrollado algoritmos para extraer información y características de dichas secuencias. Sin embargo, estos algoritmos bioinformáticos tienen limitaciones. Una alternativa es utilizar herramientas propias del procesamiento digital de señales (DSP) adaptadas a secuencias genómicas (procesamiento de señales genómicas - GSP). El presente trabajo versa sobre el análisis de los cuatro primeros momentos centrales (media, desviación estándar, asimetría y curtosis) y dos momentos estadísticos (mediana y varianza) de los espectros frecuenciales de las 15 Regiones Reguladoras (RRs) de la base de datos ENCODE con el objetivo de estudiar diferencias estadísticas y frecuencias características. La base de datos seleccionada es "mapeada". Luego, la FFT es calculada a estas señales genómicas y finalmente los momentos estadísticos son implementados. Los resultados mues tran la existencia de 3 grupos de RRs utilizando la media, mediana y curtosis. La desviación estándar y la varianza, parecen no resaltar información importante. Finalmente, la asimetría revela un comportamiento homogéneo ante la presencia de valores atípicos en algunas RRs. Estas observaciones permiten inferir que la periodicidad dentro de la secuencia está relacionada o podría determinar la función biológica que desempeña la misma secuencia.

ABSTRACT Nowadays, new genomic databases (DNA sequences) are available to the whole scientist community for its analysis. The bioinformatics has developed algorithms to extract information and features of the sequences. However, the bioinformatics algorithms have restrictions. An alternative is the use of digital signal processing (DSP) tools adapted to genomic sequences (genomic signal processing - GSP). This work analyzes the first four statistics moments (mean, standard deviation, skewness and kurtosis) and other two moments (median and variance) of the frequency spectra of 15 regulatory regions (RRs) in ENCODE database with the main objective of studying the statistics di fferences and frequency features. The selected database is mapped. Then, the FFT is calculated to these genomic signals and finally the statistic moments implemented. The results show a three-group behavior in the RRs with the mean, median and kurtosis. The deviations standard and the variance do not show important behavior. Finally, the skewness shows a homogeneous behavior with the lack of atypical values in some RRs. These observations support the idea of the presence of periodicities in a sequence that may be related or may determine the biological function that a sequence may perform.

Parasites of cetaceans stranded on the Pacific coast of Costa Rica.

Information regarding parasitic fauna of cetaceans from Costa Rica is provided for the first time. A total of 25 stranded dolphins and whales were examined between 2001 and 2009, including striped dolphin (Stenella coeruleoalba) (n=19), pantropical spotted dolphin (S. attenuata) (n=2), spinner dolphin (S. longirostris) (n=1), bottlenose dolphin (Tursiops truncatus) (n=1), dwarf sperm whale (Kogia sima) (n=1) and Cuvier's beaked whale (Ziphius cavirostris) (n=1). Pathological findings associated with the parasites are also presented. In the most representative dolphin species, S. coeruleoalba, the prevalence of parasites was 89.5%; moreover, all examined specimens of S. attenuata, S. longirostris, T. truncatus and Z. cavirostris presented parasites. No parasites were recovered from K. sima. Fourteen helminth taxa were identified, including six species of cestodes (Strobilocephalus triangularis, Tetrabothrius forsteri, Trigonocotyle sp., Phyllobothrium delphini, Monorygma grimaldi, Tetraphyllidea gen. sp. plerocercoid), four digeneans (Nasitrema globicephalae, Brachycladium palliatum, B. pacificum and Oschmarinella albamarina) and four nematodes (Anisakis spp., Halocercus lagenorhynchi, Halocercus sp. and Crassicauda anthonyi). A commensal crustacean, Xenobalanus globicipitis, was also identified. All identified parasites representing new geographic records for the Pacific coast of Central America and new host records are presented. Parasitological information is valuable for conservation of cetaceans in Pacific coast of Costa Rica.

Pathology of striped dolphins (Stenella coeruleoalba) infected with Brucella ceti.

Seventeen striped dolphins (Stenella coeruleoalba) displaying swimming disorders compatible with neurological syndromes were investigated for Brucella infection. Sixteen dolphins had meningoencephalomyelitis. Serum antibody against Brucella antigen was detected in all 14 animals tested and Brucella ceti was isolated from eight out of nine animals. Brucella antigen was detected in the brain by immunofluorescence, but not by immunohistochemical labelling. By contrast, Brucella antigen was demonstrated by immunohistochemistry in the trophoblast of animals with severe placentitis and in the mitral valve of animals with myocarditis. The microscopical lesions observed in the tissues of the infected dolphins were similar to those of chronic brucellosis in man. The severity of brucellosis in S. coeruleoalba indicates that this dolphin species is highly susceptible to infection by B. ceti.

Francisella sp., an emerging pathogen of tilapia, Oreochromis niloticus (L.), in Costa Rica.

Francisella sp. is an emergent bacterial pathogen that causes acute to chronic disease in warm and cold water cultured and wild fish species. During the past 3 years, the bacterium has been detected in tilapia, Oreochromis niloticus, cultured in Costa Rica. Infected fish presented non-specific clinical signs, such as erratic swimming, anorexia, anaemia, exophthalmia and high mortality. Upon macroscopic and microscopic examination, several internal organs (mainly spleen and kidney) were enlarged and contained white nodules. Histological examination revealed the presence of multifocal granulomatous lesions, with the presence of numerous small, pleomorphic, cocco-bacilli. The bacteria were isolated from infected tilapia on selective media and grown on several media with and without antibiotics. Specific PCR primers to the Francisella genus were used to confirm the preliminary diagnoses. In comparison with several bacterial 16S rRNA sequences, our isolate was found to share 99% identity with other Fransicella spp. isolated from fish, and more than 97% identity to the human pathogen Francisella tularensis. Koch's postulates were fulfilled after experimental intraperitoneal and gill exposure challenges.

Low frequency of human papillomavirus DNA in breast cancer tissue.

Human papillomavirus (HPV) is considered the aetiological agent for cervical cancer. Several reports have addressed a relationship with HPV and breast cancer, as different HPVs have been identified. The purpose of this study was to detect HPV DNA in 67 breast cancer patients and 40 non-malignant disease breast tissues by means of Polymerase Chain Reaction with consensus primers. The frequency of HPV in the cases group were 4.4% (3/67) and no positive samples among the reference group were identified. From the 3 positive samples, HPV types 16, 18 and 33 were identified by restriction patterns and direct sequencing. The high diversity among detection in the related studies shows that population genomic heterogeneity plays an important role in the disease. The low frequency detected in the present study suggests that HPV does not play an important role in breast cancer.

Isolation of Toxoplasma gondii from the keel-billed toucan (Ramphastos sulfuratus) from Costa Rica.

Pectoral muscles from a captive keel-billed toucan (Ramphastos sulfuratus) from Costa Rica were fed to a Toxoplasma gondii-free cat, and the cat shed oocysts. Laboratory mice fed these oocysts developed antibodies to T. gondii in their sera and T. gondii tissue cysts in their brains. The DNA extracted from the brains of infected mice was characterized using 10 polymerase chain reaction-restricted fragment length polymorphic markers (SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and Apico). The isolate designated TgRsCrl was found to be non-clonal with Type I, II, and III alleles at different loci. This is the first isolation of T. gondii from this host.

Mast cell tumour in a giant Galapagos tortoise (Geochelone nigra vicina).

A well-differentiated cutaneous mast cell tumour was diagnosed in a subadult female giant Galapagos tortoise. The tumour was a pedunculated, verrucose mass located near the base of the neck. The histological features, which were diagnostic for a mast cell tumour, included abundant intracytoplasmic granules that were stained metachromatically with Giemsa and toluidine blue stains. Mast cell tumours are rare in reptiles, and this is the first description of a mast cell tumour in a chelonian.

Spirorchiidiosis (Digenea: Spirorchiidae) and lesions associated with parasites in Caribbean green turtles (Chelonia mydas).

Forty-seven nesting green turtles (Chelonia mydas) were examined for parasitic pathogens. Four species of cardiovascular flukes (Digenea: Spirorchiidae), Learedius learedi, Hapalotrema postorchis, Monticellius indicum and Amphiorchis solus were collected from 39 of 40 of the turtles, and Neospirorchis species were identified in seven of the 47 by histological examination. The pathological changes associated with the spirorchiids and their eggs included aneurysms, arteritis of great vessels with dark nodular thickenings, endocarditis, haemorrhagic lesions, thrombi, vasculitis, and granulomatous reactions in all the turtles. Ozobranchus branchiatus (Hirudinea: Ozobranchidae) leeches were found on the skin of 27 of the 47 turtles and were associated with traumatic purulent and ulcerative dermatitis. Oesophagitis associated with Rameshwarotrema uterocrescens (Digenea: Pronocephalidae) was recorded in 11 of 43 of the turtles. Petechial haemorrhages and unspecific crater-like ulcerated lesions on the gastric mucosa associated with Charaxicephaloides species and Charaxicephalus robustus (Digenea: Pronocephalidae) were observed in four of 40 of the turtles. Cholycystitis and ectasia of mucosal glands associated with Rhytidodoides similis (Digenea: Rhytidodidae) were observed in one of the 47 turtles. Fibropapillomas were observed on the skin of one turtle and fibromas on the skin of two of them.

HLA-DRB1*0101 is associated with the genetic susceptibility to develop lichen planus in the Mexican Mestizo population.

The etiology of lichen planus (LP) is still unknown and previous studies have found an association between LP and HLA-DR1, DR2, DR3, DR9 and DR10 in different populations. The aim of this study was to analyze the distribution of the HLA-DRB1 alleles in Mexican Mestizo patients with LP. The aim of this study was to determine the gene frequency of HLA-DR locus in Mexican Mestizo patients with LP. We studied 20 patients with LP and 99 healthy Mexican Mestizo controls. HLA-DRB1 was performed by PCR-SSO reverse dot blot hybridization. High resolution HLA typing was performed by PCR-SSP. The HLA-DRB1*0101 allele was associated significantly in LP patients compared with healthy controls (pC = 0.0007, OR = 5.46, 95% CI = 1.86-16.06). HLA-DRB1*0101 is a marker for the development of LP in Mexican Mestizo population, yet another gene or HLA marker within MHC region may be the causatively associated gene.

Isolation and genetic characterization of Toxoplasma gondii from striped dolphin (Stenella coeruleoalba) from Costa Rica.

Toxoplasma gondii infection in marine mammals is of interest because of mortality and mode of transmission. It has been suggested that marine mammals become infected with T. gondii oocysts washed from land to the sea. We report the isolation and genetic characterization of viable T. gondii from a striped dolphin (Stenella coeruleoalba), the first time from this host. An adult female dolphin was found stranded on the Pacific Coast of Costa Rica, and the animal died the next day. The dolphin had a high (1:6400) antibody titer to T. gondii in the modified agglutination test. Severe nonsuppurative meningoencephalomyelitis was found in its brain and spinal cord, but T. gondii was not found in histological sections of the dolphin. Portions of its brain and the heart were bioassayed in mice for the isolation of T. gondii. Viable T. gondii was isolated from the brain, but not from the heart, of the dolphin. A cat fed mice infected with the dolphin isolate (designated TgSdCol) shed oocysts. Genomic DNA from tachyzoites of this isolate was used for genotyping at 10 genetic loci, including SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and Apico, and this TgSdCo1 isolate was found to be Type II.

A new pronocephalid, Pleurogonius tortugueroi n. sp. (Digenea), from the intestine of green sea turtles (Chelonia mydas) in Costa Rica.

A new species of trematode, Pleurogonius tortugueroi n. sp. (Digenea: Pronocephalidae) is described from the lower intestine of green sea turtles (Chelonia mydas) from Tortuguero National Park, Costa Rica. The new species differs from all other species of Pleurogonius by having a short oesophagus and oval testes close to lateral posterior limit of the body. It differs from all other species, except P. malaclemys Hunter 1961, by having an ovary between the testes; moreover the latter species is a parasite of freshwater turtles. All others members of the genus have a long oesophagus, testes placed to some distance from the posterior end, and the ovary is pretesticular. The new species appears most closely related to P. linearis Looss, 1901 but differs from it by having a different body shape, lappets of the head collar close at the cecal bifurcation level, a longer vitellarian field, different testis shape and position, ovary intertesticular, and different egg size.

Biologic and genetic characteristics of Toxoplasma gondii isolates in free-range chickens from Costa Rica, Central America.

The prevalence of Toxoplasma gondii in free-ranging chickens is a good indicator of the prevalence of T. gondii oocysts in the soil because chickens feed from the ground. The prevalence of T. gondii in 144 free-range chickens (Gallus domesticus) from Costa Rica was determined. Antibodies to T. gondii were assayed by the modified agglutination test (MAT), and found in 60 (40.1%) of 144 chickens with titers of 1:5 in 16, 1:10 in 5, 1:20 in 2, 1:40 in 3, 1:80 in 5, and 1:160 or higher in 29. Tissues of all chickens were bioassayed for T. gondii in mice or cats. Hearts and brains of 52 chickens with titers of 1:5 or higher and 16 chickens with doubtful titers were pooled and bioassayed in mice. Tissues from 76 chickens with MAT titers of 1:10 or less were pooled and fed to three T. gondii-free cats. Fecal floats of cats were bioassayed orally in mice but were negative for T. gondii oocysts. T. gondii was isolated by bioassay in mice from 32 chickens with MAT titers of 1:10 or higher. All infected mice from 4 of the 32 isolates died of toxoplasmosis. Genotyping of these 32 isolates using polymorphisms at the loci SAG1, SAG2, SAG3, BTUB and GRA6 revealed five genotypes. Five isolates had type I alleles and one isolate had type III alleles at all loci. The rest 26 isolates contained the combination of type I and II or I and III alleles and were divided into three genotypes. None was found to have genotype II alleles at all five loci. This is the first report of genetic characterization of T. gondii isolates from Costa Rica, Central America.

HLA-DRB1 alleles encoding the "shared epitope" are associated with susceptibility to developing rheumatoid arthritis whereas HLA-DRB1 alleles encoding an aspartic acid at position 70 of the beta-chain are protective in Mexican Mestizos.

The risk to develop rheumatoid arthritis (RA) has been associated with the presence of HLA-DRB1 alleles encoding the "shared epitope" (SE). Additionally, HLA-DRB1 alleles encoding an aspartic acid at position 70 (D70+ ) have been associated with protection against the development of RA. In this study we tested the association between either SE or D70+ and rheumatoid arthritis in Mexican Mestizos. We included 84 unrelated Mexican Mestizos patients with RA and 99 unrelated healthy controls. The HLA-typing was performed by PCR-SSO and PCR-SSP. We used the chi-squared test to detect differences in proportions of individuals carrying at least one SE or D70+ between patients and controls. We found that the proportion of individuals carrying at least one HLA-DRB1 allele encoding the SE was significantly increased in RA cases as compared to controls (p(c) = 0.0004, OR = 4.1, 95% CI = 2.2-7.7). The most frequently occurring allele was HLA-DRB1*0404 (0.161 vs 0.045). Moreover, we observed a significantly increased proportion of HLA-DRB1 SE+ cases with RF titers above the median (p = 0.005). Conversely, the proportion of individuals carrying at least one HLA-DRB1 allele encoding the D70+ was significantly decreased (p(c) = 0.004, OR = 0.4, 95% CI 0.2-0.7) among RA patients compared with controls. In conclusion, the SE is associated with RA in Mexican Mestizos as well as with the highest titers of RF.

HLA class II genotypes in Mexican Mestizo patients with myasthenia gravis.

Myasthenia gravis is an autoimmune, heterogeneous disorder, characterized by the presence of antibodies against acetylcholine receptors at the neuromuscular junction. There is a strong evidence that an individual's genetic composition is an important predisposing factor for the development of the disease. To correlate HLA class II genotypes with thymic pathology in Mexican Mestizo patients who had been subjected to thymectomy. HLA class II genes were analyzed in 60 patients and in 99 healthy ethnically matched controls. Thymic hyperplasia, atrophy, thymoma, and normal histology were encountered in 56, 33, 8 and 2% of patients, respectively. HLA-DR11 was significantly increased in patients with thymoma compared with healthy controls (pC = 0.001, OR = 13.35, 95% CI 3.5-51.3), compared with the subgroup of hyperplasia patients (pC = 0.005, OR = 15.5, 95% CI 2.78-95.58) and with the atrophy subgroup (pC = 0.04, OR = 10.5, 95% CI 1.75-70.95). This study provides the evidence of an association between HLA class II alleles with clinical and genetic heterogeneity in myasthenia gravis, particularly in those with thymoma (HLA-DR11).