Impact of tuberculosis treatment length and adherence under different transmission intensities.

Tuberculosis (TB) is a leading cause of human mortality due to infectious disease. Treatment default is a relevant factor which reduces therapeutic success and increases the risk of resistant TB. In this work we analyze the relation between treatment default and treatment length along with its consequence on the disease spreading. We use a stylized model structure to explore, systematically, the effects of varying treatment duration and compliance. We find that shortening treatment alone may not reduce TB prevalence, especially in regions where transmission intensity is high, indicating the necessity of complementing this action with increased compliance. A family of default functions relating the proportion of defaulters to the treatment length is considered and adjusted to a particular dataset. We find that the epidemiological benefits of shorter treatment regimens are tightly associated with increases in treatment compliance and depend on the epidemiological background.

Assessment of Hippocampal-Related Behavioral Changes in Adolescent Rats of both Sexes Following Voluntary Intermittent Ethanol Intake and Noise Exposure: A Putative Underlying Mechanism and Implementation of a Non-pharmacological Preventive Strategy.

Ethanol (EtOH) intake and noise exposure are particularly concerning among human adolescents because the potential to harm brain. Unfortunately, putative underlying mechanisms remain to be elucidated. Moreover, implementing non-pharmacological strategies, such as enriched environments (EE), would be pertinent in the field of neuroprotection. This study aims to explore possible underlying triggering mechanism of hippocampus-dependent behaviors in adolescent animals of both sexes following ethanol intake, noise exposure, or a combination of both, as well as the impact of EE. Adolescent Wistar rats of both sexes were subjected to an intermittent voluntary EtOH intake paradigm for one week. A subgroup of animals was exposed to white noise for two hours after the last session of EtOH intake. Some animals of both groups were housed in EE cages. Hippocampal-dependent behavioral assessment and hippocampal oxidative state evaluation were performed. Results show that different hippocampal-dependent behavioral alterations might be induced in animals of both sexes after EtOH intake and sequential noise exposure, that in some cases are sex-specific. Moreover, hippocampal oxidative imbalance seems to be one of the potential underlying mechanisms. Additionally, most behavioral and oxidative alterations were prevented by EE. These findings suggest that two frequently found environmental agents may impact behavior and oxidative pathways in both sexes in an animal model. In addition, EE resulted a partially effective neuroprotective strategy. Therefore, it could be suggested that the implementation of a non-pharmacological approach might also potentially provide neuroprotective advantages against other challenges. Finally, considering its potential for translational human benefit might be worth.

Prediction of intestinal absorption and metabolism of pharmacologically active flavones and flavanones.

Three glycosilated flavonoids (diosmin, hesperidin and naringin) and respective aglycones were characterized in terms of their apparent ionisation constants and bidirectional permeability using the cellular model Caco-2 as well as the artificial membrane model PAMPA. Ionisation curves were established by capillary electrophoresis. It was confirmed that significant amounts of the aglycones are ionised at physiological pH whereas the glycosides are in the neutral form. Permeation was not detected for the glycosides in either the apical-to-basolateral or basolateral-to-apical directions confirming the need for metabolism before absorption through the intestinal membrane. The aglycones permeated in both directions with apparent permeabilities (P(app)) in the range of 1-8x10(-5) cm/s. The results from both in vitro methods correlated providing some evidence of passive transport; however, the hypothesis of active transport cannot be excluded particularly in the case of diosmetin. Metabolism of the aglycones was detected with the cell model, more extensively when loading in the apical side. Some of the metabolites were identified as glucuronide conjugates by enzymatic hydrolysis.

Epoetin alfa resistance in hemodialysis patients with chronic kidney disease: a longitudinal study.

Anemia is an inevitable complication of hemodialysis, and the primary cause is erythropoietin deficiency. After diagnosis, treatment begins with an erythropoiesis-stimulating agent (ESA). However, some patients remain anemic even after receiving this medication. This study aimed to investigate the factors associated with resistance to recombinant human erythropoietin therapy with epoetin alfa (αEPO). We performed a prospective, longitudinal study of hemodialysis patients receiving treatment with αEPO at our reference hospital from July 2015 to June 2016. Clinical data was collected, and the response to αEPO treatment was evaluated using the erythropoietin resistance index (ERI). The ERI was defined as the weekly weight-adjusted αEPO dose (U/kg per week)/hemoglobin level (g/dL). A longitudinal linear regression model was fitted with random effects to verify the relationships between clinical and laboratory data and ERI. We enrolled 99 patients (average age, 45.7 (±17.6) years; male, 51.5%; 86.8% with hypertension). The ERI showed a significant positive association with serum ferritin and C-reactive protein, percentage interdialytic weight gain, and continuous usage of angiotensin receptor blocker (ARB) hypertension medication. The ERI was negatively associated with serum iron and albumin, age, urea reduction ratio, and body mass index. Our findings indicate that resistance to αEPO was related to a low serum iron reserve, an inflammatory state, poor nutritional status, and continuous usage of ARBs.

Sex ratio and dynamic behavior in populations of the exotic blowfly Chrysomya albiceps (Diptera, Calliphoridae).

Sex ratio is an essential component of life history to be considered in population growth. Chrysomya albiceps is a blowfly species with a naturally biased sex ratio. In this study, we evaluated the impact of changes in sex ratio on the dynamic behavior of C. albiceps using a density-dependent mathematical model that incorporated demographic parameters such as survival and fecundity. These parameters were obtained by exponential regression, with survival and fecundity being estimated experimentally as a function of larval density. Bifurcation diagram of the results indicated the evolution of stable equilibrium points as a function of sex ratio. A continually increasing sex ratio yielded a hierarchy of bifurcating stable equilibrium points that evolved into a chaotic regime. The demographic parameters obtained by exponential regression were also changed to maximum and minimum values in order to analyze their influence on dynamic behavior with sex ratio being considered as an independent variable. Bifurcations with periodicity windows between chaos regimes were also found.

Stem cell-like transcriptional reprogramming mediates metastatic resistance to mTOR inhibition.

Inhibitors of the mechanistic target of rapamycin (mTOR) are currently used to treat advanced metastatic breast cancer. However, whether an aggressive phenotype is sustained through adaptation or resistance to mTOR inhibition remains unknown. Here, complementary studies in human tumors, cancer models and cell lines reveal transcriptional reprogramming that supports metastasis in response to mTOR inhibition. This cancer feature is driven by EVI1 and SOX9. EVI1 functionally cooperates with and positively regulates SOX9, and promotes the transcriptional upregulation of key mTOR pathway components (REHB and RAPTOR) and of lung metastasis mediators (FSCN1 and SPARC). The expression of EVI1 and SOX9 is associated with stem cell-like and metastasis signatures, and their depletion impairs the metastatic potential of breast cancer cells. These results establish the mechanistic link between resistance to mTOR inhibition and cancer metastatic potential, thus enhancing our understanding of mTOR targeting failure.

What do we need to learn to optimize the SLIT alternative?

Allergen-specific immunotherapy (IT) is the only treatment of allergy in adults and children capable of modifying the immune response at early steps. As a consequence, IT improves symptoms, prevents the onset of new sensitizations, reduces the risk of developing asthma and its clinical efficacy lasts many years. The main rationale for administering sublingually IT (SLIT) is to reduce the occurrence of side effects, still yet preserving the immunological effects. SLIT with the most common allergens have been used in many studies with significant clinical effectiveness in both asthma and rhinitis. The pharmacokinetic of allergens administered through non injection routes is complex. Peptide absorption across oral mucosa occurs mainly by passive diffusion but delivery of proteins has some limitations. Moreover, the molecular mechanisms responsible for the efficacy of SLIT are poorly defined. In this review we focus on the anatomy/histology of the oral cavity as well as on the associated immunological structures to envisage what may happen when an allergen is kept in the mouth. Moreover, the induction of immune responses in this particular immunological environment is also discussed.

Organotin speciation in environmental matrices by automated on-line hydride generation-programmed temperature vaporization-capillary gas chromatography-mass spectrometry detection.

In the present contribution, a new automated on-line hydride generation methodology was developed for dibutyltin and tributyltin speciation at the trace level, using a programmable temperature-vaporizing inlet followed by capillary gas chromatography coupled to mass spectrometry in the selected ion-monitoring mode acquisition (PTV-GC/MS(SIM)). The methodology involves a sequence defined by two running methods, the first one configured for hydride generation with sodium tetrahydroborate as derivatising agent and the second configured for speciation purposes, using a conventional autosampler and data acquisition controlled by the instrument's software. From the method-development experiments, it had been established that injector configuration has a great effect on the speciation of the actual methodology, particularly, the initial inlet temperature (-20 degrees C; He: 150 ml/min), injection volume (2 microl) and solvent characteristics using the solvent venting mode. Under optimized conditions, a remarkable instrumental performance including very good precision (RSD < 4%), excellent linear dynamic range (up to 50 microg/ml) and limits of detection of 0.12 microg/ml and 9 ng/ml, were obtained for dibutyltin and tributyltin, respectively. The feasibility of the present methodology was validated through assays upon in-house spiked water (2 ng/ml) and a certified reference sediment matrix (Community Bureau of Reference, CRM 462, Nr. 330 dibutyltin: 68+/-12 ng/g; tributyltin: 54+/-15 ng/g on dry mass basis), using liquid-liquid extraction (LLE) and solid-phase extraction (SPE) sample enrichment and multiple injections (2 x 5 microl) for sensitivity enhancement. The methodology evidenced high reproducibility, is easy to work-up, sensitive and showed to be a suitable alternative to replace the currently dedicated analytical systems for organotin speciation in environmental matrices at the trace level.

In vitro induction of primary, antigen-specific CTL from human peripheral blood mononuclear cells stimulated with synthetic peptides.

A protocol for in vitro induction of primary, antigen-specific CTL from human peripheral blood mononuclear cells (PBMCs) was developed. Antigen presenting cells (APCs) consisted of Staphylococcus aureus Cowan-I (SAC-I) activated PBMCs treated with a citrate-phosphate buffer at pH 3 to release endogenous peptides bound to surface MHC. This treatment resulted in transient expression of empty class I molecules which could be subsequently stabilized with peptide and beta 2-microglobulin (beta 2m). SAC-I activated PBMCs from HLA-A2.1 normal donors loaded with HBV core 18-27 peptide following acid treatment were used to stimulate PBMCs depleted of CD4+ T cells, in the presence of recombinant interleukin-7 (rIL-7). After 12 days, cells were restimulated with autologous, peptide-pulsed, adherent cells and tested for CTL activity 7 days later. In 23 independent experiments from 13 different HLA-A2.1 donors, this protocol resulted in induction of primary CTL more than 90% of the time. As indicated by both the frequency and magnitude of the response against peptide-sensitized target cells, SAC-I activated PBMCs treated with acid were the most efficient stimulator APC. Thirteen per cent of the cultures generated were capable of lysing target cells transfected with the HBV core antigen and, in general, these CTL cultures exhibited high avidity for the HBV core peptide. This protocol is generally applicable to different antigens and class I alleles, and thus, may be utilized to screen large numbers of peptides to identify human CTL epitopes.

On the Cornea of Healthy Merino Sheep: A Detailed Ex Vivo Confocal, Histological and Ultrastructural Study.

Our study performed qualitative and quantitative studies on the corneal ultrastructure of healthy female Merino sheep of ages 4 months and 6 years old from the Argentinean Pampa. The corneas were evaluated using ex vivo laser-scanning confocal microscopy, light microscopy and transmission electron microscopy. Those studies allowed us to obtain detailed images of the corneal layers as well as quantitative data of the cellular and sub-basal nerve densities in the cornea from sheep of different ages. The density of the corneal cells was significantly different in the anterior versus the posterior epithelium and stroma. Moreover, the density of the epithelial, stromal cells and endothelial cells, as well as the sub-basal nerve density were significantly lower in adult than in young animals. Our work provided a wide-ranging description of the corneal ultrastructure of healthy female Merino sheep, which adds to the current knowledge about the ophthalmological aspects of this species and undoubtedly benefits veterinarians.

Effects produced by different types of laser in cornea of Guinea pigs: Identification of a laser capable of producing superficial lesions without leaving scars.

PURPOSE: Climatic droplets keratopathy (CDK) is closely associated with superficial corneal erosions and lack of protective mechanisms against the harmful effects of ultraviolet radiation (UVR) during a prolonged period of time. One of the difficulties in studying the pathogenic mechanisms involved in this human disease is the lack of an experimental animal model. In this paper, a study is conducted on the effects of 4 types of lasers at various powers and time conditions on the normal guinea pig corneas in order to select only one laser condition that reversibly injures the epithelium and superficial stroma, without leaving scarring. METHODS: Damage was induced in the cornea of Guinea pigs using different powers and exposure times of 4 types of laser: argon, CO2, diode and Nd-Yag, and any injuries were evaluated by biomicroscopy (BM) and optical microscopy. Corneas from other normal animals were exposed to argon laser (350 mW, 0.3s, 50 µm of diameter), and the induced alterations were studied at different times using BM, optical coherence tomography (OCT) and transmission electron microscopy (TEM). RESULTS: Only argon laser at 350 mW, 0.3s, 50 µm of diameter produced epithelium and superficial stroma lesions. Some leukomas were observed by BM, and they disappeared by day 15. Corneal thickness measured by OCT decreased in the eyes treated with argon laser during the first week. Using TEM, different ultra structural alterations in corneal epithelium and stroma were observed during the early days, which disappeared by day 15. CONCLUSIONS: It was possible to develop reproducible corneal epithelium and anterior stroma injuries using Argon laser at 350 mW, 0.3s, 50 µm of diameter. In vivo and in vitro studies showed that injured corneas with these laser conditions did not leave irreversible microscopic or ultra structural alterations. This protocol of corneal erosion combined with exposure to UVR and partial deficiency of ascorbate in the diets of the animals for an extended period of time has been used in order to try to develop an experimental model of CDK.

Valores típicos de dosis para tomografía computada de cerebro en pacientes adultos / Typical dose values for brain computed tomography in adult patients

Resumen: Debido al aumento en el uso de la Tomografía Computada (TC), y en consecuencia, la probabilidad de generar un incremento progresivo de la dosis recibida por los paciente y su relación en el potencial riesgo de los efectos de las radiaciones ionizantes, es importante implementar el uso de niveles de referencia diagnóstico (DRLs) en TC, como herramienta fundamental dentro de un programa de control de calidad que permita la evaluación y optimización de las dosis entregadas a los pacientes según la tarea clínica deseada. Con el objetivo de establecer valores típicos de dosis en TC de cerebro en Clínica Bupa Reñaca, se estudió una muestra de 73 informes dosimétricos generados en un equipo TC Toshiba Aquilion 64, en términos de indicadores de dosis para TC: Índice de dosis en TC por volumen (CTDIvol) y Producto dosis longitud (DLP). Con los datos obtenidos, se estimó el valor del percentil 50 (p50) para cada indicador de dosis, y se determinaron los valores típicos de dosis en cada grupo estudiado según sexo, edad e indicación clínica. Se logró definir y establecer una metodología que permitió la obtención de los valores típicos de dosis para TC de cerebro, optimizando las dosis sin producir una disminución en la calidad de la imagen necesaria para nuestro propósito clínico.

Abstract: Due to the increase in the use of Computed Tomography (CT), and consequently, the probability of generating a progressive increase in the dose received by the patient and its relationship in the potential risk of the effects of ionizing radiation, it is important to implement the use of diagnostic reference levels (DRLs) in CT, as a fundamental tool within a quality control program that allows the evaluation and optimization of the doses delivered to patients according to the desired clinical task. In order to establish typical dose values in brain CT at Bupa Reñaca Clinic, a sample of 73 dosimetric reports generated on a Toshiba Aquilion 64 CT unit was studied, in terms of dose indicators for CT: Dose rate on CT by volume (CTDIvol) and Dose Length Product (DLP). With the data obtained, the value of the 50th percentile (p50) for each dose indicator was estimated, and typical dose values were determined in each group studied according to sex, age and clinical indication. It was possible to define and establish a methodology that allowed obtaining typical dose values for brain CT, optimizing the doses without producing a decrease in the image quality necessary for our clinical purpose.

Evaluation of the immunosuppression in an experimental model of autoimmunity: suppressor activity of spleen cells from cyclophosphamide-treated rats.

Rats immunized with chemically modified rat male accessory glands (MRAG) and injected 3 days later with cyclophosphamide (CY) were unable to develop humoral and cellular immune response to the autoantigen of MRAG. The present report demonstrates that the spleen mononuclear (SpM) cells transference from rats injected with CY 3 days after the antigen to normal male or female syngeneic animals before immunization with MRAG did not suppress the immune response to this antigen, whereas the transference of SpM cells from suppressed animals to animals previously immunized, depressed the delayed type hypersensitivity (DTH) response against MRAG (suppression of the expression) only in male rats. Similar results were obtained by transference of purified T cells. SpM cells did not suppress an established humoral immune response induced in male or female rats. The results suggest that non-adherent cells present in the spleen of male suppressed rats might be one of the responsible mechanisms for suppression of the efferent phase of the cellular autoimmune response to MRAG.

Apparent loss of 2H4+ T cells in peripheral blood lymphocytes of normal donors: a 2H4-specific artefact unique to T cells.

The antigenic cluster designated CD45R is recognized by a family of monoclonal antibodies. However, one of those most frequently used because of its commercial availability is 2H4. In a series of experiments we show that 2H4 immunofluorescence is almost completely lost from CD4+ or CD8+ T cells of human origin if they are fixed after staining with 2H4. Loss of fluorescence intensity of the total population of 2H4+ lymphocytes and nearly complete quantitative loss of CD45R+ T cells is observed after fixation. This apparent loss of CD45R as detected by 2H4 is not seen if other CD45R-specific monoclonal antibodies are used. Fixation of B cells previously stained with 2H4 gives rise to slightly diminished fluorescence intensity, but no reduction in number of 2H4+ B cells. This appears to be due to a lower antigen density on T cells as compared to B cells. The effect is unique to 2H4, as other monoclonal antibodies recognizing CD45R, while exhibiting a decrease in fluorescence intensity after fixation, still unequivocally detect CD45R+ CD4+ or CD8+ T cells. The effect is restricted to human T cells, as primate lymphocytes, fixed after staining with 2H4, show no loss in fluorescence intensity. We conclude that if it is necessary to fix human T cells after fluorescent staining, the use of CD45R-specific antibodies other than 2H4 is mandatory.

Prevalence of antibody to human T cell lymphotropic virus types 1/2 among aboriginal groups inhabiting northern Argentina and the Amazon region of Peru.

We carried out a seroepidemiologic survey to define the prevalence of human T cell lymphotropic virus types 1/2 (HTLV-1/2) infections among aboriginal populations from isolated regions of northern Argentina and the Amazon region of Peru. Antibodies against HTLV were measured with agglutination tests and confirmed with by an immunofluorescence assay (IFA) and Western blotting. Five (6.94%) of 72 samples from the Tobas Indians in Argentina were positive by the IFA; two samples were typed as HTLV-1 (2.78%), two as HTLV-2 (2.78%), and one (1.39%) could not be typed because it had similar antibody titers against both viruses. No positive samples were found among 84 Andinos Puneños and 47 Matacos Wichis Indians. Seroprevalences of 2.50% (1 of 40) and 1.43% (1 of 70) for HTLV-1 were observed among Wayku and San Francisco communities in the Amazon region of Peru, and seroprevalences of 4.54% (1 of 22) and 2.38% (1 of 42) for HTLV-2 were observed among Boca Colorada and Galilea communities. No serologic evidence of human immunodeficiency virus (HIV) infection was found among the Indians tested. These results indicated the presence of HTLV-1 and HTLV-2 in the indigenous populations of Argentina and Peru. Moreover, the lack of HIV infection indicates that the virus has probably not yet been introduced into these populations.

Seroprevalence of human herpesvirus 6 in Andino Puneños (Argentina).

We carried out a seroepidemiological survey to define the prevalence of human herpesvirus 6 (HHV6) infection in an aboriginal population (Andino Puneños) from a remote region in north-west Argentina. Antibodies against HHV6 (total IgG and the 4 subclasses of IgG) were studied in 84 serum samples (collected in 1995 and stored at -70 degrees C), using core blood mononuclear cells infected with HHV6 in an immunofluorescence assay. Of the 84 samples, 70 (83%; 95% confidence interval, 75-91%) exhibited IgG antibodies against HHV6. No significant differences in the frequency of humoral immunity were found among the 4 age-groups studied (mean ages 13, 31, 47 and 70 years) namely, 75%, 89.7%, 79.2% and 100%, respectively. HHV6-specific IgG1 was found in all the positive serum samples tested but none of them contained specific IgG2, IgG3 and IgG4. These results confirmed a high rate of infection with HHV6 within this aboriginal group in Argentina and an IgG1 anti-HHV6 activity compatible with a maintenance of immunity.

Antibody isotypes profiles against Trypanosoma cruzi acidic antigens in two Amerindian populations from a Chagas' disease endemic area.

The isotype distribution of the antibody response against one Trypanosoma cruzi antigenic fraction, FIV, and the putative association to heart disease were analyzed in patients of two apparently genetically distinct Amerindian populations, Mataco (M) and Toba (T), infected with this parasite. The isotypes profiles were analyzed by ELISA, and the antigen specificity of IgG immune response was determined by the immunoblot method. The percentages of infected individuals with abnormal electrocardiograms (GII) were 50% for population M and 10% for population T. Many individuals from both populations had measureable IgG2, IgG3 and IgG4 antibodies to FIV, but the level and frequency (%) of positive sera in population T was considerably higher than in population M (70% vs 15% for IgG2; 75% vs 40% for IgG3; 85% vs 20% for IgG4). The level and frequency of IgG1 reactivity against FIV were similar in the two populations. When the sera were titrated, the most remarkable difference in isotype levels between populations T and M was seen for IgG2 and IgG4, the T population showing the highest titer. No association between clinical state and a particular isotype profile was found by ELISA in any population. When the antigen specificity of antibody response was determined by immunoblot, the antigen patterns recognized by sera from the two clinical groups showed some differences only in population M. All sera assayed from GII of population M fixed more IgG than those with normal electrocardiograms (GI). Two bands of 36 and 43 kD were revealed only in GII of this population. Similar antigenic patterns between the two clinical groups from population T were observed, and they were comparable with those obtained with GI from population M.

Injury prevention: is there a role for out-of-hospital emergency medical services?

OBJECTIVE: To review the literature for options for integrating injury prevention into the role of out-of-hospital emergency medical services (EMS). DATA SOURCES: Computerized searches of the English-language literature from 1966 through 1994 were conducted using the MEDLINE and National Association of EMS Physicians (NAEMSP) databases. These were supplemented by hand searches of pertinent journals not indexed on MEDLINE or by NAEMSP and the reference lists of retrieved articles. Key words searched included emergency medical services, accident, injury, prevention, and safety. ARTICLE SELECTION: The review included all articles that described the experience of EMS organizations or individuals providing primary injury prevention (PIP) services or that proposed EMS PIP activities. SYNTHESIS: PIP EMS experiences and PIP activities proposed for EMS included: preventing injuries in EMS providers, serving as role models, identifying persons at risk for injury, providing prevention counseling, collecting injury data, surveying residences and institutions for injury risks and hazards, conducting educational programs and media campaigns, and advocating legislative changes that promote injury prevention. Few studies have evaluated the effectiveness of EMS PIP activities. CONCLUSION: As changes in the market compel health care systems to focus more on prevention, EMS organizations and individual providers may be assuming new injury prevention roles. Some EMS systems in many parts of the country have incorporated PIP into their work. It is necessary, however, to determine which PIP roles are effective and how they will be supported.

Clinical signs and symptoms are not enough for the correct diagnosis of allergic conjunctivitis.

BACKGROUND: Allergic Conjunctivitis (AC) has a high incidence in the general population and sometimes it is difficult to make a correct diagnosis, distinguish among the different subtypes of AC, and therefore, to indicate the suitable therapy. OBJECTIVE: To determine the best way to carry out an appropriate diagnosis of AC. METHODS: Thirty-one patients with clinical manifestations of AC and eleven controls were studied by measuring allergic and immunologic parameters. Only those patients confirmed as having AC were treated with ketotifen fumarate and further evaluated. RESULTS: According to allergic and immunological parameters, patients were divided into two groups. Group I patients had positive prick test toward at least one allergen, 60% exhibited high levels of tear-IgE, and only 36% conjunctival eosinophils. By contrast, patients from Group II had negative prick tests and laboratory findings similar to the control group. In group I there was a good correlation between levels of tear-IgE and eosinophils (r = 0.55; p = 0.009); key symptoms and signs and prick test (r = 0.52; p = 0.015), and prick test and eosinophils (r = 0.50 p = 0.022). The cardinal signs and symptoms scores dropped significantly in Group I as a consequence of the treatment (p < 0.0001). CONCLUSION: In order to have a reliable AC diagnosis, allergen-skin prick test, IgE in tears, and conjunctival eosinophils must be studied. Serum IgE is less important.

Molecular diagnosis of Huntington disease in Brazilian patients.

Huntington disease (HD) is a progressive neurodegenerative disorder with autosomal dominant inheritance, characterized by choreiform movements and cognitive impairment. Onset of symptoms is around 40 years of age and progression to death occurs in approximately 10 to 15 years from the time of disease onset. HD is associated with an unstable CAG repeat expansion at the 5' and of the IT15 gene. We have genotyped the CAG repeat in the IT15 gene in 44 Brazilian individuals (42 patients and 2 unaffected family members) belonging to 34 unrelated families thought to segregate HD. We found one expanded CAG allele in 32 individuals (76%) belonging to 25 unrelated families. In these HD patients, expanded alleles varied from 43 to 73 CAG units and normal alleles varied from 18 to 26 CAGs. A significant negative correlation between age at onset of symptoms and size of the expanded CAG allele was found (r=0.6; p=0.0001); however, the size of the expanded CAG repeat could explain only about 40% of the variability in age at onset (r2=0.4). In addition, we genotyped 25 unrelated control individuals (total of 50 alleles) and found normal CAG repeats varying from 16 to 33 units. The percentage of heterozigocity of the normal allele in the control population was 88%. In conclusion, our results showed that not all patients with the "HD" phenotype carried the expansion at the IT15 gene. Furthermore, molecular diagnosis was possible in all individuals, since no alleles of intermediate size were found. Therefore, molecular confirmation of the clinical diagnosis in HD should be sought in all suspected patients, making it possible for adequate genetic counseling.