Development of a pediatric differentiated thyroid carcinoma registry within the EuRRECa project: rationale and protocol.

Background: Although differentiated thyroid carcinoma (DTC) is the most frequent endocrine pediatric cancer, it is rare in childhood and adolescence. While tumor persistence and recurrence are not uncommon, mortality remains extremely low. Complications of treatment are however reported in up to 48% of the survivors. Due to the rarity of the disease, current treatment guidelines are predominantly based on the results of small observational retrospective studies and extrapolations from results in adult patients. In order to develop more personalized treatment and follow-up strategies (aiming to reduce complication rates), there is an unmet need for uniform international prospective data collection and clinical trials. Methods and analysis: The European pediatric thyroid carcinoma registry aims to collect clinical data for all patients ≤18 years of age with a confirmed diagnosis of DTC who have been diagnosed, assessed, or treated at a participating site. This registry will be a component of the wider European Registries for Rare Endocrine Conditions project which has close links to Endo-ERN, the European Reference Network for Rare Endocrine Conditions. A multidisciplinary expert working group was formed to develop a minimal dataset comprising information regarding demographic data, diagnosis, treatment, and outcome. We constructed an umbrella-type registry, with a detailed basic dataset. In the future, this may provide the opportunity for research teams to integrate clinical research questions. Ethics and dissemination: Written informed consent will be obtained from all participants and/or their parents/guardians. Summaries and descriptive analyses of the registry will be disseminated via conference presentations and peer-reviewed publications.

First Look Into the Ambrosia Beetle-Fungus Symbiosis Present in Commercial Avocado Orchards in Michoacán, Mexico.

Most beetle-fungus symbioses do not represent a threat to agricultural and natural ecosystems; however, a few beetles are able to inoculate healthy hosts with disease-causing fungal symbionts. Here, we report the putative nutritional symbionts associated with five native species of ambrosia beetles colonizing commercial avocado trees in four locations in Michoacán. Knowing which beetles are present in the commercial orchards and the surrounding areas, as well as their fungal associates, is imperative for developing a realistic risk assessment and an effective monitoring system that allows for timely management actions. Phylogenetic analysis revealed five potentially new, previously undescribed species of Raffaelea, and three known species (R. arxi, R. brunnea, R. fusca). The genus Raffaelea was recovered from all the beetle species and across the different locations. Raffaelea lauricola (RL), which causes a deadly vascular fungal disease known as laurel wilt (LW) in Lauraceae species, including avocado, was not recovered. This study points to the imminent danger of native ambrosia beetles spreading RL if the pathogen is introduced to Mexico's avocado orchards or natural areas given that these beetles are associated with Raffaelea species and that lateral transfer of RL among ambrosia beetles in Florida suggests that the likelihood of this phenomenon increases when partners are phylogenetically close. Therefore, this study provides important information about the potential vectors of RL in Mexico and other avocado producing regions. Confirming beetle-fungal identities in these areas is especially important given the serious threat laurel wilt disease represents to the avocado industry in Mexico.

Supporting international networks through platforms for standardised data collection-the European Registries for Rare Endocrine Conditions (EuRRECa) model.

Rare endocrine pathology is manifested by either a deficiency or excess of one or more hormones. These conditions can be life-threatening and are almost universally associated with long-term morbidity. Understanding the aetiology of these conditions requires multicentre collaboration and expertise, most often across national boundaries, with the capacity for long-term follow-up. The EuRRECa (European Registries for Rare Endocrine Conditions) project ( www.eurreca.net ), funded by the EU Health Programme, aims to support the needs of the wider endocrine community by maximising the opportunity for collaboration between patients, health care professionals and researchers across Europe and beyond. At the heart of the EuRRECa collaboration is a Core Endocrine Registry that collects a core dataset for all rare endocrine conditions that are covered within Endo-ERN. The registry incorporates patient reported markers of clinical outcome and will signpost participants to high-quality, disease-specific registries. Furthermore, an electronic surveillance programme (e-REC) captures clinical activity and epidemiology for these rare conditions. EuRRECa receives guidance compliant with the highest ethical standards from Expert Working Groups that align with the Main Thematic Groups of Endo-ERN. Security, data quality and data governance are cornerstones of this platform. Clear policies that are acceptable to patients, researchers and industry for data governance coupled with widespread dissemination and knowledge exchange through closely affiliated stakeholders will ensure sustainability beyond the current lifetime of the project. This paper describes the infrastructure that has been developed, stakeholder involvement, the data fields that are captured within the registry and details on the process for using the platform.

Gene expression profile following an oral unsaturated fat load in abdominal obese subjects.

PURPOSE: Our aim was to evaluate the postprandial effect of an oral fat load test (OFLT) rich in unsaturated fatty acids on gene expression profile in peripheral blood mononuclear cells (PBMC) from subjects with abdominal obesity as an insulin resistance model and controls. METHODS: A total of 20 controls and 20 abdominal obese patients were studied. Metabolic parameters and oxidative stress markers were measured with standardized protocols. The whole gene expression at fasting state and after the OFLT (0, 4 and 8 h) was analysed using human HT-12-v4 expression beadchips, from Illumina. RESULTS: We found a significant decrease in plasma glucose, insulin and oxidative stress markers in abdominal obese patients and controls. We found beneficial metabolic postprandial gene expression in three genes: FKBP5, DDIT4 and DHRS9. Following an OFLT, the postprandial mRNA expression of FKBP5, and DDIT4 was downregulated while that of DHRS9 was overexpressed, both in nondiabetic normolipidemic subjects and in insulin-resistant subjects with abdominal obesity. CONCLUSIONS: Our results suggest that an OFLT rich in unsaturated fatty acids downregulates the expression of FKBP5, coding for the glucocorticoid receptor pathway, and that of DDIT4, involved in the oxidative stress response. These changes could favourably influence the insulin resistance and oxidative stress status in the postprandial state.

Assessment of genetic relationship in Persea spp by traditional molecular markers.

Currently, the reclassification of the genus Persea is under discussion with molecular techniques for DNA analysis representing an alternative for inter- and intra-specific differentiation. In the present study, the traditional random-amplified polymorphic DNA (RAPD) and the inter simple sequence repeat (ISSR) markers were used to determine the genomic relationship of different species and hybrids representative of the subgenera Eriodaphne and Persea in a population conserved in a germplasm bank. The data were analyzed statistically using multivariate methods. In the RAPD analysis, a total of 190 polymorphic bands were produced, with an average of 23.7 bands per primer, the percentage contribution of each primer was from 7.66 to 19.63; the polymorphic information content (PIC) ranged from 0.23 to 0.45, with an average of 0.35. In the ISSR analysis, a total of 111 polymorphic bands were considered, with an average of 18.5 bands per primer, the percentage contribution of each was from 11.83 to 19.57; the PIC ranged from 0.35 to 0.48, with an average of 0.42. The phenograms obtained in each technique showed the relationship among the accessions through the clusters formed. In general, both the techniques grouped representatives of the Persea americana races (P. americana var. drymifolia, P. americana var. guatemalensis, and P. americana var. americana). However, it was not possible to separate the species of Persea used as reference into independent clades. In addition, they tended to separate the representatives of subgenera Eriodaphne and Persea.

Morphometric analysis of nodules in human onchocerciasis collected in communities of the southern Chiapas focus, México.

Human onchocerciasis is a disease that remains as an important public health problem. The morphometric and physical characteristics of 363 Onchocerca volvulus nodules collected in the major endemic focus of onchocerciasis in Southern Chiapas (Soconusco), was assessed. In the present work we found that treatment the morphometry of 363 onchocercal nodules preserved in a 67% glycerol solution was determined by measuring the length, width and thick of each nodule with a Vernier caliper. The mass was determined with an analytical balance and the volume by measuring the water displacement, while the specific gravity was calculated by dividing mass over the volume. Statistical analysis was calculated for each parameter. The results showed that the nodules were rather longer than wider or thicker. Morphometric characteristics were 9.87 +/-3.70 (mean +/- standard deviation), 7.52 +/- 2.81, and 4.62 +/-+/- 2.06 mm for length, width and thick respectively. In regard to the shape, 62.81% of the nodules showed a lenticular shape, while 18.18% were spherical and 19.01% were ovoid. Based on the distribution of frequencies of the length, the nodules were classified in three groups: the "small" (5.77 +/- 0.73 mm; n = 104, 28.65%), the "medium" group (9.86 +/- 2.05 mm; n = 203 nodules, 55.92%), and the group of the "big" ones (16.03 +/- 1.91 mm; n = 56, 15.43%). Moreover, the physical characteristics were: for the mass 0.33 +/- 0.24 g, the volume of displaced water was 0.28 +/- 0.26 ml, and the specific gravity was 1.10 +/- 0.55 g/ml. The results indicated that most of the Mexican Onchocerca nodules have a lenticular shape with average size of 10x7x5 mm, which is useful in the knowledge of the genus biodiversity and can be taken as a parameter in clinical or epidemiological trials, where onchocerciasis remains as a public health problem.

Plasma homocysteine levels are associated with ulceration of the foot in patients with type 2 diabetes mellitus.

BACKGROUND: To examine the association of biochemical markers of risk (plasma Hcy, microalbuminuria, lipoprotein (a)(Lp(a)) and diabetic dyslipidaemia) with the prevalence of diabetic foot ulceration in type 2 diabetic patients. METHODS: Case/control study conducted in 198 type 2 diabetic patients. 89 patients have foot ulcers and 109 have no foot ulcers (control group), in order to establish ORs for diabetic foot ulceration. In all subjects plasma Hcy, Lp(a), total cholesterol, triglycerides, low-density lipoprotein cholesterol, high-density lipoprotein cholesterol, apolipoprotein B, HbA(1c) and microalbuminuria were measured using standard procedures. RESULTS: Plasma Hcy, microalbuminuria, HbA(1c) and apolipoprotein B levels were significantly higher in patients with foot ulceration compared with the control group. Plasma lipids, Lp(a), vitamin B12 and folic acid values were similar in both groups. In the logistic regression model, plasma Hcy (OR 1.09; 95% CI 1.04-1.69), microalbuminuria (OR 1,01; 95% CI 1.01-1.17) and HbA(1c) levels (OR 1.33; 95% CI 1.04-1.69) were independent risk factors for the presence of diabetic foot ulceration. CONCLUSIONS: In our study, for each micromol increase in plasma Hcy levels there was a 10% increase in the risk of diabetic foot ulceration. In addition, plasma homocysteine, HbA(1c) and microalbuminuria accounted for 50% prevalence risk of diabetic foot ulceration. Further prospective studies should be conducted to confirm the association of plasma Hcy levels with the risk of foot ulceration.

Increased plasma xanthine oxidase activity is related to nuclear factor kappa beta activation and inflammatory markers in familial combined hyperlipidemia.

BACKGROUND AND AIMS: Xanthine oxidase (XO) has been described as one of the major enzymes producing free radicals in blood. Oxidative stress and inflammatory processes have been implicated in the pathogenesis of endothelial dysfunction and the progression of atherosclerosis but until now, there is little data about the influence of vascular prooxidant systems and inflammation in familial combined hyperlipidemia (FCH). Our goal was to evaluate whether XO activity was altered in FCH and if it was related to the inflammatory process represented by NFkB, IL-6 and hsCRP, and assessing the correlation between XO activity and insulin resistance (IR). METHOD AND RESULTS: 40 Non-related subjects with FCH and 30 control subjects were included, all of them non-diabetic, normotensive and non-smokers. We measured lipid profile, glucose, insulin, uric acid, XO activity, malondialdehyde (MDA), IL-6 and hsCRP in plasma and NFkB activity in circulating mononuclear cells. Patients with FCH showed significantly higher levels of uric acid, XO activity, MDA, NFkB activity, IL-6 and hsCRP than controls. XO activity was independently related to NFkB activity with an odds ratio of 4.082; to IL-6 with an odds ratio of 4.191; and to IR with an odds ratio of 3.830. Furthermore, mean NFkB activity, IL-6 levels, and IR were highest in the highest percentile of XO activity. CONCLUSIONS: Subjects with FCH showed increased XO and NFkB activities and low grade inflammatory markers related to atherosclerosis. XO activity was correlated with higher inflammatory activity and IR. These data could explain, in part, the high cardiovascular disease risk present in these patients.

Influence of different processing procedures on the reproductive capacity of Trichinella spiralis in pork meat.

The aim of this work was to determine the influence of different processing procedures and preparations on the viability and infectivity of Trichinella spiralis ML. The muscles of limbs tongue and masseters of pigs experimentally infected were collected, splitted to pieces, and pooled. Five batches were used for the following processing procedures: (1) seasoning with "adobo", commercially acquired chilli and several other spices, (2) "wet-curing" by immersion of meat pieces in 3% brine during 24 hours, (3) cold storage without any further processing or preparation, (4) freezing to -20 degrees C and, (5) drying for 24 hours at 60 degrees C. Samples were stored at 4 degrees C for 15, 45, 60, 75, 90, 105 or 266 days after preparation. At the last-mentioned dates, ML were recovered and used to determine the reproductive capacity by infecting naïve mice. The state of meat conservation or spoilage respectively was tested by visual and tactile examination. In samples treated by freezing or drying no motile larvae were found after artificial digestion and, following inoculation of mice with larvae recovered from these groups, no ML were founded after 40 days of infection. After the artificial digestion of the cold stored samples, the ones seasoned with "adobo" and "wet-cured", a number of motile ML were consistently obtained. Initial reproductive capacity index was as of 80+/-0.5, then rates decreased to 60 - 70 between days 15 and 105 PT and dropped to 40+/-6.7 at day 266 for seasoned, 33+/-2.7 for cold-stored and 33+/-2.5 for cured samples. The influence of storage time (p=0.000005; factorial ANOVA) but not for processing procedure (p=0.724; factorial ANOVA) were statistically significant. The sensorial examination of the meat samples showed severe changes caused by spoilage in odour, texture and colour from day 45 of storage. Data reported from this trial proves that curing or flavoring do not inactivate the Trichinella Mexican strain, although cold storage for more than three months led to a partial decrease of the reproductive capacity. Freezing and drying seemed to be effective measures to eliminate the ML infectivity.

Study of the reproductive capacity of Trichinella spiralis recovered from experimentally infected mice under-dosed with albendazole or mebendazole.

The reproductive capacity of Trichinella spiralis recovered from experimentally infected mice under-dosed with albendazole (ALB) or mebendazole (MEB) was studied. Different groups of male C57/BL mice were infected with 10 +/- 0.5 muscular larvae (ML) per gram of body weight and treated with a single dose by oral (20 mg/kg) of ALB, MEB or praziquantel (PZQ) given at 5th day post infection (DPI), during the intestinal phase of infection. In other group of mice, treatment with the same drugs and dosage was for seven days, starting at day 45 PI through the stage of encapsulating larvae (parenteral phase of infection). A reduction of 72.9 to 89.9% in the parasitic load was observed in ALB or MEB treated groups but not in mice untreated or administered with PZQ. The recovered larvae were used to infect naïve mice and, after 45 DPI, a similar Reproductive Capacity Index (RCI) was observed between the different groups (P=0.323, one-way ANOVA), either from mice infected with larvae recovered from the intestinal treatments (RCI-ALB = 51.6 +/- 12.1 and RCI-MEB = 49.2 +/- 14.) or from the parenteral ones (RCI-ALB = 52.2 +/- 14.0 and RCI-MEB = 51.9 +/- 11.8). The RCI of non-treated ML was 59.5 +/- 7.7 and 57.9 +/- 15.9 for PZQ. This information is significant for practical strategies when under-dosage is dispensed.

Searching for antibodies against Trichinella spiralis in the sera of patients with fever of unknown cause.

Human cases of trichinellosis are often difficult to identify because the signs and symptoms of the disease, if the infection produces any at all, are non-specific, being similar to those observed in several other infectious diseases. In an investigation of Mexican patients with fever of unknown aetiology, attempts were made to develop a serodiagnostic test for the detection of antibodies specific for Trichinella spiralis. The excretory and secretory products of T. spiralis larvae (from the muscle tissue of experimentally infected rats) were used as the antigens in an enzyme-linked immuno-electrotransfer blot assay. The sera tested came from patients with fever of unknown cause (N=250), patients confirmed to have infectious or parasitic diseases other than trichinellosis (N=134) and 168 apparently healthy subjects. Overall, 4% of the samples from the febrile group, 1.8% of those from the healthy subjects but none of the sera from those with 'other diseases' reacted with the antigens of interest (of 45, 49 and 55 kDa). The results not only confirm that human infection with T. spiralis may be asymptomatic but also indicate that such infection may be mis-diagnosed.

[Insulin resistance quantification by fasting insulin plasma values and HOMA index in a non-diabetic population]. / Cuantificación de insulinorresistencia con los valores de insulina basal e índice HOMA en una población no diabética.

BACKGROUND: Our goal was to determine the prevalence of insulin resistance (IR) by means of fasting insulin plasma values and the HOMA index,and to analyse the relation of these parameters with other components of the metabolic syndrome. SUBJECTS AND METHOD: We studied 292 non-diabetic individuals, aged between 20 and 65 years, randomly selected from subjects attending an outpatient care center (Valencia-Spain-metropolitan area). 97 subjects who lacked clinical and biological criteria of IR were selected. In addition to calculating the HOMA index, anthropometric parameters, plasma lipids values, fasting glucose and insulin plasma levels were measured by standard methods in all subjects. RESULTS: In the subgroup of 97 subjects without clinical and biological criteria of IR, we established a diagnosis of IR when the fasting plasma insulin value was >= 16.7 mU/l or the HOMA index was >= 3.8. Cut-off points of the percentil 90th of this subgroup were taken into account. In the whole group,the prevalence of IR (HOMA >= 3.8) was 31.8%, with a higher frequency in men compared to women. CONCLUSIONS: In addition to fasting plasma insulin values and HOMA index, the best clinical and biochemical indicators of IR were fasting plasma glucose levels, BMI and triglycerides (TG) plasma values. Thus, the odds ratio for insulin resistance was 5.9 for a glycemia >= 110 mg/dl, 2.6% for BMI >= 25 kg/m(2) and 2.2 for triglycerides >= 150 mg/dl.

Detection of circulating and fecal Trichinella spiralis antigens during experimental infection using monoclonal antibodies against the new born larvae.

Different assays to detect antigens of Trichinella spiralis during current infection have been standardized, although sensitivity values have been the main limitation to use them as routine diagnostic test. We report the production and use of monoclonal antibodies (MAbs) against the new born larvae which recognize both somatic and metabolic antigens from adult and muscular larvae (ML). We used two IgG3 MAbs (4B1, 4B2) and two IgG2a (2D3, 2D4) to detect antigens during experimental infection. All MAbs detect fecal antigens starting the second to third week post infection (wpi), although it was less clear with MAbs 4B1 and 4B2, while circulating antigens were detected from third to fourth wpi. Thus, the recognition of shared antigens among T. spiralis developmental stages can be used for early diagnosis of trichinellosis.

Serological evaluation of thin-layer immunoassay-enzyme-linked immunosorbent assay for antibody detection in human trichinellosis.

A new immunoenzymatic test, named the thin-layer immunoassay-enzyme-linked immunosorbent assay (TIA-ELISA), was evaluated for antibody detection in human trichinellosis using excretion and secretion products prepared from Trichinella spiralis muscle larvae. Serum samples from people with positive muscle biopsies or symptoms compatible with the disease (n = 8 or 26, respectively), all reactive in enzyme-linked immunoelectrotransfer blot assay (EITB), as well as 67 serum samples from healthy, EITB-negative people, were tested in an ELISA and TIA-ELISA. TIA-ELISA was performed in polystyrene plastic petri dishes by adding dots of 10 microl each of antigen (7 microg/ml) followed by adding diluted serum and the conjugate. Finally, the substrate mixed with agar was added to develop the reaction. Enzymatic by-products were easily detected by the naked eye as defined dots. Sensitivity and specificity were 76 and 94% for ELISA, and both parameters were 91% for TIA-ELISA. The kappa correlation indices for both tests in relation to EITB were 0.73 and 0.80, respectively. The TIA-ELISA can be carried out with common laboratory equipment in 3 h and uses lower quantities of antigen than EITB and ELISA. Since TIA-ELISA is easy to perform, cheap, sensitive, and specific, the test could be an acceptable alternative to use in clinical laboratories lacking specialized equipment needed for ELISA and EITB and in field studies for antibody detection in human trichinellosis.

[Search for antibodies against Trichinella spiralis in free roaming rodents caught in a zoological park from Mexico City]. / Búsqueda de anticuerpos contra Trichinella spiralis en roedores de libre desplazamiento capturados en un zoológico de la Ciudad de México.

A serological survey to search for antibodies against T. spiralis was performed in free roaming rats (n = 64) and mice (n = 35) caught in a zoological park from Mexico City. Serum samples were analyzed by ELISA and immunoelectrotransfer blot assay (EIBT). None serum show positive absorbance values in ELISA nor recognized T. spiralis specific antigenic fractions in EIBT. However, two rat samples recognized three antigens of 31, 37 y 55 kDa, while one of them reacted with two additional antigens of 64 and 67 kDa. As it is known that the antigen epitope profiles varied among Trichinella species, it could be possible that in rats, there is 3% of antibody prevalence against Trichinella sp.; however, due that other organisms could induce the production of cross-reacting antibodies, such conclusion can not be supported at all. These results suggest that T. spiralis was not part of helminthological fauna in these rodents.

Insulin resistance in patients with familial combined hyperlipidemia and coronary artery disease.

The minimum model modified by the administration of insulin provides an objective and relatively easily measured index of peripheral sensitivity to insulin which was significantly lower (p <0.02) in familial combined hyperlipidemia (FCH) with ischemic heart disease (IHD) than in FCH without IHD and in control subjects (1.2 +/- 0.6, 1.9 +/- 1.0, 2.9 +/- 1.2 x 10(-4) mU/L/ min, respectively). In patients with FCH, insulin resistance explains, at least in part, their metabolic alterations (hypertension, abnormal glucose tolerance, hyperinsulinemia) and elevated IHD.

[Changes in insulin secretion in familial combined hyperlipemia]. / Alteración de la secreción de insulina en la hiperlipemia familiar combinada.

BACKGROUND: We have studied the abnormalities in glucose and insulin metabolism in a group of nondiabetic subjects with familial combined hyperlipidemia (FCH) in order to ascertain the contribution of metabolic risk factors to the elevated coronary heart disease incidence observed in FCH. PATIENTS AND METHODS: The study includes 42 non-diabetic subjects (25 male and 17 women, mean age 49.1 +/- 9.3 years), diagnosed with FCH by clinical and analytical studies of the probands and first degree relatives. Forty two control subjects of similar age, sex and body weight were also studied. In both groups plasma lipids and lipoproteins, plasma glucose and insulin basal and after oral glucose tolerance test (OGTT) were studied. RESULTS: The mean age, BMI and the separation by gender was similar in the two groups. The mean systolic and diastolic blood pressures were higher (p < 0.01) in the FCH group compared with controls (145.4/90.1 and 131.5/76.3 mmHg, respectively). The levels of lipids and apo B were also higher in the FCH group. The plasma glucose values were significantly higher at 30, 60, 90 and 120 minutes during OGTT and the plasma insulin at 0, 60, 90 and 120 minutes of OGTT in FCH respect to controls. The area under the curve of the secretion of insulin was 11652.0 +/- 2281.1 and 7205.4 +/- 2289.1 pmol/l/min in FCH and controls (p < 0.01), respectively. The percentage of subjects with basal hyperinsulinemia was 66.6% in the FCH group and 9.5% in the controls (p < 0.01); at 2 hours OGTT, 78.5% and 9.5% in FCH and controls, respectively (p <0.01). The insulin secretion was significantly related with the plasma triglycerides levels, cholesterol bourded to very low density lipoproteins and systolic and diastolic blood pressure. CONCLUSIONS: Hyperinsulinism is a frequent finding in non-diabetic subjects with FCH, both with normal and abnormal glucose tolerance and could contribute to the high incidence of cardiovascular risk in these patients.

DIG-ELISA test in onchocerciasis. Serum antibodies and probability of infection.

An immunoenzymatic test (DIG-ELISA) was serologically evaluated for the serodiagnosis of onchocerciasis. Control and infected sera from the onchocerciasis endemic area of Mexico was collected and the donors assessed for onchocerciasis according to parasitological, clinical, and epidemiological data. The sera were submitted to the DIG-ELISA test using a crude extract prepared from O. volvulus adult worms which had been preserved in nodules in 67% glycerol. The test showed a 100% sensitivity with sera from 38 microfilariae carriers and 96% specificity with sera from 133 non-infected people living outside the endemic zone. In addition, seropositivity was 52.9% with samples from non-onchocercotic people living inside the endemic area, while 82.5 and 90.5% of sera from patients with clinical symptoms and subcutaneous nodules, respectively, were positive. A high rate (30%) of cross-reactivity with serum samples from people infected with Wuchereria bancrofti or Brugia timori was obtained, which is in contrast with the low seropositivity rates (7.4%) obtained with sera from patients infected with other parasites. These results suggest that DIG-ELISA test may be a useful serological test for antibody detection in onchocerciasis, especially for epidemiological surveillance of disease, but because of the high cross-reactivity observed, its use must be limited to areas where this parasitic infection does not coexist with other human filariasis. Finally, the arrangement of sera in groups according to the relative likelihood to have onchocerciasis seems to be a useful procedure for evaluation of serological tests.

Detection of serum antibodies in human Hymenolepis infection by enzyme immunoassay.

Although hymenolepiasis is the commonest cestode infection of man, there are no data available on the human immune response to this parasite. Thus, in order to determine if infection induces antibodies against Hymenolepis nana antigens, sera from 52 infected children were initially studied on Ouchterlony plates and then by enzyme-linked immunosorbent assay (ELISA), using a crude antigenic extract prepared from scolex and neck regions of adult worms. In addition, sera from persons with cysticercosis, taeniasis and other parasitoses, and normal human sera, were studied. Only one serum from the Hymenolepis group showed precipitin antibodies against H. nana antigen, while several were positive by ELISA. The sensitivity of the ELISA was 84.62% and its specificity was 100%. Very high cross-reactivity rates were obtained with taeniasis (70.6%) and cysticercosis (75%) sera. These results show that Hymenolepis infection in man induces a low but detectable humoral immune response. Although not useful for diagnostic purposes, this may be relevant to the serodiagnosis of other tissue cestode infections of man, since antibodies detected in serological tests used for the diagnosis of cysticercosis, and probably hydatidosis, could be induced by H. nana instead of Taenia solium or Echinococcus larvae.

Specific detection of human antibodies to Onchocerca volvulus.

Specific diagnosis of antibodies to Onchocerca was achieved through (1) the construction of direct and indirect ELISA systems, and (2) restricting ELISA assays to the IgG4 class. The direct ELISA was based on the isolation of a surface derived, low molecular weight surface antigen preparation containing two main antigens (M. wt. 16.2 and 12.8 kDA) as defined by Western blot analysis. The direct ELISA system detected antibodies in children of six years old, and may therefore be applicable to detecting reinvasion in OCP areas of Onchocerca volvulus control. The indirect ELISA system was a competitive binding ELISA-based assay using a monoclonal antibody recognising two Onchocerca components (M. wts. 15.6 and 25.9) on a Western blot. The direct and indirect ELISA systems were similarly specific and sensitive when evaluated in a preliminary survey. The direct ELISA system yielded a specificity and sensitivity of: 100% and 100% respectively, using Mexican endemic and Mexican intestinal nematode infection sera as positive and negative controls respectively: 91% and 96% respectively, using Venezuelan endemic and Venezuelan Mansonella ozzardi infection sera as positive and negative controls, respectively: 87% and 93% respectively, using African endemic and Papuan (New Guinea) Wuchereria bancrofti infection sera as positive and negative controls respectively: 93% and 93% respectively, using African endemic and Indian W. bancrofti infection sera as positive and negative controls respectively. Similar specificity and sensitivity levels were obtained when the same comparisons were made using the indirect (inhibition) ELISA assay. These values may be contrasted with the currently used PBS extract of O. volvulus which yielded specificities of less than 10% in all the above comparisons.(ABSTRACT TRUNCATED AT 250 WORDS)