Potential association of Trypanosoma cruzi DTUs TcV and TcVI with the digestive form of Chagas disease.

The relationship among genetic diversity of Trypanosoma cruzi and clinical forms of Chagas disease remain elusive. In order to assess the possible association between different T. cruzi Discrete Typing Units (DTUs) and the clinical pictures of the disease, 205 chronic patients from Salta province, Argentina, were analysed. One hundred and twenty-two of these patients were clinically categorized as: cardiac 38.5% (47/122), digestive 15% (18/122), cardio-digestive 16% (20/122) and asymptomatic 30% (37/122). From each patient, blood samples were taken for both, Polymerase Chain Reaction (PCR) targeting kDNA and blood culture analyses. The presence of T. cruzi kDNA was detected in 43% (88/205) of the patients. T. cruzi DTUs were identified in 74% (65/88) of the kDNA positive patients by PCR-hybridization using specific probes. We detected the presence of DTUs TcI, TcII, TcV and TcVI. Single infections (i.e. presence of only one DTU in the sample) were detected in 38.64% of the samples (34/88), while mixed infections were 35.23% (31/88). TcV was the most prevalent DTU (60.3%- 53/88). The association analyses showed, for the first time to the best of our knowledge, that TcV and TcVI were associated with the digestive form of Chagas Disease (Fisher p = .0001).

The TcTASV proteins are novel promising antigens to detect active Trypanosoma cruzi infection in dogs.

In regions where Chagas disease is endemic, canine Trypanosoma cruzi infection is highly correlated with the risk of transmission of the parasite to humans. Herein we evaluated the novel TcTASV protein family (subfamilies A, B, C), differentially expressed in bloodstream trypomastigotes, for the detection of naturally infected dogs. A gene of each TcTASV subfamily was cloned and expressed. Indirect enzyme-linked immunosorbent assays (ELISA) were developed using recombinant antigens individually or mixed together. Our results showed that dogs with active T. cruzi infection differentially reacted against the TcTASV-C subfamily. The use of both TcTASV-C plus TcTASV-A proteins (Mix A+C-ELISA) enhanced the reactivity of sera from dogs with active infection, detecting 94% of the evaluated samples. These findings agree with our previous observations, where the infected animals exhibited a quick anti-TcTASV-C antibody response, coincident with the beginning of parasitaemia, in a murine model of the disease. Results obtained in the present work prove that the Mix A+C-ELISA is a specific, simple and cheap technique to be applied in endemic areas in screening studies. The Mix A+C-ELISA could help to differentially detect canine hosts with active infection and therefore with high impact in the risk of transmission to humans.

Evaluation of recombinant antigens of Trypanosoma cruzi to diagnose infection in naturally infected dogs from Chaco region, Argentina.

Dogs are considered the main mammal reservoir of Trypanosoma cruzi in domiciliary environments. Consequently, accurate detection of T. cruzi infection in canine populations is epidemiologically relevant. Here, we analysed the utility of the T. cruzi recombinant antigens FRA, SAPA, CP1, Ag1 and a SAPA/TSSA VI mixture, in an ELISA format. We used a positive control group of sera obtained from 38 dogs from the Chaco region in Argentina with positive homogenate-ELISA reaction, all of them also positive by xenodiagnosis and/or PCR. The negative group included 19 dogs from a nonendemic area. Sensitivity, specificity, area under the curve (AUC) of the receiver operating charactheristic (ROC) curve and Kappa index were obtained to compare the diagnostic efficiency of the tests. The SAPA/TSSA VI had the highest performance, with a sensitivity of 94.7% and an AUC ROC of 0.99 that indicates high accuracy. Among individual antigens, SAPA-ELISA yielded the highest sensitivity (86.8%) and AUC ROC (0.96), whereas FRA-ELISA was the least efficient test (sensitivity = 36.8%; AUC ROC = 0.53). Our results showed that the use of SAPA/TSSA VI in ELISAs could be a useful tool to study dogs naturally infected with T. cruzi in endemic areas.

Benznidazole treatment in chronic children infected with Trypanosoma cruzi: serological and molecular follow-up of patients and identification of Discrete Typing Units.

A total of 221 children from two rural settlements in Northeast Argentina were examined for T. cruzi infection. Blood samples were taken for serology tests and PCR assays. In addition, T. cruzi Discrete Typing Units (DTUs) were determined by hybridization with specific DNA probes of the minicircle hypervariable regions (mHVR). Serological results indicated that 26% (57/215) were reactive against T. cruzi antigens. PCR analyses were performed on seropositive samples showing presence of parasite DNA in 31 out of 53 samples (58.5%). All seropositive children underwent specific chemotherapy with Benznidazole (5mg/kg/day) for a period of two months and were monitored two and five years after treatment. Overall the treatment was well tolerated and low side effects were observed. Serological conversion was observed at two years post -treatment in one child form Pampa Ávila and at five years in two children from Tres Estacas. However, at the end of the follow-up period, T. cruzi DNA could not be detected by PCR in samples from treated children, except in two cases. In addition, the results of hybridizations with specific DNA probes showed that DTU TcV was detected in 68% (21/31), TcVI in 7% (2/31) and TcV/VI in 3% (1/31) of the samples. Altogether, results of the follow-up of treated children showed a low rate of seroconversion; however trend toward seroconversion was evident at five years post-treatment. On the other hand, detection of T. cruzi DNA by PCR significantly decreased after Benznidazole treatment. The existence of data regarding serological and molecular follow-ups from controlled studies in the Chaco Region will be important for future treatment efforts against T. cruzi infection in this region. The results obtained in the present study represent a contribution in this regard.

[Manual lifting and manual transport: risk assessment and prevalence of work-related diseases in construction companies in Basilicata]. / Movimentazione manuale dei carichi e trasporto manuale: valutazione del rischio e prevalenza di patologie correlate nelle imprese edili lucane.

BACKGROUND: The Basilicata Regional Headquarters of the Italian Institute for Insurance against Occupational Accidents and Disease (INAIL) and the Basilicata association of small building enterprises (Edilcassa di Basilicata) promoted a research project to assess the risk of manual lifting and manual transport in construction enterprises in the Basilicata Region and estimate the prevalence of related diseases. METHODS: Manual lifting risk assessment was performed by calculating the VLI of 204 working days in as many building workers. Manual transport risk assessment was carried out comparing the weights transported (on the 204 days tested) with the reference values of the "Snoock and Ciriello" tables. RESULTS: Manual Ifting risk was present on 195 of the 204 days, with an average value of VLI equal to 2.1 (min 0.4, max 8.5), with higher values in the restructuring sector (VLI average of 2.3, min 0.4, max 8.5), and no significant differences between the different tasks. Manual transport risk was present on 129 of the 204 days, with average values of 1.2 (min 0.2, max 3.3), with no significant differences between the different tasks analyzed For both risks additional factors were present that were not analyzed by the methods of assessment used (for manual lifting: 8.8% of the geometries in the critical area; for manual transport: 39% of transport on shoulders, 42.5% on a route with uneven surface and 31.9% on a sloping route), so it is likely that the actual risk is greater than that indicated by the synthetic indices of exposure. The medical questionnaire showed from the case histories that 148 out of 546 subjects were positive for the threshold forpain or discomfort in the lumbosacral spine area and 99 out of 546 subjects reported suffering from an already diagnosed herniated spinal disk. Only 18% of osteoarticular diseases was reported to the Insurance Institute, al though there was widespread awareness that the diseases in question might be related to work. Diseases of the spine were responsible for 1.9% of absenteeism, equal to 30-40% of total absenteeism ofworkers enrolled in "Edilcassa di Basilicata". CONCLUSIONS: The method used provides a solid basis for evaluating the two risks in the construction industry, where employment is subject to extreme organizational, environmental and structural (machines, tools, operators involved) variability. Employment in the construction industry involves significant exposure to the two risks, counting for 30-40% of total absenteeism in this sector.

Immuno-enzymatic evaluation of the recombinant TSSA-II protein of Trypanosoma cruzi in dogs and human sera: a tool for epidemiological studies.

The rTSSA-II (recombinant Trypomastigote Small Surface II) antigen was evaluated by ELISA to detect anti-Trypanosoma cruzi antibodies in sera from naturally infected dogs and humans. For this evaluation ELISA-rTSSA-II was standardized and groups were classified according to the results obtained through xenodiagnosis, ELISA and PCR. Sensitivity (Se), Specificity (Sp), Kappa index (KI) and area under curve (AUC) were determined. The Se was determined by using 14 sera from dogs infected with T. cruzi VI (TcVI) whereas Sp was determined by using 95 non-chagasic sera by xenodiagnosis, ELISA-Homogenate and PCR. The performance of ELISA-rTSSA-II in dog sera was high (AUC=0·93 and KI=0·91). The Se was 92·85% (1 false negative) and Sp was 100%. Two sera from dogs infected with TcI and 1 with TcIII were negative. For patients infected with T. cruzi, reactivity was 87·8% (36/41), there was only 1 indeterminate, and Sp was 100%. Fifty-four sera from non-chagasic and 68 sera from patients with cutaneous leishmaniasis did not react with rTSS-II. ELISA-rTSSA-II showed a high performance when studying sera from naturally infected dogs and it also presented 100% Sp. This assay could be an important tool to carry out sero-epidemiological surveys on the prevalence of T. cruzi circulating lineages in the region.

Tissue distribution of metals in striped dolphins (Stenella coeruleoalba) from the Apulian coasts, southern Italy.

Tissue distributions of metals (mercury, lead, cadmium, zinc, copper, iron, manganese) were determined in six specimens of striped dolphins (Stenella coeruleoalba, Meyen) stranded on the Apulian coasts (Southern Italy) between February and June 1987. Methyl mercury and selenium were also determined in the liver samples. The liver accumulated the highest concentrations of metals, except for cadmium and chromium. Metal levels were higher than those found in dolphins living in the Atlantic, but lower than those recorded in the same species from the French Mediterranean coasts. Necroscopic surveys found that all specimens were affected by haemorrhagic gastritis, but the cause was not clear. While it was not possible to related the death of dolphins to a specific cause, or to contaminants, the accumulation of metals is likely to contribute to the health of the organism and represents a risk factor for dolphins.

Biological image analysis for microscopical diagnosis: the work-station S.A.M. (Shape Analytical Morphometry).

The S.A.M. (Shape Analytical Morphometry) software system and its related work-station give a generalized and easy-to-handle tool to face a classic and intriguing problem in biomedical morphological diagnosis. What is the shape of an object in a microscopic image? How can we understand the relationship between size and shape? According to Holloway: "Measurements such as length, width, height, whether in chords or arcs only describe space, ... and further run into the abyss of allometric correction ... if additional information (shape?) to size is expected, some method of allometric correction must be used." The S.A.M. software system assumes a logic architecture able to separate and to parametrize independently shape characteristics in terms of allometry and local pertubation by analytical procedures (polynomials, parabolic fitting, Fourier analysis) in addition to the classic evaluations of size and density carried out by image analyzers for microscopical diagnosis.

Analytical morphometry of sagittal fronto-facial profiles of 2nd-6th month human foetus.

A prominent problem in the morphological study of developing biological structures is shape parametrization with the aim to evaluate transformations in a non subjective way. We carried out an analytical morphometrical study on a series of human embryos, by means of the S.A.M. (Shape Analytical Morphometry) software package. After standardization and normalization of the fronto-facial sagittal profiles, the analytical procedures applied were: 1) Fourier analysis: each profile was considered as an irregular but periodic function obtained by the sum of sinusoids of increasing order. 2) Shape Asymmetry Evaluation: couples of profiles are compared by means of "Janus" procedure; by a parabolic fitting we obtained parameters able to evaluate allometric and isometric differences between the two profiles. The preliminary results of the applied procedures indicated that rate and direction of allometric and isometric growth are not constant during the time.

[HCV and HBV infection in 3 communities in the Neapolitan area]. / Infezione da HCV ed HBV in tre comunita' dell' area napoletana.

Using the antibody to HCV and HBc (anti-HCV, anti-HBc), we studied the prevalence of Hepatitis C and B virus in three groups: intravenous drug abusers, subjects lodging in huts and elderly clergymen. Statistical analysis was carried out by chi-square and Fisher's exact tests. Our results show statistical significance for anti-HCV seroprevalence among the three groups, while anti-HBc doesn't differ. These preliminary data seem to show that the two viruses have different ways of transmission.

[Quantitative evaluation of the allometric transformation of of biological structures]. / Valutazione quantitativa delle transformazioni allometriche delle strutture biologiche.

Detection and quantification of allometry is a crucial problem in understanding morphological changes, both for systematic and morphogenetic purposes. A section of S.A.M. (Shape Analytical Morphometry) software system was used for this attempt. It consists of the following steps: a) boundary detection; b) starting point detection; c) size normalization; d) extraction of the fundamental shape by Kth order polynomials; e) finding of symmetry evaluator (S.A.E.) by means of a second degree equation. This last procedure gives an arc-chord complex that expresses a vector for allometry where intercept value was for application point, first degree coefficient was for direction and second degree coefficient was for modulus and versus. The main parameters, isometry fraction and allometry fraction may be understood referring them to morphogenetic models.

Primary spontaneous mesencephalic hemorrhage.

The case histories of two patients with clinically diverse forms of the very rare entity primary, spontaneous mesencephalic hemorrhage are presented. Each exhibited characteristic mesencephalic signs and neuroimaging profiles. However, both patients differed markedly in their predisposing factors, state of arousal, memory, temporal course and functional residua. One patient was unique in displaying a normal level of alertness throughout his clinical course and an isolated retrograde amnesia. The latter sign suggests a role for mesencephalic reticular activation in long-term retrieval and illustrates the differential effects of a subtle activation deficit on memory and arousal. These findings broaden the distinctive clinico-anatomical profile of primary, spontaneous mesencephalic hemorrhage.

[Lithium therapy in Horton's neuralgia: preliminary results]. / I sali di litio nella terapia della cefalea di Horton: risultati preliminari.

Prophylactic use of Lithium salts in patient suffering from cluster headaches has been evaluated looking at the mean number of headache attacks in one critical period and the mean weekly duration of the period itself. These two elements have been compared to those observed on other drugs treatments. Plasma Lithium monitoring has been performed weekly during the trial. Authors discuss the results reported and the hypothetic basis of them.