A mouse model to study the alterations in haemostatic and inflammatory parameters induced by Lonomia achelous caterpillar haemolymph.

A mouse model was established to reproduce the haemorrhagic syndrome which occurs in humans after accidental contact with the hairs of the caterpillar Lonomia achelous (LA) and measures the haemostatic and inflammatory alterations that occur as a result of this contact. Mice were injected intradermally with different doses (0.4, 0.8 and 1.6 mg/animal) of L. achelous haemolymph (LAH). Haematological (haemoglobin, haematocrit, platelet count, differential leukocyte count), haemostatic (fibrinogen, plasminogen, factor XIII [FXIII], fibrinolytic activity) and inflammatory parameters (tumour necrosis factor alpha [TNF-α], nitric oxide [NO]) were measured at different times up to 48 h. C57BL/6 mice responded to LAH injection, in terms of these parameters, in a manner similar to that seen in humans, whereas the BALB/c mice were unresponsive. In C57BL/6 mice injected with LAH, time course measurements showed: a) a reduction in the haemoglobin, haematocrit, fibrinogen, FXIII and plasminogen levels, b) no effect on the platelet count and c) immediate leukocytosis and an increase in the fibrinolytic activity in plasma. An inflammatory response (TNF-α) was observed within 1 h post-injection, followed by a more persistent increase in serum NO. These findings suggest that C57BL/6 mice represent a useful model of the haemorrhagic syndrome observed in humans who have suffered contact with the caterpillar, permitting a deeper understanding of the role of the inflammatory response in the haematological and haemostatic manifestations of this syndrome.

Seroprevalence of Toxoplasma gondii infection in chickens in Durango State, Mexico.

Little is known concerning the seroprevalence of Toxoplasma gondii infection in chickens (Gallus domesticus) in Mexico. Antibodies to T. gondii were determined in 519 chickens in Durango, Mexico using the modified agglutination test (MAT). Two groups (A, B) of chickens were sampled. Group A chickens (n  =  51) were raised in backyards in 7 municipalities in 3 geographical regions in Durango State. Group B chickens were raised in farms in the Mexican States of Sinaloa (n  =  289) and Nayarit (n  =  179) but slaughtered in 2 abattoirs in Durango City. Overall, antibodies to T. gondii were found in 36 (6.9%) of 519 chickens, with MAT titers of 1∶25 in 22, 1∶50 in 8, 1∶100 in 2, 1∶200 in 3, and 1∶400 in 1. Seroprevalence of T. gondii increased significantly with age and was significantly higher in Group A chickens than in Group B chickens. In Group A chickens, a 25.5% seroprevalence of T. gondii infection was found. Seropositive chickens were found in all 7 municipalities sampled. In Group B chickens, the seroprevalence of T. gondii infection was 4.9%. This is the first report of T. gondii infection in chickens in Durango State, Mexico.

Seroprevalence of Toxoplasma gondii infection in domestic pigs in Durango State, Mexico.

Little is known concerning the prevalence of Toxoplasma gondii infection in pigs in Mexico. Accordingly, antibodies to T. gondii were determined in 1,074 domestic pigs in Durango, Mexico using the modified agglutination test. Two groups (A, B) of pigs were sampled: Group A pigs (n  =  555) were raised in 3 geographical regions in Durango State and Group B pigs (n  =  519) were from Sonora State but slaughtered in Durango City. Overall, antibodies to T. gondii were found in 136 (12.7%) of 1,074 pigs with titers of 1∶25 in 29, 1∶50 in 23, 1∶100 in 18, 1∶200 in 22, 1∶400 in 12, 1∶800 in 8, 1∶1,600 in 2, and 1∶3,200 or higher in 22. Of the pigs raised in Durango State, seroprevalence varied with age, management, and the geographic region; pigs raised in backyards in the mountainous region had a significantly higher seroprevalence (32.1%) than those raised in the valley (13.0%) and the semi-desert regions (14.0%). In Group A pigs from Durango, seroprevalence of T. gondii infection was significantly higher in pigs older than 8 mo (19.5%) than in younger pigs (10.9%). In the whole pig population (Groups A and B together), seroprevalence was higher in pigs raised in Durango (16.0%) than in those raised in Sonora (9.1%) and higher in mixed-breed pigs (15.7%) than in pure-bred pigs (10.3%). This is the first, in-depth study on the seroprevalence of T. gondii infection of pigs in Mexico and the first report on pigs from Durango State, Mexico. Results indicate that infected pork is likely an important source of T. gondii infection for humans in Durango State.

Progesterone receptor isoforms expression in the prepuberal and adult male rat brain.

Progesterone receptor (PR) isoforms expression was determined in several regions of the prepuberal and adult male rat brain by using reverse transcription coupled to polymerase chain reaction. Rats under a 14:10-h light-dark cycle, with lights on at 0600 h were used. We found that in the hypothalamus of prepuberal animals the expression of both PR isoforms was similar, whereas PR-A expression was higher than that of PR-B in adults. In the cerebellum PR-B expression was predominant in both prepuberal and adult rats. In both ages PR-A and PR-B exhibited a non-significant tendency to be predominant in the hippocampus and the preoptic area respectively. In the frontal cortex and the olfactory bulb PR isoforms were expressed at a similar level. These results indicate a differential expression pattern of PR isoforms in the male rat brain and suggest that the tissue-specific expression of PR-A and PR-B is important for the appropriate response of each cerebral region to progesterone.

Induction of p53 protein expression by sodium arsenite.

Arsenic is carcinogen for humans and has been shown to act as an enhancer in initiated animal models. In a previous work we found impairment of lymphocyte proliferation in arsenic-exposed individuals and in vitro we obtained dose-related inhibition of mitotic response and lymphocyte proliferation. Intrigued by these effects and based on the role of p53 on cell proliferation, we tested different concentrations of sodium arsenite for their ability to induce the expression of tumor suppressor gene p53 in different cell lines (HeLa, C-33A. Jurkat) and a lymphoblast cell line transformed with Epstein-Barr virus (LCL-EBV). We also evaluated changes in their viability after 24 h arsenic treatment; C-33A cells showed the higher sensitivity to arsenic treatment while HeLa, Jurkat and LCL-EBV cells showed similar cytotoxicity curves. Immunoblots showed an increased expression of p53 gene with 1 microM sodium arsenite in Jurkat cells and 10 microM sodium arsenite in HeLa and LCL-EBV cells. In addition, we transfected Jurkat cells and human lymphocytes with wild-type and mutated p53 genes; lymphocytes and Jurkat cells that received the mutated p53 showed increased sensitivity to arsenic cytotoxicity. Data obtained indicate that arsenic induces p53 expression and that cells with a functional p53 contend better with damage induced by this metalloid.

Genotoxic effects of metronidazole.

Metronidazole (MTZ) is an effective agent used in the treatment of parasitic infections. Its genotoxic effects have been shown in a variety of prokaryotic systems; however, negative results have been reported in human in vivo studies. Due to its wide spread use, a study was performed to evaluate the chromosomal aberration frequencies in peripheral blood lymphocyte cultures from 10 individuals, before and after metronidazole treatment. A significant increase in the percentage of cells with chromatid and isochromatid breaks was observed after metronidazole treatment (1500 mg per day for 10 days). The percentages of cells with aberrations did not correlate with the levels of MTZ found in plasma. Individual variability was observed with respect to both the induction of aberrations and the concentration of MTZ in plasma. They could represent differences at the metabolic level, since metronidazole is known to be biotransformed by a polymorphic P450 cytochrome, and its metabolites have shown mutagenic activity.

Genotoxic monitoring of workers at a hazardous waste disposal site in Mexico.

Chromosomal aberration and sister chromatid exchange (SCE) frequencies were determined in lymphocytes cultured from 12 high-risk individuals working at a landfill for hazardous waste disposal. Cell proliferation kinetics (CPK) was also determined. Compared with 7 control individuals, no effects were observed with respect to SCE nor on CPK. However, the workers exhibited significantly higher frequencies of chromatid and chromosomal deletions, the magnitude of which was related with exposure time. This study suggests that when high-risk exposure is suspected, determining biomarkers of genotoxic damage (e.g., chromosomal aberrations), is useful for risk assessments.

Localization of peripheral pulmonary nodules for thoracoscopic excision: value of CT-guided wire placement.

OBJECTIVE: One of the indications for the rapidly expanding use of thoracoscopic surgery as an alternative to thoracotomy is the excision of peripheral lung nodules. Nodules judged too small or too far from the pleural surface to be seen or palpated during thoracoscopy must be localized beforehand. The purpose of this study was to evaluate the feasibility and effectiveness of percutaneous placement of spring hookwires to localize such nodules before video-assisted thoracoscopy. SUBJECTS AND METHODS: Under CT guidance, 17 nodules in 14 patients were preoperatively localized with the Kopans breast lesion localization system. Three patients who had solitary nodules had thoracoscopic resections for diagnosis because a previous transthoracic needle or transbronchial biopsy had been unsuccessful. Four patients who had lesions less than 8 mm in diameter had thoracoscopic biopsies because transthoracic fine-needle aspiration biopsy was not likely to be diagnostic. Seven patients, who had a total of 10 nodules, had therapeutic wedge resections of either limited metastases or a second bronchogenic carcinoma. Mean nodule diameter was 10 mm (range, 3-20 mm). The mean distance from nodule to costal pleura was 9 mm (range, 0-25 mm). At the end of the procedure, wire placement was confirmed by CT scanning. After thoracoscopy, the surgeons were questioned about the stability and utility of each hookwire localization. RESULTS: In all 17 procedures, a hookwire was placed successfully. In one case, the wire dislodged before thoracoscopy (after a 6-hr preoperative delay and severe bending of the wire during induction of anesthesia). In 16 of the 17 resections, the surgeon thought that thoracoscopic identification of the lesion would not have been possible without hookwire localization. Only one localization, across a major fissure, required placement of a second wire to localize a nodule. Wire-related complications included two instances of serious pain, five cases of clinically insignificant pneumothorax, and one large pneumothorax requiring drainage before a second nodule in the same lung was localized. CT scanning showed presumed local pulmonary hemorrhage in six cases without hemoptysis or hemothorax. CONCLUSION: CT-guided hookwire localization is easily and safely performed and permits thoracoscopic resection of lung nodules, which might otherwise be impossible.