Activities 2007

During the year 2007, the recently created respiratory pathology research unit carried out the following investigative studies:

• A retrospective study of the presence of monocytes in cytological sputum smears. 1,234 smears were examined, of which 101 (16.3%) contained monocytes. In 73 of this latter group (72.2%) protozoal forms were observed

• A study on immunosupression and respiratory pathology. Three patients with AIDS and characteristic respiratory clinical condition were studied. In bronchial aspiration and sputum samples the presence of flagellated protozoa was observed. This study was presented at the XXXIII European Congress of Cytology, held in Madrid (14-17 October).

• A study based on a bibliographical review of medical literature dealing with protozoa and the respiratory apparatus. 109 publications were examined, with the assistance of Prof. Guillermo Esteban, of the Parasitology Department of the University of Valencia . This study will be the basis for a future contribution to a specialist journal.

Activities 2006

Briefly described are the activities carried out by the working group that is studying the repercussion of protozoa forms in the respiratory system:

• On the basis of works published that describe the immunoreactivity of Trichomona vaginalis with p16 (Pantanowitz et al, 2005), we carried out various tests on fresh sputum extrusions and extrusions stained with the Papanicolaou technique, both positive for protozoa forms. Positive immunostaining results were not obtained.

• Transmission Electron Microscope (TEM) studies in dust mites. Of the semi-fine cuts performed in 2005, only two sections were good for ultramicrotomy. The first ultra-fine cuts were performed in the first quarter of 2006. In the images obtained with TEM, structures compatible with protozoa forms have not been recognised. During the final quarter of 2006 a new analysis was tried, succeeding in including 20 specimens in resin for semi-fine and ultra-fine cuts.

• 22 sputum smears, fixed with aerosol, corresponding to 11 patients, were received from the work group established with the Department of Epidemiology, Statistics and Public Health of the University of Cardiff. The smears were stained using the Papanicolaou method and examined by two observers. Only 5 samples were positive for protozoa forms.

• Tests were continued with the process of liquefying fresh sputum samples, previously contaminated with dust mites, with sodium hypochlorite. In various types of expectoration (serous, mucous, purulent and hemoptoic), effectiveness in the recovery of dust mites of 80% was achieved.

• A study has been initiated into the presence of monocytes in sputum smears stained with the Papanicolaou technique. As well as characterising these cells morphologically and by immunostaining with CD68, this study will attempt to determine whether there exists a relationship between the presence of monocytes and that of protozoa forms observed in the cytological smears.

• Studies have been carried out on samples (broncho-alveolar lavages and tracheo-bronchial aspirations) from immunodepressed patients, with severe respiratory failure, admitted to the Intensive Care Unit. The samples received were examined under the microscope, fresh and Papanicolaou stained, and the presence of protozoa forms was observed.

ACTIVITIES 2005

We shall proceed to briefly describe the activities that have been carried out by the workgroup studying the repercussion of protozoa forms on the respiratory apparatus:

• In relation with the study of certain types of cancer in cytological sputum smears and the presence of protozoa forms, a total of 402 samples have so far been analysed. Of these, 172 corresponded to scaly carcinoma, 131 to microcytic carcinoma, 90 to adenocarcinoma and 9 to large-cell anaplasic carcinoma. Although we observed a slight increase in protozoa forms in sputa corresponding to adenocarcinoma (68% as against 52 % for the rest), his increase was not statistically significant.

• Various trials have been performed with the fluorescent compound Ethydium Bromide, with the aim of specifically staining protozoal forms. The results have been negative for fluorescence positivity of protozoa forms in sputa extrusions.

• The ability to detect foreign bodies, including dust mites, in sputum smears has been improved by using a secretion liquefaction process, subsequent rinsing with sodium hypochlorite at differing concentrations, and fresh samples microscope observation. We would like to stress here the simplicity of the method and its low economic cost

• A workgroup has been set up with the Department of Epidemiology, Statistics and Public Health of the University of Cardiff (UK), coordinated by Dr Hugo van Woerden. The title of the project is: "Investigation of the presence of micro-organisms in sputum". The intention of this study is to detect the existence, in a certain population group, of dust mites, DerP1 protein and protozoa forms.

• Cooperation continues, on certain research projects, with professors Dr José-Guillermo Esteban (Department of Parasitology of the Faculty of Pharmacy at the University of Valencia ) and Dr Rafael Tormo Molina (Department of Biology and Vegetal Production at the University of Extremadura ).

• The study of the presence of protozoal forms in the intestinal content of dust mites continues, attempting their visualisation by electronic transmission microscopy. So far, the selection of dust mites, their fixation in paraformaldehyde-gluteraldehyde, post-fixation with osmium, inclusion in resin and semi-fine cuts have been performed. Dr Ricardo Andrade, of the General High-Resolution Analytical Microscopy in Biomedicine Service of the University of the Basque Country is collaborating in this study.

Furthermore, the lung cancer research group carries out systematic cell culture of dried broncho-pulmonary pieces. The ex-plant cultures of all the cell types (epidermoid, adenocarcinoma, anaplasic and neuroendocrine), have grown without failures. The rapid growth achieved allows these cultures to be used for future advances in the field of chemotherapy.