Lines of Research

The Inclinica Foundation works on two lines of research: broncho-pulmonary cancer and protozoa in airways.

LUNG CANCER

The work group investigating lung cancer varies out systematic cell cultivation of dried samples of broncho-pulmonary cancer. Ex-plant cultures have been grown, without failure, from all the cell types cultivated: epidermoid, adenocarcinoma, anaplasic and neuroendocrine.

The maintenance of rapid-growth cell cultures, about one week, allows for study of the cytolytic activity of the chemotherapy agents, with immediate clinical application, as well as opening other lines of research. The rapid rate of growth achieved facilitates the use of such cultures in future advances in the field of chemotherapy.

Microcytic lung carcinoma (2000x).

PROTOZOA IN AIRWAYS

Three types of study are indicated for this significant area of research:

1. Characteristics of the micro-organisms found in the respiratory pathways.

2. Repercussion of the protozoal forms in certain diseases of the respiratory apparatus.

3. The implication of certain arthropods (dust mites and cockroaches) in respiratory allergies.

1. Characteristics of the micro-organisms found in the respiratory pathways (morphology, study of the life cycle, cultivation and attempt at taxonomic classification of the protozoa forms).

The purpose, through various techniques, is to study the morphology, life cycle, cultivation and attempt at taxonomic classification of the protozoal forms observed in the secretions of the airways.

One of the key aims of this research is to establish a difference, based on a series of morphological criteria using conventional microscopy, between, on the one hand, what may be considered new protozoa forms, and, on the other, cytoplasmic remains of bronchial ciliate cells (CCT). It has to be acknowledged that the dilemma between the possible existence of ciliate parasites and the presence of cytoplasmic remains of bronchial ciliate cells (CCT) is not a new problem as, since the first mention (Papanicolaou, 1956; Pierce and Hirsch, 1958) of certain alterations in the bronchial ciliate cells, known as "Ciliocytophthoria", numerous works have been published on the subject (Papanicolaou et al, 1961; Rosenblatt et al, 1963; Kim et al, 1964; Monteforte and Maida, 1968; Papaioanou et al, 1969; Hollander and Gupta, 191974: Muller and Beyer-Boon, 1975; Huisjes, 1976; Rilke and Alasio, 1976; Jabamoni et al, 1977; Hollander and Gupta, 1979; Husami et al. 1981; Coleman, 1986; Poropatich and Ehya, 1986; Roxby et al, 1986; Sidaway et al, 1987; Kuritzkes et al, 1988; Sasaki, 1988; Ashfaq-Drewett et al, 1990; Hubel et al, 1990; Nosanchuk, 1993; Mahoney et al, 1993; Stanley et al, 1995; Cormiter at al, 1995; Ribas and Mosquera, 1998; Hadziyannis et al, 2000; Ribas et al, 2003).

• Protozoa forms:

Images in movement. The two videos shown were obtained with an immersion lens, and belong to fresh sputa samples from asthmatic patients.

( Video download time will depend on the characteristics of your connection )

- See video 1 (3.3 Mb)

- See video 2 (4.4 Mb)

Fresh sputa. The images were obtained with an immersion lens (1000x).

Bronchial asthma sputa. All the images presented (one of them under ultraviolet light) were obtained at 1000x and belong to Papanicolaou-stained cytological extrusions.

Sputa in AIDS. All the images obtained were obtained at 1000x. Six of them belonged to Papanicolaou-stained cytological smears and the remaining five to Wheatley's trichromic.

Allergic rhinitis. The images corresponding to nasal extrussions and obtaining with x1000. Two of them has been stained with the Papanicolaou method, and the remaining with the Wheatley´s trichromic.

• CCT and bronchial ciliate cells:

Fresh sputa. The images were obtained with an immersion lens (1000x).

Pap. Extrusions. The images correspond to Papanicolaou-stained sputum extrusions and were obtained at 1000x.

2. Repercussion of the protozoal forms in certain diseases of the respiratory apparatus.

Our research is designed to demonstrate that a causal-effect relationship may exist between multiflagellated protozoa and certain diseases of the respiratory apparatus such as bronchial asthma, allergic rhinosinusitis, idiopathic interstitial pneumonias, etc.

In addition of the studies realized in asthmatic patients, allergic rhinitis or AIDS, we are carrying a follow-up of immunodepressed patients who are admitted in the ICU with severe respiratory failure.

3. The implication of certain arthropods (dust mites and cockroaches) in respiratory allergies.

Since the relationship between household dust mites and respiratory allergy was first shown (Voorhorst et al, 1964), more than three thousand papers have been published on the subject.

In its study and analysis of household dust, our team has made a finding that we believe may be of the greatest importance. Structures with filiform projections have been observed, compatible with protozoa forms (see images), in the intestinal content of dust mites (the species studied is Dermatophagoides pteronyssinus ). This finding is described for the first time in the literature. Its significance is due to the great similitude existing between the protozoal forms observed in the sputa of asthmatic patients and those observed in the intestinal content of dust mites, which strengthens the relationship between mites and bronchial asthma.

Furthermore, similar structures have also been observed in the intestinal content of cockroaches (the species studied was Blatella germanica).

The images correspond to two dust mites (Dermatophagoides pteronyssinus) observed, in accordance with our handling technique, in saline solution (200x), and to different protozoa forms found in extracts of the intestinal content of the dust mites observed in fresco (1000x).

• Dust mites in sputum.

Presence of mites in the sputum is documented in the literature (Farley et al. 1989, Ryu et al. 2003). Besides its inhalation (Nadchatram 2005), it has been postulated on the presence of mites into airways and its relation with bronchial asthma (van Woerden 2004).

We have described an easy procedure, trough the use of bleach to provoke the sputum liquefaction, in order to identify dust mites in samples from expectorations (see works published).

The images corresponding to several dust mites, previously introduced in samples from expectorations, such as they are observed under the microscope in fresh sputum smears (x200) and stained with the Papanicolaou method (x200).

The images corresponding to two dust mites (lateral and dorsal view), such as they are observed under the microscope, after the liquefaction of the expectorations with bleach as they have been previously introduced.