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Biomedical subjects

J Guillot

Publications and source records attributed to J Guillot.

At least 19 recordsLinked to original sources

Potential anti-inflammatory effects of Melaleuca alternifolia essential oil on human peripheral blood leukocytes.

The fungicidal and bactericidal actions of the essential oil (EO) of Melaleuca alternifolia seem well established, but their anti-inflammatory and antioxidative effects remain unclear. This study investigated in vitro the possible role of whole Melaleuca alternifolia EO as a modulator of the inflammatory/non-specific immune response by exploring the chemotaxis and kinetic radical oxygen species (ROS) production of leukocytes and cytokine secretion in peripheral blood mononuclear cells (PBMCs) in humans. The influence of Melaleuca alternifolia EO on the chemotaxis under agarose of isolated neutrophils (PMNs) was evaluated. The kinetics of ROS production by stimulated total circulating leukocytes was followed over 2 h by recording the fluorescence intensity of oxidized dihydrorhodamine 123. The effects of this EO on pro-(interleukin IL-2) and anti-(IL-4 and IL10) inflammatory cytokine secretions were determined by ELISA following incubation of PBMCs with the EO for 24 h. Melaleuca alternifolia EO was inefficient on the chemotaxis of PMNs. It exerted an antioxidant effect, reducing ROS production throughout the kinetic study. Melaleuca alternifolia EO inhibited PBMC proliferation, as revealed by a reduction in IL-2 secretion by stimulated lymphocytes. This EO at 0.1% directly increased the secretion of the anti-inflammatory cytokine IL-4 compared with IL-4 secretion without EO (18.5 +/- 10.0 vs 3.3 +/- 1, p < 0.05), and also increased IL-10 secretion at 0.01% (94.9 +/- 38.7 vs 44.1 +/- 18, ns). Melaleuca alternifolia EO may not only act as an anti-inflammatory mediator through its antioxidant activity but may also efficiently protect the organism by reducing the proliferation of inflammatory cells without affecting their capacity to secrete anti-inflammatory cytokines.

Anti-Inflammatory Agents↗

Evolution of the environmental contamination by thermophilic fungi in a turkey confinement house in France.

Fungal species constitute a major part of environmental contaminants in facilities where animals are housed. The present investigation was aimed at describing the relative abundances of fungal species and their concentrations in a turkey confinement house in France. Fungal cultures from poultry feed, litter, and air were undertaken every week throughout the 16-wk period of breeding. The incubation temperature of 40 degrees C was selected to isolate thermophilic fungal species (especially Aspergillus spp. and Candida albicans) that are potentially pathogenic for birds. The 2 species Aspergillus fumigatus and Aspergillus flavus were recovered at a mean of 10.5 and 37.0 cfu/m(3) of air sampled, respectively. Individual samplings yielded concentrations of up to 150.0 cfu/m(3) for A. flavus in the first weeks of the investigation. Other fungal species were recovered at a mean of 18.9 cfu/m(3) (maximum 36.3 cfu/m(3)) in the air. The yeast C. albicans was first detected at wk 4 from litter samples and at wk 7 from poultry feed. Densities of C. albicans remained very high in litter samples (63.2 cfu/g) even after new litter was added at wk 10. To analyze the genetic polymorphism of A. fumigatus, the most pathogenic mold in birds, a total number of 198 isolates (134 from air, 34 from litter, and 30 from feed samples) were genotyped using 2 polymorphic microsatellite markers. More than half (42 out of 73, 57.5%) of the genotypes were detected only once. This finding suggests that the contamination of the breeding environment is not due to a single source and confirms the very high genetic diversity of environmental A. fumigatus isolates. As during the study period, no outbreak of fungal infections occurred; the levels of fungal contaminations reported here do not seem sufficient, at least alone, to trigger fungal infections.

Air Microbiology↗

Study of the expression of Tn antigen in different types of human breast cancer cells using VVA-B4 lectin.

VVA-B4 lectin was used to investigate the differences in Tn antigen expression in tissues of different types of human breast cancer (benign lesions, carcinoma in situ, invasive carcinoma) and in normal tissues neighboring lobular carcinoma. Locations in which Tn antigen was expressed were identified using the avidin-biotin-peroxidase labeling system. Tissues collected during cosmetic procedures and classified as normal were completely negative, except for one case. Benign proliferative changes including fibroadenoma, apocrine and cylindrical metaplasia showed a very weak positive reaction, although strongly positive cells were also observed. The reaction in non-invasive cases of atypical hyperplasia was diversified depending on site. Intralobular hyperplasia was characterized by a particularly high percentage of labeled cells. A majority (up to 80%) of ductal and lobular carcinoma in situ showed very strong or moderate staining. In invasive cancers, there were conspicuous differences between stage of cancer development and tendency towards a decrease in intensely labeled cell count in the most advanced stages. In normal tissues in the direct neighborhood of carcinoma in situ, the cytoplasm of 40% of cells was strongly labeled. However, the findings for normal tissues in the close vicinity of invasive cancer were the most surprising, since there was either no or only very weak positive reaction. It can be concluded that glycosylation modifications during carcinogenesis, as demonstrated by the presence of Tn epitope, develop very early, before any destructive changes in proliferation/apoptosis or cell differentiation become discernible.

Antigens, Tumor-Associated, Carbohydrate↗

Leptin: a proliferative factor for breast cancer? Study on human ductal carcinoma.

Mammary adipose tissue is an important source of paracrine mitogens and anti-mitogens, including insulin-like growth factor, transforming growth factors, and cytokines (especially, TNFalpha and IL-1beta). Nevertheless, it is also an important source of the adipocytokine, leptin. Recently, leptin was reported to stimulate the proliferation of various cell types (pancreatic beta cells, prostate, colorectal, lung, etc.) as a new growth factor. It was also shown to stimulate the proliferation of breast cancer cell lines. In this study, we conducted an immunohistochemical analysis of leptin expression in normal tissue and benign and malignant ductal breast cell, representing the different states of the invasion process. We determined for the first time that leptin is expressed both by ductal breast tumors and by benign lesions as atypical hyperplasia. This suggests that leptin may be taken up or synthesized by all modified ductal breast cells, and may prove a proliferative factor. Moreover, leptin is unexpressed by normal tissue in the healthy breast but is exhibited by the normal tissue in near vicinity of the malignant ductal breast lesions. We also postulated that leptin may be a prognostic or diagnostic factor for ductal breast cancer. These putative hypotheses require further study.

Adult↗

Disseminated acute concomitant aspergillosis and mucormycosis in a pony.

A 6-year-old female pony died after 2 days of prostration. Clinical signs included hyperthermia and abnormal pulmonary auscultation sounds. Necropsy revealed diffuse severe necrohaemorrhagic colitis and splenitis, multiple visceral ecchymoses, petechial haemorrhages in the brain and lungs. Microscopical examination showed acute necrohaemorrhagic colitis, encephalitis, pneumonia and splenitis associated with fibrinoid vasculitis, thrombosis and fungal hyphae within and around vessels. Immunohistologically, concomitant aspergillosis (caused by Aspergillus fumigatus) and mucormycosis (causde by Absidia corymbifera) were identified in the colonic and pulmonary lesions, whereas pure mucormycosis was observed in cerebral and splenic lesions. Dual mycotic infections are very rarely described, and the present case emphasizes the need of immunohistochemistry in order to obtain a clear-cut diagnosis of mixed fungal infections.

Acute Disease↗

Anti-inflammatory effects of Melaleuca alternifolia essential oil on human polymorphonuclear neutrophils and monocytes.

OBJECTIVE AND DESIGN: The fungicidal and bactericidal actions of the essential oil (EO) of Melaleuca alternifolia seem well established, but their anti-inflammatory and anti-oxidative effects remain unclear. In this study, we investigated in vitro the possible role of whole M. alternifolia EO as a modulator of the oxidative response, i.e. reactive oxygen species (ROS) production, of leukocytes (monocytes and polymorphonuclear neutrophils (PMNs)) in humans. METHODS: Whole blood leukocytes from healthy human volunteers (n = 7), isolated from erythrocytes by haemolytic shock, were incubated for 30 min with M. alternifolia EO (0-0.1%) to determine their ROS production by flow cytometry with or without stimulation of cells. We compared the effects of 3 different stimulating agents acting differently on transductional pathways to stimulate the ROS production: a phorbol ester (PMA), formyl-methionyl-leucyl-phenylalanine (fMLP) and opsonised zymosan (OZ). RESULTS: As attested by the Krüskall-Wallis test, M. alternifolia EO at 0.1% directly stimulated ROS production by PMNs (x 8.7 vs. 0% EO, p < 0.05) and increased the intracellular ROS produced by monocytes. Whichever the stimulating agent used (PMA, fMLP or OZ), M. alternifolia EO decreased the intracellular ROS production at the dilution of 0.1% by PMNs and monocytes, more so with PMNs. CONCLUSION: M. alternifolia EO may be both a direct active mediator of the bactericidal action of the circulating leukocytes and may be efficient in protecting the organism from an excess of ROS, through an anti-oxidant and radical scavenging activity.

Anti-Inflammatory Agents↗

Eumycetoma caused by Cladophialophora bantiana in a dog.

We report a case of eumycetoma due to Cladophialophora bantiana in a 3-year-old male Siberian Husky living in France. The dog presented a tumefaction on the thorax and deformity of the second and third subjacent ribs, which were surgically removed. Macroscopic black granules were visible on the ribs, and direct microscopic examination revealed their fungal origin. Cultures yielded pure colonies of C. bantiana. The identification of the causative agent was confirmed after amplification and sequence analysis of fungal internal transcribed spacers 1 and 2 and 5.8S ribosomal DNA regions. Surgery and antifungal treatment with oral itraconazole associated with flucytosine allowed apparent cure after a 10-month follow-up. Envenomation with pine processionary caterpillars (Thaumetopoea pityocampa) and subsequently intensive corticotherapy were considered as possible predisposing factors. This is, to the best of our knowledge, the first case in which C. bantiana is identified as the causative agent of eumycetoma.

Animals↗

Fatal systemic phaeohyphomycosis in a cat due to Cladophialophora bantiana.

We report a case of fatal systemic mycosis in a 9-year-old cat. Diagnosis of phaeohyphomycosis was made by histology. Morphological and molecular identification of the fungus isolated from the lesions yielded the species Cladophialophora bantiana. This species is well known to be a neurotropic fungus causing cerebral pyogranulomatous lesions in humans. In this case, the lesions were widespread, distributed without the involvement of central nervous system. The origin of systemic manifestation is still unknown and no evidence of immunosuppression was found. It is the first feline case of C. bantiana infection reported in Europe.

Animals↗

Pneumocystis jirovecii dihydropteroate synthase genotypes in French patients with pneumocystosis: a 1998-2001 prospective study.

Dihydropteroate synthase gene (DHPS) mutations at codons 55 and 57 have been associated with sulfa/sulfone resistance in Pneumocystis jirovecii strains from patients who previously received prophylaxis. To evaluate the prevalence of these mutations, a portion of P. jirovecii DHPS gene was analysed using PCR combined with restriction fragment length polymorphism (RFLP) analysis in 92 bronchoalveolar fluid samples collected between January 1998 and September 2001 from French patients with pulmonary pneumocystosis (PCP). Seventy-six samples contained the wild-type DHPS genotype (82.6%) and 16 contained a mutant genotype (17.4%). Twelve out of the 16 isolates with a mutant DHPS genotype corresponded to patients who had never received sulfa or sulfone prophylaxis, suggesting that DHPS mutants may be acquired de novo. There was no significant difference in favourable or adverse outcome in PCP caused by the wild or mutant DHPS genotypes (P = 0.34).

DNA, Fungal↗

Evaluation of the efficacy of oral lufenuron combined with topical enilconazole for the management of dermatophytosis in catteries.

The efficacy of oral lufenuron, a chitin synthetase inhibitor, combined with topical enilconazole, was evaluated for the management of Microsporum canis infection in 100 cats housed in two catteries in France. The cats were treated with weekly rinses with enilconazole (0.2 per cent) for four weeks and, in each cattery, one group (A) was also treated with micronised griseofulvin (25 mg/kg administered orally twice a day for five weeks) and a second group (B) was treated with 60 mg/kg lufenuron administered orally once on day 0 and again after 30 days. All the cats were examined individually for cutaneous lesions and mycological cultures were made when the treatment began and after 15, 30, 60 and 90 days. In the first cattery, the cats' clinical scores after 30 and 60 days were significantly lower in group B than in group A. In both catteries and both treatment groups, the mean number of fungal colonies decreased rapidly during the first 15 days of treatment, remained stable for the following 45 days but increased from day 60 to the end of the experiment on day 90.

Administration, Cutaneous↗

Comparison of two sampling techniques to assess quantity and distribution of Malassezia yeasts on the skin of Basset Hounds.

Cytological examination using the tape-strip technique and fungal culture using contact plates with modified Dixon's medium were compared to evaluate the carriage of Malassezia yeasts on four cutaneous sites (left pinna, umbilical region, axilla and perianal area) in adult Basset Hounds. Twenty animals were included in the study. High numbers of Malassezia were isolated from at least one area in 100% of the animals. The frequencies of isolation and population sizes differed significantly according to anatomical location. They were greater on the pinna, followed by the umbilical area, axilla and perianal area. Fungal culture was more sensitive than cytology for the isolation of Malassezia yeasts. Frequencies of isolation were greater using this method, but population sizes were constantly smaller than with cytology.

Anal Canal↗

Absence of mutations associated with sulfa resistance in Pneumocystis carinii dihydropteroate synthase gene from non-human primates.

The dihydropteroate synthase (DHPS) gene from Pneumocystis carinii isolated from non-human primates was amplified using a polymerase chain reaction (PCR) and sequenced to analyse point mutations associated with sulfa resistance. P. carinii DHPS gene amplification was obtained from eight lung samples from five New World primate species and one Old World primate species. None of the animals had been exposed to sulfa drugs and only the wild-type P. carinii DHPS sequence at codons 55 and 57 was observed. These data support the hypothesis that high rates of DHPS mutants in P. carinii f. sp. hominis have arisen with increased use of sulfa drugs for P. carinii pneumonia prophylaxis.

Amino Acid Sequence↗

Ultrastructural localization of binding sites for PNA and VVA-B(4) lectins in human breast cancer cell lines detected by confocal fluorescence microscopy.

Three cancer cell lines (MCF-7, HBL-100, MDA-MB 231) and subnormal breast epithelial cell line MCF-10A were labeled with FITC-conjugated VVA-B4 lectin, specific for D-GalNAcalpha-O-ser/thr, matching the structure of Tn antigen sugar residues, and with RTIC-conjugated PNA lectin, specific for DGalbeta1-3GalNAc-O-ser/thr, corresponding to the structure of T antigen. Simultaneous expression of Tn and T antigens on the same cells (but in widely differing proportions) led to their large heterogeneity and occurrence of numerous cell subpopulations within each of the studied cell lines. This observation proved that the changes leading to the formation of Tn antigen are not caused by an irreversible genetic mutation of beta1-3-galactosyltransferase. Expression of Tn antigen on MCF-10A cells with normal (or subnormal) karyotype suggests that the process of malignant transformation of the cell begins with the changes in molecular structure of glycoconjugates.

Antigens, Tumor-Associated, Carbohydrate↗

Evaluation of the dermatophyte test medium RapidVet-D.

The performance of the dermatophyte test medium (DTM) RapidVet-D was assessed using hair samples collected from experimentally infected guinea pigs. Three dermatophyte species were included in the study: Microsporum canis, Trichophyton mentagrophytes and Trichophyton equinum. DTM substrates were inoculated with infected hairs and scales, incubated at 18, 21, 24, 27 or 37 degrees C and examined daily for 15 days. The rapidity of colour change was clearly related to the incubation temperature and to the number of infected hairs deposited on the reactive substrates. With the optimum incubation temperature 27 degrees C, a systematic colour change could be observed only a few days post-inoculation: 3 days with M. canis infected hairs, 4 days with T. equinum and 5 days with T. mentagrophytes.

Animals↗