Israela Berdicevsky

Oral side effects of head and neck irradiation: Correlation between clinical manifestations and laboratory data

Salivary flow rate and composition, oral microflora and clinical manifestations of radiation damage were studied in 32 patients treated with external irradiation to head and neck areas. Several parameters were investigated: field arrangement, amount of salivary glands irradiated, clinical manifestations such as dryness of the mouth, taste impairment, dysphagia, salivary secretion and composition, and oral yeast flora. The salivary glands have a greater sensitivity to radiation damage than the gustatory tissues. The decrease in salivary secretion is accompanied by a rise in salivary sodium concentration, and in oral yeast flora. The clinical symptomatology was correlated with the amount of salivary glands irradiated. We found that most of the parotids have to be outside of the treated volume, when the rest of the major salivary glands are irradiated, to prevent severe dryness phenomena.

In vitro microbial inhibition and cellular response to novel biodegradable composite wound dressings with controlled release of antibiotics

About 70% of all people with severe burns die from related infections, despite advances in treatment regimens and the best efforts of nurses and doctors. Although silver-eluting wound dressings are available for addressing this problem, there is growing evidence of the deleterious effects of such dressings in delaying the healing process owing to cellular toxicity. A new concept of antibiotic-eluting composite wound dressings is described here. These dressings are based on a polyglyconate mesh coated with a porous poly-(dl-lactic-co-glycolic acid) matrix loaded with antibiotic drugs. The effect of antibiotic release on bacterial inhibition was studied, and cell cytotoxicity was examined. The dressings resulted in a 99.99% decrease in the viable counts of Pseudomonas aeruginosa and Staphylococcus albus at very high initial inoculations of 10⁷-10⁸ CFU ml⁻¹ after only 1 day, while such a decrease in Staphylococcus aureus was obtained within 3 days. Bacterial inhibition zones around the dressing material were found to persist for 2 weeks, indicating a long-lasting antimicrobial effect. Despite severe toxicity to bacteria, the dressing material was found to have no toxic effect on cultured fibroblasts, indicating that the new antibiotic-eluting wound dressings represent an effective option for selective treatment of bacterial infections.

Infection stages of the dermatophyte pathogen Trichophyton: microscopic characterization and proteolytic enzymes

Dermatophytes are pathogenic fungi that infect human skin, nails and hair and cause dermatophytosis. Trichophyton mentagrophytes is one of the most widespread species that belong to this group. Infection of the skin tissues include several stages, i.e., adhesion to the surface of the skin, invasion into the sublayers by the penetration of fungal elements and secretion of enzymes that degrade the skin components. In this study we have followed the morphology of the fungal elements, such as arthroconidia and hyphae, during the adhesion and invasion stages. Skin explants were inoculated with the dermatophyte and observed by scanning electron microscopy (SEM) and transmission electron microscopy (TEM). Skin explants were also inoculated with a transgenic isolate of T. mentagrophytes expressing the green fluorescent protein (GFP). The infected sublayers were investigated by confocal scanning laser microscopy (CSLM). As an adaptation to the tissue environment, the dermatophyte produced long fibrils when it is on the open surface of the stratum corneum, while short and thin fibrils are produced inside the dense sublayers. The short and long projections might have a role in adhesion. Invasion may be produced by mechanical and biochemical means. Invasion of the tissue showed hyphal branching and growth in multiple directions. The proteolytic profile was assayed by substrate gel and proteolytic activity. Two serine proteases of similar molecular weight were secreted during growth on the epidermal matrix components keratin and elastin. The dermatophyte may use the proteolytic enzymes to invade the surface and also the deep layer of the skin in immunocompromised patients. Dermatophytes, which are well adapted infectious agents, seem to use their mechanical and biochemical capabilities to invade the skin tissue effectively.

Susceptibility of different yeast species to environmental toxic metals.

The purpose of the study reported here was to investigate the relative resistance of yeast species to various metallic and metalloid ions, with a view to gaining more knowledge on this subject, as resistant species may become dominant in habitats contaminated with the relevant metals. Saccharomyces cerevisiae, Candida albicans and Candida tropicalis were grown in media containing different concentrations of mercury (as HgCl(2)), cadmium (as CdCl(2)), lead (as Pb(CH(3)COO)(2)), arsenic (as Na(2)HAsO(4)) and selenium (as Na(2)SeO(3)) for various intervals. Invariably, the two Candida species turned out to be more resistant to all the metals studied than S. cerevisiae. The metal showing the highest toxicity for these species was mercury, with cadmium being the second, lead, the third and arsenic and selenium being the least toxic elements. Strains showing resistance to mercury were isolated, even in the case of S. cerevisiae.

Incidence of Candida in psoriasis: a study on the fungal flora of psoriatic patients

Summary.  The presence of Candida albicans and other Candida species in saliva and faeces of 50 psoriatic patients compared with a control group of 50 healthy donors was examined quantitatively. The quantity of Candida in saliva and faeces of the psoriatics proved to be significantly higher than in the controls. Candida was detected in 78% of the saliva samples of the psoriatics but in only 50% of the controls, and in the faeces samples in 72% of the psoriatics, but in only 46% of the controls. Qualitative analysis revealed a predominance of Candida albicans (saliva, 77%; faeces, 64%) and Candida rugosa (saliva, 28%; faeces, 28%). We did not find a correlation between the severity of the psoriasis according to the Psoriasis Area and Severity Index and the amount of Candida in the saliva or in the faeces. Our results reinforce the hypothesis that C. albicans is one of the triggers to both exacerbation and persistence of psoriasis. We propose that in psoriatics with a significant quantity of Candida in faeces, an antifungal treatment should be considered as an adjuvant treatment of psoriasis.

Oral Candida carriage and blood group antigen secretor status

Summary. Oral Candida carriage and blood group antigen secretor status were examined in 92 healthy, young volunteers. Candida was isolated from 61 of 92 saliva samples (66%Candida carriage). In 76% of cases this was Candida albicans. Oral Candida carriage was found to be significantly associated with non‐secretion of blood group antigens (P< 0.05). However, the numbers of Candida were higher in the saliva of secretors than of non‐secretors (P< 0.01). A higher percentage of Candida carriage was observed in individuals with blood group O. Thus, the finding of higher carrier frequency in the non‐secretors and in blood group O subjects is confirmed.

Markers for Host-Induced Gene Expression in Trichophyton Dermatophytosis

ABSTRACT Dermatophytes are adapted to infect keratinized tissues by their ability to utilize keratin as a nutrient source. Although there have been numerous reports that dermatophytes like Trichophyton sp. secrete proteolytic enzymes, virtually nothing is known about the patterns of gene expression in the host or even when the organisms are cultured on protein substrates in the absence of a host. We characterized the expression of an aminopeptidase gene, the Trichophyton mentagrophytes homolog of the Trichophyton rubrum Tri r 4 gene. The T. rubrum gene was originally isolated based on the ability of the protein encoded by it to induce immediate and delayed-type hypersensitivity in skin tests. T. mentagrophytes Tri m 4 is closely related to Tri r 4 (almost 94% identity at the protein level). Tri m 4 resembles other protease-encoding genes thought to be virulence factors (for example, DPP V of Aspergillus fumigatus). The Tri m 4 protein was detected immunochemically both in fungal extracts and in the culture medium. Expression of the Tri m 4 gene was induced severalfold when T. mentagrophytes was grown on keratin and elastin. Ex vivo, strong induction was observed after culture on blood plasma, but the use of homogenized skin did not result in a significant increase in Tri m 4 transcript levels. In order to identify additional genes encoding putative virulence factors, differential cDNA screening was performed. By this method, a fungal thioredoxin and a cellulase homolog were identified, and both genes were found to be strongly induced by skin extracellular matrix proteins. Induction by superficial (keratin) and deep (elastin) skin elements suggests that the products of these genes may be important in both superficial and deep dermatophytosis, and models for their function are proposed. Upregulation of several newly identified T. mentagrophytes genes on protein substrates suggests that these genes encode proteins which are relevant to the dermatophyte-skin interaction.

Novel biodegradable composite wound dressings with controlled release of antibiotics: Results in a guinea pig burn model

Approximately 70% of all people with severe burns die from related infections despite advances in treatment regimens and the best efforts of nurses and doctors. Silver ion-eluting wound dressings are available for overcoming this problem. However, there are reports of deleterious effects of such dressings due to cellular toxicity that delays the healing process, and the dressing changes needed 1-2 times a day are uncomfortable for the patient and time consuming for the stuff. An alternative concept in wound dressing design that combines the advantages of occlusive dressings with biodegradability and intrinsic topical antibiotic treatment is described herewith. The new composite structure presented in this article is based on a polyglyconate mesh and a porous poly-(dl-lactic-co-glycolic acid) matrix loaded with gentamicin developed to provide controlled release of antibiotics for three weeks. In vivo evaluation of the dressing material in contaminated deep second degree burn wounds in guinea pigs (n=20) demonstrated its ability to accelerate epithelialization by 40% compared to an unloaded format of the material and a conventional dressing material. Wound contraction was reduced significantly, and a better quality scar tissue was formed. The current dressing material exhibits promising results, does not require frequent bandage changes, and offers a potentially valuable and economic approach to treating the life-threatening complication of burn-related infections.

Soy protein films for wound-healing applications: antibiotic release, bacterial inhibition and cellular response

Use of naturally derived materials is becoming widespread in the biomedical field. Soy protein has advantages over the various types of natural proteins employed for biomedical applications, due to its low price, non‐animal origin and relatively long storage time and stability. In the current study, soy protein isolate (SPI) was investigated as a matrix for wound‐dressing applications. The antibiotic drug gentamicin was incorporated into the matrix for local controlled release and thus continuous bactericidal effect. Homogeneous high‐quality films were cast from aqueous solutions and tested for the effects of gentamicin release on bacterial inhibition. The cytotoxicity and in vitro biocompatibility of these films were also examined. The gentamicin release profiles exhibited a moderate burst effect followed by a decreasing release rate, which was maintained for at least 4 weeks, thus enabling a suitable bacterial inhibition effect. The materials released from the films during an indirect cytotoxicity test were found to be safe, except for a slight inhibitory effect in the presence of high concentrations of glycerol. The biocompatibility test showed confluent cell cultures in close proximity to the SPI films. It is clear that these new antibiotic‐eluting SPI films exhibit a high potential for use as wound dressings. Copyright

CD4+ and CD8+ T cells mediated direct cytotoxic effect against Trichophyton rubrum and Trichophyton mentagrophytes.

Background  The cellular immune system is the most dominant factor in curing acute dermatophytosis. However, the exact immune mechanisms involved in generating this defense are complex and still obscure. The aim of this study was to investigate the fungicidal mechanism of T cells in the normal population versus patients with chronic fungal infections.