What is the best strategy for enhancing the effects of topically applied ozonated oils in cutaneous infections?

Owing to diabetes, atherosclerosis, and ageing, there are several million patients undergoing skin lesions degenerated into infected ulcers with very little tendency to heal and implying a huge socioeconomical cost. Previous medical experience has shown that the daily application of ozonated oil eliminates the infection and promotes a rapid healing. The purpose of the study is the optimization of the antimicrobial effect of ozonated oils by testing in vitro four bacterial species and one yeast without or in the presence of different amounts of human serum. The results obtained suggest that a gentle and continuous removal of debris and exudate is an essential condition for the potent bactericidal effect of ozonated oils. In fact, even small amounts of human serum inactivate ozone derivatives and protect bacteria. The application of ozonated oil preparations is very promising in a variety of skin and mucosal infections. Moreover, ozonated oils are far less expensive than antibiotic preparations.

Tinea capitis due to Trichophyton soudanense mimicking bacterial folliculitis.

We report the case of a 36-year-old Senegalese male with non-scarring alopecia of the scalp, including nodules and pustules, diagnosed as tinea capitis caused by Trichophyton soudanense. This dermatophyte is endemic in Central Africa and is becoming more frequent in Europe because of immigration. It has seldom been isolated in Italy. Tinea capitis is common in childhood and it is rare in adults, in which female sex is preferred. In adults, alopecic patches have to be distinguished from those due to other dermatoses inducing alopecia.

The aberrant spermatogenesis of the Haplothrips simplex (Buffa) (Thysanoptera): ultrastructural study.

The aberrant spermatogenesis of the haploid insect Haplothrips simplex (Thysanoptera) is described. The process, which occurs in the pupal instars, is characterized by two mitotic divisions, the second of which gives rise to two different-sized spermatids: the larger spermatids have a nucleus with diffuse chromatin and proceed into spermiogenesis, while the small spermatids have pycnotic nuclei and degenerate. Both types of spermatids contain two centrioles parallely rather than orthogonally oriented. The occurrence of two centrioles supports a close relationship between Thysanoptera and Phthyraptera. Before the beginning of spermiogenesis, however, the functional spermatids show the unusual presence of a third parallel centriole which is formed by the duplication of one of the two pre-existing centrioles.

Localisation of heavy metals in the midgut epithelial cells of Xenillus tegeocranus (Hermann, 1804) (Acari: Oribatida).

Sites of intracellular metal deposition in the midgut ventriculus and in the proventricular glands of Xenillus tegeocranus (Hermann, 1804) (Acari: Oribatida) were studied by TEM. The study aimed to obtain new information on the ultrastructural features of heavy metal compartmentalisation and elimination mechanisms in oribatid mites. Specimens of X. tegeocranus were collected from an abandoned mining and smelting area and from an unpolluted site. A large number of electron-dense granules (EDGs) were detected: concentric spherocrystals were observed mainly in the epithelium of the midgut ventriculus, while homogeneous dark granules were found exclusively in proventricular gland cells. The elemental composition of EDGs, studied by X-ray microanalysis, showed that midgut cells of X. tegeocranus can store metals (Fe, Mn, Zn, Ni, and Cu) in granules. The chemical composition of EDGs seems to be influenced by the presence and bioavailability of heavy metals in soil, with different kinds of metals accumulating in different types of granules.

Mating of Xenos vesparum (Rossi) (Strepsiptera, Insecta) revisited.

The controversial mating of the strepsipteran Xenos vesparum was studied to investigate the possible sperm routes for fertilization. The female, which is a neotenic permanent endoparasite of Polistes wasps, extrudes only its anterior region, the "cephalothorax," from the host abdomen. This region has an opening where both mating and larval escape occur. Observations with scanning and transmission electron microscopy revealed spermatozoa not only in the hemocoel, but also in the "ventral canal" (an extragenital duct peculiar to strepsipteran females) and in the "genital ducts" (ectodermal invaginations connecting the ventral canal to the hemocoel) of recently mated females. Xenos vesparum spermatozoa can reach the oocytes either through the hemocoel as a result of a hypodermic insemination, or by moving along the extragenital ducts, which are later used by first instar larvae to escape. The hypothesis of hypodermic insemination is reconsidered in the light of behavioral and ultrastructural evidence.

Effect of hyaluronic acid (MW 500-730 kDa) on proteoglycan and nitric oxide production in human osteoarthritic chondrocyte cultures exposed to hydrostatic pressure.

OBJECTIVE: This study investigated the in vitro effects of hyaluronic acid (HA) of molecular weight (MW) 500-730 kDa on human articular chondrocytes cultivated for 48 h in the presence of interleukin-1beta (IL-1beta) with and without hydrostatic cyclical pressure. DESIGN: The effects of 10 and 100 microg/ml HA with and without IL-1beta were assessed in the culture medium of cells exposed to pressurization cycles in the form of sinusoidal waves (minimum pressure 1MPa, maximum pressure 5MPa) at a frequency of 0.25Hz for 3h, by the immunoenzymatic method on microplates for the quantitative measurement of human proteoglycans (PG) and by the Griess method for nitrites (NO). Morphological analyses were performed by transmission electron microscopy (TEM) and scanning electron microscopy (SEM). RESULTS: The presence of IL-1beta determines a significant decrease in PG and a significant increase in NO concentrations measured in the culture medium. When the cells are cultured in the presence of IL-1beta and HA at the two concentrations, a statistically significant restoration of PG and a decrease in NO levels are observed. Under pressurization conditions, we observed that the PG concentration in the medium of cells presented a very significant increase in all the conditions used in the study, except for IL-1beta alone. NO production decreased very significantly in the presence of IL-1beta+HA 10 and IL-1beta+HA 100. The results of metabolic evaluation are confirmed by morphological findings obtained by TEM and SEM. CONCLUSIONS: These in vitro studies confirm both the protective role of HA (MW 500-730 kDa), which counteracts the IL-1beta-induced effects, and the importance of pressure on chondrocyte metabolism and morphology.

Fine structure of the midgut and Malpighian papillae in Campodea (Monocampa) quilisi Silvestri, 1932 (Hexapoda, Diplura) with special reference to the metal composition and physiological significance of midgut intracellular electron-dense granules.

The fine structure of the midgut and the Malpighian papillae in Campodea (Monocampa) quilisi Silvestri, 1932 (Hexapoda, Diplura) specimens was described. We observed the presence of electron-dense granules (EDGs) in the midgut epithelial cells, similar in genesis, structure and aspect to the type A spherocrystals described in the midgut epithelium of Collembola and Diplopoda. Energy-dispersive X-ray microanalysis was used to detect the chemical composition of the granules and to relate it to the concentrations of some potential toxic heavy metals (Pb, Cu, Zn) in soil and litter. Chemical composition of the granules seems strongly influenced by the presence and bioavailability of heavy metals in the external environment. Specimens from a contaminated abandoned mining and smelting area (Colline Metallifere, southern Tuscany) were able to accumulate Fe, Mn, Zn, Pb and Cu in their midgut EDGs. In addition, we observed that C. (M.) quilisi was able to excrete the metal-containing granules into the external medium by the moulting of the intestinal epithelium. This confirms that the process of ionic retention of midgut cells is particularly significant in animals lacking Malpighian tubules.

Onychomycosis due to Ulocladium botrytis.

A case of disto-lateral onychomycosis of the third toe of the right foot of a 45-year-old man is reported. The agent of infection was Ulocladium botrytis, a mycete not hitherto reported to infect humans. The patient underwent mycological examination consisting of direct microscope observation and culture of pathological material. Diagnosis was based on the macro- and microscopic characteristics of the colonies. Fragments of the latter were also examined by scanning electron microscope. Clinical and mycological recovery was achieved after 3 months of topical therapy with ciclopiroxolamine.

Fine structure of the Nassonow's gland in the neotenic endoparasitic of female Xenos vesparum (Rossi) (Strepsiptera, Insecta).

Nassonow's gland consists of a number of cells with ducts that open on to the ventral surface of the brood canal in the cephalothoracic region of a neotenic female strepsipteran. The structural organization of the gland is reminiscent of the class 3 of the epidermal gland cells as defined by Noirot and Quennedey [Ann. Rev. Entomol. 19 (1974) 61], which consists of secretory and duct forming cells. The ultrastructure of the Nassonow's gland is described in female Xenos vesparum (Rossi) parasitic in the social wasp Polistes dominulus Christ. The large secretory cells are clustered in groups of three to four, rich in smooth endoplasmic reticulum and produce a secretion made up of lipids. In young females, just before mating, the ultrastructure of the cells and their inclusions indicate that they are active. In old-mated females the Nassonow's gland degenerates. Microvilli line an extracellular cavity and there are pores present in the irregularly thick cuticle of the efferent duct. The small duct forming cells, intermingle with epidermal cells, overlap secretory cells and produce a long efferent duct, the cuticle of which becomes thick close to its opening in the brood canal. Nassonow's gland could be the source of a sex pheromone, which might be capable of attracting the free-living male to a permanently endoparasitic female.

Case report. Onychomycosis due to Microsporum canis.

A case of distal subungual onychomycosis of the big toe due to Microsporum canis is reported in a 69-year-old male asthma patient who had been treated with systemic corticosteroids for the last 3 years. The nail infection was contracted from a cat who was a healthy carrier. The patient was treated successfully with intermittent itraconazole therapy.

Onychomycosis caused by Alternaria spp. in Tuscany, Italy from 1985 to 1999.

Cutaneous phaeohyphomycosis due to Alternaria spp. is reported with increasing frequency, especially in patients with immune deficiency. Onychomycosis caused by this mould is still rarely observed. Here we report nine cases observed in Tuscany in the period 1985-99; the agent was Alternaria alternata in eight cases and Alternaria chlamidospora in one. Diagnosis was made on the basis of repeated direct microscopic mycological examination and culture, confirmed by scanning electron microscope observation of fragments of colonies. In most cases, the clinical manifestations were dystrophy and distal subungual hyperkeratosis of one or two nails of the feet or hands. Seven cases were treated with oral itraconazole, successfully in six cases, as clinical and mycological recovery was confirmed at follow-up 1 year later.

Effect of hydrostatic pressure on morphological and ultrastructural aspects of normal and osteoarthritic human articular chondrocytes.

In this work we studied the morphological and ultrastructural aspects of normal and osteoarthritic (OA) human articular chondrocytes cultivated in alginate gel for 48 hours. After this period the chondrocytes in Petri dishes were exposed to cyclic pressurization (minimum pressure 1 MPa and maximum pressure 5 MPa) at 0.25 Hz frequency for three hours. In other loading procedures the cells were exposed to continuous pressure (24 MPa) for three hours. Some dishes were not pressurised and these served as controls. The cells were then fixed for transmission electron microscopy (T.E.M.) and for scanning electron microscopy (S.E.M.). No ultrastructural changes were observed in normal chondrocytes exposed at physiological pressure. OA cells placed under physiological pressure showed a partial recovery on morphological and ultrastructural aspects. Normal and OA samples exposed to continuous pressure (24 MPa) showed a morphological worsening in both T.E.M. and S.E.M. studies.

Studies on the biological effects of ozone: 9. Effects of ozone on human platelets.

During the course of ozonated autohaemotherapy (O3-AHT) using heparin as an anticoagulant, it was occasionally observed that a few clots were retained in the filter during blood reinfusion. This observation prompted an investigation on the effect of ozone (O3) on human platelets. We have now shown, both by biochemical and morphological criteria, that heparin in the presence of O3 can promote platelet aggregation. In contrast, after Ca(2+) chelation with citrate, platelet aggregation is much reduced. The potential role of the transient formation of hydrogen peroxide (H2O2) in the presence of Ca2+ with the possible expression of adhesion molecules is briefly discussed.

Studies on the biological effects of ozone: 7. Generation of reactive oxygen species (ROS) after exposure of human blood to ozone.

The acceptance of any complementary medical approach is conditioned by the results obtained after the same scientific scrutiny applied in orthodox medicine. Otherwise any claim of efficacy remains in the realm of fiction. In the case of ozone therapy, the mechanisms of action have remained nebulous and in a series of publications we are trying to present the biochemical, immunological and morphological evidence in favour or against ozone therapy. We have now shown that ozone (O3) dissolved in the water of either plasma or serum or physiological saline generates reactive oxygen species (ROS), of which hydrogen peroxide (H2O2) can be unequivocally demonstrated by using specific methods for its detection. Lipids present in plasma preferentially those present in lipoproteins, undergo peroxidation that is somewhat O3-dose dependent and can be observed by the measurement of thiobarbituric acid reactive substances (TBARS). While the generation of H2O2 is crucial in activating both biochemical (hexose monophosphate shunt) and immunological (via the transcription factor NF-kB) mechanisms, the role of lipid oxidation products (LOP) remains to be investigated. We have shown here that there is a small but consistent induction of some cytokines (TNF-alpha, IFN-gamma and IL-2) when human blood is directly exposed to O3 concentrations up to 100 micrograms/ml per g of blood. On the other hand, isolated blood mononuclear cells (PBMC) in tissue culture medium are far more sensitive to the oxidant action of O3 as shown by a progressive reduction of the proliferation index with comparatively far lower O3, concentrations. On the whole, these results support the concept that much of the O3 toxicity is neutralized by the powerful antioxidant system of blood. The minimal hemolysis supports this idea but as far as platelets are concerned, we must mention that they tend to aggregate in heparinized blood, even when it is exposed to an O3 concentration of 40 micrograms/ml. In spite of the lack of side-effects after autohemotherapy, this drawback must be kept in mind and avoided in clinical practice.

HIV-1 gp120 increases the permeability of rat brain endothelium cultures by a mechanism involving substance P.

OBJECTIVE: To analyse whether an HIV-1 envelope protein might play a role in damaging the blood-brain barrier as a fundamental step in the early invasion of the central nervous system by HIV-1. DESIGN: Analysis of permeability of rat brain endothelium cultures to albumin, to assess the functional integrity of the vascular component of the blood-brain barrier. METHODS: Rat brain endothelium cultures prepared by cerebral microvessels were exposed to recombinant gp120IIIB on microporous membranes and passage of biotin-labelled albumin was analysed. Scanning electron microscopy was used to analyse cell culture morphology. Some cultures were preincubated with N-nitro-L-arginine methyl ester (L-NAME), a selective inhibitor of nitric oxide synthase, or with spantide, a selective substance P antagonist. RESULTS: HIV-1 gp120 increased the permeability of rat brain endothelial cells to albumin in a dose-dependent manner. Scanning electron microscopy revealed profound gp120-induced alterations in cell morphology accounting for the increased permeability to macromolecules. These alterations were neutralized by anti-gp120 monoclonal antibody but not by isotype control antibody or L-NAME. By contrast, spantide and anti-substance P polyclonal antibody completely blocked the gp120-induced increase in albumin permeability. Control cultures exposed to measles virus nucleoprotein showed an increase in permeability that was not blocked by spantide. Brain endothelial cells, exposed to gp120, displayed cell surface immunoreactivity for substance P, suggesting that substance P is secreted by brain endothelium in response to gp120 stimulation and binds to brain endothelial cells through a receptor-mediated mechanism. CONCLUSIONS: These findings suggest a role for substance P in the gp120-induced increase in permeability of brain endothelium.

The behaviour of Sarcoptes scabiei var. hominis in human skin: an ultrastructural study.

The biology of Sarcoptes scabiei var. hominis is poorly understood because of the lack of an in vivo or in vitro propagation system. To obtain more information on the mite behaviour in its natural habitat we conducted an ultrastructural study of burrows in a number of patients with common scabies. Scanning electron microscopy furnished attractive images of the tunnel, parasite body and eggs architecture and demonstrated the presence of holes in the tunnel roof probably representing aeration structures. Transmission electron microscopy showed a marked keratinocyte damage around burrowing mites, well evident ahead of the mite capitulum also. Faecal pellets containing keratinocyte micro-organelles (melanosomes and mitochondria) were documented in posterior midgut. For the first time we disclosed the adhesion mechanism of eggs to the burrow floor. We showed that the typical finger-like projections of the outer layer of the egg shell gradually disappear where the eggs are in contact with the tunnel floor. This allows the inner layer of the egg shell to fuse and stick with the damaged keratinocytes lining the tunnel floor. Our observation substantiates that Sarcoptes scabiei produces a proteolytic substance (salivary secretions?) that has a key role in its life cycle allowing burrowing, feeding and eggs-burrow adhesion.

Two cases of cutaneous phaeohyphomycosis by Alternaria alternata and Alternaria tenuissima.

Two cases of cutaneous phaeohyphomycosis, one with a nodular appearance and the other with an erythematous infiltrating patch, are reported in immunocompromised patients. Diagnosis was based on histological examination, which revealed hyphae and round-shaped fungal cells in a granulomatous dermal infiltrate, and on identification of the moulds when biopsy fragments were cultured on Sabouraud-dextrose agar without cycloheximide. The pathogens were Alternaria tenuissima in the first case and A. alternata in the second. The fungi were examined by scanning electron microscopy. The patients were checked for bone and lung involvement and were then treated with surgical excision and itraconazole, and itraconazole only, respectively, with clinical and mycological resolution.

[An original proposal of a new partially crystalline glass material for dental use. Structural notes on SEM and the microscopic effects of surface acid treatment]. / Proposta originale di un nuovo materiale vetroso parzialmente cristallino di uso odontoiatrico. Note strutturali al SEM ed effetti microscopici del trattamento acido di superficie.

The authors propose a glass-ceramic material which is completely new for using in restorative dentistry. The surface etching of this material just after fusion transforms its optical characteristics and produces a nice translucency. This treatment, extremely fast, simple and cheap, takes the place of complex annealings and of expensive formulations which can create solidification germs. Our methodological choice is oriented to eliminate rather than to increase the orderly component of the composite structure (as shown by the etching treatment of the surface). Our aim is to show that this component is not formed by genuine crystals but by weak crystal-like structures, held together by Van der Waals links.