Mucosal Candida infection and colonisation as well as associated risk factors in solid organ transplant recipients.

More detailed information on Candida colonisation and infection of the mucous membranes in organ transplant recipients (OTR) is of particular interest. Therefore, this issue was prospectively evaluated in 400 different OTR in different posttransplantation periods as well as in 405 healthy age- and sex-matched controls. In addition, possible risk factors and the clinical condition in the OTR were evaluated. Independent of the transplanted organ there is a statistically significant decrease in the number of positive culture results, of symptomatic candidiasis and an increase of isolated non-albicans Candida species corresponding to length of the posttransplantation period. No significant differences could be observed in the OTR in association with different immunosuppressive regimen; however, higher dosages of corticosteroids and tacrolimus correlated with symptomatic candidiasis. As Candida spp. may also cause systemic infection and dissemination, additional knowledge about cofactors and associated strains may have an impact on therapeutic decisions.

Characterization of a cDNA clone, encoding a 70 kDa heat shock protein from the dermatophyte pathogen Trichophyton rubrum.

Trichophyton rubrum is an anthropophilic fungus causing up to 90% of chronic cases of dermatophytosis. To characterize T. rubrum proteins at the molecular level, we established a cDNA library of this pathogen. Here we describe a recombinant cDNA clone identical to eukaryotic 70kDa heat-shock proteins (HSPs). Western blot analysis using an anti HSP70 monoclonal antibody detected a recombinant fusion protein in Escherichia coli transformed with the expression vector containing the cloned cDNA insert. Southern blot analysis of T. rubrum genomic DNA detected no other members of the HSP70 gene family. Further analysis revealed the presence of two introns within the ORF of the HSP70 gene. In Northern blot analysis, the cDNA clone was hybridized to a RNA species of about 3.5kb which was constitutively expressed by cells cultured at 27 degrees C and was strongly up-regulated after culture at 37 degrees C. In summary, we have cloned the first member of the HSP family of dermatophytes and characterized it as a member of the Dnak subfamily of 70kDa HSPs.

Heterogeneity of Borrelia burgdorferi in the skin.

The reliability of various in vitro techniques to identify Borrelia burgdorferi infection is still unsatisfactory. Using a high-power resolution videomicroscope and staining with the borrelia genus-specific monoclonal flagellar antibody H9724, we identified borrelial structures in skin biopsies of erythema chronicum migrans (from which borrelia later was cultured), of acrodermatitis chronica atrophicans, and of morphea. In addition to typical borreliae, we noted stained structures of varying shapes identical to borreliae found in a "borrelia-injected skin" model; identical to agar-embedded borreliae; and identical to cultured borreliae following exposure to hyperimmune sera and/or antibiotics. We conclude that the H9724-reactive structures represent various forms of B. burgdorferi rather than staining artifacts. These "atypical" forms of B. burgdorferi may represent in vivo morphologic variants of this bacterium.

Lymphoproliferative responses to Borrelia burgdorferi in circumscribed scleroderma.

Humoral immune responses to Borrelia burgdorferi (Bb) have been reported to occur in certain patients with circumscribed scleroderma (CS) (morphoea). Together with the isolation of spirochaetes from CS skin biopsies, this finding was taken to suggest Bb as the aetiological agent of CS. Since there is cellular immunoreactivity to Bb in patients with chronic Lyme borreliosis (LB), Bb-specific lymphocytic responses were tested in patients with CS. For this purpose, peripheral blood mononuclear cells from CS patients and, as controls, from patients with various manifestations of LB, and from healthy volunteers without any evidence of Bb infection, were exposed to Bb organisms for 5 days and then assayed for DNA synthesis. Stimulation indices (SI) > 10 were scored positive. By performing lymphocyte proliferation tests we found: (i) that not only patients with various manifestations of LB but also a considerable percentage of seropositive (five of 13 = 38%) and seronegative (six of 26 = 23%) CS patients exhibit an elevated Bb-induced lymphocyte proliferation; (ii) that the magnitude of the cellular response seen in CS patients is comparable to that encountered in patients with established Bb manifestations; and (iii) that, within a given patient, antibiotic therapy can result in a significant reduction of this response. These results support a causative role of Bb in at least some CS patients. Bb-induced lymphocyte responses were also seen in both seropositive and seronegative erythema chronicum migrans patients. These findings show that the pattern of Bb-specific immune responses is more complex than previously thought, and underscore the importance of lymphocyte function assays in evaluating the diagnosis of potential Bb infection in seronegative patients.

Effects of penicillin, ceftriaxone, and doxycycline on morphology of Borrelia burgdorferi.

Antibiotic therapy with penicillin, doxycycline, and ceftriaxone has proven to be effective for the treatment of Lyme borreliosis. In some patients, however, it was noticed that borreliae can survival in the tissues in spite of seemingly adequate therapy. For a better understanding of this phenomenon, we investigated the different modes of degeneration of Borrelia burgdorferi suspensions during a 96-h exposure to various antibiotics. By dark-field microscopy and ultrastructural investigations, increasing blebbing and the gradual formation of granular and cystic structures could be followed during the exposure time. Although antibiotic concentrations at the MIC at which 90% of organisms are inhibited after 72 h were 80% or even greater, motile organisms were still present after incubation with penicillin and doxycycline but not after incubation with ceftriaxone. By transmission electron microscopy, intact spirochetal parts, mostly situated in cysts, were seen up to 96 h after exposure with all three antibiotics tested. According to experiences from studies with other spirochetes it is suggested that encysted borreliae, granules, and the remaining blebs might be responsible for the ongoing antigenic stimulus leading to complaints of chronic Lyme borreliosis.

Immunohistochemical study of in vivo and in vitro IgA coating of candida species in vulvovaginal candidiasis.

OBJECTIVE: To evaluate whether quantitative or qualitative IgA deficiencies in cervicovaginal secretions can be identified in patients with recurrent vulvovaginal candidiasis. DESIGN: Prospective and controlled study. SETTING: Department of Dermatology, University of Vienna. SUBJECTS: 30 patients with symptomatic and recurrent vulvovaginal candidiasis at the time of their presentation. 30 healthy women as a control group. INTERVENTION: Blood samples were drawn for measurement of serum IgA levels. Smears of the cervix and vagina were taken for direct microscopy and microbiological culture. Lavage of the vagina and ectocervix was performed with sterile saline solution for measurement of cervicovaginal IgA levels. MAIN OUTCOME MEASURES: IgA levels of serum and cervicovaginal secretion evaluated by Single Radial Immunodiffusion. IgA labelling was demonstrated on fungal elements in vaginal smears and subcultured blastospores after incubation with vaginal secretions by immunohistochemistry. RESULTS: We could not find any significant difference of IgA levels in serum and cervicovaginal secretions between the symptomatic group and healthy controls (p value for serum = 0.5796, p value for secretion = 0.2381). In vaginal smears yeasts revealed IgA coating on their surfaces, whereas three of the 61 subcultures were negative. Negative subcultures were assigned to three patients with recurrent candidiasis. No correlation was found between IgA levels of cervicovaginal secretions and staining intensity of subcultured blastospores after incubation with vaginal secretions (r = -0.0578). IgA levels of serum and vaginal secretion showed no correlation (r = -0.00012). CONCLUSION: Recurrent vulvovaginal candidiasis cannot be attributed to IgA deficiency. In some cases an IgA coating defect of yeasts might be involved. In addition inactivation of the IgA molecule by candida proteases might be of pathogenetic importance.

[Aztreonam, a new approach in the therapy of gonorrhea]. / Aztreonam, eine neue Möglichkeit in der Gonorrhoe-Therapie.

The isolated gonococcal strains cultured from 99 patients (64 men and 35 women) with uncomplicated genital gonorrhoea were tested with regard to their susceptibility to aztreonam. No resistant strain was found. The bacteriological evaluation of 50 of these strains showed a minimum inhibitory concentration (MIC) of 0.0075 to 3.906 mcg/ml. Four of the tested strains were penicillinase-producing strains of N. gonorrhoeae. 95 patients (95.9%) were cured after administration of a single dose of 1 g aztreonam by intramuscular injection. It was not possible to exclude reinfection of the 4 remaining patients. No side effects were reported.

Cutaneous aspergillosis.

We report a cutaneous infection by Aspergillus flavus in a tetraplegic but otherwise healthy young man. He presented with multiple erythematous to violaceous indurated papules and plaques which had progressed to central ulcers or black eschars. Lesional scrapings, biopsies and cultures demonstrated a fungus, Aspergillus flavus, as the aetiological agent. As the patient had to leave for his home country before diagnosis had been established, we cannot report on the results of therapy in this case.

Netilmicin: new approach to treating gonorrhoea.

Resistance and side effects to antibiotics or other chemotherapeutic agents are the main limiting factors in the treatment of gonorrhoea. As no such side effects have been reported with a new aminoglycoside, netilmicin, the therapeutic efficiency of this substance was evaluated in 1200 patients (690 women, 510 men) with uncomplicated gonorrhoea, who each received a single dose of 300 mg netilmicin by intramuscular injection. Efficacy of treatment was assessed by smear microscopy and culture, which gave results in all 1200 patients. No serious side effects were reported. These results indicate that netilmicin is an extraordinarily safe and efficient agent for treating uncomplicated gonorrhoea.

[Isolation of Neisseria meningitidis from the urethra and cervix]. / Isolation von Neisseria meningitidis aus Urethra und Zervix.

Meningococcal infections of the urogenital tract were considered rare until recently, but over the past few years an increasing number of cases of this hitherto uncommon infection have been reported. The rising incidence of N. meningitidis isolated from these sites has important epidemiological implications. The need for careful diagnostic procedures is emphasized.

[Critical remarks on gonorrhea therapy: netilmicin]. / Kritische Bemerkungen zur Gonorrhoe-Therapie: Netilmicin.

Several failures of gonorrhea treatment due to neisseria producing penicillinase (PPNG), resistance against other antibiotics, side effects like allergies, gastrointestinal problems, and other factors are the reasons for continuing trials with new drugs in the treatment of gonorrhea. With regard to modern aminoglycosides, the development of early resistance has not been observed so far. Especially with Netilmicin, no such disadvantages have been reported in the literature. Therefore a single dose schedule was introduced with Netilmicin given to 600 patients suffering from uncomplicated gonorrhea in order to evaluate the efficacy and safety of the drug. 300 mg Netilmicin were administered intramuscularly in a one shot therapy. With all patients, the diagnosis had been confirmed by culture. The efficacy of therapy was controlled by a second culture, which proved to be negative in all 600 cases. Gonorrhea was also clinically cured in all patients.

[External erythromycin therapy of acne]. / Externe Erythromycintherapie bei Akne.

24 patients suffering from papulo-pustular acne were treated with mono-therapy with 2% erythromycin-containing alcoholic solution for eight weeks. Pustular lesions responded promptly, followed by inflammatory papules. Microbiological investigations revealed a lack of propionibacteria as well as reduction of pathogenic staphylococci. Patients after extended local erythromycin treatment did not show any erythromycin serum levels.

[The effect of spermicides on physiological and pathogenic genital flora]. / Der Einfluss von Spermiziden auf die physiologische und pathogene Genitalflora.

In vitro-studies concerning the influence of commercially available spermicides on lactobacillus acidophilus as well as on pathogenic organisms of the genital tract revealed, that the preparations had only weak antimicrobial effect on Döderlein's bacteria and pathogenic fungi. However, all four preparations tested, revealed a good inhibition effect on neisseria gonorrhoeae, treponema pallidum and trichomonas vaginalis. Irreversible damage to gonococci and trichomonas vaginalis. Irreversible damage to gonococci and trichomonas was subject to considerable variation, probably due to the different chemical composition of the commercially in Austria available spermicides.

[The role of clothing in the transmission of pathogenic fungi ]. / Die Rolle der Bekleidung für die Ubertragung pathogener Pilze.

Laboratory studies were made to examine the possibility of transmission of pathogenic fungi through textiles. 1. Dermatophytes were proven to be present in clothing and leather articles worn over areas infected by fungi. In patients known to be infected with Pityriasis versicolor, contaminated scales were obtained from their textiles, the fungi could be demonstrated. 2. Artificially infected textiles were found to harbor dermatophytes and pathogenic yeasts for a number of weeks without limiting the survival capacity of the fungi. 3. At low temperatures, fungi were not affected by various detergents. Under conditions simulating machine washings (30 minutes exposure time) at a temperature of 60 degrees C any washing agent was sufficient to cause irreversible damage. At higher temperatures the time for killing the fungi was diminished. These results were confirmed by experiments with contaminated material from patients (scales, hairs, nail particles).

[Nosologic position of demodicidosis in humans]. / Nosologische Stellung der Demodicidosis biem Menschen.

In 20 patients with rosacea and rosacea-like eruptions Demodex folliculorum mites were found in high numbers within lesional skin. In one patient Demodex folliculorum could be demonstrated in pits and burrows in the epidermis of the scalp. In other instances the mouth parts of the mites were closely related to the follicular epithelium. Recent parasitologic findings showed that in spite of their name (Demodex = "sebaceous worm") the mites derive their nutritive requirements from cellular proteins obtained by epithelial destruction. To clarify the pathogenetic significance of hair follicle mites in man as suggested by these histological findings as well as their known ability to form organisational centers of foreign body granulomas, further epidemiologic and immunologic studies are needed. Topical antiparasitic agents did not offer any therapeutic advantage over traditional preparations used for rosacea and seborrheic dermatitis.

[Effect of nitroimidazole therapy on the demonstration of gonococci]. / Einfluss der Nitroimidazol-Therapie auf den Gonokokken-Nachweis.

The minimal inhibitory concentration of metronidazole and tinidazole lies between 4000 micrograms/ml and 7.8 micrograms/ml. Drug levels are present in this range for 2--5 hours in genital discharge following single dose oral therapy with 2 g tinidazole or 1.5 g metronidazole. Therefore, 5-nitroimidazole derivatives exert a bacteriostatic effect in the genital region at least on the more sensitive strains. As shown in our experiments, otherwise reliable laboratory procedures for the demonstration of gonococci may yield false negative results, if specimens for culture are obtained at an inappropriate time.

[Experimental contribution to the theme of pharyngeal gonorrhea]. / Experimenteller Beitrag zum Thema Rachengonorrhoe.

Experimental inoculations of the pharynx with gonococci were undertaken. In seven volunteers gonococci could be cultivated and identified from 16 hours to 8 weeks following infection. In three cases, it was tried to infect the female genital tract using positive pharyngeal smears. This was successful in one instance and the diagnosis was again confirmed by culture and sugar fermentation.