Systemic Aspergillosis Research Articles

Abstracts

Need for annual revaccination of dogs against CPV and CDV McCaw, D. L., Thompson, M., Tate, D., Bonderer, A. & Chen, Y-J. (1998) Journal of the American Veterinary Medical Association...

High dose chemotherapy with thiotepa, carboplatin, VP16 and autologous stem cell transplantation in treatment of malignant brain tumors with poor prognosis. Results of a mono-center pilot study

More than half of the children and adolescents with malignant brain tumors will relapse following initial therapy. Irrespective of the therapeutic modalities the prognosis of patients with recurrent or metastatic brain tumors is still poor. New strategies such as high dose chemotherapy (HDCT) with autologous blood stem cell transplantation (ABSCT) offer the possibility to improve the longterm prognosis of these patients. Following conventional chemotherapy with carboplatin/etoposide and after achieving complete or partial remission (CR or PR) 10 patients aged from 3.2 to 25.5 years (median, 10.3 years) with refractory or recurrent malignant brain tumors (anaplastic astrocytoma/glioblastoma, n = 2; medulloblastoma/PNET, n = 6; ependymoma, n = 1; plexus carcinoma, n = 1) received in a pilot study one course of HDCT with ABSCT. The consolidation regimen consisted of thiotepa (400-600mg/m2/d, i.v. 6 h, d-9), carboplatin and etoposide (500mg/m2/d, CVI 24h, d-8 to d-5, respectively) and was followed by the retransfusion of autologous blood stem cells on day 0. Before starting HDCT 6 patients showed CR and 4 patients had PR or stable disease (SD). Following the HDCT 3 of the 4 patients with residual tumor had CR or PR. 6 patients have remained in continuous CR or SD 8 to 41 months (median 17.2 months) after the HDCT. 2 patients relapsed 8.5 and 9.5 months after HDCT and died from progressive disease. Two patients died therapy-related from systemic aspergillosis and were not evaluable for response. Hematological recovery with an absolute neutrophile count of > 0.5 x 10(9)/l and a platelet count of > 30 x 10(9)/l was reached on days +11 (median; range, +9 to +14) and +16 (median; range, +6 to +47), respectively. The main nonhematological toxic effects were infections, severe mucositis, and hyperbilirubinemia. Although the long-term efficacy of HDCT with ABSCT is still not evaluable and the toxicity of this regimen is high, a multicenter phase II trial seems to be justified in view of the poor prognosis of recurrent or refractory brain tumors in children and adolescents.

Mammary and systemic aspergillosis in dairy sheep.

Mammary aspergillosis was diagnosed in four flocks of dairy sheep, comprising a total of 1,750 ewes. These animals had been treated prophylactically by intramammary infusion with cloxacillin 5 months prior to lambing. Mammary aspergillosis with concomitant spread to the regional lymph nodes was present in these flocks in a percentage ranging from 2% to 36.4% of treated sheep. Pathologic, bacteriologic, and mycologic studies were performed in seven of the affected ewes. Some of them also had lung, kidney, and liver involvement. The pathologic reaction within lesions ranged from the acute to subacute type, dominated by necrosis and vasculitis with thrombosis, to the chronic granulomatous type, with macrophages and giant cells. The distribution of lesions and the presence of a remarkable vasculitis with fungal thrombi in the mammary gland suggested a hematogenous dissemination of the infection from this organ. Immunologic staining with monoclonal antibody MAb-WF-AF-1, which reacts specifically with Aspergillus hyphae, identified the causative agent in histologic sections of the different affected tissues. The etiologic diagnosis was further supported by the isolation of Aspergillus fumigatus in pure culture from affected tissues and from eight samples of mammary secretions.

Outbreak of systemic aspergillosis in a neonatal intensive care unit

Small pre-term neonates are susceptible to cutaneous aspergillosis because of their immature immune system and because of the vulnerability of their skin. In addition, the common therapy with broad-spectrum antibiotic drugs and corticoids creates a favourable milieu for fungal superinfections. We present four pre-term neonates who succumbed to cutaneous aspergillosis that subsequently developed into a systemic infection. The source of the infection proved to be contaminated latex finger stalls. Three of the four patients died. The poor prognosis for systemic aspergillosis can only be improved by an early therapy with amphotericin B, possibly in liposomal form.

Systemic aspergillosis caused by an aflatoxin-producing strain of Aspergillus flavus

Systemic aspergillosis caused by an aflatoxin-producing strain of Aspergillus flavus

Systemic aspergillosis caused by an aflatoxin-producing strain ofAspergillus flavus

The first case of human systemic infection by an Aspergillus flavus isolate demonstrated to produce aflatoxins in vitro and in vivo is described. The patient, a 41-year-old man with acute myelogenous leukaemia, developed a complication of suspected pulmonary Aspergillus infection during remission induction therapy. Antifungal chemotherapy brought about a considerable degree of improvement, but remission of the underlying disease was not attained. Bone marrow transplantation was also not effective. The patient showed recovery from neutropenia but died despite aggressive antifungal chemotherapy. The autopsy revealed lesions in the lungs, myocardium, kidneys, brain, thyroid gland and skin due to a suspected Aspergillus sp. A fungus isolated from the right lung and the skin lesions was identified as A. flavus. Aflatoxins B1, B2 and M1 were detected in culture filtrates of the isolated A. flavus, and in an extract of lung lesions. These aflatoxins are considered to have played an important role in damaging the immune system of the patient through their toxic effects.

Systemic Aspergillosis Caused by an Aflatoxin-producing Strain of Aspergillus in a Post-Bone Marrow Transplant Patient with Acute Myeloid Leukemia

Systemic Aspergillosis Caused by an Aflatoxin-producing Strain of <i>Aspergillus</i> in a Post-Bone Marrow Transplant Patient with Acute Myeloid Leukemia

Endogenous aspergillus endophthalmitis: Clinical features and treatment outcomes

This study evaluated the clinical features and treatment outcomes in patients with endogenous Aspergillus endophthalmitis. The study design was a multicenter retrospective chart review. Ten patients (12 eyes) with culture-proven endogenous Aspergillus endophthalmitis treated by 1 of the authors were studied. Intravitreous amphotericin B injection, pars plana vitrectomy, systemic amphotericin B therapy, and oral anti-fungal therapy were performed. Elimination of endogenous Aspergillus endophthalmitis and Snellen visual acuity, best corrected, were measured. All patients had a 1- to 3-day history of pain and marked loss of visual acuity in the involved eyes. Varying degrees of vitritis was present in all 12 eyes. In 8 of 12 eyes, a central macular chorioretinal inflammatory lesion was present. Four patients (six eyes) had associated pulmonary diseases and were receiving concurrent steroid therapy. One of these patients with chronic asthma also was abusing intravenous drugs. Overall, six patients (six eyes) had a history of intravenous drug abuse, whereas a seventh patient (one eye) was suspected of abusing intravenous drugs. Blood cultures and echocardiograms were negative for systemic aspergillosis. Management consisted of a pars plana vitrectomy in 10 of 12 eyes. Intravitreous amphotericin B was administered in 11 of 12 eyes. Systemic amphotericin B therapy was used in eight patients. One patient was treated with oral antifungal agents. In three eyes without central macular involvement, final visual acuities were 20/25 to 20/200. In eight eyes with initial central macular involvement, final visual acuities were 20/400 in three eyes and 5/200 or less in four eyes. Two painful eyes with marked inflammation, hypotony, and retinal detachment were enucleated. Endogenous Aspergillus endophthalmitis usually has an acute onset of intraocular inflammation and often has a characteristic chorioretinal lesion located in the macula. Although treatment with pars plana vitrectomy and intravitreous amphotericin B is capable of eliminating the ocular infection, the visual outcome generally is poor, especially when there is direct macular involvement.

FUNGAL DISEASE IN THE IMMUNOCOMPROMISED HOST

A variety of superficial and deep mycoses may affect the immunocompromised patient. Among the superficial mycoses, candidal infections are common in all groups, but dermatophyte and pityrosporum infections may also be found. Although not primarily dangerous, they may lead to secondary bacterial infections and morbidity. Of the systemic mycoses, candidiasis, aspergillosis, and mucor-mycosis are frequently lethal and require early diagnosis and aggressive antifungal treatment. Endemic mycoses, such as histoplasmosis and coccidioidomycosis, may result in severe and often fatal infections in those patients with cellular immune alterations. The identification and prophylaxis of high-risk patients and the development of more effective antifungal therapies are beginning to have an impact on the control of fungal disease in this population.

Pulmonary bipartitioning and lobar transplantation: A new approach to donor organ shortage

The scarcity of small donors has significantly limited lung transplantation for pediatric and small adult patients. Use of single lobes procured from size-unmatched donors has overcome this difficulty, but only in a few selected cases and, in addition, it represents a waste of lung tissue. In an animal model we have shown that it is possible to divide one lung with careful partitioning of the vascular and bronchial structures and thus obtain two viable lobar grafts suitable for bilateral implantation in a smaller animal. We have now applied this procedure clinically in seven patients operated on between May 1993 and November 1994. The indications were cystic fibrosis in three children, primary pulmonary hypertension in two adults, bronchiectasis in one, and idiopathic pulmonary fibrosis in one. There were three children aged 13 to 17 years (median 14) and four adults aged 40 to 53 years (median 45). There was a 46% to 50% discrepancy for weight between recipient and donor and a 12% to 17% discrepancy for height. The surgical technique consisted of careful partitioning of the left donor lung, bilateral anterior thoracotomy in the recipient, and, with the use of cardiopulmonary bypass, implantation of the lower lobe in the left hemithorax and the upper lobe in the right hemithorax. Vascular and bronchial connections were facilitated by leaving a long pedicle on the recipient side. The pulmonary artery anastomosis for the donor left upper lobe was done with the “fissure” side of the artery to ensure an anastomosis without tension. An end-to-end bronchial anastomosis overcame the problem of size discrepancy. Six patients are alive and well 10 to 27 months (median 19) after operation. One patient with cystic fibrosis died of systemic aspergillosis infection. All were discharged from the hospital within the first or second postoperative month. No technical problems were identified: repeated bronchoscopy has demonstrated satisfactory healing without early stricture formation. All patients remain well subjectively with good exercise tolerance and all patients achieve greater than 70% of predicted values of forced expiratory volume in 1 second. Perfect adaptation of the transplanted lobes to the recipient pleural space has been demonstrated by postoperative computed tomographic scan. In conclusion, bilateral lobar transplantation from a single donor lung is possible in small adults or children when there is a large size discrepancy with the donor. This may help resolve the problem of donor availability in the pediatric population. (J Thorac Cardiovac Surg 1997;113:529-37)

Lipopeptide inhibitors of fungal glucan synthase.

The echinocandins and pneumocandins are lipopeptide antifungal agents that inhibit the synthesis of 1,3-beta-D-glucan, an essential cell wall homopolysaccharide found in many pathogenic fungi. Compounds with this fungal-specific target have several attractive features: lack of mechanism-based toxicity, potential for fungicidal activity and activity against strains with intrinsic or acquired resistance mechanisms for existing antimycotics. Semi-synthetic analogues of naturally occurring lipopeptides are currently in clinical trials with the aim of treating systemic candidiasis and aspergillosis. Thus a fuller understanding of the target enzyme and its inhibition by these compounds should be useful for epidemiological and other clinical studies. Although it has been long known that lipopeptides inhibit fungal glucan synthase activity both in cell extracts and in whole cells, the genetic and biochemical identification of the proteins involved has been accomplished only recently. We now know that in Saccharomyces cerevisiae, glucan synthase is a heteromeric enzyme complex comprising one large integral membrane protein (specified by either FKS1 or by FKS2) and one small subunit more loosely associated with the membrane (specified by RHO1). Additional components may also be involved. The heteromeric enzyme complex containing Fks1p constitutes the majority of the activity found in vegetatively growing cells in this organism. The FKS2 gene product is needed for sporulation. Lipopeptides affect the function of the Fksp component from either FKS gene. The current model for interaction and regulation of these components in S. cerevisiae and the application to Candida albicans and other pathogenic fungi are discussed in this review.

In vitro and in vivo antifungal activities of ER-30346, a novel oral triazole with a broad antifungal spectrum

ER-30346 is a novel oral triazole with a broad spectrum of potent activity against a wide range of fungi. ER-30346, with MICs at which 90% of the strains tested are inhibited (MIC90s) ranging from 0.025 to 0.78 microgram/ml, was 4 to 32 times more active than itraconazole, fluconazole, and amphotericin B against Candida albicans, Candida parapsilosis, and Candida glabrata. Against Candida tropicalis, ER-30346, with an MIC90 of 12.5 micrograms/ml, was 2 to > 8 times more active than itraconazole and fluconazole, but was 16 times less active than amphotericin B. ER-30346 (MIC90, 0.78 microgram/ml) was four to eight times more active than fluconazole and amphotericin B and had activity comparable to that of itraconazole against Trichosporon beigelli. The MIC90s of ER-30346 were 0.10 microgram/ml for Cryptococcus neoformans and 0.39 microgram/ml for Aspergillus fumigatus. ER-30346 was 2 to 8 times more active than itraconazole and amphotericin B and 32 to > 256 times more active than fluconazole. ER-30346 also showed good activity against dermatophytes, with MICs ranging from 0.05 to 0.39 microgram/ml, and its activity was comparable to or 2 to 16 times higher than those of itraconazole and amphotericin B and > 32 times higher than that of fluconazole. In vivo activity was evaluated with systemic infections in mice. Against systemic candidiasis and cryptococcosis, ER-30346 was comparable in efficacy to fluconazole and was more effective than itraconazole. Of the drugs tested, ER-30346 was the most effective drug against systemic aspergillosis. We studied the levels of ER-30346 in mouse plasma. The maximum concentration of drug in plasma and the area under the concentration-time curve for ER-30346 showed good linearity over a range of doses from 2 to 40 mg/kg of body weight.

Synthesis and antifungal activity of new azole derivatives containing an oxathiane ring

A series of azole derivatives containing an oxathiane ring, which were designed to simulate the D ring of lanosterol, were synthesized and evaluated for their antifungal activity. 3,3-Dimethyl derivatives showed potent activity against murine systemic candidosis and aspergillosis, whereas (2R ∗,3R ∗)-3- monomethyl derivatives showed only weak activity in both in vitro and in vivo.

Detection of galactomannan and complement activation in the pregnant mouse during experimental systemic aspergillosis.

Foeto-placental infections are obtained when pregnant mice are challenged intravenously with conidial suspensions of Aspergillus fumigatus. This experimental model, which is used without any immuno-suppressive pretreatment, can be employed to screen for differences in foeto-placental infectivity of A. fumigatus strains when the number of colony-forming units (CFUs) in conidial suspensions used for infection is from 1 x 10(5) to 1 x 10(6). In the foeto-placental unit, hyphal growth was initiated at the periphery of the placental disc from which infection spread to the central parts of the placenta, the extrafoetal membranes and the foetus. Complement activation was noticed as a consequence of pregnancy and conidial inoculation, but was neither dose-dependent nor related to the extent of infection. Galactomannan was present in the plasma of infected mice and, in contrast to the situation in bovine placental aspergillosis, can be used as a good marker of foeto-placental aspergillosis.

Development of murine monoclonal antibodies for the immunohistochemical diagnosis of systemic bovine aspergillosis.

Murine monoclonal antibodies (MAbs) against water-soluble somatic antigens (WSSA) and the wall fraction (WF) from Aspergillus fumigatus were produced by fusion of splenocytes from immunized BALB/c mice with mouse myeloma X63-Ag 8.653 cells. The supernatants of in vitro cultured hybridomas were initially screened for reactivity with the WSSA and the WF from A. fumigatus and WSSA of other fungi in an enzyme-linked immunosorbent assay (ELISA). Supernatants reacting only with A. fumigatus antigens were subsequently screened for homologous and heterologous reactivity with immunohistochemical techniques using formalin-fixed, paraffin-embedded tissues from experimentally infected mice. Because of a high immunohistochemical reactivity with homologous fungi, 4 MAbs raised against A. fumigatus WSSA and WF were selected for a further evaluation of cross-reactivity (diagnostic specificity) in immunohistochemical and immunoblotting assays. In immunohistochemical assays, all MAbs raised against WSSA cross-reacted heavily with a number of other fungal species. All 4 MAbs (MAb-WF-AF-1-4) raised against the WF reacted strongly with hyphae of Aspergillus spp.; hyphae of Scedosporium apiospermum were also strongly labeled by MAb-WF-AF-3 and -4. The 2 specifically reacting MAbs (MAb-WF-AF-1 and -2) were of the IgM biotype and were precipitating, and in immunoblotting experiments both bound to a 106-kD antigen of the WF, whereas they did not bind to WSSA of A. fumigatus. One of the 2 aspergillosis-specific MAbs (MAb-WF-AF-1) was used to screen 145 mycotic lesions of cattle. The diagnoses on bovine lesions obtained by MAb-WF-AF-1 were compared with results based on reactivity with heterologously absorbed polyclonal antibodies and, for some lesions, to culture results. In the vast majority of lesions (n = 133), the MAb-WF-AF-1 and the polyclonal anti-Aspergillus antibodies reacted in a similar pattern, i.e., positively in 41 aspergillosis lesions and negatively in 92 zygomycotic lesions. Hyphae in 3 of 12 lesions that were not stained by the polyclonal antibodies reacted with the specific MAb-WF-AF-1; i.e., aspergillosis was diagnosed. The characteristics of the 2 MAbs (MAb-WF-AF-1 and -2) raised against the WF of A. fumigatus in ELISA and immunoblotting and immunohistochemical assays justify their application for the in situ diagnosis of systemic aspergillosis of cattle.

Structure-activity relationships of 3-methyl and 3,3-dimethyl analogs of 2-(2,4-difluorophenyl)-3-(omega-substituted alkyl)sulfonyl-1-(1H-1,2,4-triazol-1-yl)-2-propanols.

3-Methyl and 3,3-dimethyl analogs of 2-(2,4-difluorophenyl)-3-(omega-substituted alkyl)sulfonyl-1-(1H-1,2,4-triazol-1-yl)-2-propanols were synthesized and evaluated for their antifungal activities against Candida albicans and Aspergillus fumigatus. The 3,3-dimethyl analogs were found to have more potent activity both in vitro and in vivo than the corresponding 3-mono-methyl analogs. The prophylactic efficacy of the lead compounds against murine systemic candidiasis and aspergillosis was improved significantly by dimethylation of the 3-position.

Route of infections in bovine aspergillosis.

Repeated DNA sequences were used to fingerprint strains of Aspergillus fumigatus isolated from a cow with disseminated systemic aspergillosis, cows with single aspergillosis lesions, calves aborted due to bovine aspergillosis, mothers of those calves, and cattle without aspergillosis. The analysis of the Southern blot hybridization patterns obtained suggested that: (i) the portal of entry of aspergillosis in cattle is the gastrointestinal tract, and (ii) infection of aborted calves is due to maternally derived strains. Cattle from the same farm slaughtered on the same day harbour the same strain, suggesting a contamination from feed material.

Protective effect of human macrophage colony-stimulating factor on fungal infection (2). In vitro effect of human macrophage colony-stimulating factor on systemic aspergillosis and in vitro effect on the activities of macrophage

We studied the protective effect of human macrophage colony-stimulating factor (M-CSF) of fungal infection due to systemic aspergillosis in normal mice. We also examined the effect of M-CSF against the activities of mouse peritoneal macrophage which were relating to the phagocytosis, the killing, the production of superoxide after contacting with phorbol myristate acetate and the production of nitric oxide after contacting with interferon-gamma in vitro. M-CSF improved the median survival time and the survival rate of systemic aspergillosis. Combination therapy with M-CSF and amphotericin-B (AMPH-B) showed the therapy with either M-CSF or AMPH-B alone. M-CSF enhanced the activities of phagocytosis and the killing of ingested Candida albicans H and spores of Aspergillus fumigatus K by macrophage. Furthermore, M-CSF promoted the production of superoxide and nitric oxide in macrophage. These results indicate that M-CSF can enhance the fungicidal activity of macrophages by activation in vivo, thereby preventing the dissemination of fungal infection.

An analysis of antibodies against Aspergillus jumigatus in bovine serum by iminunoblotting and enzyme‐linked immunosorbent assays

Serum samples from 20 cows with spontaneously acquired systemic aspergillosis and serum collected consecutively from a cow with experimentally induced aspergillosis were assayed for antibodies against Aspergillus fumigatus in immunoblotting and enzyme-linked immunosorbent assays. Purified antigens (18 kDa protein and 20 kDa galactomannan) from A. fumigatus as well as complex preparations of metabolic and somatic antigens were used. Antibodies against A. fumigatus were found in both infected and non-infected cows. High titers and especially an increase in titer in consecutive samples signified infection. Reactivity with the 22 and 32 kDa antigens in preparations of exocellular antigens and the 19, 20, and 39 kDa antigens from intracellular antigens was seen with sera from infected cows only, and is therefore indicative of infection.

An autopsy case of acquired immune deficiency syndrome (AIDS) with preceding aplastic anemia

A case of acquired immunodeficiency syndrome (AIDS) with preceding aplastic anemia is reported. The patient was a 36 year old female who had been diagnosed as having aplastic anemia 10 years before and thereafter had received multiple transfusions. Human immunodeficiency virus (HIV)-seropositivity was revealed 10 months prior to her death, but no particular clinical signs indicating HIV infection, pre-AIDS or onset of AIDS were recognized before serological diagnosis, although the slow progression of leukopenia was noted along with thrombocytopenia. Her general condition deteriorated during the last 10 months accompanied by an acute decrease in the CD4/CD8 ratio. Autopsy revealed full-blown AIDS: systemic aspergillosis, progressive multifocal leukoencephalopathy, Epstein-Barr virus-related B cell lymphoma arising in the diaphragm and severe lymphocyte depletion in the lymph nodes and spleen. Markedly hypoplastic bone marrow was considered to be primarily attributable to the aplastic anemia but the affection of AIDS was not excluded. The possible transmission route of HIV and the effect of the preceding aplastic anemia on the infection and clinical course of AIDS are discussed.