Increase of Candida cell virulence by anticancer drugs and irradiation.

The influence of anticancer drugs and irradiation on Candida cell proliferation, adherence to HeLa cells and susceptibility to antifungal drugs (amphotericin B and miconazole) and neutrophils were examined using two Candida albicans strains. After treatment with 5-fluorouracil (25 microg/ml to 250 microg/ml), cis-diammine-dichloroplatinum (10 microg/ml to 100 microg/ml), peplomycin (0.5 microg/ml to 5 microg/ml) or 137Cs (20 Gy to 40 Gy) for 3 days or more, surviving Candida cells proliferated more rapidly than did untreated control cells. Anticancer agent-pretreated Candida cells revealed an increased adhesion to HeLa cells corresponding to an increase of binding to the lectins. The concentration of half limited colony formation (IC50) of amphotericin B and miconazole was increased to near two-fold that of the control by pretreatment of Candida cells with the anticancer agents, except peplomycin, which only weakly increased IC50. In addition, the enolase and Candida acid proteinase activities in the culture supernatants were increased by pretreatment with the drugs and irradiation. Correspondingly, surviving Candida cells after these treatments were resistant to neutrophils, with a reduction to half of the killing. These results indicate that anti-cancer drugs and irradiation potentiate the virulence of Candida cells, or they eliminate Candida cells with low virulence, thereby enhancing the risk of oral and systemic candidiasis.

Influence of aging on candidal growth and adhesion regulatory agents in saliva.

Although oral candidiasis is frequently seen in the elderly, the factors determining candidal growth have insufficiently been explored. Hence, we examined the influence of aging on candidal adhesion and growth-inhibitory agents in saliva in 45 healthy volunteers and 60 patients with oral candidiasis. Both non-stimulated and stimulated salivary flow rates (SFRs) in the healthy controls decreased with aging. A gradual decrease of SFRs with aging was also observed in the patients, and the SFR levels were markedly lower than those in the controls. Although the salivary glucose levels were almost constant in all age groups, secretory immunoglobulin A and lactoferrin levels in saliva were significantly decreased statistically with age, and a marginal age-associated decrease in transferrin levels was also observed. In addition, the generation of superoxide from neutrophils in saliva and their Candida killing activity decreased with age, and these phenomena were more apparent in the patients. Furthermore, a larger number of Candida adhered to oral keratinocytes obtained from the elderly healthy controls than to those obtained from young controls. Correspondingly, keratinocytes from the aged controls showed more concanavalin-A binding sites than those from the young controls. However, oral Candida did not increase with increasing age in the controls, although an age-associated increase of oral Candida was observed in the patients. Taken together, these results indicate that the decreases of SFRs and salivary anti-candidal factors, suppression of salivary neutrophil function and the increase of candidal adhesion sites on keratinocytes predispose elderly individuals to oral candidiasis.

Lactoferrin peptide increases the survival of Candida albicans-inoculated mice by upregulating neutrophil and macrophage functions, especially in combination with amphotericin B and granulocyte-macrophage colony-stimulating factor.

To develop a new strategy to control candidiasis, we examined in vivo the anticandidal effects of a synthetic lactoferrin peptide, FKCRRWQWRM (peptide 2) and the peptide that mimics it, FKARRWQWRM (peptide 2'). Although all mice that underwent intraperitoneal injection of 5 x 10(8) Candida cells with or without peptide 2' died within 8 or 7 days, respectively, the survival times of mice treated with 5 to 100 microg of intravenous peptide 2 per day for 5 days after the candidal inoculation were prolonged between 8.4 +/- 2.9 and 22.4 +/- 3.6 days, depending on the dose of peptide 2. The prolongation of survival by peptide 2 was also observed in mice that were infected with 1.0 x 10(9) Candida albicans cells (3.2 +/- 1.3 days in control mice versus 8.2 +/- 2.4 days in the mice injected with 10 microg of peptide 2 per day). In the high-dose inoculation, a combination of peptide 2 (10 microg/day) with amphotericin B (0.1 microg/day) and granulocyte-macrophage colony-stimulating factor (GM-CSF) (0.1 microg/day) brought prolonged survival. With a combination of these agents, 60% of the mice were alive for more than 22 days. Correspondingly, peptide 2 activated phagocytes inducing inducible NO synthase and the expression of p47(phox) and p67(phox), and peptide 2 increased phagocyte Candida-killing activities up to 1.5-fold of the control levels upregulating the generation of superoxide, lactoferrin, and defensin from neutrophils and macrophages. These findings indicated that the anticandidal effects of peptide 2 depend not only on the direct Candida cell growth-inhibitory activity, but also on the phagocytes' upregulatory activity, and that combinations of peptide 2 with GM-CSF and antifungal drugs will help in the development of new strategies for control of candidiasis.

Pulmonary infiltration with eosinophilia (PIE) syndrome induced by antibiotics, PIPC and TFLX during cancer treatment.

Drugs induce a variety of pulmonary diseases including pulmonary infiltration with eosinophilia (PIE) syndrome. We report a case of PIE syndrome which was observed after neck dissection. An 83-year-old male patient attended our clinic complaining of upper neck swelling and was diagnosed as advanced lymph node metastasis related to previously resected oral carcinoma and underwent neck dissection. Despite administration of antibiotics (piperacillin sodium, PIPC; and tosufloxacin tosilate, TFLX), fever and an elevation of the c-reactive protein (CRP) level with neutrophilia appeared, and an infiltration shadow was observed in the right lower pulmonary field. With the suspicion of pneumonia, the antibiotics were exchanged for panipenem/betamipron. However, the pulmonary infiltration spread widely, CRP increased to 12.9 mg/dl and severe eosinophilia (23%) was observed a few days after changing the antibiotics. PIE syndrome was suspected, and the patient underwent steroid mini-pulse therapy consisting of methylprednisolone sodium succinate (500 mg) and prednisolone (60 mg). After steroid therapy, the pulmonary condition largely improved. However, about 2 weeks after the start of steroid administration, a fever and a further elevation of CRP were observed with an increase of beta-D-glucan in serum. Roentgenography revealed diffuse infiltration shadows throughout the lungs, and the patient died about 3 weeks after the onset from respiratory distress. In vitro, blastogenesis of patient's peripheral blood lymphocytes was strongly enhanced by PIPC and TFLX, and they generated a large amount of interleukin-5 in the presence of PIPC or TFLX. The clinical course and laboratory examination results revealed that PIE syndrome may have been induced by PIPC and TFLX and that PIE syndrome should be suspected in treatment of carcinomas when dyspnea and pulmonary infiltration are complicated with eosinophilia.

A novel bovine lactoferrin peptide, FKCRRWQWRM, suppresses Candida cell growth and activates neutrophils.

To identify potent new antifungal agents, the Candida cell growth inhibitory activities of six lactoferrin (Lf) peptides consisting of 6-25 amino acid residues (peptide 1, FKCRRWQWRMKKLGAPSITCVRRAF lactoferricin B; peptide 2, FKCRRWQWRM; peptide 2', FKARRWQWRM; peptide 3, GAPSITCVRRAF; peptide 4, RRWQWR; and peptide 5, RWQWRM) were examined. Of these, peptide 2 strongly suppressed the multiplication of Candida cells, but other peptides showed only weak activities. In two strains of C. albicans, the minimum inhibitory concentration 100 of peptide 2 (17.3+/-2.2 microM and 17.5+/-2.4 microM) was close to that of miconazole (13.0+/-1.7 microM and 13.1+/-1.6 microM) but markedly different from that of amphotericin B (0.52+/-0.09 microM and 0.56+/-0.11 microM). The suppression of Candida cell growth was additively increased by a combination of peptide 2 with amphotericin B and miconazole. Peptides 1, 3, 4 and 5 and Lf suppressed iron uptake by Candida cells, inversely correlated with their Candida cell growth inhibition activities. However, iron uptake was not inhibited by peptide 2. In addition, peptide 2 upregulated Candida cell killing activity of polymorphonuclear leukocytes (PMN) increasing their superoxide generation, protein kinase C activity, p38 MAPK activity and the expression of p47phox. These results indicated that the main antimicrobial activity of the Lf peptides is dependent on the N-terminal half of Lf and that the PMN upregulatory activity of peptide 2 and additive function of peptide 2 with antifungal drugs are useful for prophylaxis and control of candidiasis.

[New approaches for excellent operative field during tracheobronchial anastomoses in sleeve pneumonectomy: cardiopulmonary bypass and diseased lung ventilation].

Cardiopulmonary bypass (CPB) and diseased lung ventilation were presented as new methods of oxygenation during carinal reconstruction. Two cases of left sleeve pneumonectomy (SP) were performed for bronchial gland carcinoma through clamshell incisions. Adequate oxygenation and excellent operative fields were provided by CPB in both patients. The postoperative courses of these patients were uneventful, although they developed temporary pulmonary edema that needed mechanical ventilation and appropriate diuretics for several days. One case of patient with squamous cell carcinoma after chemotherapy was successfully treated by right SP through midline sternotomy. Adequate oxygenation and excellent operative fields was obtained by diseased right lung ventilation in this patient. The postoperative course of the patient was uneventful without mechanical ventilation support. These procedures of oxygenation in this paper are considered to be safe and effective methods for carinal reconstruction.

Allosteric silver(I) ion binding with peripheral pi clefts of a Ce(IV) double decker porphyrin.

[reaction: see text] A cerium(IV) double decker porphyrin (1) bearing four 4-methoxyphenyl groups was synthesized. Compound 1 shows a positive, homotoropic allosteric effect in metal recognition of Ag(+) ion, and the peripheral pi clefts of 1 act as effective binding sites for Ag(+) ion.

Regulation of Candida albicans growth and adhesion by saliva.

To examine the local regulation of oral Candida albicans growth, we examined non-stimulated and stimulated salivary flow rates (SFRs) and the C. albicans growth and adhesion inhibitory activities of saliva in 60 patients with oral candidiasis (divided into two groups: 25 patients with oral candidiasis only (group OC) and 35 patients with oral candidiasis and systemic diseases (group CS)) and 30 healthy control subjects. Both non-stimulated and stimulated SFRs in patients, especially in group CS; were decreased in comparison with those in the healthy control subjects. The levels of secretory immunoglobulin A (sIgA) in group OC and group CS and the lactoferrin level in group CS were decreased as compared with those in control individuals, although there were no differences in transferrin and total secretory component (SC) levels between the three groups. The secretion amounts (microg/min) of these proteins were statistically significantly decreased in the patients, especially in group CS. Saliva from the patients showed a lesser inhibitory effect on C. albicans growth and adhesion to HeLa cells than did saliva from the control subjects. In addition, polymorphonuclear leukocytes (PMNs) in patients' saliva generated smaller amounts of superoxide than did those in control subjects' saliva, and phagocytic and C. albicans killing activities were suppressed in the patients. These results indicate that the decreases in SFR, secretion of antimicrobial proteins in saliva, and salivary PMN activity are risk factors for oral candidiasis associated with aging and systemic diseases.

[Clinical analysis of patients referred for lung transplantation in eastern Japan].

We investigated the indications for lung transplantation (LTx) in Japan and discussed questions regarding the current Japanese criteria of LTx. This study examined 37 patients referred to our institute for LTx. Among these patients, there were 11 cases of primary pulmonary hypertension (PPH), 6 of lymphoangiomyomatosis (LAM), 4 of Eisenmenger's syndrome, 3 of bronchiolitis obliterans, 2 each of bronchiectasis, idiopathic pulmonary fibrosis (IPF), and pneumoconiosis, and 7 of other diseases. The most frequent age group was 30-39 years (12 cases, 32.4%), and more than half of the patients were female (22 cases, 59.5%). The regions of residence of the patients were Tohoku (19 patients), Kanto (16), and Chubu (2). Patients were referred from all over eastern Japan. On evaluation, LTx was contraindicated in 5 cases, in 4 of which mechanical ventilation had been installed before consultation with us. Six patients died before registration on the Japan Organ Transplant Network (JOT). 6 transplant candidates were registered with JOT, but one (PPH) died after 6 months of waiting, and one (LAM) received a lung transplant from a brain dead cadaver donor. We propose to reform and clarify the Japanese criteria for LTx-both indications and contraindications, to include such indications as a survival probability of less than two years, and such contraindications as mechanical ventilation. Early consultation at LTx centers was hoped for, in accordance with the International Guidelines and a transplant window.

Recombinant adenovirus expressing wild-type p53 is antiangiogenic: a proposed mechanism for bystander effect.

Angiogenesis is required for the growth and progression of malignancies. Recent studies have demonstrated that genetic alterations may accompany acquisition of the angiogenic phenotype. The tumor suppressor gene p53 is most frequently mutated in human cancers and is also known to be a transcriptional regulator of a variety of genes. Here, we investigated the antiangiogenic effect of the wild-type p53 (wt-p53) gene transfer on a human non-small cell lung cancer cell line. Mutant p53-expressing H226Br non-small cell lung cancer cells were transduced with the wt-p53 gene using a recombinant adenoviral vector (Ad5CMVp53) and applied to semiquantitative reverse transcription-PCRs for the detection of altered mRNA expression of angiogenic and/or antiangiogenic factors. In vivo neovascularization assay of Ad5CMVp53-infected cells was then performed using a membrane-diffusion chamber system s.c. transplanted in nu/nu mice. We also evaluated the effect of Ad5CMVp53-infected H226Br cells on nontransduced tumor cells in vivo by s.c. inoculating mixture of cells into nu/nu mice. Ad5CMVp53 infection markedly inhibited the expression of an angiogenic factor, vascular endothelial growth factor, and increased the expression of a novel antiangiogenic factor, brain-specific angiogenesis inhibitor 1, resulting in reduced neovascularization in vivo. Mixing experiments showed that tumor cells transduced with the wt-p53 gene inhibited the in vivo tumor growth of adjacent nontransduced cells. Our data suggest that a recombinant adenovirus expressing the wt-p53 gene is antiangiogenic, which may explain, in part, the mechanism of the bystander effect induced by the wt-p53 gene transfer on adjacent tumor cells.

Human colon cancer cells express the functional Fas ligand.

Fas ligand (FasL) belongs to the TNF superfamily. It is induced in activated lymphocytes and eliminates Fas-positive lymphocytes, resulting in the down-regulation of immune responses. FasL has also been detected in tissues other than lymphoid cells. We investigated the expression and function of FasL on human colon cancer cells. FasL mRNA was detected by RT-PCR in all six colon cancer cell lines tested and was not found on fibroblasts. FasL protein was detected in DLD-1, LoVo, HCT-116 and RPMI 4788 cells by immunohistochemical staining. DLD-1, LoVo and WiDr were cytotoxic against mouse T lymphoma cells which were transfected with human Fas receptor cDNA. The cytotoxicity was significantly enhanced by phorbol 12-myristate 13-acetate (PMA) and ionomycin. Our data suggest that the FasL expressed in human colon cancer cells may be regulated by endogenous factors in the microenvironment of the host and facilitates the escape from the host immune system.

Novel combination therapy for human colon cancer with adenovirus-mediated wild-type p53 gene transfer and DNA-damaging chemotherapeutic agent.

Alteration of the wild-type (wt) p53 gene by mutation, deletion or re-arrangement is a major factor in the development of human colon cancer. Recent studies have demonstrated that p53 might be an essential component of the apoptotic pathway triggered by DNA-damaging stimuli such as chemotherapeutic agents and ionizing radiation. We examined the anti-tumor effects of adenovirus-mediated wt-p53 gene transfer in combination with a chemotherapeutic drug on the human colon cancer cell line WiDr, which is homozygous for a mutation in the p53 gene. Treatment with the chemotherapeutic drug cisplatin following infection with a replication-deficient, recombinant adenoviral vector expressing wt-p53 (termed AdCMVp53) significantly suppressed the growth of WiDr cells compared to single treatments alone. To evaluate the in vivo efficacy of AdCMVp53 and cisplatin given sequentially, WiDr cells were inoculated s.c. in nu/nu mice. After 3 days, AdCMVp53 was injected s.c. into the area where tumor cells were implanted, followed by i.p. administration of cisplatin. Analysis of initial growth inhibition at 21 days demonstrated a profound therapeutic cooperativity, though administration of either AdCMVp53 or cisplatin alone was followed only by a slowing of growth. Our results suggest that gene therapy using wt-p53-expressing adenovirus in combination with a chemotherapeutic DNA-damaging drug could be a useful strategy for treating human colon cancer.

[A case of bronchial stenosis due to postoperative inflammation treated with expandable metallic stent].

We presented a 61-year-old man who had undergone a left sleeve upper resection because of a squamous cell carcinoma of the upper lobe of the left lung. At 5 weeks after the operation, the anastomosis between the left main bronchus and the left lower bronchus became stenotic, therefore pneumonitis occurred below the anastomosis. Because of the inflammatory granulo stenosis for short time, we used an expandable metallic stent to save a residual lung function of the operated side. The anastomosis between the left main bronchus and the left lower bronchus was kept open satisfactorily, and in the late postoperative periods the residual lung function recovered until the levels of predicted residual lung function.

[Cost effectiveness analysis of FDG-PET in the differential diagnosis and staging of lung cancer in Japan].

UNLABELLED: Several studies have shown that FDG-PET is more accurate than CT for the differential diagnosis and for the staging of lung cancer. We have analyzed potential effect of FDG-PET on the medical cost for the management of patients suspected of lung cancer. In the differential diagnosis, chest CT plus FDG-PET protocol reduced the number of bronchofiberscope (BFS) and biopsy by one fourth of that in the conventional protocol using CT alone. PET protocol reduced unnecessary examinations for the patients of benign disease, however, it increased the total cost of examinations by 25% due to the higher cost of PET than that of BFS and biopsy in Japan. In the staging of lung cancer, PET protocol improved accuracy of staging, reduced unnecessary surgery by 67%, and showed a saving of the cost of examination by 5%, and the total medical cost by 2.5% compared to that in the conventional protocol using CT, brain MRI, and bone scan. CONCLUSION: Use of FDG-PET for the staging may contribute to the improvement of patient management of lung cancer patients also to the saving of the medical cost.

A clinicopathological and immunohistochemical study of osteofibrous dysplasia, differentiated adamantinoma, and adamantinoma of long bones.

A clinicopathological and immunohistochemical study of 12 cases of osteofibrous dysplasia (OFD), two cases of differentiated adamantinoma, and five cases of adamantinoma of long bones is presented. Although OFD and differentiated adamantinoma showed similar radiologic findings, differentiated adamantinoma was more likely to be a recurrent lesion than osteofibrous dysplasia and seemed to require a more extensive surgical procedure. Immunohistochemically, cytokeratin- and vimentin-positive cells were seen in both OFD and differentiated adamantinoma. The positive cells were scattered in the former, and were both scattered and nest-like in the latter. Both these lesions, however, were negative for epithelial membrane antigen. Excluding two cases of Ewing-like adamantinoma, the other three cases of adamantinoma were also positive for cytokeratin and vimentin. These results suggest that these three lesions share the same histogenetic origin. The two cases of Ewing-like adamantinoma differ from tibial adamantinoma in their radiological, histological and immunohistochemical aspects, and seem to constitute a distinct variant of adamantinoma with a different histogenesis.

[An adult case of virus-associated hemophagocytic syndrome (VHAS) and a review of this syndrome in adults in Japan].

An adult case of Virus-associated hemophagocytic syndrome (VAHS) was reported and a review of this syndrome in adults in Japan was also made. A 79 year-old woman was referred to our hospital for detailed examination for sustained generalized fatigue lasting for about two weeks. Other clinical manifestations of this patient included fever, generalized lymphadenopathy, hepatosplenomegaly, anemia and mild liver dysfunction. The biopsy of the lymph node revealed hyperplasia of histiocyte with hemophagocytosis. There was also an elevation of IgG antibody against EB virus and the patient was therefore diagnosed to have VAHS. The prednisolone therapy was then initiated and the patient responded to this treatment very well. By the review of the Japanese literatures, seven adult cases of VAHS were found. Based on the descriptions on these cases, the prognosis of this syndrome appeared to be extremely poor which is totally different from VAHS in children. Our case showed a very favourable clinical course following steroid therapy and this suggested that steroid therapy should be considered even at the early stage of this syndrome in adults.

Chondroblastoma of temporal bone.

The case of a 55-year-old female with chondroblastoma arising from the left temporal bone is presented. Although 10 cases of temporal chondroblastoma have been reported, this is the first in which plain radiography, pluridirectional tomography, computed tomography (CT) and angiography were performed. We discuss the clinical and radiological aspects of this rare tumor.