Evaluation of amphotericin B and micafungin combination against clinical isolates of Aspergillus species.

We aimed to evaluate the efficacy of a combination of amphotericin B (AMB) and micafungin (MCFG) against 25 clinical isolates of Aspergillus species in vitro. We examined fungal growth in the presence of these drugs using a checkerboard method with the tetrazolium salt: 2,3-bis (2- methoxy-4-nitro-5-sulphophenyl)-2H-tetrazolium-5-carboxyanilide inner salt (XTT) to determine the efficacy of an AMB/MCFG combination in inhibition of filamentous fungal growth, evaluated based on 50% reduction of metabolic activity. The fractional inhibitory concentration index showed that the drugs synergistically inhibited 36% of the isolates. Activity was judged as indifferent for 64% isolates; antagonistic interaction was not detected. The AMB/MCFG combination was more effective than AMB alone when sub-inhibitory concentrations of AMB were used. This report demonstrates the efficacy of AMB/MCFG combination for inhibiting the growth of Aspergillus species in vitro, warranting the extension of such studies to animal models.

Preliminary analysis of length and GC content variation in the ribosomal first internal transcribed spacer (ITS1) of marine animals.

Length and guanine-cytosine (GC) content of the ribosomal first internal transcribed spacer (ITS1) were compared across a wide variety of marine animal species, and its phylogenetic utility was investigated. From a total of 773 individuals representing 599 species, we only failed to amplify the ITS1 sequence from 87 individuals by polymerase chain reaction with universal ITS1 primers. No species was found to have an ITS1 region shorter than 100 bp. In general, the ITS1 sequences of vertebrates were longer (318 to 2,318 bp) and richer in GC content (56.8% to 78%) than those of invertebrates (117 to 1,613 bp and 35.8% to 71.3%, respectively). Specifically, gelatinous animals (Cnidaria and Ctenophora) were observed to have short ITS1 sequences (118 to 422 bp) with lower GC content (35.8% to 61.7%) than the other animal taxa. Mollusca and Crustacea were diverse groups with respect to ITS1 length, ranging from 108 to 1,118 and 182 to 1,613 bp, respectively. No universal relationship between length and GC content was observed. Our data indicated that ITS1 has a limited utility for phylogenetic analysis as obtaining confident sequence alignment was often impossible between different genera of the same family and even between congeneric species.

In vitro combined effects of cefozopran/teicoplanin and cefozopran/vancomycin on methicillin-resistant Staphylococcus aureus.

We determined the in vitro effects of teicoplanin (TEIC) or vancomycin (VCM) added to cefozopran (CZOP) on 50 methicillin-resistant Staphylococcus aureus (MRSA) strains, using a modified checkerboard method with serial 1.25-fold dilutions, and assessed the time-kill curve. CZOP + TEIC was synergistic (fractional inhibitory concentration index < or = 0.5) against 98% and CZOP + VCM against 20% of strains. Both drug combinations were additive against the remaining strains. A comparison of the fractional bactericidal concentration indices for 32 strains showed synergistic bactericidal effects for CZOP + TEIC in 88%, and in 48% for CZOP +VCM, confirming that CZOP + TEIC is superior to CZOP + VCM. The time-kill curve confirms that the bactericidal potency of these drugs is increased through combined use with CZOP. These results suggest that treatment using TEIC or VCM with CZOP may be useful in treating MRSA infections, including polymicrobial infections and those involving Gram-negative rods.

Dietary n-3 polyunsaturated fatty acid and status of immunocompetent cells involved in innate immunity in female rats.

The aim of this study was to estimate the contributions of dietary n-3 polyunsaturated fatty acid (PUFA), a representative dietary immunosuppressant, to the activity of both alveolar macrophages (AM) and natural killer (NK) cells, and compare them to those of n-6 PUFA. Twelve 5-week-old female Sprague-Dawley rats were divided into two dietary groups, one fed a 10% fat diet for 9 weeks enriched with n-3 PUFA (n-3 diet) and the other an n-6 PUFA (n-6 diet). AM reduced the release of nitric oxide, monocyte chemoattractant protein 1 and tumor necrosis factor alpha in the rats fed the n-3 diet, compared with rats fed the n-6 diet. NK cell activity was reduced by consumption of the n-3 diet. This study suggests that consumption of n-3 PUFA can ameliorate pulmonary inflammatory disorders which are affected by the reduction of not only proinflammatory cytokines but also chemokine released from AM.

Dietary docosahexaenoic acid can alter the surface expression of CD4 and CD8 on T cells in peripheral blood.

The effect of dietary docosahexaenoic acid (DHA) on T cell states in peripheral blood was investigated. Weanling male C57Bl/6N mice were kept on one of three 10% fat diets containing various amounts of DHA and linoleic acid for 4 weeks. Changing the concentration of dietary DHA did not alter the proportion of T cells expressing CD4 or CD8. However, increasing the concentration of dietary DHA lowered the expression of CD4 and CD8 on the cell surface. The decreased expression of these surface molecules involved in T cell proliferation has serious implications in the role of DHA as an immunosuppressant.

[Examination of the inhibitor to factor VIII in non-haemophilic patient].

We examined an inhibitor to factor VIII in non-haemophilic patient who had been developed widely spread ecchymosis and intramuscular bleeding. He had no previous personal or family history of abnormal bleeding tendency. His laboratory data was all normal except examination for blood coagulation. Coagulation studies showed prolonged activated partial thromboplastin time (48 sec) and decreased factor VIII activity (8%). The activity of the inhibitor to factor VIII was demonstrated to be 4.0 Bethesda unit. By the studies of dilution and time response curve, this inhibitor was found to inhibit up to 90% of factor VIII activity but not 100%. This inhibitor was shown to be IgG by protein-A affinity chromatography. In addition, bleeding time was prolonged in the patient. The value of von Willebrand factor antigen was 200%, but that of Ristocetin cofactor was 93%. Since the gel filtration analysis indicated that this inhibitor also suppressed Ristocetin cofactor activity, the relatively low value of Ristocetin cofactor might occur through the action of the inhibitor. These data suggest that patient's inhibitor react to factor VIII high molecular subunit.

Serous cystadenoma of the esophagus.

A 77-year-old man found to have an esophageal polyp underwent polypectomy. Histopathological examination revealed that the polyp consisted of multiple microcystic structures similar to those observed in cystadenoma of the pancreas. This type of tumor is very rare in the esophagus, and is suggested to originate from esophageal glands or remnants from the embryonal state.

Early pancreatic duct adenocarcinoma.

A 68-year-old Japanese woman presented with complaints of appetite and weight loss. Early pancreatic duct adenocarcinoma was diagnosed by endoscopic retrograde cholangiopancreatography (ERCP), which revealed partial obstruction of the main pancreatic duct. Pancreatoduodenectomy with lymph node dissection was performed. Macroscopically, no pancreatic tumor was detected. Histologically, the pancreatic lesion showed continuous changes consisting of duct epithelial cell hyperplasia, carcinoma in situ, and invasive carcinoma. Immunohistochemical stains for carcinoembryonic antigen and CA19-9 were positive in cancer cells.

The prognostic value of serum immunosuppressive effect in patients with ovarian cancer.

Sixteen patients who underwent surgery and postoperative chemotherapy for carcinoma of the ovary were studied. Eleven survived more than 18 months without evidence of recurrence, and 5 died more than 5 months after surgery. Serial determinations were made preoperatively and postoperatively of the immunosuppressive effect of the serums of these patients on the response of normal lymphocytes to phytohemagglutinin stimulation. These determinations were plotted for each patient; the shapes of the curves for patients who survived are markedly different from those of patients who died. The prognostic value of these findings is discussed.

Immunosuppressive effect of serum in patients with ovarian carcinoma.

Cell-mediated immune response was measured in 23 patients with ovarian cystadenocarcinoma, 38 patients with benign ovarian tumor, and 44 healthy volunteers. The method used two indexes: the lymphocyte response per unit volume of peripheral blood to phytohemagglutinin (PHA) and the immunosuppressive effect of serum on the response of normal lymphocytes to PHA stimulation. The lymphocyte response per 50 microliter peripheral blood did not differ significantly between patients with ovarian cancer and healthy volunteers. The serum effect, in contrast, differed significantly between malignant and benign ovarian tumors, and was found to increase significantly even when the cancer masses were as small as about 5 x 5 x 5 cm in size, ie, in FIGO Stage I. It is our belief that the measurement of the serum effect in patients with any ovarian tumor enables the early detection of ovarian cancer.