[3 cases of polymyalgia rheumatica (PMR), in which serum amyloid A was a useful index of the disease activity].

We encountered 3 cases of polymyalgia rheumatica (PMR), in which serum amyloid A (SAA) levels were correlated with clinical pictures after normalization of ESR and CRP levels. Therefore, it is suggested that SAA may be a useful index for evaluating the severity of intractable PMR. Case 1: The patient was a 75-year-old man. Although ESR and CRP levels were normalized after the administration of PSL (20 mg/day), myalgia persisted. When the dose of PSL was reduced, PMR recurred, which was relieved by administering 15 mg/day of PSL. However, myalgia recurred again when the dose of PSL was reduced thereafter. The elevated SAA level (33.0 micrograms/ml) was normalized by continuous administration of PSL without reducing the dose, resulting in the relief of myalgia. Case 2: The patient was a 65-year-old woman. The administration of PSL was initiated at a dose of 15 mg/day. Although myalgia was relieved, the symptom and elevated SAA levels persisted for approximately 3 months. Thereafter, PMR recurred, and SAA levels were markedly increased to 78.2 micrograms/ml. However, the symptom of PMR was eliminated by continuously administering PSL without reducing the dose. Although the dose of PSL was then reduced after the decrease in SAA levels, PMR did not recur. Case 3: The patient was an 88-year-old woman. Although the symptom of PMR was relieved by administering 15 mg/day of PSL, myalgia persisted. Since SAA levels were increased to 106 micrograms/ml, PSL was continuously administered without reducing the dose, resulting in the disappearance of the symptom and normalization of SAA levels approximately 3 months later. Although the dose of PSL was then reduced to 12.5 mg/day, PMR did not recur.

Hydrogen peroxide-induced deacetylation of acetyl resorufin as a novel indicator reaction for fluorometric detection of glucose using only glucose oxidase.

Hydrogen peroxide (H2O2)-induced deacetylation of non-fluorescent acetyl resorufin (1) to fluorescent resorufin (2) as a novel indicator reaction for fluorometric detection of glucose using only glucose oxidase (GOD) is described. When a 1:1:1 mixture of 1 (in CH3CN), glucose, and GOD (each in pH 7.4 phosphate buffer) was incubated at 25 degrees C under aerobic conditions, the resulting solution turned yellow to fluorescent pink due to 2. The formation of 2 was markedly retarded on incubation under anaerobic conditions. When a mixture of 1 and H2O2 was incubated under aerobic conditions, the formation of 2 was noted as in the case of the enzymatic reaction of 1. These results demonstrated that the observed color change is brought about through deacetylation of 1 to 2 induced by H2O2 generated in GOD-catalyzed oxidation of glucose. With regard to the fluorometric traces of the enzymatic reaction with 1 (0.2 mM), GOD (0.5 mg/ml), and glucose at 25 degrees C, fluorescence intensity exhibited a linear relationship against glucose concentration between 0.2 and 2.0 mm, with a correlation coefficient of 0.997. Neither ascorbic acid, uric acid, nor bilirubin significantly interfered with the transformation of 1 to 2 through GOD-catalyzed oxidation of glucose.

Japanese spotted fever associated with multiorgan failure.

A 49-year-old man was admitted to our hospital, with a diagnosis of multiple organ failure, on June 10, 2000. Physical examination revealed high fever, generalized maculopapular erythema, and an eschar on his lower leg. Laboratory findings revealed severe renal and liver dysfunction, disseminated intravascular coagulation (DIC), and markedly elevated soluble interleukin 2-receptor (sIL2-R) level (>10 000 U/ml). Administration of minocycline was started immediately, with a diagnosis of rickettsial infection. Simultaneously, anti-thrombin III and heparin were started to treat the DIC, and hemodialysis was also initiated. However, the day after admission, his consciousness level lapsed, to the level of coma, and blood pressure was less than 60 mmHg, indicating shock. Therefore, 500 mg of methylprednisolone was administered once; as a result, rapid pyretolysis and improvement of consciousness disturbance were achieved. Laboratory data indicative of inflammation gradually improved after a few days. Hemodialysis was required ten times. During the recovery period, the level of specific IgM antibody against Rickettsia japonica increased to x2560, and he was diagnosed as having Japanese spotted fever. On July 11, he was discharged without sequelae. The course in our patient was very severe, and treatment with minocycline alone may have resulted in a fatal outcome. The level of sIL2-R, which is produced by activated lymphocytes, was markedly increased. Therefore, markedly elevated lymphocyte activation and hypercytokinemia may have been present on admission. The short-term steroid therapy may have been effective in inhibiting the excessive activation of lymphocytes in the critical stage. In the severe form of Japanese spotted fever with organ failure, combination therapy with minocycline and short-term steroids may be very useful.

Resorufin as an electron acceptor in glucose oxidase-catalyzed oxidation of glucose.

Resorufin (1) has been found to act as an electron acceptor in glucose oxidase (GOD)-catalyzed oxidation of glucose. When a 1: 1: 1 mixture of solutions of 1 (5.0 microM), glucose, and GOD (4.0 mg/ml) in phosphate buffer (pH 7.4, 0.1 M) was incubated at 36 degrees C under aerobic conditions and the reaction was followed by a measurement of changes in fluorescence intensity due to 1, only two types of fluorometric traces were observed: (1) when a glucose solution of less than 0.7 mM was subjected to the enzymatic reaction, no consumption of 1 was observed; (2) the reaction with glucose at more than 1.0 mM always consumed 1, affording a regression fluorometric curve, and yet the obtained fluorometric traces could be almost superimposed on one another with no dependence on the glucose concentration. The reasons for the observed phenomena are discussed.

[Effects of gadobenate dimeglumine on MRI of mouse liver metastasis model constructed by orthotopic transplantation of highly metastatic murine colon carcinoma (C38-OT7): comparison with gadopentetate dimeglumine].

Twelve mice with metastatic liver tumors were divided into two groups of six, with one group administered contrast medium at 0.1 mmol/kg, and the other at 0.2 mmol/kg. Contrast medium, gadobenate dimeglumine (GD) or gadopentetate dimeglumine (GP), was administered at 0.1 or 0.2 mmol/kg via the tail vein to each animal in each group. Using a Signa Horizon 1.5-Tesla MRI unit, spin-echo transverse and coronal T1-weighted sections were obtained every 15 minutes until 2 hours after administration. The numbers of liver tumors detected on films were counted before and after the administration of contrast medium, and the liver/tumor contrast-to-noise ratio (CNR) was calculated. After the completion of MRI, livers were removed, and the number of metastatic nodules on the liver surface were counted as the number of tumors. Liver/tumor CNR rose after administration in both of the GD groups. In the GP group, liver/tumor CNR remained almost constant throughout the observation period. Relative to the number of tumors detected at optical microscopy, approximately 80% and 100% of tumors were detected at MRI after the administration of GD at 0.1 and 0.2 mmol/kg, respectively. On the other hand, approximately similar numbers of tumors were detected at MRI before and after the administration of GP. These results suggest that GD administered by intravenous injection was transported promptly to the liver, increased liver/tumor CNR, and enhanced detection performance for metastatic liver tumor.

Enhanced antitumor activity of a combination treatment with a mouse/human chimeric anti-MK-1 antibody and lymphokine-activated killer cells in vitro and in a severe combined immunodeficient mouse xenograft model.

Mouse monoclonal antibody FU-MK-1, raised against a human gastric adenocarcinoma, recognizes a glycoprotein antigen (termed MK-1 antigen) present on most carcinomas and seems to be valuable in immunodiagnosis and immunotherapy of various cancers. In a recent study, we constructed a mouse/human chimeric antibody, designated Ch FU-MK-1, by fusing the FU-MK-1 V(H) and Vkappa genes to the human Cgamma1 and Ckappa genes, respectively. In the present study, we tested combination immunotherapy of Ch FU-MK-1 with human lymphokine-activated killer (LAK) cells in vitro and in mice with severe combined immunodeficiency (SCID) bearing human MK-1-expressing tumors. In in vitro experiments, Ch FU-MK-1 effectively mediated antibody-dependent cell-mediated cytotoxicity (ADCC) against MK-1-expressing MKN-74 cells, which was completely blocked by an anti-FcR antibody. Since the apoptotic pathway as well as the necrotic pathway have been shown to be utilized in various cytotoxic effector mechanisms, we investigated the role of apoptosis in ADCC mediated by LAK cells and Ch FU-MK-1 against MKN-74 cells. The implication of the apoptosis during ADCC was demonstrated by means of both a terminal-deoxynucleotidyltransferase-mediated dUTP-biotin nick-end-labeling assay and a propidium iodide staining method. In vivo antitumor activity of combination treatment with LAK cells and Ch FU-MK-1 was estimated using SCID mice inoculated s.c. with MKN-74 cells. The i.v. administration of LAK cells and i.p. administration of Ch FU-MK-1 and interleukin-2 (IL-2) produced a marked growth inhibition of MKN-74 tumors in SCID mice. When the actual tumor weights were measured 16 days after initiation of treatment, more than 70% reduction was observed in the group receiving LAK cells plus Ch FU-MK-1 plus IL-2 as compared to the control untreated group. Together these results suggest that Ch FU-MK-1 may serve as a potentially useful immunotherapeutic reagent for human MK-1-expressing tumors.

[A clinicobacteriologic study on clavulanic acid/amoxicillin in pediatric acute otitis media].

We carried out clinical and bacteriological studies on clavulanic acid/amoxicillin and amoxicillin in pediatric acute otitis media at 14 general practice settings. The results are summarized as follows. 1. The major isolated organisms from content of middle ear effusion were Streptococcus pneumoniae 31.8%, Haemophilus influenzae 35.8% and Moraxella subgenus Branhamella catarrhalis 1.5%. Similar results were observed for the major isolates organisms from content of nasopharynx Streptococcus pneumoniae 31.1%, Haemophilus influenzae 33.9% and Moraxella subgenus Branhamella catarrhalis 19.2%. 2. 42.2% of S. pneumoniae isolated from middle ear effusion were drug resistant S. pneumoniae (PISP, PRSP) and they were increasing year by year. 3. 46.7% of S. pneumoniae isolated from nasopharyngeal swab were drug resistant S. pneumoniae (PISP, PRSP) and they were increasing year by year. The incidence of drug resistant S. pneumoniae isolated from all cases and organisms were 26.3% and 14.5%, respectively. 4. On MIC90, antimicrobial activity of CVA/AMPC against Streptococcus pneumoniae, Haemophilus influenzae and Moraxella subgenus Branhamella catarrhalis was superior to SBTPC. 5. In the evaluation of clinical efficacy, bacteriological efficacy and utility, CVA/AMPC-treated group was significantly superior to AMPC-treated group. 6. Adverse reactions were observed in 22% of CVA/AMPC-treated group, involving diarrhea and loose stool.

[A clinicobacteriologic study on clavulanic acid/amoxicillin in pediatric sinusitis].

We carried out clinical and bacteriological studies on clavulanic acid/amoxicillin and amoxicillin in pediatric sinusitis at 11 general practice settings. The results are summarized as follows. 1. The major isolated organisms from content of middle meatus were Streptococcus pneumoniae 32.2%, Haemophilus influenzae 32.0% and Moraxella subgenus Branhamella catarrhalis 25.1%. Similar results were observed for the major isolates from nasopharynx. 2. 62.1% of S. pneumoniae isolated were drug resistant S. pneumoniae (PISP, PRSP) and they were increasing year by year. 3. Drug resistant S. pneumoniae was isolated from 38.6% of all cases. 4. Regarding MIC90, CVA/AMPC showed superior antimicrobial activity against Streptococcus pneumoniae, Haemophilus influenzae and Moraxella subgenus Branhamella catarrhalis. 5. The clinical efficacy, bacteriological efficacy and utility of CVA/AMPC-treated group were 78%, 58% and 72.8%, respectively, and they were significantly superior to AMPC-treated group. 6. Adverse reactions were observed in 11.2% of CVA/AMPC group, involving diarrhea and stool loose and there was no statistical deference from those of AMPC group.

Nonspecific cross-reacting antigen-50/90 (NCA-50/90) as a new tumor marker.

BACKGROUND: The nonspecific cross-reacting antigen-50/90 (NCA-50/90) is a glycoprotein antigen which shares some antigenic determinants with carcinoembryonic antigen (CEA). No definite clinical value has been established for the measurement of NCA-50/90 in cancer patients. METHODS: We established and evaluated a chemiluminescent enzyme immunoassay (CLEIA) specific for NCA-50/90 using two monoclonal antibodies. RESULTS: No significant reactivity with 4 purified related antigens including CEA was found in the NCA-50/90 CLEIA. Serum samples (n = 572) from patients with malignant (n = 326) as well as from healthy individuals (n = 246) were analyzed by the NCA-50/90 CLEIA and by the established ACCESS CEA assay. The average sensitivity of NCA-50/90 for malignant disease was 40.8%, compared to 45.4% of CEA. Relatively high positive rates of NCA-50/90 were observed in sera from patients with lung cancer (72.0%), hepatoma (62.5%), pancreatic cancer (47.6%), breast cancer (35.6%), and colorectal cancer (34.5%). About 15% of patients with malignant disease were positive only for NCA-50/90. The levels of NCA-50/90 and CEA in sera from patients with malignant disease correlated only poorly. CONCLUSIONS: The present study suggests that increases in blood levels of NCA-50/90 occur in a population of cancer patients which is different from those with elevated levels of CEA and that NCA-50/90 might be useful for NCA-50/90-positive, but CEA-negative patients.

Tumor growth suppression by a mouse/human chimeric anti-CEA antibody and lymphokine-activated killer cells in vitro and in SCID mouse xenograft model.

The mouse/human chimeric antibody Ch F11-39, recently generated by ourselves, shows the same high specificity and affinity for carcinoembryonic antigen (CEA) as those of its parental mouse monoclonal antibody. Ch F11-39 is capable of mediating antibody-dependent cell-mediated cytotoxicity (ADCC) with human peripheral blood lymphocytes (PBL). Interleukin-2 (IL-2) modulates the function of immunocytes, in particular inducing lymphokine-activated killer (LAK) cells and enhancing ADCC. In the present study, we therefore tested the combination immunotherapy of Ch F11-39 with LAK cells in vitro and in severe combined immunodeficiency (SCID) mice bearing human CEA-producing tumors. In vitro experiments using human gastric tumor cell lines, Ch F11-39 effectively mediated ADCC against CEA-positive MKN-45 cells, but not against CEA-negative cells. The specificity of ADCC for Ch F11-39 was demonstrated by experiments with irrelevant target cells or irrelevant antibody. ADCC activity of PBL with Ch F11-39 was enhanced by double after preincubation with IL-2 at 10 U/ml. The concentration of Ch F11-39 required for 50% maximal cell killing was about 0.25 microgram/ml at 10 U/ml of IL-2. Increasing ADCC was triggered by IL-2 earlier (1 day) than the generation of LAK cells (3 days). Control human IgG blocked the ADCC, suggesting that the enhancement of ADCC by IL-2 may be caused by activation of effector cells expressing Fc receptors. In vivo anti-tumor activity of combined immunotherapy was estimated using SCID mice inoculated s.c. with 1 x 10(7) MKN-45 cells. The i.v. administration of LAK cells and i.p. administration of Ch F11-39 and IL-2 produced a marked growth inhibition of MKN-45 tumors in SCID mice (about 50% reduction in tumor size as compared to the control untreated group, measured 15 days after treatment). In summary, the enhanced antitumor activity of Ch F11-39 with LAK cells suggests that it might be a useful immunotherapeutic reagent for CEA-expressing tumors.

Establishment and evaluation of a new chemiluminescent enzyme immunoassay for carcinoembryonic antigen adapted to the fully automated ACCESS system.

We have established a new chemiluminescent enzyme immunoassay for carcinoembryonic antigen (CEA), designated ACCESS CEA, which is adapted to the fully automated ACCESS immunoassay analyzer. The assay is based on a one step sandwich-type method using two monoclonal antibodies, one of which is immobilized on micrometer-size paramagnetic particles and the other is conjugated to alkaline phosphatase. Ten microliters of calibrators or sera are incubated for 5 minutes at 37 degrees C with the particles and with the alkaline phosphatase conjugate. The particles are then magnetically separated and washed to remove unbound components. Time needed to obtain the first result is less than 15 minutes. The assay range was 0.04-1000 micrograms/l of CEA, and the possible high-dose hook effect was prevented at CEA concentrations up to 100000 micrograms/l in this working range. The coefficient of variation (CV) for intra-assay precision was 3.0 to 4.7%, and inter-assay CV was 3.4 to 5.6%. The sample carryover was less than 0.001%. The analytical recovery ranged from 98 to 104% and a dilution linearity was demonstrated. No interference was detected in any sample with levels up to 300 mg/l for bilirubin, 12000 mg/l for haemoglobin, 50000 mg/l for human serum albumin, 8500 mg/l for triacylglycerol, and 500000 IU/l for rheumatoid factor. The ACCESS CEA assay also showed very homogeneous reactivity with purified CEA preparations from different tumours and could discriminate CEA from four CEA-related normal antigens tested. Serum samples (n = 362) from patients with malignant or non-malignant disease, as well as from healthy individuals, were analyzed by the ACCESS CEA assay and by the established IMx CEA assay. The CEA values determined by the ACCESS CEA assay were in good agreement with those determined by the IMx CEA assay, and the ACCESS CEA assay significantly increased the sensitivity and specificity of tumour diagnosis as compared with the IMx CEA assay.

Cloning and sequencing of the VH and V kappa genes of an anti-CD3 monoclonal antibody, and construction of a mouse/human chimeric antibody.

Mouse monoclonal antibodies against CD3 on human T lymphocytes have been used for therapy in organ-transplant patients as a potent immunosuppressive agent or for treatment of cancer as a potent T cell activating agent. However, an inherent problem in their in vivo application is the human anti-mouse antibody response. In this study, we cloned and sequenced the variable region genes of the heavy and light chains (VH and V kappa) of a mouse anti-human CD3 monoclonal antibody (OKT3) using the reverse transcription-polymerase chain reaction method. Then, we constructed a mouse/human chimeric antibody, designated as Ch OKT3, by fusing the OKT3 VH and V kappa genes to the human heavy and light chain constant region genes (C gamma 1 and C kappa) derived from a human plasma cell leukemia line (ARH77), respectively. The chimeric gene constructs were sequentially co-transfected into mouse non-Ig-producing hybridoma cells (Sp2/0) by electroporation. The Ch OKT3 antibody thus prepared bound to human peripheral blood mononuclear cells and competitively inhibited the binding of the parental MAb OKT3 to the blood mononuclear cells, indicating that this chimeric antibody seems to be suitable for in vivo therapeutic approaches.

A mouse/human-chimeric bispecific antibody reactive with human carcinoembryonic antigen-expressing cells and human T-lymphocytes.

A mouse/human-chimeric bispecific antibody, designated CBA-CEACD3, with dual specificities for carcinoembryonic antigen (CEA) and CD3, was generated by chemical cross-linking of a chimeric antibody specific for CEA to another chimeric antibody against CD3. Flow cytometric analysis showed that CBA-CEACD3 can bind specifically to cells expressing CEA and to normal human peripheral blood mononuclear cells (HPBMCs) bearing CD3, respectively. Furthermore, a cell to cell adhesion analysis by a colorimetric assay using the dye 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) demonstrated that CBA-CEACD3 is able to bind CEA-producing cells to CD3-expressing cells, suggesting that both arms of CBA-CEACD3 are simultaneously working and can retarget T-cells to the tumor. In an additional colorimetric assay using MTT, this antibody was shown to effectively mediate CEA-expressing tumor cell killing by freshly isolated HPBMCs. Together these results demonstrate that this chimeric bispecific antibody may serve as a potentially useful immunotherapeutic reagent for human CEA-producing cancers.

Tumor-specific accumulation of 125I-labeled mouse-human chimeric anti-CEA antibody in a xenografted human cancer model demonstrated by whole-body autoradiography and immunostaining.

Whole-body autoradiography (WBAR) was used to study the biodistribution of 125I-labeled mouse-human chimeric antibody (Ch F11-39) to carcinoembryonic antigen (CEA) in athymic nude mice bearing the CEA-producing MKN-45 human gastric carcinoma xenografts. Significantly high uptake of 125I-Ch F11-39 in the tumors obtained by tissue-counting technique was confirmed by WBAR of mice of 12, 24, 48, and 96 h postinjection of 125I-Ch F11-39. When compared with histochemical or immunohistochemical staining results of the tumor tissue sections, imaging profiles of 125I-Ch F11-39 obtained by WBARs were topographically correlated with histopathological findings of tissues and immunohistochemical localization of CEA in the tumor tissues, indicating that the accumulation of 125I-Ch F11-39 at the tumor site is based on its specificity for CEA. These results demonstrate that this chimeric antibody may serve as a potential useful diagnostic and/or therapeutic reagent for human CEA-producing cancers.

Binding reactivity of monoclonal anti-carcinoembryonic antigen (CEA) antibodies with cell membrane-bound CEA and with free CEA in solution.

Binding reactivities of 62 anti-CEA MAbs from 10 different research groups with cell membrane-bound CEA and with free CEA in solution were compared by inhibition of MAb binding to CEA-expressing tumor cells by free CEA. Bindings of 30 MAbs to the cell membrane-bound CEA (280 ng CEA/2 x 10(5) cells) were inhibited by approximately equal amounts of free CEA, indicating that binding affinities of about half the MAbs for cell membrane-bound CEA are similar to those for free CEA, respectively. Bindings of 15 MAbs to the cell membrane-bound CEA were easily inhibited by free CEA of less than half the amount of the cell membrane-bound CEA, while inhibition of bindings of the remaining 17 MAbs required twice more free CEA than the amount of cell membrane-bound CEA, showing that about one-fourth of the MAbs have higher affinities for free CEA and the remaining about one-fourth of the MAbs possess higher affinities for cell membrane-bound CEA. These results help form the basis for selecting the anti-CEA MAbs for use in clinical applications, such as serum CEA assay, tumor imaging and immunotherapy.

Reducing interference from heterophilic antibodies in a two-site immunoassay for carcinoembryonic antigen (CEA) by using a human/mouse chimeric antibody to CEA as the tracer.

To reduce heterophilic antibody interference in a two-site immunoassay for carcinoembryonic antigen (CEA), we utilized a human/mouse chimeric antibody to CEA as the tracer. One mouse monoclonal antibody (MAb), F82-61, which reacts with an epitope present on the domain N of CEA, was immobilized on 96-well polystyrene microtiter plates. A human/mouse chimeric antibody (Ch F11-39), which recognizes an epitope present on the domain B3 of CEA, was biotinylated for the tracer (Ch F11-39 system). Another MAb F11-39, the parental MAb of Ch F11-39, was also biotinylated and used as the control tracer (F11-39 system). For a fair comparison, the same 503 serum samples from healthy individuals were simultaneously assayed in the present study. When a tentative common reference limit of 5 ng/ml was used, the false positive rate with the Ch F11-39 system was only 2.8% (14/503) and that with the F11-39 system was 29.0% (146/503). Adding normal mouse serum (NMS; 1%) or a mixture of purified mouse IgG subclasses (heterophilic blocking reagent (HBR, 15 micrograms/test)) to the F11-39 system reduced the false positive rate from 29.0% to 6.2% (31/503) or 4.8% (24/503), respectively, suggesting that heterophilic antibodies reactive with mouse IgG gave rise to the high positive rate in normal populations with the F11-39 system. On the other hand, the false positive rate with the Ch F11-39 system was only slightly reduced from 2.8% to 2.6% (13/503) or to 2.0% (10/503) by adding NMS or HBR to the Ch F11-39 system. The false positive rates with two commercially available assay systems, CEA Roche EIA.DM or Abbott IMx CEA, were 5.4% (27/503) and 5.8% (29/503), respectively, which both corresponded roughly to that with the F11-39 system including NMS or HBR. These results indicate that the application of human/mouse chimeric antibodies in two-site immunoassays is more effective for reducing interference from heterophilic antibodies than the adding of NMS or purified mouse IgG in the assay using conventional MAbs.

Studies on thermophile products. VIII. Isolation of Bacillus stearothermophilus UBT8038, a component that inhibits antigen presentation on mouse macrophages.

We have found a new compound from thermophile extracts which inhibits antigen presentation on mouse macrophages. The substance inhibits the expression of the class II major histocompatibility molecule (Ia). It was extracted from Bacillus stearothermophilus UBT8038 and purified by silica gel column chromatography. The isolated inhibitor, Fr. 8-A, was a phosphatidylethanolamine with isofatty acids and chemically different from any of the natural or synthetic products which have been reported to modify Ia expression. Fraction 8-A inhibits Ia expression by mouse peritoneal macrophages induced by the supernatant from concanavalin A stimulated spleen cell cultures in a dose-dependent fashion over the range 0.1-10 micrograms/ml. This fraction also exhibited inhibitory effects on antigen presentation by splenic macrophages in vitro and on the mixed leukocyte reaction. The present results show that Fr. 8-A has a unique inhibitory effect on antigen presenting cells.

[Combined CDDP-THP-PEP chemotherapy (PTP therapy) for head and neck squamous cell carcinoma--a group study].

A group study of chemotherapy for head and neck squamous cell carcinoma, using CDDP, THP and PEP, was carried out on 32 cases at the department of Otolaryngology, University of Tokyo, and 7 affiliated hospitals. The combined chemotherapy, which we call "PTP therapy", consisted of 30 mg/m2 of THP on day 1, 70 mg/m2 of CDDP on day 2, and 5 mg/body/day of PEP on 4 successive days. The overall response rate was 78.1% (CR: 15.6%; PR: 62.5%). No major side effect was observed in any case. PTP therapy is useful in neoadjuvant chemotherapy.

Bilateral internal auditory canal enlargement without acoustic nerve tumor in von Recklinghausen neurofibromatosis.

The common (and definitive) intracranial tumors in patients with one form of neurofibromatosis (NF), bilateral acoustic NF (NF-2), are bilateral acoustic neuromas. In this study, we present 2 cases with what appears to be another form of NF, namely, NF-1, who showed bilateral enlargement of the internal auditory canals. Bilateral caloric responses were remarkably impaired in both cases; a unilateral sensorineural hearing loss was observed in 1. No tumor was demonstrated in the enlarged internal auditory canals in either case. Although the pathophysiology remains uncertain, some patients with NF-1 show abnormal cochleovestibular function in association with bilateral internal auditory canal enlargement without an acoustic tumor.