[Surgical treatment for stage IV lung cancer].

OBJECTIVE: To find out the optimal surgical indication in stage IV lung cancer patients, we evaluated them retrospectively. METHODS & RESULTS: From 1975 to 2005, 62 patients without multiple metastases were operated at our hospital. The most common histological type was adenocarcinoma (67.7%). The metastatic lesions were lung (33.9%), brain (24.2%), liver, bone, adrenal gland and so on. The overall survival rate of stage IV lung cancer was 10.4% at 5-year. Five-year survival for patients with lung or brain metastasis who had no lymph node metastasis were significantly more superior than those with lymph node metastasis (p=0.0389, 0.0021). Four of 62 patients had 5-year survival. Two were lung and the others were brain and adrenal gland metastasis without lymph node metastasis. CONCLUSION: Stage IV lung cancer with lung or brain or adrenal gland metastasis without lymph node metastasis should be resected.

Identification of lactoferrin as an essential growth factor for human lymphocytic cell lines in serum-free medium.

A serum-free medium supplemented with growth factor(s) was devised to grow human lymphocytic cell lines. The medium was developed using human lymphocytic cell line, Bri 7 cells. In the process of constructing the medium, human lactoferrin was found to be an essential growth factor for the cell line. Human lactoferrin has higher growth stimulatory activity than human transferrin, and was sensitive to heat. Long-term cultivation of the cells was achieved in the defined medium supplemented with human lactoferrin only. The defined medium specifically supported the growth of various other human B- and T-lymphocytic cell lines but not the growth of various mouse lymphocytic cell lines. In lactoferrin-supplemented medium, the growth of some human cell lines were further stimulated by the addition of a combination of insulin, ethanolamine and selenium, or another combination of 2-mercaptoethanol and the above three factors. Bovine lactoferrin could be substituted for human lactoferrin.

Incorporation of isoleucine into protein by a soluble fraction from spermatozoa.

A heat-labile, non-dialyzable factor(s) in soluble fractions from porcine, bull, rabbit and cock spermatozoa was found to incorporate the radioactivity of [14C]isoleucine into a 95 degrees C CCl3COOH-insoluble fraction. The incorporation required ATP, Mg2+, casein and 2-mercaptoethanol. Trypsin and alpha-chymotrypsin inhibited the incorporation, while RNase A and DNase I did not. A mixture of 19 amino acids other than isoleucine had no effect on the incorporation. The reaction product was identified as protein. The incorporated moiety was the isoleucyl moiety of isoleucine and it retained a free alpha-amino group in the product protein. Some other characteristics of this incorporation are also described.

TRPV1 activation and induction of nociceptive response by a non-pungent capsaicin-like compound, capsiate.

Capsiate is a capsaicin-like ingredient of a non-pungent cultivar of red pepper, CH-19 sweet. To elucidate the mechanisms underlying the non-pungency of capsiate, we investigated whether capsiate activates the cloned capsaicin receptor, TRPV1 (VR1). In patch-clamp experiments, capsiate was found to activate TRPV1 expressed transiently in HEK293 cells with a similar potency as capsaicin. Capsiate induced nociceptive responses in mice when injected subcutaneously into their hindpaws with a similar dose dependency as capsaicin. These data indicate that the non-pungent capsiate is an agonist for TRPV1 and could excite peripheral nociceptors. In contrast to this, capsiate did not induce any significant responses when applied to the skin surface, eye or oral cavity of mice, suggesting that capsiate requires direct access to nerve endings to exhibit its effects. Capsiate was proved to have high lipophilicity and to be easily broken down in normal aqueous conditions, leading to less accessibility to nociceptors. Another highly lipophilic capsaicin analogue, olvanil, was similar to capsiate in that it did not produce irritant responses when applied to the skin surface, although it could activate TRPV1. Taken together, high lipophilicity and instability might be critical determinants for pungency and so help in understanding the effects of capsaicin-related compounds.

Accumulation enhancement of human monoclonal antibody HB4C5 to lung tumor xenografts by N-deglycosylation.

UNLABELLED: The fractional uptake of intact monoclonal antibodies by tumors is relatively low. Various methods to alter the molecular structure have been used to augment tumor uptake. These chemical manipulations, however, may alter the specificity of antibody binding. METHODS: Comparative studies of biodistribution, radioimmunoimaging and macroautoradiography in LC-6 xenografted mice were conducted with the 125I-labeled intact and N-terminal deglycosylated monoclonal antibodies to evaluate the effect on deglycosylation on antibody binding. RESULTS: The removal of N-glycosyl residues from this monoclonal antibody significantly enhanced specific localization of the radioactivity to the tumor, especially to its necrotic fraction. Nonspecific accumulation of radioactivity to the necrotic fraction of the tumor was excluded by biodistribution studies demonstrating selective accumulation of 125I-labeled monoclonal antibody after coadministration of 125I-monoclonal antibody (intact or N-deglycosylated) with 131I-labeled control IgM. CONCLUSION: The lung cancer-associated human monoclonal antibody HB4C5, which recognizes histone H2B as the antigen, accumulates specifically to the necrotic fraction of tumor. The uptake is enhanced by removal of N-terminal glycosyl residues from the antigen-binding site of the light chain.

Incorporation of glutamic acid into protein by a soluble system.

A heat-labile, non-dialyzable factor(s) in soluble fractions from Escherichia coli strains and Bacillus subtilis was found to incorporate the radioactivity of [14C]glutamic acid into 95 degrees C CCl3COOH-insoluble fraction. Incorporation catalyzed by a partially purified factor from E. coli B required ATP, Mg2+, tRNA, casein, carbonate, and 2-mercaptoethanol. A mixture of nineteen amino acids other than glutamic acid had no effect on the incorporation. Heparin, spermine and monovalent cations were inhibitory. Incorporation proceeded via glutamyl-tRNA. The incorporation from [14C]glutamyl-tRNA required Mg2+, casein, carbonate, and 2-mercaptoethanol, and there was no incorporation from [14C]aspartyl-tRNA. The reaction product was identified as protein. The incorporated moiety was the glutamyl moiety of glutamic acid and it retained a free alpha-amino group in the product protein. The incorporating factor of E. coli B was demonstrated to be glutamyl-tRNA synthetase.

Human cell clones, RSa and UVr-1, differing in their capability for UV-induced virus reactivation and phenotypic mutation.

UVr-1 is a human cell clone established as a variant with increased resistance to cell killing by ultraviolet light (UV, principally 254 nm wavelength) from a UV-sensitive cell clone, RSa. Both cells have been characterized to have much the same capacity of UV-induced DNA repair synthesis in whole cells, and the parent RSa cells were recently found to be hypermutable. In the present study UVr-1 cells were characterized in comparison RSa cells with respect to UV-induced virus reactivation and phenotypic mutation. Survival levels of UV-irradiated vaccinia virus and herpes simplex virus type 1 (HSV-1) were much the same in logarithmically proliferating UVr-1 and RSa cells. Correlated with these host cell reactivation levels, the same extent of UV-induced DNA repair replication synthesis was observed in isolated nuclei of the two cell clones. Enhancement of survival levels of UV-irradiated HSV-1 was detected when proliferating RSa cells were irradiated with UV prior to the virus infection. In contrast, this enhanced virus reactivation (EVR) was not detected in similarly irradiated and infected UVr-1 cells. As for phenotypic mutation frequencies assessed by the cloning efficiency of cells with increased resistance to ouabain cell killing (OuaR), OuaR mutants were not obtained from UVr-1 cells either with or without UV irradiation. When the proliferation of cells was synchronized, both EVR and OuaR mutations were detected in RSa cells irradiated with UV at any cell cycle phase, being greatest in the later half of the G1 phase. However, there was no detectable EVR or mutation in any phase of synchronous UVr-1 cells. The hypomutability of UVr-1 cells and hypermutability of RSa cells in a G1 cell cycle phase was also found even if 4-nitroquinoline 1-oxide was used as a mutagen or mutant cells with increased resistance to 6-thioguanine cell killing were estimated.

Molecular cloning of yeast cytochrome c-like polypeptide expressed in human lung carcinoma: an antigen recognizable by lung cancer-specific human monoclonal antibody.

We previously determined the amino acid sequence to the epitope (ATLFKTR) of cytochrome c from Candida krusei, which is cross-reactive to the lung cancer-specific human monoclonal antibody HB4C5. Here we report that an antigen messenger RNA, which codes for a structure similar to the cytochrome c epitope, is expressed in the human lung adenocarcinoma A549. Sequencing analysis has revealed that this messenger RNA encodes a novel 190 amino acid polypeptide of 21-kDa containing an amino acid sequence (ALLFFT) similar to the cytochrome c epitope, although the total messenger RNA sequence is apparently different from the cytochrome c messenger RNA. Western analysis indicated that an antibody-recognizable 21-kDa antigen which has the same molecular weight as the predicted polypeptide is expressed in the A549 adenocarcinoma. The in vitro translated product of the antigen messenger RNA and synthesized ALLFFT peptide were both shown to be reactive with the monoclonal antibody, indicating that this protein contains the epitope which enables A549 cells to specifically react with the antibody. The antigen mRNA was not expressed in non-transformed fibroblasts, suggesting that the antigen mRNA expression was associated with cellular transformation. Also in part of the antigen nucleotide sequence, there was a segment that had about 90% homology to the long terminal repeat sequence (no. 297-475) of the human endogenous retrovirus HERV-K10, which was related to the mouse mammary tumor virus.

Class-switching of the IgM type anti-adenocarcinoma human antibody HB4C5 into an IgG1 type resulted in the loss of the antigen binding ability.

The heavy and light chain genes (mu and lambda) of the human anti-adenocarcinoma monoclonal antibody HB4C5 (MAb-C5) were cloned from the human-human hybridoma cell line HB4C5 and co-expressed in Chinese hamster ovary (CHO) and COS-7 cells. ELISA assay and the RI imaging of the cancer tissue xenografted into nude mice showed that the recombinant MAb-C5 retained the original antigen binding activity. We then replaced the IgM constant region of the MAb-C5 heavy chain gene with the human IgG1 constant region gene and co-expressed it with the light chain gene. This recombination was confirmed by a complete DNA sequencing and Western-blot analysis. Despite the fact that the DNA sequence, the expressed protein size, and the assembly of heavy and light chains were indicated to be normal, the IgG1 type MAb-C5 could not bind to the original antigen. This result suggest that this antibody alters its antigen binding property upon class-switching.

Elevated expression levels of the 14-3-3 family of proteins in lung cancer tissues.

Immunochemical staining of lung cancer sections with a murine monoclonal anti-14-3-3 antibody showed a sharp discrimination of the cancer tissue from neighboring normal counterparts in 88 of 121 primary lung cancer tissue specimens of all four major lung cancer histologies; specifically, 32 of 48 adenocarcinomas, 36 of 44 squamous cell carcinomas, 10 of 13 large cell carcinomas, and 10 of 16 small cell carcinomas, respectively, were stained positively. Sets of the 10,000 x g supernatants of normal and cancerous lung tissue homogenates, each set prepared from surgically dissected tissues of the cancer and its surrounding normal part, were assayed for 14-3-3 proteins by the sandwich enzyme-linked immunosorbent assay using two different monoclonal antibodies to 14-3-3 proteins. The results of the assay demonstrated 7.2 times higher 14-3-3 protein content in the lung cancer tissue (378 +/- 200 ng ml-1) as compared with the normal lung (54 +/- 35 ng ml-1). These results indicate that the 14-3-3 family of proteins can be an effective marker for lung cancer diagnosis such as sputum cytodiagnosis and that 14-3-3 proteins might be involved in the development of lung cancers.

Cytokeratin 8 and 19 as antigens recognized by adenocarcinoma-reactive human monoclonal antibody AE6F4.

The human monoclonal antibody (MAb) AE6F4 is secreted by a human-human hybridoma line established from the in vitro immunization of normal human peripheral blood lymphocytes with the human lung adenocarcinoma cell line, A549. This MAb is strongly reactive to lung cancer tissues. In the previous study, the antigens recognized by the MAb AE6F4 were purified from A549 cells and identified as 14-3-3 protein and 31 kDa cytosolic phospholipase A2 (cPLA2). The MAb AE6F4 also binds two kinds of antigens (53 kDa and 40 kDa), which are not related to 14-3-3 protein or 31 kDa cPLA2, in the human breast adenocarcinoma cell line, MCF-7. We purified a 38 kDa antigen, which is a degradation product of 53 kDa antigen from breast adenocarcinoma MCF-7 cells using ion-exchange and hydroxyapatite column chromatography. Two partial amino acid sequences of the purified 38 kDa antigen showed 95-100% homology to human cytokeratin 8 (CK8). Two-dimensional gel electrophoresis and immunoblot analysis of intermediate filament fraction separated from MCF-7 cells demonstrated that the 53 kDa and 40 kDa antigens were CK8 and CK19, respectively. Antigenic determinants on CK8 and CK19 recognized by the MAb AE6F4 were resistant to sodium periodate treatment, although antigenic determinant on 31 kDa antigen (14-3-3 protein and(or) cPLA2) was sensitive to this treatment. These results suggest that the MAb AE6F4 reacts with both carbohydrate and peptide antigenic determinants.

Recombinant light chain of human monoclonal antibody HB4C5 as a potentially useful lung cancer-targeting vehicle.

Recombinant lambda light chain of lung cancer-reacting human monoclonal antibody HB4C5 was expressed in Escherichia coli. Expression in bacteria ensured the generation of homogeneous light chain species devoid of activity-hampering N-linked glycosylation usually found in the light chain CDR-1 of HB4C5. Molecular engineering was also employed to eliminate the C-terminal two amino acid residues, i.e., Cys and Ser, to prevent the formation of lambda light chain dimers which are less reactive than the monomeric form. The lambda light chain was overexpressed in E. coli as inclusion bodies, which were solubilized, refolded, and treated with Aeromonas proteolytica aminopeptidase to remove the N-terminal Met with subsequent natural cyclization of the penultimate Gln residue to pyroglutamate, the same N-terminal end as that of naturally occurring lambda light chain in HB4C5. Monomeric recombinant lambda light chains, both before and after removal of the N-terminal Met residue, were 40 times more immunoreactive than the parent HB4C5. The immunostaining of lung cancer tissue sections with the recombinant lambda light chain indicated cancer-specific reactions to all specimens of adenocarcinoma, squamous cell carcinoma and large cell carcinoma histologies, but did not react with small cell carcinoma. Tumor radioimmunoimaging experiments in LC6 (lung squamous cell carcinoma line)--xenografted nude mice by the i.p. injection of 125I-labeled recombinant lambda light chain and 125I-labeled human lambda light chain control gave tumor-specific and recombinant lambda light chain-dependent images on day 5 postinjection, and images were also detectable on day 3. Biodistribution studies with 125I-labeled recombinant lambda light chain demonstrated that the lambda light chain could penetrate better into the tumor sites, both at the necrotic and solid parts of the xenograft, as compared to our previous results with 125I-labeled HB4C5 which could localize to the necrotic part only. These results suggest that the recombinant lambda light chain is potentially useful as a lung cancer-targeting vehicle, for such as radioimmunoimaging and radioimmunotherapy, with least possible adverse immunogenic effects.

Progress with inactivated poliovirus vaccines derived from the Sabin strains.

To produce a safe and effective inactivated poliovirus vaccine (IPV), we have developed S-IPV using Vero cells infected with the Sabin strains in a semi-production scale. All production steps including virus culture on microcarrier beads were highly reproducible. Mean recovery percents of infectious viruses or D-antigens during all processes for concentration, purification and inactivation were 30-50% in the three types. The S-IPV potency was adjusted for D-antigen content as determined by in-house ELISA and was comparable to WHO reference IPV derived from the virulent strains in immunogenicity tests in rats. Antibody development in more than 30 seronegative infant volunteers after two shots of S-IPV at four-week interval were 100% without notable adverse reactions. The mean antibody titres (log2) to Sabin 1, 2 and 3 viruses were 11.1, 8.3 and 8.9, respectively. The antibodies neutralized the Mahoney, MEF-1, and Saukett virulent strains with slightly inferior titres to those of the Sabin strains. D-antigens for each type of S-IPV were stable at 4 degrees C without any significant decrease over more than two years.

[Poliovirus (Sabin strain) multiplication in human intestinal tract after oral polio vaccination].

Fecal specimens from a baby vaccine were collected every day from 1 to 51 days after primary vaccination and from 0 to 15 days after secondary vaccination. Polioviruses were isolated with GMK-2 cell line from 10% emulsion of the feces and titrated the virus contents in the emulsion of the feces. The isolated viruses were tested the reproductive capacity at 39.0 degrees C and 39.5 degrees C by the plaque method with primary cynomologous monkey kidney cells. Viruses were isolated from the feces during 28 days for type 1, 39 days for type 2 and 36 days for type 3 after primary vaccination, however, only type 1 viruses were isolated during 7 days after secondary vaccination. The multiplication of type 3 viruses in the intestine were increased after diminished the multiplication of type 1 and type 2. In plaque formation capacity at 39.0 degrees C and 39.5 degrees C, the isolates had shown to differ clearly among the types of poliovirus. After primary vaccination, type 1 isolates were not produced the plaques at 39.0 degrees C and 39.5 degrees C. Although type 2 isolates were not formed the plaques until the 14th day at 39.5 degrees C, the plaque formation capacity of the these isolates were increased gradually i.e.; on the 20th day (10(0.88) PFU/ml), the 26th day (10(2.00) PFU/ml) and the 39th day (10(2.63) PFU/ml) at 39.5 degrees C, and all of type 2 isolates tested were showed the plaque formation capacity (10(2.88 approximately 10(3.76) PFU/ml) at 39.0 degrees C. Type 3 isolates were formed plaques at 39.0 degrees C and 39.5 degrees C from the 7th day. After the secondary vaccination, type 1 isolates (7th day) was a little changed them. Neutralizing antibody titers were shown that type 1 was 320, type 2 was 110 and type 3 was 60 after 1 year of the second administration. These titers were closely similar the geometric mean titers of 2 year old babies in Japan.

[Detection of Chlamydia trachomatis antigen for diagnosis of respiratory infection I. Cross reaction with bacteriae consisting normal flora respiratory tract].

We examined the cross reaction of Chlamydia trachomatis (C. trachomatis) by antigen detection method using EIA against the aerobic bacteria which colonize in the respiratory tract of children. Chlamydiazyme showed a cross reaction among 5 out of 7 species of Gram negative bacteria (= 71.4%) and Neisseria and Branhamella catarrhalis showed a cross reaction even in low concentrations of 1 X 10(3-4) CFU/ml. There were no Gram positive bacteria which showed cross reaction. IDEIA Mark III did not react to Gram negative bacteria even in high concentrations of 1 X 10(7-10) CFU/ml. One strain of coagulase positive staphylococcus (CPS) showed positive at a concentration of 1 X 10(8): however, other Gram positive bacteria including two other strains of CPS exhibited no cross reaction. The detectable concentration of C. trachomatis (D/UW 3 strain) using IDEIA Mark III was 1 X 10(3) IFU/ml. Low cross reaction rate and sensitivity suggest IDEIA Mark III is preferable for diagnosis of respiratory infection of C. trachomatis though further clinical studies are necessary.

[Polypeptide profiles of Chlamydia pneumoniae strains isolated from Japan and western blot analysis using patients' sera].

Polypeptide profiles were compared among Chlamydia pneumoniae strain of AC-43, which was isolated from a Japanese child, and other strains of C. pneumoniae. There was no distinctive differences among these strains. All strains showed marked band on 39.5 KDa area, which is equivalent with the major outer membrane protein (MOMP). Patients infected with C. pneumoniae were analyzed by Western blot using AC-43. Reaction for 98, 68, 60 KDa proteins were recognized among sera which showed positive results for anti-C. pneumoniae IgG antibody. There were two family members from whom C. pneumoniae was not isolated and anti-C. pneumoniae IgG antibody was negative. Sera obtained from them also showed weak reaction on 98 KDa protein area. Strong reaction on MOMP area was demonstrated in one patient from whom C. pneumoniae was isolated. Monoclonal antibody produced from AC-43 strain showed no reaction against homologous antigen. Reaction to 98 KDa protein was detected in all of the patients infected with C. pneumoniae. However, recognition to MOMP of C. pneumoniae seems to be different among patients. This result might suggest the presence of subtype among C. pneumoniae strains.

[A report of 4 cases from whom Chlamydia pneumoniae was isolated].

Chlamydia pneumoniae (C. pneumoniae) was isolated from respiratory tract of 4 girls. Case 1 is 5-year-old, diagnosed as pneumonia, had had a prolonged productive cough. She was treated with RKM with improvement of symptoms, however, C. pneumoniae was isolated repeatedly and was not deleted. Neither anti-mycoplasmal nor viral antibodies were positive and no significant pathogens were cultured from nasophrayngeal swabs. Case 2, the sister of case 1, is a 3-year-old girl with acute bronchitis treated with EM. C. pneumoniae was negative on the tenth day after treatment. Case 3, a 5-year-old girl, had a fever and was diagnosed as bronchopneumonia with a mild attack of bronchial asthma. She was initially treated with CFIX followed by therapy including EM. Her symptoms had disappeared after treatment and anti-mycoplasmal antibody was 1:320. Case 4 was an asymptomatic carrier of C. pneumoniae. Specimen was obtained at regular health examinations of junior high school. C. pneumoniae was isolated from a 14-year-old girl without respiratory symptoms. Clinical figures of C. pneumoniae infections varies from asymptomatic carrier to pneumonia. Pathogens other than C. pneumoniae could modify symptoms of infections. Precise examinations of these cases would establish a proper management of a C. pneumoniae infection.

[Antibacterial effects of cacao mass on enterohemorrhagic Escherichia coli O157:H7].

The antimicrobial activities of aqueous cacao mass extract against enterohemorrhagic Escherichia coli (EHEC) O157:H7 006 strain were studied. Hot water extract of cacao mass (cocoa extract) was shown to inhibit the growth of EHEC O157:H7 006 strain in PBS or CAYE medium. In addition, the production of verotoxins (types 1 and 2) of EHEC O157:H7 006 strain was significantly inhibited by 8.0% cocoa extract. The cocoa extract did not neutralize the cytotoxity of verotoxins, but had inhibitory effect on adhesion of verotoxins to the target Vero cells. These results demonstrate that cacao mass has antimicrobial effects on EHEC O157:H7.

Assessment of Chlamydia trachomatis-specific IgA and IgG serum antibodies in genitourinary Chlamydia trachomatis infection--comparative study between HITAZYME and IPAzyme.

We assessed the C. trachomatis antibody assay kit HITAZYME (Hitachi Chemical Co., Ltd.) using clinical specimens. This kit is based on an enzyme immunoassay (EIA) which utilized purified Chlamydia trachomatis outer membrane antigen as the solid phase antigen. Twenty-nine untreated male urethritis patients, 816 pregnant housewives, 188 cervicitis patients, and 76 pelvic inflammatory disease patients were tested. Agreement between the HITAZYME test and antigen detection in infected area was assessed, and a comparison was made with IPAzyme (a commercially available indirect immunoperoxidase assay kit). 1) Summary of HITAZYME and IPAzyme IgA: Agreement between the two assays was relatively good, i.e., 82.6% (916/1109). However, 5.5% (61/1109) were HITAZYME (-), IPAzyme (+), and 11.9% (132/1109) were HITAZYME (+), IPAzyme (-). Thus, in quite a few cases the results did not agree. IgG: Agreement between the two assays was 73.7% (817/1109). Agreement was relatively low, 24.4% (271/1109) were HITAZYME (-), IPAzyme (+). 2) In the cases of disagreement, more specific Western blot analysis was performed to check the reactivity of the anti-C. trachomatis antibody. When IgA was used, agreement between HITAZYME and Western blot analysis was 69.6% (16/23), and agreement between IPAzyme and Western blot analysis was 30.4% (7/23), whereas when IgG was used, agreement between HITAZYME and Western blot analysis was 80.0% (12/15), and agreement between IPAzyme and Western blot analysis was 20.0% (3/15). There was significantly greater agreement with HITAZYME than with IPAzyme. In other words, HITAZYME had greater specificity when reacted with C. trachomatis antigen than IPAzyme. 3) The IgA antibody-positive rate in antigen (+) cases (male urethritis: 72.7%, pregnant housewives: 65.7%, cervicitis: 70.3%, pelvic inflammatory disease: 70.0%) was significantly (p < 0.01) higher than in antigen (-) cases (male urethritis: 16.7%, pregnant housewives: 13.6%, cervicitis: 22.6%, pelvic inflammatory disease: 30.4%). Therefore, IgA antibody can serve as a suitable indicator for active infection. 4) The IgG antibody-positive rate in antigen (-) female cases was 15.5% using HITAZYME and significantly (p < 0.01) lower than with IPAzyme. HITAZYME had greater specificity than IPAzyme. In conclusion, HITAZYME has relatively good sensitivity and specificity. Moreover, since it is an EIA assay, it allows objective evaluation of results. It permits processing of a large number of specimens because it is easy to perform. Thus, HITAZYME is a superior antibody assay for C. trachomatis. It can be used when antigen tests are difficult to perform. It is strongly anticipated that HITAZYME will be able to be used clinically as a screening test.

[Clinical observation of non-gonococcal urethritis].

Sixty-one male urethritis cases, 28 gonococcal urethritis and 33 nongonococcal urethritis (NGU), were out-patients at the Department of Urology, Asahi General Hospital, during the 4 months, Oct. 1, 1984-Jan. 31, 1985. Thirteen of the 33 NGU patients (39.4%) were infected with C. trachomatis (CT). The efficacy of Doxycycline and the route of infection was studied in the cases of CT-positive CT-negative-NGU. CT infection from prostitutes was not so frequent as in the other pathogen infection of NGU. The efficacy of Doxycycline (100 mg b.i.d. for 2 weeks) against CT infection was excellent in the disappearance of subjective complaints and that of white blood cells in the urethral discharge in the CT positive-NGU group (13/13), in comparison with CT negative-NGU group (7/18). The efficacy of Doxycycline against CT was also confirmed from the follow-up study by the isolation of CT and by detection of CT antigen from urethral swabs using FITC conjugated monoclonal antibody against CT antigen.