Clinical outcomes of tracheoesophageal diversion for intractable aspiration.

OBJECTIVES: The purpose of the present study was to examine the clinical outcomes of using tracheoesophageal diversion for preventing intractable aspiration. METHOD: We retrospectively reviewed 25 patients who underwent tracheoesophageal diversion from 2003 to 2009 at our hospital (median age, 25 years; range, 0-78 years). End-to-side anastomosis was used in 16 cases and side-to-side anastomosis was used in 9. RESULTS: The average operative time was 141 minutes for end-to-side anastomosis and 191 minutes for side-to-side anastomosis. Peri-operative complications were observed in only two (8 per cent) cases: one with infection and one with haematoma. No fistulas were observed. Aspiration was prevented in all cases, but the nutritional route depended on the swallowing function of the patient. Oral feeding was the main nutritional route after surgery in only four patients (16 per cent). CONCLUSION: This procedure is well suited to patients who lack speech communication and are at high risk of aspiration.

New technique for laryngomicrosurgery: narrow band imaging-assisted video-laryngomicrosurgery for laryngeal papillomatosis.

OBJECTIVE: In the present report, we describe the use of narrow band imaging during video-laryngomicrosurgery for laryngeal papillomatosis. CASE REPORT: It is difficult to peri-operatively locate all the superficial papillomatous lesions when the disease is widespread, which then results in tumor recurrence. Therefore, we have constructed a narrow band imaging video-laryngomicrosurgery system, which we have used for two cases of laryngeal papillomatosis. CONCLUSION: Our narrow band imaging-assisted video-laryngomicrosurgery system to visualise superficial laryngeal papillomatosis more clearly.

Stimulating effect of Japanese herbal (kampo) medicine, hochuekkito on upper respiratory mucosal immune system.

Japanese herbal (Kampo) medicine, Hochuekkito (Bu-Zhong-Yi-Qi-Tang in Chinese, TJ-41) and Juzentaihoto (Shi-Quan-Da-Bu-Tang in Chinese, TJ-48) are well-known Kampo formulas used as tonic. Although these medicines have separately been applied to the patients clinically depending on their symptoms, the differences of the pharmacological activities for these medicines have not been fully understood. TJ-48 and TJ-41 were compared for their effects on antibody response in upper respiratory mucosal immune system in vivo. Oral administration of TJ-41 (100 mg kg(-1) per day) to early aged BALB/c mice, which were nasally sensitized with influenza hemagglutinin vaccine, significantly enhanced influenza virus-specific IgA and IgG antibody titers in nasal cavity and sera, respectively. However, oral administration of TJ-48 (100 mg kg(-1) per day) failed to show the enhancing activity. TJ-41 increased not only influenza virus-specific IgA antibody titer but also total IgA antibody titer in nasal cavity. The stimulating activity of TJ-41 disappeared after treatment with methotrexate. The present study strongly suggests that TJ-41 can stimulate the mucosal immune system of upper respiratory tract, and results in enhancement of antigen-specific antibody response in upper respiratory mucosal and systemic immune systems.

Screening of Thai medicinal plant extracts and their active constituents for in vitro antimalarial activity.

To discover antimalarial substances from plants cultivated in Thailand 80%-EtOH extracts from selected plants were screened for in vitro antimalarial activity against the drug resistant K1 strain of Plasmodium falciparum. In total, 86 Thai medicinal plant samples representing 48 species from 35 genera in 16 families were screened and two species (Polyalthia viridis and Goniothalamus marcanii) were found to show notable antimalarial activity (IC50: 10.0 and 6.3 microg/mL). Marcanine A and 16-hydroxycleroda-3,13(14)Z-dien-15,16-olide were identified as the respective major active constituents in P. viridis and G. marcanii, respectively.

The presence of natural human antibodies reactive against pharmacologically active pectic polysaccharides from herbal medicines.

Direct ELISA was performed using normal human sera and human colostrum, to analyse the presence of antibodies which react with pharmacologically active pectic polysaccharides isolated from plants used in traditional Japanese herbal (Kampo) medicine. All sera and colostrum were shown to contain IgM, IgG, IgA and secretory IgA class antibodies which react with the active pectic polysaccharides to different degrees. The reacting IgG antibody in normal human serum recognized the ramified regions (rhamnogalacturonan core with carbohydrate side-chains) of the pharmacologically active pectic polysaccharides as the active sites for complement-activating activity. Correlation analysis indicated that a significant and positive correlation was observed between reactivity with the reacting antibody of IgG class and the degree of complement-activating activity of the active polysaccharides. The reacting IgG class antibody, which was purified from normal human serum by affinity chromatography on bupleuran 2IIc (a pharmacologically active pectic polysaccharide from the roots of Bupleurum falcatum)-immobilized Sepharose, showed cross-reactivity not only with some other pharmacologically active pectic polysaccharides from other medicinal herbs but also with autoantigens such as single-strand DNA, myosin and tublin from mammals.

Induction of NGF synthesis in astrocytes by onjisaponins of Polygala tenuifolia, constituents of kampo (Japanese herbal) medicine, Ninjin-yoei-to.

The effect of a kampo medicine, Ninjin-yoei-to (NYT; Ren-shen-yang-rong-tang in Chinese) on nerve growth factor (NGF) secretion from the cultured rat astrocytes was examined in vitro. When rat embryo astrocytes were cultured in the presence of NYT for 24 h, the amount of NGF in the medium was significantly increased in a dose dependent manner. Among 14 kinds of component herbs in NYT, the roots of Polygala tenuifolia and roots of Panax ginseng extracts increased NGF levels from the astrocytes. Saponin fraction from the roots of P. tenuifolia enhanced the production of NGF, however phenolic glycoside fraction showed no effect. Onjisaponins A, B, E, F and G as major saponins of the root of P. tenuifolia strongly increased the NGF level, whereas ginsenosides Rb1 and Rg1 did not affect the NGF level. Onjisaponin F also induced ChAT mRNA level in rat basal forebrain cells. These results indicate the possibility that NYT and/or onjisaponins in P. tenuifolia may have potential therapeutic effects for the treatment of Alzheimer disease patients.

Intestinal immune system modulating polysaccharides in a Japanese herbal (Kampo) medicine, Juzen-Taiho-To.

A commercially available dried extract (TJ-48) of Japanese herbal (Kampo) prescription, Juzen-Taiho-To has been found to enhance functions of Peyer's patch cells (intestinal immune system modulating activity) in vitro and in vivo. When TJ-48 was fractionated, the dialyzable fraction (F-3) and the polysaccharide fraction (F-5) expressed the in vitro activity. Oral administration of F-5 (150 mg/kg/day) to mice showed the intestinal immune system modulating activity. When the galacturonan moiety of pectic polysaccharides in F-5 was degraded enzymatically by endo-polygalacturonase, the digestion products significantly increased the activity of F-5. Purification the polygalacturonase-digested F-5 indicated that the active substances were composed mainly of the enzyme-resistant or undigestable polysaccharide molecules. Gel filtrations and anion-exchange chromatographies of F-5 gave 12 kinds of polysaccharides, and among them, 7 polysaccharides had significant intestinal immune system modulating activity. Component sugar analysis suggests that some active polysaccharides are grouped into pectic polysaccharides containing arabinogalactan or heteroglycan chains. Single radial gel diffusion analysis using beta-D-glucosyl-Yariv antigen indicated that some of the active polysaccharides comprised arabino-3,6-galactan moiety, and this moiety was suggested to play an important role partly for expression of the activity by single linear regression analysis between degree of the activity and reactivity with beta-D-glyucosyl-Yariv antigen.

Onjisaponins, from the root of Polygala tenuifolia Willdenow, as effective adjuvants for nasal influenza and diphtheria-pertussis-tetanus vaccines.

Active substances from hot water extracts from 267 different Chinese and Japanese medicinal herbs were screened for mucosal adjuvant activity with influenza HA vaccine in mice. The extract from the root of Polygala tenuifolia was found to contain potent mucosal adjuvant activity. The active substances were purified and identified as onjisaponins A, E, F, and G. When each onjisaponin (10 microg) was intranasally (i.n.) inoculated with influenza vaccine (10 microg) in mice, serum hemagglutination-inhibiting (HI) antibody titers increased 3-14 times over control mice administered vaccine alone after 4 weeks. When each onjisaponin (10 microg) was i.n. inoculated with the vaccine (10 microg) followed by i.n. vaccination of the vaccine alone after 3 weeks, serum HI antibody titers increased 27-50 fold over those mice given i.n. vaccinations without onjisaponins. These same conditions also significantly increased nasal anti-influenza virus IgA antibody titers. Two inoculations with onjisaponin F (1 microg) and influenza HA vaccine (1 microg) at 3 weeks intervals, significantly increased serum HI antibody and nasal anti-influenza virus IgA and IgG antibody titers after only 1 week over mice given HA vaccine alone after the secondary vaccination. Intranasal vaccination with onjisaponin F inhibited proliferation of mouse adapted influenza virus A/PR/8/34 in bronchoalveolar lavages of infected mice. Separate intranasal vaccinations with onjisaponins A, E, F, and G (10 microg) each and diphtheria-pertussis-tetanus (DPT) vaccine (10 microg) of mice followed by i.n. vaccination with DPT vaccine alone after 4 weeks showed significant increases in serum IgG and nasal IgA antibody titers after 2 weeks following secondary vaccination over mice vaccinated with DPT vaccine alone. All onjisaponins showed little hemolytic activity at concentrations up to 100 microg/ml. The results of this study suggest that onjisaponins may provide safe and potent adjuvants for intranasal inoculation of influenza HA and DPT vaccines.

Acidic polysaccharides from rhizomes of Atractylodes lancea as protective principle in Candida-lnfected mice.

Prophylactic effects upon imunnosuppressed mice lethally infected by Candida albicans were examined in fractions prepared from a constituent herb of Juzen-taiho-to (TJ-48, Si-Quan-Da-Bu-Tang), rhizomes of Atractylodes lancea DC. The oral administration of water extract obtained from a residue after MeOH extraction of rhizomes significantly prolonged the survival period of the infected mice at a dose of 140 mg/kg/day compared with control mice, while the MeOH extract did not. In the crude polysaccharide fraction (F-2) obtained by EtOH precipitation of the water extract, a significant life-prolonging effect was observed by the administration of 70 mg/kg/day. F-2 was further fractionated, and the resulting strongly acidic polysaccharide fraction, F-2-2, had a protective effect at a dose of 17.5 mg/kg/day. This fraction mainly consisted of acidic pectic polysaccharides containing about 80% galacturonic acid. The protective activity of F-2-2 was lost by periodate oxidation, but not by protease digestion, suggesting that the polysaccharide component of F-2-2 plays a major role in the protective activity against Candida-infected mice.

[Prophylactic effect of pirarubicin (THP) on postoperative recurrence of superficial bladder cancer in terms of intravesical retention time].

In order to determine the modality of prophylactic intravesical instillation of pirarubicin (THP = tetrahydropyranyladriamycin) following transurethral resection (TUR) of superficial bladder cancer, a prospective randomized study was performed. A total of 79 patients were randomized into "2-hour instillation" (A), "5-min instillation" (B) and "control" (C) groups. Prophylactic efficacy and side effects were analyzed in each group. In groups A and B, 20 mg of THP was first dissolved in 10 ml of distilled water, adjusted to 40 ml with saline and was administered intravesically once a week for 10 weeks, starting from 1 week after TUR. The recurrence-free rate was calculated in 65 evaluable patients. The one-year recurrence-free rate was 70.2% in group A, 62.8% in group B and 52.1% in group C. The one-year recurrence-free rate was significantly higher in group A than in group C. Adverse effects were observed in 21.4% of the patients in group A and 40.7% in group B. There was no significant difference in the occurrence rate of side effects between these two groups. Taking the prophylactic efficacy and side effects into consideration, "2-hour instillation" seemed to be better than "5-min instillation".

[Clinical efficacy of treatment with low-dose flutamide in maximum androgen blockade therapy].

To prevent treatment withdrawal due to flutamide-induced liver dysfunction, we performed maximum androgen blockade (MAB) therapy by combining a luteinizing hormone-releasing hormone agonist or orchiectomy with low-dose flutamide (125 mg x 2/day) in patients with prostate cancer. In this study, the efficacy, adverse effects such as hepatotoxicity, and compliance were compared retrospectively between 35 patients who received low-dose flutamide therapy (1995-1999) and 27 patients who received flutamide at its ordinary dose (125 mg x 3/day). No significant difference was observed in the response rate (> or = PR) as determined from the prostate-specific antigen parameter (p = 0.6211) or the incidence of hepatotoxicity based on the aspartate aminotransferase and alanine aminotransferase levels. However, flutamide withdrawal due to liver dysfunction was less frequent in the low-dose group (2.9%) than in the ordinary dose group (18.5%) (p = 0.0386). MAB therapy using low-dose flutamide is expected to prevent the reduction in the compliance due to side effects and to improve the long-term prognosis in patients with prostate cancer, who are mostly elderly individuals.

Lignin-carbohydrate complexes: intestinal immune system modulating ingredients in kampo (Japanese herbal) medicine, juzen-taiho-to.

An extract preparation (TJ-48) of a Japanese herbal (Kampo) medicine, Juzen-Taiho-To has been found to show an enhancing activity on proliferation of bone marrow cells mediated by Peyer's patch cells (intestinal immune system modulating activity) (1). When TJ-48 was fractionated by MeOH- and water-extractions, EtOH precipitation and dialysis, water-soluble dialyzable (F-3) and polysaccharide fractions (F-5) both showed a significant intestinal immune system modulating activity in vitro; other fractions had no activity. Oral administration of F-3 (150 mg/kg) also showed the intestinal immune system modulating activity in C3H/HeJ mice. Two active substances were purified by gel filtration from F-3. Chemical analysis indicated that the active substances comprised lignin as well as a carbohydrate consisting mainly of arabinose, galactose, glucose and galacturonic acid. Activity was significantly reduced not only by NalO4 oxidation but also by NaClO2 treatment. Standard lignin showed no activity. Carbohydrate and lignin in the active substances were co-eluted by hydrophobic-interaction chromatography on phenyl-Sepharose, and the stimulating effect and lignin content of the substances did not change even after the hydrophobic-interaction chromatography. These results suggest that lignin-carbohydrate complexes are involved in intestinal immune system modulation by TJ-48.

Orally administered decoction of Kampo (Japanese herbal) medicine, "Juzen-Taiho-To" modulates cytokine secretion and induces NKT cells in mouse liver.

The effects of orally administered decoction of Juzen-Taiho-To (JTT; Si-Quan-Da-Bu-Tang in Chinese) on cytokine production in hepatic lymphocytes were studied in mice. JTT was found to increase interferon gamma (IFN-gamma), as well as interleukin-4 (IL-4), IL-5 and IL-6 secretion from stimulated hepatic lymphocytes, whereas IL-2 secretion was reduced. The number of IFN-gamma- and IL-4-spot forming cells (SFC) were not changed by administration of JTT. These results suggest that modulation of cytokine secretion by JTT might not be due to changes in the number of cytokine secreting cells within liver lymphocytes. CD4/CD8 ratio and alphabeta/gammadelta T cell receptor (TCR) ratio in hepatic lymphocytes were not changed. However, flow cytometric analysis revealed that the population of CD3 positive intermediate cells in NK positive cells (NKT cells) was increased after oral administration of JTT. The population of CD3int IL-2Rbeta+ cells was also increased. The induction of NKT cells by JTT was reduced by injection of 2-chloroadenosine. JTT enhanced transcription of IL-12 mRNA in liver. From these results, it may be concluded that a rise in NKT cell population contributes, at least partially, to the modulating effect of JTT on cytokine production in liver lymphocytes, and macrophages. The production of IL-12 in liver may also contribute to this NKT induction.

PcpA, which is involved in the degradation of pentachlorophenol in Sphingomonas chlorophenolica ATCC39723, is a novel type of ring-cleavage dioxygenase.

The pentachlorophenol (PCP) mineralizing bacterium Sphingomonas chlorophenolica ATCC39723 degrades PCP via 2,6-dichlorohydroquinone (2,6-DCHQ). The pathway converting PCP to 2,6-DCHQ has been established previously; however, the pathway beyond 2,6-DCHQ is not clear, although it has been suggested that a PcpA plays a role in 2, 6-DCHQ conversion. In this study, PcpA expressed in Escherichia coli was purified to homogeneity and shown to have novel ring-cleavage dioxygenase activity in conjunction with hydroquinone derivatives, and converting 2,6-DCHQ to 2-chloromaleylacetate.

Isolation and characterization of a thermophilic Bacillus sp. JF8 capable of degrading polychlorinated biphenyls and naphthalene.

Bacillus sp. strain JF8, which was isolated from compost, utilizes naphthalene and biphenyl as carbon sources at 60 degrees C. Biphenyl grown cells of strain JF8 barely degraded naphthalene while naphthalene grown cells did not degrade p-chlorobiphenyl, suggesting the existince of two independent degradation pathways. Isolation of JF8N, a mutant strain which can not utilize biphenyl as a carbon source while retaining the ability to utilize naphthalene, supports this hypothesis. Biphenyl grown cells of strain JF8 can degrade several polychlorinated biphenyl congeners including tetra- and pentachlorobiphenyl. bph and nah probes from mesophilic organisms failed to hybridize to strain JF8 DNA.

B-cell proliferation activity of pectic polysaccharide from a medicinal herb, the roots of Bupleurum falcatum L. and its structural requirement.

Pectic polysaccharide fraction (BR-2) containing pharmacologically active pectic polysaccharide, bupleuran 2IIc, which was prepared from a medicinal herb, the roots of Bupleurum falcatum L., was administered orally to C3H/HeJ mice for 7 consecutive days. Proliferative responses of spleen cells were enhanced in the presence of the purified pectic polysaccharide, bupleuran 2IIc, but another B-cell mitogen, lipopolysaccharide (LPS) did not give a similar effect. In vitro studies using spleen cells showed that bupleuran 2IIc also stimulated lymphocytes, depleted of adherent cells or T cells. Bupleuran 2IIc treatment increased subpopulation of CD25+ and surface immunoglobulin M-positive (sIgM+) lymphocytes. Non-specific immunoglobulin secretion of spleen cells treated with bupleuran 2IIc was increased according to the culture time, and coexistence of interleukin-6 (IL-6) enhanced the secretion more than that of bupleuran 2IIc alone. These results suggest that bupleuran 2IIc proliferates B cells in the absence of macrophages, and the resulting activated B cells are then induced into antibody-forming cells in the presence of IL-6. Among the structural region of bupleuran 2IIc, ramified region (PG-1), which consists of rhamnogalacturonan core rich in neutral sugar chain, showed the potent mitogenic activity suggesting it to be an active site. Mitogenic activity of bupleuran 2IIc was reduced in the presence of antipolysaccharide antibody (antibupleuran 2IIc/PG-1-IgG), which recognizes the ramified region of bupleuran 2IIc as the antigenic epitope. Mitogenic activity of bupleuran 2IIc was also reduced by the addition of beta-d-GlcpA-(1-->6)-beta-d-Galp-(1-->6)-d-Galp or beta-d-GlcpA-(1-->6)-d-Galp, which are a part of the epitopes of antibupleuran 2IIc/PG-1-IgG. These results suggest that the epitopes in bupleuran 2IIc act as active sites of the polysaccharide during mitogenic activity.

[ACTH-independent bilateral macronodular adrenocortical hyperplasia (AIMAH): a case report].

We report a case of adrenocorticotropic hormone (ACTH)-independent macronodular adrenocortical hyperplasia (AIMAH). A 54-year-old Japanese man was admitted to our hospital for further examination of obesity and hypertension. Endocrinological studies showed that plasma cortisol was high (22.5 micrograms/dl) without diurnal rhythm, and plasma ACTH was low. Two or 8 mg of dexamethasone did not suppress the plasma cortisol levels. Abdominal computed tomography revealed nodular hyperplasia of bilateral adrenal glands. Adrenal scintigraphy showed the positive uptake of 131I-adosterol to bilateral adrenal glands. Brain magnetic resonance imaging revealed no abnormalities. He was diagnosed as having Cushing's syndrome with bilateral adrenal hyperplasia, and bilateral adrenalectomy was performed. Left and right adrenal glands were 52 g and 35 g, respectively, and were occupied by yellow nodular lesions. Histologically, hyperplastic lesions were composed of clear cells. Finally he was diagnosed with AIMAH.

[Choroid plexus metastasis of renal cell carcinoma causing intraventricular hemorrhage: a case report].

We report a rare case of choroid plexus metastasis of renal cell carcinoma causing intraventricular hemorrhage. A 75-year-old female was admitted to our hospital for SAH. Preoperative examination evoked suspicion of an anterior communicating aneurysm as a cause of SAH. Furthermore, there were lesions on the tuberculum sellae and in the left trigone of the lateral ventricle, which were enhanced by Gd-DTPA on MRI. The patient was operated on via the pterional approach on December 3, 1996, but no aneurysm was found. She underwent total removal of the tuberculum sellae mass, which was postoperatively proved to be a meningioma by histological examination. The intraventricular tumor was supposed to be a meningioma, but it was not treated surgically. Two months later, the patient presented hematuria and was diagnosed as having a right renal cancer and underwent right nephrectomy on March 18, 1997. However, postoperatively, disturbance of consciousness continued. A CT scan revealed intraventricular hemorrhage around the tumor in the trigone of the lateral ventricle on March 21. An emergency operation for tumor and clot removal succeeded in improving the patient's condition. Histological examination of the tumor revealed clear-cell type renal cell carcinoma. Solitary choroidal plexus metastasis from renal cell carcinoma is quite rare: only 5 cases have been reported. But only our case was accompanied by intraventricular hemorrhage. Renal cell carcinomas are divided into two types: a slowly progressive type and a rapidly progressive type. Four cases among the reported 5 cases were the slowly progressive type, and our case was regarded as being of the same type.

Cloning and sequence analysis of hydroxyquinol 1,2-dioxygenase gene in 2,4,6-trichlorophenol-degrading Ralstonia pickettii DTP0602 and characterization of its product.

A gene encoding hydroxyquinol 1,2-dioxygenase was cloned from 2,4,6-trichlorophenol-degrading Ralstonia (Pseudomonas) pickettii strain DTP0602. Cell-free extracts of Escherichia coli containing a cloned 1.4-kb StuI-XhoI DNA fragment of R. pickettii DTP0602 hydroxyquinol 1,2-dioxygenase converted hydroxyquinol into maleylacetate and also degraded 6-chlorohydroxyquinol. The 1.4-kb DNA fragment contained one open reading frame (designated hadC) composed of 948 nucleotides. The molecular mass of 34,591 deduced from the gene product (HadC) was in agreement with the size (35 kDa) of the purified HadC protein determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The amino acid sequence of HadC exhibited high homology to that of the hydroxyquinol 1,2-dioxygenase of 2,4,5-trichlorophenoxyacetic acid-degrading Burkholderia cepacia AC1100 (Daubaras, D. L. et al., Appl. Environ. Microbiol., 61, 1279-1289, 1995). The active enzyme had a molecular mass of 68 kDa, suggesting that it is functional as a homodimer. The enzyme also catalyzed the oxidation of pyrogallol and 3-methylcatechol, possible intermediates in the degradation of 2,4,6-trichlorophenol, in addition to 6-chlorohydroxyquinol and hydroxyquinol. The dioxygenase catalyzed both ortho- and meta-cleavage of 3-methylcatechol.

Nucleotide sequences and characterization of genes encoding naphthalene upper pathway of pseudomonas aeruginosa PaK1 and Pseudomonas putida OUS82.

A 12,808-nucleotide containing DNA fragment cloned from naphthalene-utilizing (Nah+) Pseudomonas aeruginosa PaK1 was analyzed and compared with the genes (pah(OUS)) of a 14,462-nucleotide DNA fragment from Pseudomonas putida OUS82. The DNA sequence analyses demonstrated that the naphthalene upper-pathway genes and their deduced enzymes were very similar between the two bacteria: nucleotide similarities, 83-93%; amino acid similarities, 79-95%. These genes were also similar to those of the nah operon of plasmid NAH7; in particular, the OUS82 genes were similar to the nah genes, whereas the PaK1 genes were almost identical to the dox genes of Pseudomonas sp. C18. A region homologous with the 84-bp repeated sequence that Eaton (J. Bacteriol., 176, 7757-7762, 1994) has found at a site upstream of he nah operon was found only in a region downstream of the pah(PaK) gene cluster in PaK1 and on both sides of the pah(OUS) gene cluster in OUS82. A PaK1 gene, corresponding to an unknown gene (nahQ) in the nah operon, is located between the 1,2-dihydroxynaphthalene dioxygenase gene and the trans-o-hydroxybenzylindenepyruvate (tHBP A) hydratase-aldolase gene (nahE), and was suggested to be involved in the conversion of naphthalene to salicylate. Just downstream of the pah(PaK) gene cluster, a portion of a region was identical to one-third of the transposase gene (tnpA) in a phenol-catabolic plasmid pEST1226.