Facial emphysema caused by cheek bite.

Biting of the buccal mucosa is very frequent injury, whereas facial emphysema caused by cheek bite is rare. We report a case of facial emphysema causing puffing of the cheek through a self-inflicted bite of the buccal mucosa.

Purification of two DNA-dependent adenosinetriphosphatases having DNA helicase activity from HeLa cells and comparison of the properties of the two enzymes.

DNA-dependent ATPase activities in crude extracts prepared from HeLa cells were separated into five peaks designated Q1 to Q5 by FPLC Mono Q column chromatography. In our previous study, we observed that crude extracts prepared from xeroderma pigmentosum complementation group C (XP-C) cells contained no DNA-dependent ATPase activity at the peak position of Q1 and exhibited a broader peak with higher activity than normal Q2 at the peak position of Q2 [Yanagisawa, J., Seki, M., Ui, M., & Enomoto, T. (1992) J. Biol. Chem. 267, 3585-3588]. We have purified two DNA-dependent ATPases Q1 and Q2 from HeLa cells and characterized their properties in order to obtain a means to discriminate ATPase Q1 from Q2 in XP-C cells. The apparent molecular masses of Q1 and Q2 on SDS-polyacrylamide gel electrophoresis were 73 and 100 kDa, respectively. The two enzymes required a divalent cation for activity. DNA-dependent ATPase Q1 hydrolyzed ATP and dATP and Q2 hydrolyzed ATP preferentially among the nucleotides tested. Both enzymes preferred single-stranded DNA as a cofactor. The DNA-dependent ATPase activity of Q2 was inhibited by 90% in the presence of 200 mM NaCl, whereas that of Q1 was not affected by NaCl at concentrations up to 200 mM. Both enzymes had DNA helicase activity, that of Q1 being more resistant to NaCl than that of Q2. The DNA helicase activity of Q2 was about 150-fold higher than that of Q1, when compared with units of ATPase activity.(ABSTRACT TRUNCATED AT 250 WORDS)

A long-term survival case of multiple hepatocellular carcinoma with metachronous lymph node metastasis.

A long-term survival case of multiple hepatocellular carcinoma (HCC) with metachronous metastasis to a lymph node is reported. The patient, a 66-year-old woman, had two primary HCC nodules, one each in the left and right hepatic lobes, which were resected. She developed a lymph node lesion and a secondary HCC 45 and 62 months after the first operation, respectively. She has been well for the 7 years since the first operation despite undergoing hepatic resection for HCC twice as well as lymph node resection. Clonal analysis, based on the methylation pattern of the X chromosome-linked androgen receptor gene, suggested that the two primary tumors were multicentric and that the lymph node lesion had arisen by metastasis from the primary tumor in the right hepatic lobe.

Enlarged focal nodular hyperplasia of the liver under the influence of oral contraceptives.

Two cases of focal nodular hyperplasia of the liver in mature women treated with the oral contraceptive are described. Radiological investigations in one case revealed the typical findings of focal nodular hyperplasia with computed tomography and magnetic resonance imaging demonstrating central scar structures while spoke-wheel appearance was evident on arteriography, in the other case however findings were atypical. Routine investigations including liver function tests and alpha-fetoprotein levels were normal while hepatitis B surface antigen and hepatitis C virus antibody were negative. The lesions of these two cases enlarged significantly during the follow-up and they were therefore surgically resected. Pathological features of both resected specimens, such as hepatocellular hyperplasia, bile duct proliferation and vascular abnormalities, were compatible with focal nodular hyperplasia. It has been suggested that tumor growth may be augmented by sex hormone stimulation and therefore estrogen and progesterone receptor expressions in the resected tumors were determined by immunocytochemistry. Interestingly, stainings for both receptors were negative. In case 2, the tumor was enlarging although oral contraceptive use had been discontinued for the past 7 years. These results suggest that there is no direct relationship of focal nodular hyperplasia with oral contraceptives. The role of sex hormones in focal nodular hyperplasia of the liver merits further study.

Effects of heparin dosage and sperm capacitation time on in vitro fertilization and cleavage of bovine oocytes matured in vitro.

The present study was conducted to clarify the effect of heparin dosage and sperm capacitation time on in vitro fertilization (Experiment 1) and cleavage (Experiment 2) rates of bovine oocytes matured in vitro. For in vitro fertilization, seven dosages of heparin (0, 5, 10, 25, 50, 100 and 200 microg/ml) and nine incubation periods (0, 5, 15, 30, 45, 60, 120, 180 and 240 min) in a capacitation medium were examined, using 6,634 oocytes. The mean proportions of fertilized oocytes in 25, 50 and 100 microg/ml of heparin were significantly (P<0.05) higher (53 to 59%) than in the other dosages (3 to 44%). Incubation with heparin for longer than 60 min lowered the frequencies of fertilization (20 to 36%) compared with the shorter incubation periods (38 to 49%). Higher proportions of fertilized oocytes were obtained by 5, 15, 30 or 45 min of incubation (42 to 49%) than by the other time periods (20 to 38%). Cleavage rates were found by using 2,098 oocytes in a factorial study (4x4x15: dosages -25, 50, 100 and 200 mug/ml; incubation periods -0, 15, 30 and 60 min; and replicates). The incubation periods and replicates resulted in highly significant differences (P<0.001) in development rates to eight-cell stage, but the four dosages of heparin showed no significant differences. The present results indicate that heparin dosage and sperm capacitation time are important factors influencing in vitro fertilization and cleavage rates. Optimal heparin dosages for the capacitation of bull spermatozoa ranged from 25 to 100 microg/ml; optimal incubation periods ranged from 5 to 60 min.

[Phase I clinical studies of S6472 (sustained release preparations of cefaclor)].

Current (regular) preparation of cefaclor (CCL) require the 3-time-a-day administration. S6472 (sustained release preparation) which can be used with the twice-a-day administration in the morning and the evening is capsule and granule preparations consisting of 40% of nonenteric and 60% of enteric coated granules of CCL. Phase I clinical studies of S6472 were conducted in 12 nonfasted healthy adult male volunteers with cross over method using a single dose of 375 mg in capsule and granule forms of S6472, and 250 mg in capsule form of regular CCL as a control drug. The volunteers received the 3 preparations at 1-week interval. The summary of the results from the above studies is as follows: Grouping of the volunteers. The 12 volunteers were divided into 3 groups (each group consists of 4 volunteers) and there were no significant differences between each group regarding background factors of the volunteers. Tolerance. None of the volunteers who received the 3 preparations at 1-week interval complained of subjective abnormalities. No abnormalities which are considered to be due to S6472 and regular CCL were found in the clinical laboratory tests carried out before the administration and 1 week after the completion of the studies. Plasma level. There were no significant differences between capsule and granule forms of S6472 regarding Cmax and AUC, and it was confirmed that bioavailability of both preparations was the same. It was also confirmed that plasma levels of the 2 preparations of S6472 were maintained for longer period of time than those of regular CCL. Urinary excretion. Mean urine levels of the 2 preparations of S6472 every 2 hours after the administration were confirmed to be maintained for longer period of time than those of regular CCL. There were no significant differences between the 2 preparations of S6472 regarding urinary recovery rate. However, the significant differences between the 2 preparations of S6472 and regular CCL were observed. Urinary recovery rate of the 2 preparations of S6472 was 87 approximately 88% of that of regular CCL.

[Absorption and excretion studies of S6472 (sustained release preparations of cefaclor)].

In order to see absorption and excretion of S6472 (sustained release preparations of cefaclor (CCL], 3 studies regarding, 1) influence of meals (single dose of 375 mg in capsule form), 2) dose response (single dose of 375 mg vs. 750 mg in granule form) and 3) continuous administration (twice-a-day administration of 750 mg in granule form for 8 days) were conducted in 15 healthy adult male volunteers using capsule and granule forms of S6472. The following is the summary of the results from the above studies: Tolerance. In the above 3 studies, none of the 15 volunteers complained of subjective abnormalities. In the clinical laboratory tests performed before the administration of S6472 and at the next day of the completion of the studies, the values from the laboratory tests were within normal range. Influence of meals. The average time for peak plasma level of CCL and decrease in the plasma level following the administration of S6472 were the fastest in the fasted volunteers, followed by the volunteers with light meals and usual meals. From this, it was confirmed that plasma levels of CCL were maintained for longer period of time in the non-fasted volunteers than in the fasted volunteers. In the volunteers who had light and usual meals, the peak plasma levels, AUC of the plasma levels and urinary recovery rate were almost the same. Influence by amount of meals was scarcely observed. Dose response. Mean serum levels and their AUC in the volunteers receiving 750 mg of S6472 were approximately twice as much as those in the volunteers receiving 375 mg of S6472, and dose response between the 2 doses was confirmed. Continuous administration. Mean serum levels and their AUC at the first dose and the 15th dose (final dose) were almost the same. In the continuous administration of S6472 for 8 days (15 doses), no accumulation of CCL in the body was observed.

[Prolonged action preparation of cefaclor].

This study was conducted to develop a prolonged action preparation of cefaclor (CCL) which can offer, with the twice-a-day administration, as much effectiveness as its conventional preparation (Kefral capsule) with the 3 times-a-day administration. Absorption site of CCL in gastrointestinal tract, preparation form (enteric coated granules) which slowly release CCL, dissolution property of the form, and mixed ratio of the form and rapid release form (nonenteric coated granules) were studied and complex granules consisting of 40% of nonenteric coated granules and 60% of enteric coated granules which dissolve at pH 6 were chosen as a prolonged action preparation of CCL. Bactericidal activity of the prolonged action preparation (S6472) was confirmed to be the same as that of the conventional preparation by comparative viable cell count study in which concentrations of CCL simulated to plasma concentrations following the administration of S6472 at the dosage of 375 mg b.i.d. and the conventional preparation at the dosage of 250 mg t.i.d. were used. From the above, S6472 is considered to be a prolonged action preparation of CCL which serve our purpose. Since S6472 can be given with the twice-a-day administration, its daytime administration is not necessary. Therefore, S6472 is considered to be much useful preparation for the patients.

[Effect of anabolic steroid on immune response].

Using lymphocyte, monocyte and eosinophil counts of the peripheral blood, PHA-blastoid transformation, immunoglobulin and beta 2-microglobulin, the influence of anabolic steroid on the immune reactivity of the host was dissected by administration of Deca-Durabolin ( nandrolone decanoate) to both tumor-bearing host and tumor-free host after operation for alimentary tract. The number of peripheral lymphocytes and monocytes, the PHA-blastoid transformation of peripheral lymphocytes and the IgG level were increased, and the beta 2-microglobulin level showed the tendency of decrease after the administration of Deca-Durabolin.

[Clonal analysis of hepatocellular carcinoma].

We investigated the cell clonality of 12 cases of female solitary hepatocellular carcinoma (HCC) that were associated with hepatitis virus infection. The clonal origin of HCC could be assessed by the method based on restriction fragment length polymorphism (RFLP) of X-chromosome-linked androgen receptor gene (AR) and phosphoglycerate kinase (PGK) gene, taking advantage of random inactivation of one of two X-chromosomes by methylation in females. We extracted DNA samples from both fresh and paraffin-embedded specimens of the same lesion as a source of DNA sample for polymerase chain reaction (PCR). Consequently, it was possible to use methylation-sensitive restriction enzymes and PCR to study differential methylation patterns among alleles of these genes for both DNA samples. The RFLPs of AR gene and PGK gene were found in eight of 12 cases and five of 12 cases, respectively. There were two cases which had no RFLPs in either AR gene or PGK gene. All cases of HCC which had RFLP in either AR gene or PGK gene demonstrated monoclonal origin of the tumor regardless of their histologic patterns.

[Locoregional adoptive immunotherapy using LAK cells and IL-2 against liver metastases from digestive tract cancer].

We investigated the clinical efficacy of locoregional and systemic adoptive immunotherapy (AIT) with or without interleukin-2 (IL-2) against solid metastatic lesions from digestive tract cancer. Eighteen patients, who were treated with more than 10(10) lymphokine-activated killer (LAK) cells, were enrolled in this study. Seven of the 18 patients received hepatic arterial infusion (HAI) of LAK cells with or without IL-2 against metastatic liver tumors (locoregional therapy group). The remaining 11 patients received systemic transfer of LAK cells with IL-2 against metastatic lesions located in organs other than the liver (systemic therapy group). Three of 7 locoregional therapy group patients showed clinically significant tumor regressions that were evaluated as being equivalent to partial response (PR). Two of the 11 systemic therapy group patients showed significant tumor regressions, but this response rate was much lower than that of the locoregional therapy group. The 2 effective cases in the systemic therapy group were esophageal cancer patients. Locoregional AIT with or without IL-2 against liver metastases from digestive tract cancer could be an effective therapeutic modality in some patients who are refractory to conventional therapies (e.g., chemotherapy and radiotherapy). It is necessary to find a new way to augment the anti-tumor effect of this therapy in combination with prior or concomitant chemotherapy.

Non-cavernomatous superior mesenteric thrombosis successfully recanalized with interventional radiological procedures carried out with a combination transmesenteric and transjugular approaches.

This is the study of a 52-year-old man with oesophageal, rectal and anal varices caused by portal hypertension with complete obstruction of the superior mesenteric vein. Treatment by two sessions of interventional radiological procedures was successful. The first was a catheter-directed thrombolysis using the transmesenteric approach. The second was percutaneous transluminal angioplasty and stent implantation for the obstructed segment of the superior mesenteric vein and the creation of a transjugular intrahepatic portosystemic shunt. In the second session, devices were advanced over a guidewire inserted from the right jugular vein and pulled out of the ileocolic vein using the pull-through technique.

Gene cloning, nucleotide sequence, and expression of a cephalosporin-C deacetylase from Bacillus subtilis.

The gene encoding a cephalosporin-C deacetylase (CAH) from Bacillus subtilis SHS 0133 was cloned and sequenced. The nucleotide sequence contained an open reading frame encoding a polypeptide consisting of 318 amino acids, the molecular weight of which was in good agreement with the value obtained by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The deduced amino acid sequence contained the common sequence Gly-X-Ser-X-Gly found in many esterases, lipases, and serine proteases. This indicates that CAH is a serine enzyme. A possible promoter sequence which is very similar to the consensus sequences of -35 and -10 regions recognized by B. subtilis RNA polymerase utilizing sigma factor H was found in the 5'-flanking region of the CAH structural gene. Two repeated A+T-rich blocks consisting of 24 bp were also found in the upstream region of the initiation codon. We constructed a series of expression plasmids by inserting the CAH gene into Escherichia coli ATG vectors. The degree of CAH gene expression depended on promoters and vector plasmids, which have different replication origins. The expressed CAH protein was an active form in the soluble fraction obtained after cell disruption. The highest expression level was accomplished with an expression plasmid, pCAH400, which has the trp promoter and the replication origin derived from pAT153. In the fermentation using a 30-liter jar fermentor, the transformant E. coli JM103(pCAH400) produced 440 U of CAH per ml of culture during a 24-h incubation. This value corresponded to 2.1 g of CAH protein in 1 liter of culture broth.

Pantoea punctata sp. nov., Pantoea citrea sp. nov., and Pantoea terrea sp. nov. isolated from fruit and soil samples.

A total of 37 bacterial strains with the general characteristics of the family Enterobacteriaceae were isolated from fruit and soil samples in Japan as producers of 2,5-diketo-D-gluconic acid from D-glucose. These organisms were phenotypically most closely related to the genus Pantoea (F. Gavini, J. Mergaert, A. Beji, C. Mielearek, D. Izard, K. Kersters, and J. De Ley, Int. J. Syst. Bacteriol. 39:337-345, 1989) and were divided into three phenotypic groups. We selected nine representative strains from the three groups for an examination of DNA relatedness, as determined by the S1 nuclease method at 60 degrees C. Strain SHS 2003T (T = type strain) exhibited 30 to 41 and 28 to 33% DNA relatedness to the strains belonging to the strain SHS 2006T group (strains SHS 2004, SHS 2005, SHS 2006T, and SHS 2007) and to the strains belonging to the strain SHS 2008T group (strains SHS 2008T, SHS 2009, SHS 2010, and SHS 2011), respectively. Strain SHS 2006T exhibited 38 to 46% DNA relatedness to the strains belonging to the strain SHS 2008T group. The levels of DNA relatedness within the strain SHS 2006T group and within the strain SHS 2008T group were more than 85 and 71%, respectively. Strain SHS 2003T, SHS 2006T, and SHS 2008T DNAs exhibited less than 18% binding to Pantoea dispersa ATCC 14589T and Pantoea agglomerans ATCC 27155T DNAs.(ABSTRACT TRUNCATED AT 250 WORDS)

Characterization of the properties of a human homologue of Escherichia coli RecQ from xeroderma pigmentosum group C and from HeLa cells.

We showed that DNA-dependent ATPase Q1 (DNA helicase Q1) from xeroderma pigmentosum complementation group C (XP-C) cells elutes from FPLC Mono Q column at higher concentrations of KCl than that from other human cells (35). We purified DNA helicase Q1 from XP-C and HeLa cells. The purified fractions of both cells contained a major polypeptide with a molecular mass of 73 kDa and had the same enzymatic properties, including salt- and temperature-sensitivity. Characterization using an anti-DNA helicase Q1 antibody indicated that this enzyme localized in the nuclei and was not modified by incorporating phosphate groups through phosphorylation and ADP-ribosylation. No interactions of DNA helicase Q1 with other proteins were indicated by immunoprecipitation of the helicase from crude extracts. No difference was observed in XP-C cells in intracellular localization of DNA helicase Q1, phosphorylation, and the interaction with other proteins as compared to HeLa cells.

Laparotomy versus interventional radiological procedures for the implantation of arterial infusion devices.

Although there have been numerous reports on implantable infusion devices for chemotherapy of patients with malignancy, we occasionally face problems with this therapy due to trouble with implantation. We performed a retrospective review of 81 implantations in 77 patients, who were treated with intraarterial chemotherapy via implanted devices from 1985 to 1993. They were divided into two groups according to the procedures: the operative procedure group (group A, n = 41) and the interventional radiological procedure group (group B, n = 36). Both groups were then analyzed regarding the respective complications. We experienced 25 complications: (a) 9 obstructions of the catheter, (b) 4 infections, (c) 4 dislocations of the catheter, (d) 3 hematomas, (e) 3 breakdowns of the device, (f) 1 pneumothorax, and (g) 1 hepatic artery occlusion. The results of a comparison of the complication rate between groups A and B were (a) 14.0%:8%, (b) 4%:0%, (c) 0%:10%, (d) 4%:2%, (e) 7%:0%, (f) 0%:2%, and (g) 2%:0%, respectively. A statistically significant difference was observed for (b) and (c) (P < 0.05). Infection occurred mainly in the cirrhotic cases of group A, but not in group B. In addition, one case fell into fatal sepsis. Based on the above findings, the interventional radiological procedure is thus considered to be the appropriate method for the prevention of infection in the case of a compromised host.

Production of 2-Keto-l-Gulonic Acid from d-Glucose by Two-Stage Fermentation.

A practical method for the production of calcium 2-keto-l-gulonate (an intermediate in the Reichstein synthesis of l-ascorbic acid) from d-glucose has been established by using a two-stage fermentation system. d-Glucose was first converted to calcium 2,5-diketo-d-gluconate by a mutant strain of Erwinia sp. in a medium containing d-glucose, corn steep liquor, (NH(4))(2)HPO(4), and CaCO(3). After a 26-h cultivation, 328.6 mg of calcium 2,5-diketo-d-gluconate per ml was obtained, with a 94.5% yield from d-glucose. This broth was used directly for the next conversion without removal of cells by treatment with sodium dodecyl sulfate. The stereospecific reduction of calcium 2,5-diketo-d-gluconate to calcium 2-keto-l-gulonate was performed with a mutant strain of Corynebacterium sp. When the cell growth reached a maximum (about 16 h) in a medium containing d-glucose, corn steep liquor, NaNO(3), KH(2)PO(4), and trace elements, NaNO(3) was added to the culture, and then the calcium 2,5-diketo-d-gluconate broth was fed over a period of about 50 h. Since the mutant strain requires a hydrogen donor for reduction, the calcium 2,5-diketo-d-gluconate broth was mixed with d-glucose before being fed. The results of four two-stage fermentations in 10-m conventional fermentors showed that an average of 106.3 mg of calcium 2-keto-l-gulonate per ml was obtained, with a 84.6% yield from d-glucose, the starting material of calcium 2,5-diketo-d-gluconate production. Calcium 2-keto-l-gulonate was stable in the broth. Neither 2-keto-d-gluconic acid nor 5-keto-d-gluconic acid was detected in the final broth.

[Two cases of rectal carcinoma that developed as a late complication of pelvic radiotherapy].

Two cases of a pelvic evisceration have been performed, due to an irradiation-induced rectal cancer. Described are the cases of two women who had been treated with irradiation for a cervical cancer following a hysterectomy, one patient being 65 years old and the other 67. After a latent period that lasted for 12 and 24 years, respectively, each had developed a rectal cancer. In each case, case excised specimen showed diffuse fibrosis and a hyaline change, reflecting the effect of radiation on the tumoral tissue. Cases of an irradiation-induced rectal cancer are uncommon, and the symptoms of enterocolitis caused by irradiation are similar to those of a colorectal cancer. The authors therefore suggest careful and long-term follow-up of patients that have received pelvic radiation.