Effects of CO2 laser irradiation of the gingiva during tooth movement.

Patients often feel pain or discomfort in response to orthodontic force. It was hypothesized that CO(2) laser irradiation may reduce the early responses to nociceptive stimuli during tooth movement. The distribution of Fos-immunoreactive (Fos-IR) neurons in the medullary dorsal horn of rats was evaluated. Two hrs after tooth movement, Fos-IR neurons in the ipsilateral part of the medullary dorsal horn increased significantly. CO(2) laser irradiation to the gingiva just after tooth movement caused a significant decrease of Fos-IR neurons. PGP 9.5- and CGRP-positive nerve fibers were observed in the PDL of all study groups. The maximum temperature below the mucosa during CO(2) laser irradiation was less than 40 degrees C. It was suggested that CO(2) laser irradiation reduced the early responses to nociceptive stimuli during tooth movement and might not have adverse effects on periodontal tissue.

Elution and adsorptive concentration of Japanese cedar (Cryptomeria japonica) pollen allergen in environmental water.

Elution of Japanese cedar pollen allergens (Cry j I and others) from pollen grains and its adsorptive concentration onto hydrophobic and hydrophilic surfaces were investigated using the surface plasmon resonance technique. Results showed that the allergen elution was obviously enhanced when the ion concentration was higher than that within the human body, indicating that the pollen tend to release its allergen in environmental water having a high ion concentration. However, higher adsorption capacity was observed on hydrophobic surface than hydrophilic surface. These results indicate that water puddles on roadsides beside heavy traffic including large amounts of ion compounds and hydrophobic diesel exhaust particles (DEPs) are a pollen allergen-DEP complex generator. DEPs are easily absorbed into the living body; therefore these mechanisms may be responsible for causing the highest incidence of pollinosis among residents living alongside roads with heavy automobile traffic.

Effects of kava (Kava-kava, 'Awa, Yaqona, Piper methysticum) on c-DNA-expressed cytochrome P450 enzymes and human cryopreserved hepatocytes.

The effects of the herbal product kava (Kava kava, 'Awa, Yaqona, Piper methysticum) on human P450 isoforms were studied in vitro using both cDNA-expressed human enzymes and cryopreserved human hepatocytes. Increasing concentrations of an ethanolic extract of dried kava root and three purified kava lactones (methysticin, desmethoxyyangonin, and yangonin) were tested for their ability to inhibit the catalytic activity of a panel of P450 isoforms (1A2, 2A6, 2C9, C2C19, 2D6, 2E1, and 3A4) present as c-DNA expressed-enzymes and in previously cryopreserved human hepatocytes. In addition, the test compounds' effect on hepatocyte viability was evaluated by measuring cellular ATP content. In both models, the kava extract and the three kava lactones were found to be potent inhibitors of CYPs 1A2, 2C9, 2C19, 2E1, and 3A4 with IC50 values of approximately 10 microM. The test compounds were also moderately cytotoxic to human hepatocytes (EC50 values of approximately 50 microM). Methysticin was the most potent enzyme inhibitor as well as the most cytotoxic, followed by (in order of potency:) the kava root extract, desmethoxyyangonin, and yangonin. Our results suggest that the drug interaction and hepatotoxic potential of kava should be further investigated.

Acceleration of superoxide generation in polymorphonuclear leukocytes and inhibition of platelet aggregation by alk(en)yl thiosulfates derived from onion and garlic in dogs and humans.

We recently identified sodium n-propyl thiosulfate (NPTS) and sodium 2-propenyl thiosulfate (2PTS) from boiled onion and garlic, respectively, as causative agents of hemolytic anemia in dogs. We present here data concerning the effects of these alk(en)yl thiosulfates on superoxide (O(2)(-)) generation in peripheral polymorphonuclear leukocytes (PMNs) and on adenosine 5'-diphosphate (ADP)-induced platelet aggregation in dogs and humans in vitro. Both NPTS and 2PTS increased O(2)(-) generation significantly (P<0.05 at 1mM NPTS, P<0.005 at 0.1 and 1mM 2PTS) and reduced its reaction time significantly (P<0.05 between 0.01 and 1mM NPTS and at 1mM 2PTS) in canine PMNs stimulated by phorbol 12-myristate 13-acetate, compared with the control without alk(en)yl thiosulfates. However, a tendency to return to the control level was observed at 10mM of the alk(en)yl thiosulfates in both O(2)(-) generation and its reaction time. Although NPTS and 2PTS did not exert any significant effect on the O(2)(-) generation in human PMNs, 2PTS reduced its reaction time significantly (P<0.05) at 1 and 10mM compared with the control, showing that 2PTS accelerated O(2)(-) generation in human PMNs. The difference in effects on O(2)(-) generation may be due to that in susceptibility to alk(en)yl thiosulfates between canine and human PMNs. On the other hand, NPTS and 2PTS were shown to significantly inhibit ADP-induced platelet aggregation at 0.01mM (P<0.01) in canine platelets and at 0.001-0.1mM (P<0.05) in human platelets. In contrast, the maximal aggregation percentage returned to the control level at 1mM of alk(en)yl thiosulfates in both canine and human platelets. From these results, we conclude that NPTS and 2PTS have the potential to promote immune functions and prevent cardiovascular diseases.

Inhibitory effect of magnolol and honokiol from Magnolia obovata on human fibrosarcoma HT-1080. Invasiveness in vitro.

We investigated the inhibitory effect of Magnolia obovata Thunb. bark ethanol extracts on human fibrosarcoma HT-1080 cells invasion in a reconstituted basement membrane [Matrigel (MG)]. We found that the effective components of the bark ethanol extracts were magnolol and honokiol, two biphenyl compounds. The extracts, magnolol and honokiol, did not affect HT-1080 cells adhesion to MG, but did inhibit HT-1080 cells migration at a high concentration (100 microM). HT-1080 cells secrete matrix metalloproteinase (MMP)-9, which degrades the extracellular matrix as a part of the invasive process. Magnolol and honokiol inhibited the activity of MMP-9, which may have been responsible, in part, for the inhibition of tumor cell invasiveness.

Microwave coagulation versus insertion of a second stent for occluded biliary metal stent.

BACKGROUND/AIMS: There is no consensus regarding optimal management of self-expandable metallic stent occlusion. We investigated the efficacy of microwave coagulation therapy for recanalization as compared to second stent placement. METHODOLOGY: Sixty patients with malignant obstruction of the common bile duct were treated with metal stent placement from January 1992 to July 1999. Of these, 13 patients subsequently developed stent occlusion due to tumor ingrowth. We compared stent patency and patient survival rates after microwave coagulation to those after insertion of a second stent. The influence of the duration of patency of the first stent on the second stent patency was also evaluated. RESULTS: Of the 13 patients with stent occlusion, 7 were treated with microwave coagulation therapy, and 6 with insertion of a second metal stent. In all cases, occluded stents were successfully recanalized without any complications. There was no significant difference in duration of first stent patency between the two groups. The median duration of second stent patency was prolonged in microwave-treated patients (152 days vs. 104 days, P > 0.05). The median duration of patient survival after last recanalizing procedure was also prolonged in microwave-treated patients (131 days vs. 78 days, P > 0.05). Microwave energy did not induce destruction of the stent filament. CONCLUSIONS: Microwave coagulation did not offer significantly longer duration of stent patency and patient survival compared to insertion of a second metal stent. However, this procedure is safe, feasible, and certainly as good as a second stent placement. It may be an alternative to insertion of a second stent within the occluded stent.

Transpapillary microwave coagulation therapy for recanalizing self-expandable metallic stents occluded by tumor ingrowth: initial experience.

Percutaneous microwave coagulation for recanalizing stents occluded by tumor ingrowth has been reported. With this technique, however, the percutaneous drain diminishes the quality of life in patients with unresectable tumors and a limited prognosis. Transpapillary microwave ablation was attempted in three patients with occluded stents. After a sheath had been inserted into the proximal hepatic duct across the occluded region, a microwave electrode was introduced into the intrahepatic duct via the sheath. We used microwave therapy with an output power of 40 W, based on our previous in vitro study. Except in one patient, the stents were successfully recanalized with one or two attempts. In one patient who underwent ablation in the intrahepatic duct, a 1.8-mm electrode enabled recanalization of the stent. In another who underwent ablation in the extrahepatic duct, however, a larger electrode was required. There were no procedure-related complications. Transpapillary microwave coagulation of occluded stents appears to be an alternative to percutaneous microwave coagulation with an electrode fitting the stent size. The technique might be easier with the use of a redesigned electrode with a guide wire lumen.

A methylotrophic pathway participates in pectin utilization by Candida boidinii.

The methylotrophic yeast Candida boidinii S2 was found to be able to grow on pectin or polygalacturonate as a carbon source. When cells were grown on 1% (wt/vol) pectin, C. boidinii exhibited induced levels of the pectin-depolymerizing enzymes pectin methylesterase (208 mU/mg of protein), pectin lyase (673 mU/mg), pectate lyase (673 mU/mg), and polygalacturonase (3.45 U/mg) and two methanol-metabolizing peroxisomal enzymes, alcohol oxidase (0.26 U/mg) and dihydroxyacetone synthase (94 mU/mg). The numbers of peroxisomes also increased ca. two- to threefold in cells grown on these pectic compounds (3.34 and 2.76 peroxisomes/cell for cells grown on pectin and polygalacturonate, respectively) compared to the numbers in cells grown on glucose (1.29 peroxisomes/cell). The cell density obtained with pectin increased as the degree of methyl esterification of pectic compounds increased, and it decreased in strains from which genes encoding alcohol oxidase and dihydroxyacetone synthase were deleted and in a peroxisome assembly mutant. Our study showed that methanol metabolism and peroxisome assembly play important roles in the degradation of pectin, especially in the utilization of its methyl ester moieties.

Purification and characterization of the allergenic components of shimeji mushroom (Tricholoma conglobatum) spore for shimeji workers' hypersensitivity pneumonitis.

Respiratory symptoms and hypersensitivity pneumonitis (HP) among mushroom workers have been well documented. Inhalation of shimeji mushroom (Tricholoma conglobatum) spore has been assumed to be the cause of HP among indoor shimeji cultivating workers. We isolated and partially characterized the allergenic components of shimeji. The sera from 9 HP patients, 10 asymptomatic shimeji workers and 15 normal individuals were examined for shimeji specific IgG and IgA antibodies by ELISA using crude shimeji extract. Shimeji specific IgG- and IgA-antibodies were higher in sera from HP patients than in sera from control subjects. Crude shimeji spore extract was separated and purified by HPLC followed by SDS-PAGE, and their antigenic activity was studied by immunoblotting with a pool of sera from patients. Sera from all HP patients showed IgG and IgA antibody activities to 21, 16 and 14 kD proteins extracted from shimeji spore. The 21 kD protein contained internal peptide amino acid sequence of Gly-Gly-Thr-Val-Ile-Asn-Leu-Leu-Gly, Gln-Arg-Phe-Glu-Glu and Gln-Gly-Ile-Tyr. These results demonstrate that shimeji spore extract contains multiple proteinous components, which have antigenic activity to react with the sera from HP patients among shimeji workers. These proteins may be the potent sensitizing allergens to cause HP among shimeji cultivating workers.

[Diagnosis of ulcerative colitis--barium enema examination].

Barium enema is useful especially for differential diagnosis of right sided type or segmental type of ulcerative colitis from other inflammatory bowel diseases. Compared with endoscopical examination, it provides more objective informations about the range of affected colon which is necessary for the choice of treatment strategy to the reluctant case. However, as preparation procedures may cause aggravation of ulcerative colitis, an unnecessary barium study should be avoided. Even the case in remission stage, it is advisable to add 40 to 60 mg of water-soluble predonisolone to barium solution. Roentogenographic diagnosis of ulcerative colitis is not difficult, in so far as two major points are properly evaluated. The first is the presence or absence of haustration, and the second is mucosal surface of the colon, namely, multiple ulcers or erosions distributing diffusely and continuously in active stage, or granular mucosa and disarray of network pattern of colonic mucosa in remission stage.

[Nutritional therapy for ulcerative colitis].

Despite its supplementary role for a remedy of the Ulcerative Colitis, nutritional therapy performs the important role as well as medicine. In the active state, the aim of nutrition therapy is to reduce the activity of the disease. In the remission state, the aim is to keep the condition and to supply elements of nutrition which the disease causes these deficiency such as vitamins or minerals. Needless to say, it is required to consider a few points; patient's condition, lesion's digestion disorder and leakage, or side effects of caused by medicines. High calorie, low fat and rich protein is ideal for the nutrition therapy for this disease, however it is suggested to satisfy patient's mental health at the same time. Nutrition therapy for Ulcerative Colitis should be regarded as the long-term treatment that requires the co-operation and comprehension between patients and medical supporting staffs.

[Treatment of mild and moderate ulcerative colitis].

Selection of treatment for ulcerative colitis is based on the definition of clinical severity. In those medication is a main course. Mild and moderate cases are treated mainly with salicylazosulfapyridine or mesalazine. Patients with proctitis-type are added betamethazone suppositories and those with left-sided and total colitis-type predonisolone enema therapy simultaneously. If there is no response, they are given additional oral predonisolone. Even after remission, patients should be carefully observed. A good patient-physician relationship, continuation of maintenance therapy, and periodic follow-up examinations using colonoscopy or barium enema are essential to successful long-term management of this disease. Patients who have repeated relapsing should receive the stronger treatment, according to the endoscopic findings and clinical course in addition to the clinical severity.

[Anesthesia in a patient with epidermolysis bullosa].

Epidermolysis bullosa hereditaria, a rare inherited disorder presents clinically with recurrent cutaneous blister formation with possible involvement of mucous membrane and organs following minimal trauma. The sequelae of this disease pose significant anesthetic problems to operating room staffs. We describe the anesthetic management of a 32-year old woman with epidermolysis bullosa hereditaria who underwent palmar skin graft using regional anesthesia, and discuss the problems associated with this disease.

Molecular cloning and expression of Xenopus laevis requiem cDNA1.

Requiem is an apoptosis-associated gene, which was originally identified in mouse (T.G. Gabig et al., J. Biol. Chem. 269 (1994) 29515-29519). In this study, we isolated five independent cDNA clones for frog Requiem (xReq) from Xenopus laevis ovary. Sequence analysis of the multiple cDNAs has suggested that Xenopus genome contains at least two non-allelic copies of the xReq gene. The amino acid sequence deduced from the cDNAs contained a single Krüppel-type zinc-finger motif and two PHD-finger motifs. Northern analysis revealed that the ovary expressed xReq transcripts with different sizes.

Cloning and expression of Shaker alpha- and beta-subunits during inner ear development.

Sensory cells of the chicken cochlea exhibit different ion channels relative to their position along the epithelium. One of these channels conducts an A-type potassium current which is found primarily in 'short' hair cells. Here, we report the first full length cloning and developmental expression of Shaker genes from this endorgan. Clones were obtained by screening a chicken (Gallus gallus) cochlea cDNA library, using probes made from RHK1 (i.e., Kvalpha1.4) cDNA, a Shaker homologue isolated from rat heart, and hKvbeta1.2 cDNA, a beta homologue isolated from human heart. Sequence analysis revealed a chick homologue of Kvalpha1.4, with a deduced amino acid similarity of 76-79% to mammalian Kvalpha1.4, and a chick homologue of Kvbeta1.1, with a similarity of 95% to mammalian Kvbeta1.1. In addition, we isolated a variant of cKvalpha1. 4 (cKvalpha1.4(m)) that differs in its untranslated regions and shows complete similarity in its coding region, except for the deletion of a single nucleotide. During development of the inner ear, reverse transcription-polymerase chain reaction (RT-PCR) studies show that the beta-subunit is expressed as early as embryonic day 3, whereas alpha- and beta-subunits are coexpressed on embryonic days 7 to 10, 14, and in adult.

Saireito (a Chinese herbal drug) decreases inhibitory effect of prednisolone and accelerates the recovery of rat hypothalamic-pituitary-adrenal axis.

Saireito, a saiko agent (a Chinese herbal drug), increases the synthesis and secretion of ACTH by stimulating hypothalamic CRH release. In the present study, we examined the effect of food containing saireito (1.5%) on the recovery of the hypothalamic-pituitary-adrenal axis after treating male rats with prednisolone (PSL, 200 microM) in drinking water for 14 days. Saireito was administered during and after PSL administration. The rats were decapitated at various times after PSL administration. Tail-pinch stress had been applied to some rats. The plasma ACTH response to tail-pinch stress in the PSL + saireito group recovered to the control level on day 1, but that in the group given PSL alone recovered on day 3. The ACTH level in the anterior pituitary and the CRH level in the median eminence of the PSL + saireito group returned to the control level on day 3, and that in the group given PSL alone returned to it on day 5. These results indicate that the administration of saireito reduces the negative feedback effect of PSL on the hypothalamus and pituitary and accelerates the recovery of the hypothalamic CRH and pituitary ACTH level after glucocorticoid treatment.

Two novel testicular serine proteases, TESP1 and TESP2, are present in the mouse sperm acrosome.

To identify a novel candidate(s) for acrosomal proteins that act on the sperm/egg interaction, a DNA fragment was PCR-amplified from a cDNA library of acrosin-deficient mouse testis and then used as a probe to screen a mouse testis cDNA library. Complementary DNA clones encoding each of two similar but different serine proteases, TESP1 and TESP2, have been identified. The nucleotide sequences of these clones indicate that mouse TESP1 and TESP2 are initially synthesized as preproproteins of 367 and 366 amino acids, respectively. Comparison of the two TESP sequences with those of typical serine proteases suggests that each TESP zymogen is probably converted into a two-chain mature enzyme consisting of light and heavy chains covalently linked by a single pre-existing disulfide bond. The conversion may be accomplished by another protease(s) with a trypsin-like cleavage specificity, since it is unlikely that the mature TESP1 and TESP2 are capable of splitting the Lys-Ile bond between the light and heavy chains. Northern blot analysis of total cellular RNA demonstrates that the TESP1 and TESP2 genes are expressed only in the testis, and the transcripts are abundantly present in the haploid round spermatids. Moreover, immunocytochemical analysis of mouse cauda epididymal sperm using affinity-purified antibodies reveals that these two TESPs are both localized in the sperm acrosome and are released during the acrosome reaction induced by calcium ionophore A23187. These findings provide additional clues for elucidating the mechanisms involved in the sperm/egg interactions, including penetration of the zona pellucida by sperm.

Importance of the cysteine-rich carboxyl-terminal half of V protein for Sendai virus pathogenesis.

The Sendai virus V protein is a nonstructural trans-frame protein whose cysteine-rich C-terminal half is fused to the acidic N-terminal half of the P protein via mRNA editing. We recently created a mutant by disrupting the editing motif, which is devoid of mRNA editing and hence unable to produce the V protein, and demonstrated that this V(-) virus replicated normally or even faster with augmented gene expression and cytopathogenicity in cells in vitro, but was strongly attenuated in pathogenicity for mice (A. Kato, K. Kiyotani, Y. Sakai, T. Yoshida, and Y. Nagai, EMBO J. 16:578-587, 1997). Thus, although categorized as a nonessential protein, the V protein appeared to encode a luxury function required for the viral in vivo pathogenesis. Here, we created another version of a V-deficient mutant, VdeltaC, encoding only the N-terminal half but not the V-specific C-terminal half, by introducing a stop codon in the trans-V frame, and then we compared its in vitro and in vivo phenotypes with those of the V(-) and wild-type viruses. The VdeltaC virus was found to be similar to the wild-type virus in vitro with no augmented gene expression and cytopathogenicity, but in vivo, it resembled the V(-) virus, displaying a similarly attenuated phenotype. Thus, the pathogenicity determinant in the V protein was mapped to the C-terminal half. The N-terminal half was likely sufficient to confer normal (wild-type) in vitro phenotypes.

GE3, a novel hexadepsipeptide antitumor antibiotic, produced by Streptomyces sp. I. Taxonomy, production, isolation, physico-chemical properties, and biological activities.

GE3, a novel cyclic hexadepsipeptide antibiotic, was isolated from the culture broth of Streptomyces sp. GE3. GE3 was weakly active against some Gram-positive and Gram-negative bacteria and showed potent cytotoxicity against human tumor cell lines. GE3 also exhibited antitumor activity against human pancreatic carcinoma, PSN-1, in vivo. GE3B, a linear peptide form of GE3, which was isolated from the same culture broth with GE3, showed no antibiotic and cytotoxic activities, suggesting the necessity of the cyclic structure of GE3 for its biological activities.