Characterization of a Constitutive β-Lactamase from a Pseudomonas Aeruginosa : a Preliminary Report

Shin, Wan-Shik,Kim, Yang-Rae,Huh, Dong-Ho,Yoo, Jin-Hong,Choi, Jung-Hyun,Kang, Moon-Won THE CATHOLIC UNIVERSITY OF KOREA 1997 Bulletin of The Catholic Research Institutes of Me Vol.25 No.-

We analyzed some physicochemical characteristics of a constitutive β-lactamase from a clinical isolate of a Pseudomonas aeruginosa which was resistant to various β-Iactams except imipenem and susceptible to aminoglycosides. The aims of this study were to identify the class of this enzyme and to estimate the proportion of its contribution to such resistance profile. Kinetics study for β-lactamase was performed by spectrophotometric assay using the crude extracts and various antimicrobial agents. Inhibition profile was done using EDTA, HgCl_2 and p-chloromercuribenzoate (pCMB). and also measured 50% inhibitory concentration(IC_50) using β-lactamase inhibitors, such as clavulanic acid, sulbactam. The strain was resistant to ceftriaxone, sulperazon, piperacillin, ceftazidime and aztreonam, while it was susceptible to gentamicin, tobramycin, amikacin and imipenem. The relative maximal hydrolysis rate for β-lactamase was calculated by setting V_max for cephaloridine at 100. The enzyme hydrolyzed cephalosporins more rapidly than penicillins. The enzyme activity was not inhibited by EDTA, and pCMB, whereas it was inhibited by HgCl_2. The IC_50 of clavulanic acid was over 100μM and that of sulbactam was 22μM. The isoelectric point(pI) was 9.3, and the molecular weight was estimated to be about 40kDa. According to its substrate profile, pi and molecular weight, the β-lactamase from a clinical isolate of P.aeruginosa was a cephalosporinase belonging to group 1 in the Bush-Jacoby-Medeiros classification scheme.

In Situ Hybridization and Immunohistochemical Study of Epidermal Growth Factor Receptor in the Human Inflamed Gingival Epithelium

Bae, Chang,Kim, Young Ho THE CATHOLIC UNIVERSITY OF KOREA 1997 Bulletin of The Catholic Research Institutes of Me Vol.25 No.-

The authors investigated the expression of EGFR in the normal and inflamed human gingival epithelium to Study the role of EGFR in the inflammation of the gingival epithelium by using in situ mRNA Hybridization and immunohistochemistry. The results showed: (1) The expression of EGFR mRNA in the normal gingival epithelium on in situ mRNA hybridization was mainly localized on the basal cell layer, and the spinous layer and the granular layer were weakly positive. The cornified layer wag negative. (2) The expression of EGFR mRNA in the inflamed gingival epithelium on in situ mRNA hybridization was evenly and homogeneously distributed with the high staining intensity in the whole layers of the gingival epithelium except the cornified layer. (3) The expression of EGFR protein in the normal gingival epithelium on immunohistochemistry wag mainly localized on the cornified layer and the granular layer, and the spinous layer was weakly positive, The basal cell layer was negative, (4) The expression of EGFR protein in the inflamed gingival epithelium on immunohistochemistry was evenly and homogeneously distributed with the high staining intensity in the whole layers of the gingival epithelium. In conclusion, the up-regulation of EGFR in the inflamed gingival epithelium can be regarded as responses to the possible damages to the oral environment to maintain the homeostatic conditions.

The Effects of Atropine and Glycopyrrolate on the Recovery from Mivacurium-Induced Neuromuscular Blockade in the Rabbit

Kim, Changsung,Kwon, Oukyoung THE CATHOLIC UNIVERSITY OF KOREA 1997 Bulletin of The Catholic Research Institutes of Me Vol.25 No.-

The neuromuscular effects of atropine and glycopyrrolate on the recovery from Mivacurium-induced neuromuscular blockade were evaluated in rabbits. The results were as follows; There were no significant differences in recovery indices and post-tetanic potentiations between the antimuscarinic groups(atropine groups, glycopyrrolate groups) and the control group. Significant differences were found in tetanic fades between the antimuscarinic groups and the control group. These results showed that atropine and glycopyrrolate hastened the recovery from mivacurium induced neuromuscular blockade.

Numerical Chromosome Aberrations in Bladder Cancer Using Non-Fluorescent in Situ Hybridization

Kim, Sae Woong,Cho, Yong-Hyun,Yoon, Moon Soo THE CATHOLIC UNIVERSITY OF KOREA 1997 Bulletin of The Catholic Research Institutes of Me Vol.25 No.-

Bladder cancers vary greatly in their biological behavior, ranging from superficial to invasive and have stepwise carcinogenetic process. By the development of cytogenetics and molecular genetics, it is generally accepted that numerical aberration and deletion of specific chromosome is closely related with carcinogenetic process. But the conventional cytogenetics have limitations in chromosome study due to metaphase analysis and limited type of probe. Recently in situ hybridization of interphase cells and loss of heterozygosity using microsatellite markers have introduced for the molecular cytogenetic study in paraffin-embedded solid tumor. We performed interphase cytogenetics by non-fluorescent in situ hybridization and microsatellite analysis of 21 cases of bladder cancer in chromosome 7, 9, 17 and long arm of chromosome 9. The results were analyzed to investigate the role of stepwise carcinogenetic process and the relationship between centromeric copy number or loss of heterozygosity and tumor of grade or stage. The results were as follows; 1. Nineteen out of twenty one patients revealed numerical aberration at one or more chromosome. The changes of chromosome number were most frequently found in chromosome 7(15 cases) followed by chromosome 9(13 cases) and 17(11 cases) 2. In the distribution of centromeric copy number for chromosome 7, trisomy was most frequently found and followed by tetrasomy, diasomy, monosomy. In chromose 9, monosomy was most frequently found and followed by diasomy, trisomy, tetrasomy. In chromosome 17, diasomy was frequently found and followed by trisomy, tetrasomy, monosomy. 3. In the relationship between centromeric copy number and tumor of grade or stage, chromosome 7 and 17 were significantely correlated with increasing tumor of grade and stage and chromosome 7 is more strongly correlated than chromosome 17 with tumor grade and stage. Chromosome 9 did not significantely correlated with either grade or stage. Monosomy 9 was even distributed at all grade and stage and polysomy 7 and 17 were frequently found at high grade and stage. Our results suggest that monosomy 9 may be early event in the oncogenesis of bladder cancer and centromeric copy number of 7 and 17 may be highly predictive of bladder tumor aggressiveness. Molecular cytogenetic analysis using in situ hybridization may offer possible role of pathogenesis in bladder cancer.

The Expression of IL-4 and Interferon-gamma Genes in Proportion to Serum IgE Levels in Peripheral Blood Mononuclear Cell of Patien

Rhyu, Mun Gan,Kim, Jin Wou,Park, Chul Jong,Kim, Chung Won,Yi, Jong Yuk THE CATHOLIC UNIVERSITY OF KOREA 1997 Bulletin of The Catholic Research Institutes of Me Vol.25 No.-

IL-4 and IFN-γ gene expressions in peripheral mononuclear cells(PBMC) were compared by semi-quantitative reverse transcription-polymerase chain reaction (semi-quantitative RT-PCR) between atopic dermatitis(AD) patients showing high serum IgE level and those showing low serum IgE level. The subjects of this study were 15 cages; 5 AD patients with high serum IgE(>2,000kU/l), 5 AD patients with low serum IgE(<100kU/l), and 5 healthy controls. Cultures of PBMC were harvested at 0 hr(unstimulated), 3 hr, 6 hr, 12 hr, and 24 hr(stimulated). Phorbol 12-myristate 13-acetate in combination with calcium ionophore A231879(PMA / CA) was used as a stimulant. Semi-quantitative RT-PCR was performed. The results were as follws; 1) IL-4 gene expression in unstimulated PBMC was higher at AD patient groups than at control group but statistically higher only at AD patient group with high serum IgE level than at control group (P<0.05). 2) IFN-γ gene expression in unstimulnted PBMC was higher at AD patient groups than at control group without statistical significance. 3) IL-4 mRNA was first detected at 3 hr after stimulation in vitro and persisted until 6 hr, and slowly decreased in all three groups. IL-4 gene expression was higher at 3 hr than at 12 hr or 24 hr in AD patient group with high IgE level(P<0.05). 4) IFN-γ gene expression was first detected at 3 hr and Persisted until 24 hr. IFN-γ gene expression was lower at 24hr than at 3 hr, 6hr, and 12hr in AD patient group with low IgE level(P<0.05). 5) IL-4 and IFN-γ gene expressions in stimulated PBMC were not different among all three groups. In light of our results, it is suggested that T cells of AD patients are more activated in vivo because IL-4 and IFN-γ gene expressions are increased at AD patient groups than at control group. Since there are no differences of IL-4 and IFN-γ gene expressions in PBMC stimulated with PMA / CA, it may be suggested that AD is not characterized by the shift in the reciprocal relationship between IL-4 and IFN-γ production when T cells are stimulated under antigen presenting cell - independent conditions.

Concurrent Chemoradiation Therapy in Stage Ⅲ Non-small Cell Lung Cancer

Song, Jung Sub,Jang, Jie Young,Shinn, Kyung Sub,Lee, Sun Hee,Choi, Ihl Bhong,Kim, In Ah,Kang, Ki Mun,Park, Jae Gil,Kuak, Mun Sub THE CATHOLIC UNIVERSITY OF KOREA 1997 Bulletin of The Catholic Research Institutes of Me Vol.25 No.-

This study was tried to evaluate the potential benefits of concurrent chemoradiation therapy(low dose daily cisplatin combined with split course radiation therapy) compared with conventional radiation therapy alone in stage Ⅲ non-small cell lung cancer. The end points of analyses were response rate, overall survival, survival without locoregional failure, survival without distant metastasis, prognostic factors affecting survival and treatment related toxicities. Between April 1992 and March 1994, 32 patients who had stage Ⅲ non-small cell lung cancer were treated with concurrent chemoradiation therapy. Radiation therapy for 2 weeks (300 cGy given 10 times up to 3000 cGy) followed by a 3 weeks rest period and then radiation therapy for 2 more weeks(250 cGy given 10 times up to 2500 cGy) was combined with 6㎎/M^2 of cisplatin. Follow-up period ranged from 13 months to 48 months with median of 24 months. Historical control group consisted of 32 patients who had stage Ⅲ non-small cell lung cancer were received conventionally fractionated(daily 170-200 cGy) radiation therapy alone. Total radiation dose ranged from 5580 cGy to 7000 cGy with median of 5940 cGy. Follow-up period ranged from 36 months to 105 months with median of 62 months. Complete reponse rate was higher in chemoradiation therapy(CRT) group than radiation thernpy(RT) group(18.8% vs. 6.3%). CRT group showed lower in-field failure rate compared with RT group(25% vs. 47%). The overall survival rate and no significant differences in between CRT group and RT group(17.5% vs. 9.4% at 2 years). The survival without locoregional failure (16.5% vs. 5.3% at 2 years) and survival without distant metastasis(17% vs. 4.6% at 3 years) also had no significant differences. In subgroup analyses for patients with good performance status(Karnofsky performance scale≥80), CRT group showed significantly higher overall survival rate compared with RT group(62.5% vs. 15.6% at 2 years). The prognostic factors affecting survival rate were performance status and pathologic subtype(sqamous cell cancer vs. nonsquamous cell cancer) in CRT group. In RT alone group, performance status and stage (Ⅲa vs. Ⅲb) were identified as a prognostic factors. RTOG/EORTC grade 2-3 nausea and vomiting(22% vs. 6%) and bone marrow toxicities(25% vs. 15.6%) were significantly higher in CRT group compared with RT alone group. The incidence of RTOG/EORTC grade 3-4 pulmonary toxicity had no significant differences in between CRT group and RT group(16% vs. 6%). The incidence of WHO grade 3-4 pulmonary fibrosis also had no significant differences in both group(38% vs. 25%). In analyses for relationship of field size and pulmonary toxicity, the patients who treated with field size beyond 200㎠ had significantly higher rates of pulmonary toxicities. Conclusions: The CRT group showed significantly higher local control rate than RT group. There were no significant differences of survival rate in between two groups. The subgroup of patients who had good performance status showed higher overall survival rate in CRT group than RT group. In spite of higher incidence of acute toxicities with concurrent chemoradiation therapy, the survival gain in subgroup of patients with good performance status were encouraging. CRT group showed higher rate of early death within 1 year, higher 2 year survival rate compared with RT group. Therefore, to evaluate the accurate effect on survival of con- current chemoradiation therapy, systematic follow-up for long termsurvivors are needed.

The Ileal Permeability Changes on Temporary Occlusion of Superior Mesentric Artery in Rats

Park, Seung Cheo,Cho, Haeng Sik,Kim, Eung Kook,Chun, Sung Won,Song, Young Tack THE CATHOLIC UNIVERSITY OF KOREA 1997 Bulletin of The Catholic Research Institutes of Me Vol.25 No.-

Ischemia is a common clinical event with potenitally serious consequence. The small intestine is one of the most sensitive tissue to ischemia-reperfusion injury. Reperfusion of intestine is often associated with increases in vascular and mucosal permeability and mucosal ulceration. Several studies have shown that relatively little injury to the intestinal mucosa occurs during the ischemic period, the majority occuring upon reperfusion. The exact mechanisms leading to mucosal injury, due to mesenteric ischemia-reperfusion, have not been fully elucidated, yet here is an increasing body of evidence to suggest a role for xanthine oxidase derived oxygen metabolites. The purpose of this study was to examine the mucosal permeability in mesenteric ischemia-peperfusion injury. More specifically, this study attempts to evaluate the effect of aminophylline, as xanthine derivatives, and allopurinol, a xanthine oxidase inhibitor, on temporary occlusion of the superior mesenteric artery of rats. The experimental rats were divided into 4 groups : group Ⅰ was sham operated control group(n=10), group Ⅱ was superior mesentric artery(SMA) clamping for 35 min(n=10), group Ⅲ was injected aminophylline (5㎎/100g B.W) into inferior vena cava 5 min before declamping of SMA(n=10), group Ⅳ was injected allopurinol(5㎎/100g B.W) intraperitoneally Ⅰ hour before clamping of SMA(n=10). Mucosal permeability was determined quantitatively from double ligated blind loop of the distal ileum, measuring the luminal-to-plasma clearance of ^51Cr-EDTA (cpm/10㎕/min) at 30 minutes after reperfusion in all groups. The results were as follows : 1. The mucogal permeability(cpm/10㎕/min) in group Ⅰ was 173.2±40.4, group Ⅱ was 415.5±49.4, group Ⅲ was 588.0±126.61 and group Ⅳ was 300.6±94.9. 2. Group Ⅱ showed significant increases in mucosal permeability compared with sham control rats(P< 0.001). 3. Aminophylline-treated rats(group Ⅲ) were more significantly increased the mucosal permeability than sham control group as 3.4 times, and than simple SMA clamping group(group Ⅱ ) as 1.4 times(P<0.002). 4. Allopurinol(group Ⅳ) significantly attenuated the mucosal permeability compared with simple SMA clamping group(P<0.005). This experimental study confirmed that the xanthine oxidase-derived oxygen metabolites play a significant role in the mucosal injury, due to increases in mucosal permeability, of mesenteric ischemia-reperfusion model of rats.

Detection of Minimal Lesion and Identification of Clonality in Malignant Lymphoma

Lee, Kyo Young,Kim, Won Il,Kim, Yong Goo,Kang, Chang Suk,Kim, Young Shin,Han, Kyungja,Shim, Sang In THE CATHOLIC UNIVERSITY OF KOREA 1997 Bulletin of The Catholic Research Institutes of Me Vol.25 No.-

The bone marrow biopsy is an integral of the staging process in patients with malignant lymphomas. Marrow involvement indicates stage Ⅳ disease, but there are always some cases in which clear distinction is not possible based on morphology alone. Additional difficulties are caused by morphologic discordance between the bone marrow and the primary lymphoma. Immunohistochemical stain, mRNA in situ hybridization and polymerase chain reaction were performed to find out the minimal lesion and the clonality in formalin fixed paraffin embedded tissues of 39 primary lymphomas and corresponding bone marrow biopsy specimens. As a result, nine morphologically negative bone marrows of 18 lymphomas were positive by PCR (Group Ⅰ). Among the 6 lymphomas with morphologically suspicious bone marrow involvement (Group Ⅱ), one was confirmed to be positive for marrow involvement by both mRNA ISH and PCR and other four cases by PCR alone. The positive bone marrows of Group Ⅰ and Ⅱ revealed gene rearrangement at the same time as the primary lesion, suggesting the same clonality. Thirteen of 15 lymphomas with morphologically positive bone marrow(Group Ⅲ) had the same clonality in the primary lymphomas and the bone marrow lesion. Three cases among the Group Ⅲ with morphologic discordance also revealed the same clonality by PCR. This study shows that combination of mRNA ISH and PCR in addition to immunohistochemical stain improve the diagnostic sensitivity in the detection of bone marrow involvement and identification of clonality, and among the three methods used, PCR is the most sensitive in detecting minimal lesion.

Frequency of Multiple Consecutive Rib Fractures After Blunt Chest Trauma

Ah, Kyung,Kim, Young Joo,Shinn, Kyung Sub,Park, Seog Hee,Son, Young Bo,Kim, Sung Hoon THE CATHOLIC UNIVERSITY OF KOREA 1997 Bulletin of The Catholic Research Institutes of Me Vol.25 No.-

Rib fractures often are not seen on initial radiographs. This study was undertaken to determine the frequency of the multiple consecutive rib fractures after blunt chest trauma. We retrospectively reviewed all the radiographs and medical charts of 87 patients with rib fractures after blunt chest trauma. Rib fractures were divided into single and multiple. The multiple rib fractures were subclassified as consecutive, interrupted continuous, or random. Thirteen cases of single and 83 cases of multiple rib fractures were present. Among the 83 cases of multiple factures, there were 73 cases(88%) of consecutive fractures, 5 cases(6%) of interrupted continuous fractures and 5 cases(6%) of random fractures. Among the multiple rib fractures consecutive rib fractures are much more common than noncontiguous rib fractures. We recommend that if one find an apparently nonfractured rib between contiguously fractured upper and lower ribs, one should meticulously search for possible fracture of an apparently normal rib with high-index of suspicion.

Alteration of Renal Parenchyma on Magnetic Resonance Imaging after Renal Artery Ligation : Experimental Study in the Rabbit

Park, Seog Hee,Kim, Young Joo,Kim, Sung Hoon,Chun, Kyung Ah,Jeon, Hae Myung,Kim, Euy Neyng,Lee, Eun Jung,Shinn, Kyung Sub THE CATHOLIC UNIVERSITY OF KOREA 1997 Bulletin of The Catholic Research Institutes of Me Vol.25 No.-

The purpose of this study is to evaluate the sequential change of magnetic resonance imaging(MRI) in the induced renal infarction after renal artery ligation. Renal infarction was induced in 10 rabbits by surgical ligation of the left renal artery. All MRIs were performed with a 1.0T MR system and, a spin-echo technique was used with varying TR and TE parameters on the transverse plane. Kidneys were evaluated before and 2 hours, 1 day, 1 week and 4 weeks after ligation of the left renal artery, and spin echo images were analyzed for the cortex-to-medulla contrast(CMC) and Tl, T2 relaxation times in the cortex and the medulla of both kidneys. Changes in renal size after ligation were also evaluated. At two hours after ligation of the left renal artery, the cortex of the left kidney on MRI showed a 25% shortening of the mean T2 relaxation times(P<0.001) and a 21% shortening of the mean 71 relaxation time(P<0.001). The relaxation times of the cortex increased relative to normal value on one day and ore week after ligation. Four weeks after ligation, relaxation time of cortex and medulla of the left kidney decreased. One day after ligation, CMC of the left kidney decreased 41% on T2-weighted image. One week after ligation, CMC of the left kidney decreased 62% on T1-weighted image(T1WI). There were no significant changes either in relaxation times of the medulla of the left kidneys or in relaxation times and CMC of the right kidneys 2 hours, one day and 1 weeks after ligation. T1 and T2 relaxation times were significantly prolonged on the right kidney in the cortex and medulla 4 weeks after ligation of the left renal artery. In conclusion, MRI is capable of detecting total renal infarction. Change of CMC in T1WI and T2-weighted image(T2WI), and T1 and T2 relaxation times of the cortex were suggested to be the useful MR findings for the diagnosis of renal infarction.