A standardized method to study immune responses using porcine whole blood

Background@#Peripheral blood mononuclear cells (PBMCs) are commonly used to assess in vitro immune responses. However, PBMC isolation is a time-consuming procedure, introduces technical variability, and requires a relatively large volume of blood. By contrast, whole blood assay (WBA) is faster, cheaper, maintains more physiological conditions, and requires less sample volume, laboratory training, and equipment. @*Objectives@#Herein, this study aimed to develop a porcine WBA for in vitro evaluation of immune responses. @*Methods@#Heparinized whole blood (WB) was diluted (non-diluted, 1/2, 1/8, and 1/16) in RPMI-1640 media, followed by phorbol myristate acetate and ionomycin. After 24 h, cells were stained for interferon (IFN)-γ secreting T-cells followed by flow cytometry, and the supernatant was analyzed for tumor necrosis factor (TNF)-α. In addition, diluted WB was stimulated by lipopolysaccharide (LPS) and polyinosinic:polycytidylic acid (poly I:C), reference strain KCTC3557 (RS), field isolate (FI), of heat-killed (HK) Streptococcus suis, and porcine reproductive and respiratory syndrome virus (PRRSV). @*Results@#The frequency of IFN-γ + CD3 + T-cells and concentration of TNF-α in the supernatant of WB increased with increasing dilution factor and were optimal at 1/8. WB TNF-α and interleukin (IL)-10 cytokine levels increased significantly following stimulation with LPS or poly I:C. Further, FI and RS induced IL-10 production in WB. Additionally, PRRSV strains increased the frequency of IFN-γ + CD4 - CD8 + cells, and IFN-γ was non-significantly induced in the supernatant of re-stimulated samples. @*Conclusions@#We propose that the WBA is a rapid, reliable, and simple method to evaluate immune responses and WB should be diluted to trigger immune cells.

¹⁸FFET PET is a useful tool for treatment evaluation and prognosis prediction of anti-angiogenic drug in an orthotopic glioblastoma mouse model / 한국실험동물학회지

O-2-¹⁸F-fluoroethyl-l-tyrosine ([¹⁸F]FET) has been widely used for glioblastomas (GBM) in clinical practice, although evaluation of its applicability in non-clinical research is still lacking. The objective of this study was to examine the value of [¹⁸F]FET for treatment evaluation and prognosis prediction of anti-angiogenic drug in an orthotopic mouse model of GBM. Human U87MG cells were implanted into nude mice and then bevacizumab, a representative anti-angiogenic drug, was administered. We monitored the effect of anti-angiogenic agents using multiple imaging modalities, including bioluminescence imaging (BLI), magnetic resonance imaging (MRI), and positron emission tomography-computed tomography (PET/CT). Among these imaging methods analyzed, only [¹⁸F]FET uptake showed a statistically significant decrease in the treatment group compared to the control group (P=0.02 and P=0.03 at 5 and 20 mg/kg, respectively). This indicates that [¹⁸F]FET PET is a sensitive method to monitor the response of GBM bearing mice to anti-angiogenic drug. Moreover, [¹⁸F]FET uptake was confirmed to be a significant parameter for predicting the prognosis of anti-angiogenic drug (P=0.041 and P=0.007, on Days 7 and 12, respectively, on Pearson's correlation; P=0.048 and P=0.030, on Days 7 and 12, respectively, on Cox regression analysis). However, results of BLI or MRI were not significantly associated with survival time. In conclusion, this study suggests that [¹⁸F]FET PET imaging is a pertinent imaging modality for sensitive monitoring and accurate prediction of treatment response to anti-angiogenic agents in an orthotopic model of GBM.

Expression analysis of ciliary rootlet coiled coil protein mRNA during Xenopus development / 대한수의학회지

Ciliary rootlet coiled coil protein (CROCC), the structural component that originates from the basal body at the proximal end of the ciliary rootlet, plays a crucial role in maintaining the cellular integrity of ciliated cells. In the current study, we cloned Xenopus CROCC and performed the expression analysis. The amino acid sequence of Xenopus laevis was related to those of Drosophila, cow, goat, horse, chicken, mouse and human. Reverse transcription polymerase chain reaction analysis revealed that CROCC mRNA encoding a coiled coil protein was present maternally, as well as throughout early development. In situ hybridization indicated that CROCC mRNA occurred in the animal pole of embryo during gastrulation and subsequently in the presumptive neuroectoderm at the end of gastrulation. At tailbud stages, CROCC mRNA expression was localized in the anterior roof plate of the developing brain, pharyngeal epithelium connected to gills, esophagus, olfactory placode, intestine and nephrostomes of the pronephric kidney. Our study suggests that CROCC may be responsible for control of the development of various ciliated organs.

Association analyses of DNA polymorphisms in immune-related candidate genes GBP1, GBP2, CD163, and CD169 with porcine growth and meat quality traits / 동물의과학연구지

Genetic polymorphisms within immunity-related candidate genes in pigs have been identified to control variations in immune functions and/or disease resistance. It has become necessary to evaluate the effects of other genetic markers of economically important traits prior to introducing them into marker-assisted selection programs. In this study, polymorphisms of porcine genes coding Interferon-induced Gunylate binding protein 1 (GBP1), GBP2, CD163, and CD169 were investigated for their association with growth and meat quality traits in a Korean native pig breed-Yorkshire inter-crossed F2 pig population (KY-F2). KY-F2 animals (n=346) have been successfully used for linkage mapping to identify quantitative loci that control meat quality, growth, and immunity traits. In our results, polymorphisms in genes GBP1 and GBP2 showed association with pig growth rate as well as meat quality traits such as crude fat, drip loss, and meat color (yellowness) in the KY-F2 population. The polymorphism in gene CD163 only showed association with crude fat, as a meat quality trait. CD169 gene was associated with pork tenderness. In conclusion, four immune-related genetic markers were validated for their association with growth and meat quality traits to gauge their potential use in a swine selection program. The results warrant further studies in other commercial pig populations.

Human Anaplasmosis in Acute Febrile Patients during Scrub Typhus Season in Korea

No abstract available.

First Report of Rhizopus oryzae as a Postharvest Pathogen of Apple in Korea

Soft rot in apple caused by Rhizopus oryzae was found for the first time in Korea. A detailed description of the specimen is given along with its internal transcribed spacer rDNA sequence. The fungus was identified as Rhizopus oryzae based on the mycological characteristics, molecular data, and pathogenicity testing.

Cases series: ultrasound-guided supraclavicular block in 105 patients / 대한마취과학회지

BACKGROUND: Multiple needle attempts to gain a muscle twitch or a paresthesia for a classical supraclavicular brachial plexus block can increase the risks of nerve damage or pain. The aims of this study were to obtain reliable clinical data on ultrasound-guided supraclavicular blocks, demonstrate the higher success rate and fewer complications, and design an injection method for patients whose brachial plexus can not be located. METHODS: 105 patients received an ultrasound-guided supraclavicular block. 40 ml of 1% mepivacaine was injected without a muscle twitch or paresthesia. The groups were divided into two groups - Group A (n = 92, patients who had visible brachial plexus) and Group B (n = 13, patients whose brachial plexus can't be located). After the blocks, the clinical characteristics such as the success rate, the time to onset, the extent of the sensory block, and occurrence of complications were evaluated. RESULTS: The Success rate of Group A (98.9%) was higher than that of Group B (84.6%) (P < 0.05). The overall success rate was 97.1%. All patients could be operated on under sedation. The time to onset of Group A (12.6 +/- 4.4 min) was shorter than that in Group B (23.1 +/- 5.1 min) (P < 0.05). The overall time to onset was 13.8 +/- 5.5 min. There were no serious complications such as pneumothorax. CONCLUSIONS: An ultrasound-guided supraclavicular block is very effective in even patients whose brachial plexus can not be located.

Comparison of a supraclavicular block showing upper arm twitching response with a supraclavicular block showing wrist or finger twitching response / 대한마취과학회지

BACKGROUND: Although an ultrasound-guided brachial plexus block has become the standard, conventional brachial plexus blocks with a paresthesia or muscle twitch are still performed. However despite eliciting a paresthesia or muscle twitch, there are some cases in whom the brachial plexus block fails. This has been attributed to the difference between the proximal response (PR) and distal response (DR). Therefore, this study compared a supraclavicular block showing a PR with that showing a DR. In addition, clinical data such as success rate, onset time, and complications were examined. METHODS: Eighty three patients received a supraclavicular block with a nerve stimulator. All blocks were performed with 1% mepivacaine 40 ml. The subjects were divided into two groups-Group PR (n = 20, contraction of triceps or biceps) and Group DR (n = 63, flexion or extension of wrist or fingers) according to the types of muscle twitch. The success rate, onset time, and complications were measured and evaluated. RESULTS: The success rate of Group DR (93.7%) was higher than that of Group PR (75.0%) (P < 0.05). The onset times of Group PR and DR were 15.3 +/- 6.7 min and 14.4 +/- 6.0 min, respectively. CONCLUSIONS: The elicitation of a DR was more effective in increasing the success rate and reducing the onset time than the elicitation of a PR in a single-injection supraclavicular block.

Inhibition of Graft Versus Host Disease Using CD4+ CD25+ T Cells Induced with Interleukin-2 in Mismatched Allogeneic Murine Hematopoietic Stem Cell Transplantation

BACKGROUND: In kidney transplantation, donor specific transfusion may induce tolerance as a result of some immune regulatory cells against the graft. In organ transplantation, the immune state arises from a relationship between the immunocompromised graft and the immunocompetent host. However, a reverse immunological situation exists between the graft and the host in hematopoietic stem cell transplantation (HSCT). In addition, early IL-2 injections after an allogeneic murine HSCT have been shown to prevent lethal graft versus host disease (GVHD) due to CD4+ cells. We investigated the induction of the regulatory CD4+CD25+ cells after a transfusion of irradiated recipient cells with IL-2 into a donor. METHODS: The splenocytes (SP) were obtained from 6 week-old BALB/c mice (H-2(d)) and irradiated as a single cell suspension. The donor mice (C3H/ He, H-2(k)) received 5x10(6) irradiated SP, and 5,000 IU IL-2 injected intraperitoneally on the day prior to HSCT. The CD4+CD25+ cell populations in SP treated C3H/He were analyzed. In order to determine the in vivo effect of CD4+CD25+ cells, the lethally irradiated BALB/c were transplanted with 1x10(7) donor BM and5x10(6) CD4+CD25+ cells. The other recipient mice received either 1x10(7) donor BM with 5x10(6) CD4+ CD25- cells or the untreated SP. The survival and GVHD was assessed daily by a clinical scoring system. RESULTS: In the MLR assay, BALB/c SP was used as a stimulator with C3H/He SP, as a responder, with or without treatment. The inhibition of proliferation was 30.0 13% compared to the control. In addition, the MLR with either the CD4+CD25+ or CD4+CD25- cells, which were isolated by MidiMacs, from the C3H/He SP treated with the recipient SP and IL-2 was evaluated. The donor SP treated with the recipient cells and IL-2 contained more CD4+CD25+ cells (5.4+/-1.5%) than the untreated mice SP (1.4+/-0.3%)(P<0.01). There was a profound inhibition in the CD4+CD25+ cells (61.1+/-6.1%), but a marked proliferation in the CD4+CD25- cells (129.8+/-65.2%). Mice in the CD4+CD25+ group showed low GVHD scores and a slow progression from the post-HSCT day 4 to day 9, but those in the control and CD4+CD25- groups had a high score and rapid progression (P<0.001). The probability of survival was 83.3% in the CD4+CD25+ group until post-HSC day 35 and all mice in the control and CD4+CD25- groups died on post-HSCT day 8 or 9 (P=0.0105). CONCLUSION: Donor graft engineering with irradiated recipient SP and IL-2 (recipient specific transfusion) can induce abundant regulatory CD4+CD25+ cells to prevent GVHD.

Mycobacterium bovis Infection in a Farmed Elk in Korea

A case of tuberculosis is reported in an eight-year-old, male, elk (Cervus elaphus nelsoni). The elk showed severe coughing, respiratory distress, abdominal breathing, anorexia, and severe progressive emaciation in the elk farm. At necropsy, the elk appeared in poor body condition. Mild enlargement of retropharyngeal and submandibular lymph node was observed in the head. Diffuse fibrinous pleuritis and purple red lobar pneumonia were found in the thorax. Well demarcated numerous dark yellow discrete or confluent nodules from 0.3 to 2 cm in diameter were scattered in the whole lung. Bronchial and mediastinal lymph nodes were also enlarged. Histopathologically, lungs had typical classical tuberculous granulomas, multiple abscesses, and numerous macrophages and Langhans giant cells infiltration in alveolar lumen. In the lymph nodes, there were small clusters of necrosis and infiltration of numerous macrophages, epithelioid cells, and Langhans giant cells. With the acid-fast staining, numerous mycobacteria were revealed in the lung and lymph nodes. According to this study, there are differences of the histopathologic lesions and the numbers of acid-fast bacilli in the lesions between this elk and cattle. Mycobacterium bovis was confirmed as a causative agent in this elk using bacterial isolation, biochemical characteristics, and PCR technique. The isolate was negative for niacin test, nitrate reductase, and pyrazinamidase. This is a first report for bovine tuberculosis of farmed elk in Asia.

Effect of Nalbuphin Dosage and Incision Length of Abdominal Wall on Return of Bowel Function after Colectomy

PURPOSE: Nalbuphin has definitive advantages over the more commonly used narcotic analgesic:a ceiling respiratory depression, little effect on the cardiovascular system and a lower incidence of nausea and vomiting. The use of a small incision results in early return of bowel function and shortening of hospital stay. Narcotic use has been felt to be proportional to the length of the abdominal incision. The aim of this study was to determine whether return of bowel function after colectomy in the postoperative period and incision length were directly proportional to the narcotics. METHODS: 38 patients undergoing colon and rectal resection for benign and malignant colorectal disease between July 2000 and April 2001 participated in this study. Nalbuphin and ketorolac was administered continually by patient controlled analgesia for 48 hours. Additional nalbuphin was used for further pain control. Patients were followed for return of bowel function as measured by first audible bowel sounds, first passage of flatus and first defecation. RESULTS: There was a significant correlation between the amount of total nalbuphin administered and return of bowel function as measured by bowel sound (r=0.89; P=0.01), time to first passage of flatus (r=0.76; P=0.01), and time to first defecation (r=0.58; P=0.05). Incision length did not show any correlation with either nalbuphin use or return of bowel function. CONCLUSIONS: There is no apparent benefit for lesser incision length. Return of bowel function is influenced by use of postoperative nalbuphin. So adequate sized abdominal incision is needed and lesser use of narcotics is more beneficial for the return of bowel function.

One Hundred Cases of HLA-matched Sibling Allogeneic bone Marrow Transplantation in Children: Single Center Study

PURPOSE: We reviewed 100 cases of HLA-matched sibling allogeneic bone marrow transplantation(allo-BMT) in children and wish to share these results. MEHTODS: One hundred children had undergone allo-BMT from HLA-identical siblings between Nov. 1983 and May 1998. There were 50 males and 50 females with a median age of 10 years and a median follow-up of 38 months. Out of 100 cases, 43 children were transplanted for severe aplastic anemia (SAA), 29 for acute myelogenous leukemia (AML), 18 for acute lymphocytic leukemia (ALL), 8 for chronic myelogenous leukemia (CML) and 2 for hemophagocytic lympho-histiocytosis (HLH). RESULTS: SAA : The 5-year event free survival (EFS) of SAA was 91%. The types of events that occurred were 3 thrombotic thrombocytopenic purpura (TTP), 2 venoocclusive disease (VOD) and 1 rejection. AML : In 25 of 29 cases, the 4-year EFS after allogeneic BMT in first remission was 71%. That of the TBI-based and Busulfan-based group was 44% and 77%, respectively. The most favorable results were observed in the Busulfan-based group in first remission with an EFS of 81% (n=18). The types of events that occurred were 4 TTP, 3 VOD, 2 rejections and 1 relapse. ALL : Five-year EFS of children with complete remission (CR; n=14, 7 CR1, 7 CR2) was 81%. CML : For the 6 children who received transplants while in the first chronic phase, the event free survival was 67%. HLH : Both of the two children with HLH survived 9 months and 24 months after BMT, respectively. Acute GVHD (> or =Grade ll) was observed in 13 children. Chronic GVHD developed in 10 children; 8 cases were localized and 2 were extensive type. CONCLUSION: Allo-BMT can cure children with refractory stem cell disorders. The most important factor that influences survival after transplantation is interval between diagnosis and transplantation for patients with severe aplastic anemia and remission state at transplantation for patients with leu-

One Hundred Cases of HLA-matched Sibling Allogeneic bone Marrow Transplantation in Children: Single Center Study

PURPOSE: We reviewed 100 cases of HLA-matched sibling allogeneic bone marrow transplantation(allo-BMT) in children and wish to share these results. MEHTODS: One hundred children had undergone allo-BMT from HLA-identical siblings between Nov. 1983 and May 1998. There were 50 males and 50 females with a median age of 10 years and a median follow-up of 38 months. Out of 100 cases, 43 children were transplanted for severe aplastic anemia (SAA), 29 for acute myelogenous leukemia (AML), 18 for acute lymphocytic leukemia (ALL), 8 for chronic myelogenous leukemia (CML) and 2 for hemophagocytic lympho-histiocytosis (HLH). RESULTS: SAA : The 5-year event free survival (EFS) of SAA was 91%. The types of events that occurred were 3 thrombotic thrombocytopenic purpura (TTP), 2 venoocclusive disease (VOD) and 1 rejection. AML : In 25 of 29 cases, the 4-year EFS after allogeneic BMT in first remission was 71%. That of the TBI-based and Busulfan-based group was 44% and 77%, respectively. The most favorable results were observed in the Busulfan-based group in first remission with an EFS of 81% (n=18). The types of events that occurred were 4 TTP, 3 VOD, 2 rejections and 1 relapse. ALL : Five-year EFS of children with complete remission (CR; n=14, 7 CR1, 7 CR2) was 81%. CML : For the 6 children who received transplants while in the first chronic phase, the event free survival was 67%. HLH : Both of the two children with HLH survived 9 months and 24 months after BMT, respectively. Acute GVHD (> or =Grade ll) was observed in 13 children. Chronic GVHD developed in 10 children; 8 cases were localized and 2 were extensive type. CONCLUSION: Allo-BMT can cure children with refractory stem cell disorders. The most important factor that influences survival after transplantation is interval between diagnosis and transplantation for patients with severe aplastic anemia and remission state at transplantation for patients with leu-

Allogeneic Bone Marrow Transplantation for Myelodysplastic Syndromes / 대한혈액학회지

BACKGROUND: The myelodysplastic syndromes (MDS) can be categorized as a group of clonal hematopoietic disorders characterized by ineffective hematopoiesis and peripheral cytopenias. Although the natural history of MDS varies, traditional treatments are not curative and allogeneic marrow transplantation offers potentially curative treatment for MDS. METHODS: In our center, 10 patients underwent allogeneic bone marrow transplantation (BMT) between December 1989 and May 1997. The minimum follow-up of 3 months was possible in 10 patients, for whom treatment-related complications and clinical outcomes were assessed. RESULTS: The median age of the 10 patients was 33 (range 20~40) years. The median time from diagnosis to BMT was 34 (3~116) months. By morphology, 5 patients had advanced MDS (i.e., RAEB, RAEB-t, CMML) and 5 patients had less advanced MDS (RA). By Bournemouth score, 8 patients had a score 2~3 and two patients had a score 4. By IPSS, 5 patients were in intermediate-1 group, 3 patients in intermediate-2 group and 2 patients in high risk group. Patients were prepared for transplant with either a total body irradiation (TBI)+cyclophosphamide (n=7), busulfan+TBI (n=2) and busulfan+cyclophophamide (n=1). All patients received CsA+short course MTX for GVHD prophylaxis. Successful engraftment was confirmed in all patients. The overall incidence of acute GVHD was noted in 70% (7/10 patients) and grade IV acute GVHD developed in 2 patients (20%). Five patients were evaluable for the development of chronic GVHD and 2 patients (40%) developed limited chronic GVHD. The duration of median follow-up was 8.1 months. At present five patients are alive and disease-free 3 to 21 months (median survival duration : 8.2 months) post-transplantation resulting in a 2-year disease-free survival of 44%. 2-year disease free survival was 63% in less advanced MDS and 25% in advanced MDS. CONCLUSION: Allogeneic BMT should be considered when any clinical evidence of disease progression to a more advanced stage becomes apparent. International prognostic scoring system (IPSS) and Bournemouth score can also be used to gauge timing for BMT. For patients were in intermediate-1 or intermediate-2 group by IPSS, BMT can be justified if the patient is young and has an HLA matched sibling donor.

Reconstitution of Bone Marrow after Allogeneic and Autologous Bone Marrow Transplantation / 대한혈액학회지

BACKGROUND: The bone marrow transplantation (BMT) is an effective treatment for patients with several hematologic diseases. However, most studies for immune reconstitution after BMT are limited to immunophenotyping of lymphocytes in the peripheral blood and little information about cell surface antigen expression and distribution of the nucleated elements in the bone marrow after BMT have been published. To set up baseline data for bone marrow recovery after BMT, we investigated reconstitution of the bone marrow at 21th day after allogeneic and autologous BMT. METHODS: The flow cytometric analyses for 17 monoclonal antibodies in the 35 bone marrow transplant recipients (allogeneic BMT 25, autologous BMT 10) and 20 healthy donors of allogeneic BMT were performed. The percentage of positive cell population was calculated in the gated region and compared data for BMT vs normal controls and allogeneic vs autologous BMT. RESULTS: CD8+ T cell and CD56+ NK cell were the first signs of immunological recovery at 21th day after BMT, whereas the CD4+ subsets were significantly decreased. These findings were prominent in the autologous BMT. After BMT, the CD2+CD5- cell population was characteristically increased. All B cell markers were decreased and most lympocytes were expressed HLA-DR. The donor's immunophenotypes before BMT did not correlate with the recipient's immunophenotypes after BMT. CONCLUSIONS: We demonstrated that the CD8+ T cells and NK cells were main cell populations in the bone marrow for primary immunological defences at 21th day after BMT and these findings were more distinct in the autologous BMT than the allogeneic BMT. It indicated that there were more rapid reconstitution in the autologous BMT. The B cell recovery was delayed rather than T cell and increased CD2+CD5- cell population was characteristic finding in post-BMT.

Lymphocyte Immunophenotype of Chronic Myelogenous Leukemia / 대한혈액학회지

BACKGROUND: Detection of bcr/abl fusion mRNA using reverse transcription polymerase chain reaction has been used for diagnosis of chronic myelogenous leukemia (CML) and monitoring after treatment. However, this conventional method is not quantitative. Therefore, new quantitative marker for CML is necessary for the follow-up of the patients after treatment including bone marrow transplantation. Whether the lymphocytes are involved in CML clone or not is still a moot question. If the lymphocytes are involved in CML clone, there could be an abnormal surface antigen expressed on these cells. We tried to find out abnormal surface antigen expression on the lymphocytes of CML. METHODS: We analyzed the immunophenotypic distribution of the bone marrow lymphocytes using flow cytometry in 22 cases of CML and 20 normal persons, and searched for characteristic abnormal immunophenotype of CML. Both peripheral blood and bone marrow samples were analyzed using dual color immunophenotying for CD19 and CD22 expression in 14 cases of CML. RESULTS: The proportion of lymphocytes with T cell antigen expression and CD10, CD19, CD20 expression were lower in CML than in normal control (P<0.05). In contrast to these antigens, the proportion of CD22 positive lymphocytes was higher in CML than in normal control (P=0.0434). And loss of correlation between CD19 and CD22 expression was observed in CML. The proportion of CD19-/CD22+ lymphocytes in the bone marrow of 14 cases of CML was higher than that of normal control (P=0.0001). The proportion of CD19-/CD22+ lymphocytes in the peripheral blood of CML was 45.0+/-22.6%, and higher than 1.0+/-0.3% of normal control (P= 0.0000). The proportion of CD19-/CD22+ lymphocytes in the peripheral blood was higher than that of the bone marrow (P=0.0018). CD22 was not coexpressed with T cell antigens (CD2, CD3) or myeloid antigen (CD14) in CML or normal control. CONCLUSION: We demonstrated that the cells, representing unusual immunophenotype of CD19-/ CD22+, are characteristically increased in the bone marrow and peripheral blood of CML. This immunophenotype could be a valuable marker for the diagnosis of CML and monitoring after treatment.

The Expression of p53, c-erbB-2 and nm23 Proteins in Breast Cancer

Recently, p53, c-erbB-2 and nm23 proteins have been studied in breast cancer. The expression of p53 protein indicates the mutation of p53 gene known as a tumor supressor gene, and c-erbB-2 gene amplification has been considered an indicator of poor prognosis and nm23 a metastsis suppressor gene. In order to elucidate the roles and relations of these proteins in the develpoment, progression and metastasis in breast cancer, we studied 89 cases of invasive breast cancer and 32 cases of lymph node metastasis for the expression of p53, c-erbB-2 and nm23 proteins using an immunohistochemical method. The results were as follows: 1) The expression rates of p53, c-erbB-2, and nm23 proteins in breast cancer were 40.4%, 34.8% and 55.1%, respectively. Co-expression of p53 protein and c-erbB-2 protein was found in 20.2% of cases, showing the highest incidence in poorly differentiated type (40%). 2) p53 protein expression was increased in poorly differentiated type but was not statistically significant. On the other hand, the expression of nm23 protein was decreased in poorly differentiated type, which was statistically significant (p<0.05). 3) The correlation of p53 protein expression with c-erbB-2 protein expression was statistically significant (p<0.05) but that with nm23 protein was not. 4) In the cases with lymph node metastasis, discordant expression of p53, c-erbB-2 and nm23 proteins between primary tumor and the lymph node metastatic tumor was found in 9.4%, 3.1% and 18.8% of cases, respectively. The above results suggest that overexpression of p53 and c-erbB-2 proteins and downregulation of nm23 protein are associated with the tumor progression in the breast cancer.

Argyrophilic Nucleolar Organizer Regions in Acute Lymphoblastic Leukemia / 대한임상병리학회지

BACKGROUND: In acute lymphoblastic leukemia (ALL), the importance of argyrophilic nucleolar organizer regions (AgNOR) is not established. METHODS: NOR silver staining was carried out in 74 ALL patients. We analyzed the AgNOR parameters ; counting parameters including number of AgNOR per cell, percentage of cells with one cluster, and area parameters including mean AgNOR area, total AgNOR area, and its percentage of nuclear area by morphometry. Cyclin A index was evaluated by immunohistochemical stain. We compared the AgNOR parameters with cyclin A index and evaluated the differences of AgNOR parameters in accordance with FAB classification, immunophenotype, a new classification of ALL (ALL with maturation), and response to induction chemotherapy. RESULTS: A positive correlation was found between cyclin A index and AgNOR area parameters and a significant negative correlation was found between mean AgNOR area and number of AgNOR per cell (r=-0.433, P=0.000). AgNOR area parameters revealed the highest value in L3. The number of AgNOR per cell in T cell ALL was higher than non-T cell ALL (P=0.011), and the percentage of cells with one cluster was lower (P=0.002). The number of AgNOR per cell in ALLm was lower (P=0.004) and the percentage of cells with one cluster was higher than in typical ALL (P=0.002). In cases achieved complete remission (CR) after induction chemotherapy, the number of AgNOR per cell was higher (P=0.005) and the percentage of cells with one cluster was much lower than in cases failed to achieve CR (P=0.013). CONCLUSIONS: Our study indicates that the AgNOR area parameters are helpful to predict the proliferating activity of leukemic blasts in ALL. It is suggested that the number of AgNOR per cell and the percentage of cells with one cluster provide a valuable information to estimate the response to chemotherapy in ALL.

Expression of Multidrug Resistant Genes in Bone Marrow Mononuclear Cells of Patients with Myeloid Leukemia / 대한암학회지

PURPOSE: Multidrug resistance mediated by several drug resistant genes impedes the successful outcome of anti-cancer chemotherapy. In this study, we investigated the expressions of drug resistant genes encoding multidrug resistance (MDR1), multidrug resistance-associated protein (MRP), topoisomerase I (Topo I), topoisomerase II g (Topo II a) in narmal volunteers (n=12) in and patients with myeloid leukemia (n=34). Material and Method: We compared the levels of their transcripts in bone matrow mononuclear cells by semiquantitative RT-PCR. The amount of specific transcripts was represented as the optical density ratio of PCR product of target gene to that of B2- microglobulin (MG). Twenty patients of acute myelogenous leukemia (eight in remission state, twelve in refractory) and fourteen patients of chronic myelogenous leukemia (nine in chronic phase and five in blastic crisis) were examined. Twelve normal healthy persons were compared with leukemic patients. RESULTS: The expression levels of all resistant genes in normal volunteers were relatively high as those of AML patients. Regardless of the disease status including remission status of AML (complete remission versus refractory) and the phase of CML (chronic phase versus blastic phase), the expression levels of all resistant genes in patients with CML were significantly lower than in the patients with AML (p < 0.05). Of interest, the patients with refractary AML did not show any statistical difference in comparison with normal controls and even the patients with AML in complete remission. Among the four drug resistant genes, the optical density ratio of MDRl was significantly lower than that of any other genes (p<0.05). Using HL-60 cell line, we compared the changes of various resistant gene expressions before and after differentiation induced by dimethylsulfoxide. The expressions of resistant genes declined in paralle1 with granulocytic differentiation, suggesting that the induction of cell differentiation might make leukemic cells susceptible to chemotherapeutic agents. CONCLUSION: It is impossibble to explain the mechanism of drug resistance by comparing the level of drug resistant gene expression between nonnal subjects and patients with myeloid leukemias. Therefore, we suppose that longitudinal study of drug resistant gene expression is necessary to demonstrate the development of drug resistant during chemotherapy.

Autologous or Allogeneic Bone Marrow Transplantation Compared with Consolidation Chemotherapy in Acute promyelocytic Leukemia / 대한암학회지

PURPOSE: Acute promyelocytic leukemia (APL) is a distinct subtype of acute myelogenous leukemia characterized by the morphology of blast cells (M3 in FAB classification), the t (15;17) translocation, and a coagulopathy combining disseminated intravascular coagulation and fibrinolysis. It has been considered to have better response to combination chemo- therapy of an anthracycline and cytosine arabinoside and a higher cure rate than other subtypes because of recent approach of differentiating leukemic blasts by all-transretinoic acid (ATRA). The role of stem cell transplantation in APL has to be determined in comparison with that of consolidation chemotherapy. MATERIALS AND METHODS: We compared the leukemia-free survival and overall survival between APL patients receiving the consolidation chemotherapy and those undergoing the allogeneic or autologous stem cell transplantation following the high-dose anticancer therapy. Of the 65 patients achieving the first complete remission from 1992 to 1997, 33 patients were treated with 3 courses of consolidation chemotherapies and 32 with the stem cell transplantation. RESULTS: With a median follow-up of 22 months (8-60), the actuarial leukemia-free survival at 3 years was significantly higher in transplantation group than in chematherapy group (73.8% versus 33.5%; P=0.0087), and the probability of leukemic relapse was considerably lower in transplantation group than in chemotherapy group (6.3% versus 57.5%; P=0.001). The treatment-related mortalities of the groups were 0% in chemotherapy group and 14.3% in transplantation group. The main cause of deaths was relapse in the consolidation chemotherapy group. CONCLUSION: These data demonstrate that the stem cell transplantation results in better leukemia-free survival than the consolidation chemotherapy for patients with APL in the first complete remission because of lower risk of relapse.