Diagnostic Utility of a Clonality Test for Lymphoproliferative Diseases in Koreans Using the BIOMED-2 PCR Assay

BACKGROUND: A clonality test for immunoglobulin (IG) and T cell receptor (TCR) is a useful adjunctive method for the diagnosis of lymphoproliferative diseases (LPDs). Recently, the BIOMED-2 multiplex polymerase chain reaction (PCR) assay has been established as a standard method for assessing the clonality of LPDs. We tested clonality in LPDs in Koreans using the BIOMED-2 multiplex PCR and compared the results with those obtained in European, Taiwanese, and Thai participants. We also evaluated the usefulness of the test as an ancillary method for diagnosing LPDs. METHODS: Two hundred and nineteen specimens embedded in paraffin, including 78 B cell lymphomas, 80 T cell lymphomas and 61 cases of reactive lymphadenitis, were used for the clonality test. RESULTS: Mature B cell malignancies showed 95.7% clonality for IG, 2.9% co-existing clonality, and 4.3% polyclonality. Mature T cell malignancies exhibited 83.8% clonality for TCR, 8.1% co-existing clonality, and 16.2% polyclonality. Reactive lymphadenitis showed 93.4% polyclonality for IG and TCR. The majority of our results were similar to those obtained in Europeans. However, the clonality for IGK of B cell malignancies and TCRG of T cell malignancies was lower in Koreans than Europeans. CONCLUSIONS: The BIOMED-2 multiplex PCR assay was a useful adjunctive method for diagnosing LPDs.

Intravascular Papillary Endothelial Hyperplasia of the Chest Wall Misdiagnosed as a Malignancy on Fine Needle Aspiration

No abstract available.

WHO Classification of Malignant Lymphomas in Korea: Report of the Third Nationwide Study

BACKGROUND: The aim of study was to determine the relative frequency of malignant lymphoma according to World Health Organization (WHO) classification in Korea. METHODS: A total of 3,998 cases diagnosed at 31 institutes between 2005 and 2006 were enrolled. Information including age, gender, pathologic diagnosis, site of involvement and immunophenotypes were obtained. RESULTS: The relative frequency of non-Hodgkin lymphoma (NHL) and Hodgkin lymphoma (HL) was 95.4% and 4.6%, respectively. B-cell lymphomas accounted for 77.6% of all NHL, while T/natural killer (T/NK)-cell lymphomas accounted for 22.4%. The most frequent subtypes of NHL were diffuse large B-cell lymphoma (42.7%), extranodal marginal zone B-cell lymphoma (MZBCL) of mucosa-associated lymphoid tissue (19.0%), NK/T-cell lymphoma (6.3%) and peripheral T-cell lymphoma (PTCL), unspecified (6.3%), in decreasing order. The relative frequency of HL was nodular sclerosis (47.4%), mixed cellularity (30.6%), and nodular lymphocyte predominant (12.1%) subtypes. Compared with a previous study in 1998, increase in gastric MZBCL and nodular sclerosis HL, and slight decrease of follicular lymphoma, PTCL, and NK/T-cell lymphoma were observed. CONCLUSIONS: Korea had lower rates of HL and follicular lymphoma, and higher rates of extranodal NHL, extranodal MZBCL, and NK/T-cell lymphoma of nasal type compared with Western countries. Changes in the relative frequency of lymphoma subtypes are likely ascribed to refined diagnostic criteria and a change in national health care policy.

Korean Pediatric/Adolescent Lymphoma: Incidence and Pathologic Characteristics

BACKGROUND: The Hematopathology Study Group of the Korean Society of Pathologists conducted a nation-wide retrospective analysis of Korean pediatric lymphoma, to provide pathologic data on pediatric/adolescent lymphoma subtypes and features. METHODS: All lymphoma cases of all age groups were collected during a recent 2 year-period (2005-2006) from 32 institutes in Korea. Among 3,686 lymphoma patients, 142 who were age 18 or less were classified according to the World Health Organization (WHO) classification. RESULTS: Among 142 pediatric/adolescent lymphoma patients, Hodgkin lymphoma accounted for 21 (14.8%) and non-Hodgkin lymphoma (NHL) for 121 (85.2%). Hodgkin lymphoma appears to be more common in the pediatric/adolescent age group than in the all-ages group (14.8% vs 4.4%). T- and natural killer cell-NHL was more common in the pediatric/adolescent age group than in the all ages group (46.3% vs 22%). The majority of Korean pediatric/adolescent NHL cases was composed of Burkitt lymphoma, T- or B-lymphoblastic lymphoma, anaplastic large-cell lymphoma, and diffuse large B-cell lymphoma. For lymphoma patients under the age of 6 years, most had B-lymphoblastic or Burkitt lymphoma, which commonly presented at extranodal sites. CONCLUSIONS: The distribution of lymphoma subtypes in the pediatric/adolescent age group is quite different from the distribution of adults, but it was quite similar to distribution in Western countries.

Perivascular epithelioid cell tumor (PEComa) of the ascending colon: the implication of IFN-alpha2b treatment / 소아과

A 7-year-old boy presented with hematochezia and abdominal pain. A 3.7-cm-sized mass was identified in the ascending colon by abdominal computed tomography and colonoscopy. The patient underwent surgical resection. Pathological examination revealed a low-grade perivascular epithelioid cell tumor (PEComa). PEComa in the colon is very rare. Only a few cases have been reported so far. An effective treatment method for this rare tumor has not been established yet. The patient received adjuvant interferon-alpha immunotherapy for 1 year. He has been tumor-free for 26 months since the initial diagnosis. This report is the first documented case of the use of interferon-alpha for pediatric PEComa of the colon.

HPV Genotyping in Squamous Cell Carcinoma of Upper Aerodigestive Tract

BACKGROUND: Smoking and alcohol consumption are the main risk factors for squamous cell carcinoma of the upper aerodigestive tract (SCCUAT). However, human papillomavirus (HPV) has been etiologically linked with tonsillar squamous cell carcinoma (TSCC). Therefore, we investigated the etiologic role of HPV in the context of SCCUAT in Korea. METHODS: Archival paraffin block samples from 136 cases previously diagnosed as SCCUAT were randomly selected. A commercial HPV DNA chip was used for HPV genotyping. RESULTS: One hundred and seventeen cases were available after checking beta-globin (47 cases of tonsil and 70 of non-tonsil). A HPV-positive result (HPV 16 and 18) occurred in 13 cases of SCCUAT, and 12 cases were tonsil (25.5%, 12/47). Among the 12 HPV-positive patients with TSCC, nine were non-smokers and non-drinkers. Most HPV-negative patients with TSCC had a history of alcohol drinking and smoking (32/35, 91.4%). HPV infection status was not significantly associated with histological grade, clinical stage, or survival in patients with TSCC. CONCLUSIONS: HPV infection was significantly higher in patients with TSCC among those with SCCUAT. HPV may be independent risk factor in development of TSCC, such as smoking and alcohol drinking.

Analysis of HPV-other Samples by Performing HPV DNA Sequencing

BACKGROUND: HPV-other samples are designated as being positive on HPV-PCR, but negative when using specific HPV hybridization probes. We wanted to determine the types on the HPV-other samples by performing sequencing, and to know the pathologic status of the uterine cervix according to the HPV type detected on sequencing. METHODS: For HPV genotying, we used the commercially available HPV DNA Chip test, which contains 15 types of high-risk HPV and 9 types of low-risk HPV. The HPV DNA sequencing was performed for the HPV-other samples of 209 patients who subsequently underwent cervical biopsy. RESULTS: For 204 of the 209 samples, the HPV types detected by sequencing were absent types at used HPV DNA chip. For the remaining 5 samples, sequencing was impossible due to mixed peaks. HPV-81 (19.6%), HPV-61 (18.6%), HPV-62 (16.7%) and HPV-84 (13.9%) were frequently detected. For the HPV-81, -62, -71, and -72 samples, most of the samples displayed normal or LSIL. However, HPV-84 and -61 were more associated with HSIL or worse, as compared to the other types. Conclusion: HPV-81, -61, -62 and -84 were frequently found on sequencing analysis of the HPV-other samples. The pathologic status was diverse, according to the HPV type detected on sequencing.

Cytologic Diagnosis of Malignant Pleural Effusion in Multiple Myeloma: Two Case Reports

Malignant pleural effusion in multiple myeloma (MM) is extremely rare and is associated with poor prognosis. We experienced two cases of MM IgA type with malignant pleural effusion. The diagnoses were based on characteristic cytology and CD138 immunocytochemistry. The patients received several cycles of combination chemotherapy, since symptoms were more aggressive with an uncontrolled pleural effusion. We review the clinical features of these cases and literature concerning myelomatous pleural effusion.

Changes of antioxidant levels in the uterine venous plasma amniotic fluid, and chorioamnion after maternal supplementation with vitamin C and vitamin E / 대한주산의학회잡지

PURPOSE:To investigate the change in the antioxidant vitamin levels in maternal uterine venous plasma (MUVP), amniotic fluid (AF), and chorioamnion after vitamin C and E supplementation during pregnancy. METHODS:Thirty pregnant women who were scheduled for elective cesarean section between 37 and 39 gestational weeks were randomized in this study. Fifteen women were given a daily oral dose of vitamin C 1,000 mg and vitamin E 400 IU from 33~34 gestational weeks to delivery. The other fifteen women were not given, as a control group. Maternal uterine venous blood, AF, and chorioamnion were obtained after cesarean section. Lipid peroxides and oxygen-radical absorbance capacity value were measured by thiobarbituric acid reaction and Cao's method respectively. Ascorbic acid, uric acid, beta-carotene, retinol, alpha-tocopherol, and gamma-tocopherol were measured by high performance liquid chromatography. The tissue sections of chorioamnion were stained with hematoxylin-eosin and Masson-trichrome stain, and immunohistochemical stain for collagen type IV was also performed. RESULTS:The lipid peroxide levels in MUVP of the study group were significantly lower than those of the control group but in contrast, ORAC (Oxygen-radical absorbance capacity) values were lower in the control group. The alpha-tocopherol levels in MUVP, AF, and chorioamnion study group were significantly higher than those of the control group. Amniotic membrane and subepithelial stromal tissue in the study group were thicker than those in the control group. And subchorionic type IV collagen of placenta tissue in the study group was more stained than that of the control group. CONCLUSION:Maternal vitamin C and E supplementation may be beneficial in the prevention of diseases caused by oxidative stress such as preeclampsia and PROM and in increasing fibrin and type IV collagen in chorioamnion.

Solitary Splenic Metastases from Uterine Cervical Cancer: Case Reports and Review of the Literature

Splenic metastasis from gynecologic tumors is extremely rare, especially in the absence of apparent disease at other sites. We report two patients that underwent splenectomy for a solitary splenic metastasis from uterine cervical carcinoma. In case 1, a 54-year-old woman with FIGO Stage IIb squamous cell carcinoma of the uterine cervix treated with radiotherapy and chemotherapy developed a solitary splenic metastasis 10 months after initial treatment. In case 2, a 46-year-old woman with FIGO Stage IIb adenocarcinoma of the uterine cervix treated with radiotherapy and chemotherapy was found to have a solitary splenic metastasis 11 months after treatment. Thus all abdominal organs including the spleen must be evaluated for metastases during follow-up of gynecologic tumors.

Tetranucleotide Repeat Microsatellite Instability in Uterine Cervical Carcinomas

BACKGROUND: Elevated levels of microsatellite alterations at selected tetranucleotide repeat regions (EMAST) have been recently described, and they are a distinct type of microsatellite instability (MSI). We investigated the prevalence of EMAST in squamous cell carcinoma (SCC) of the uterine cervix and we determined the correlation between EMAST and the clinicopathologic parameters, HPV infection and the p53 mutation. METHODS: We examined the 3 mono-, 3 di-, and 5 tetranucleotide repeat markers in 47 cases of SCC, and we performed immunohistochemical staining for p53. HPV detection and genotyping was performed using a commercially available HPV DNA chip. RESULTS: Thirteen out of 47 cases (27.7%) were EMAST(+) with at least one of five tetranucleotide repeat markers. However, MSI at mono- and dinucleo- tide markers was noted in only one case (2.1%). EMAST was not related with stage, size, lymph node metastasis, vascular/lymphatic invasion or the depth of invasion. Positive immunostaining for p53 was significantly more common in EMAST(+) tumors than in the EMAST(-) tumors (p=0.04). HPV-infection was positive in 32 cases. EMAST was not correlated with the state of HPV infection state or the HPV genotype. CONCLUSIONS: 27.7% of the invasive SCCs of the uterine cervix exhibited EMAST, and EMAST in the SCC of the uterine cervix was significantly associated with the p53 mutation.

Changes of Interleukin-6, C-reactive Protein, and Lipid Peroxide Levels in the Umbilical Venous Plasma of Preterm Birth with or without Chorioamnionitis / 대한주산의학회잡지

OBJECTIVE:To investigate interleukin-6 (IL-6), C-reactive protein (CRP), and lipid peroxide levels in the umbilical venous plasma of preterm birth with or without histologic chorioamnionitis and to evaluate their roles in the pathophysiology in preterm labor and perinatal outcome. METHODS:This cohort study included 66 cases of preterm delivery with preterm labor and intact membranes (PTL) (n=39) and preterm premature rupture of membranes (PPROM) (n=27). The umbilical venous blood samples were collected at the time of delivery. IL-6, CRP, and lipid peroxide levels were measured by ELISA Kit, latex agglutination assay, and thiobarbituric acid reaction. Histologic chorioamnionitis was diagnosed by the presence of neutrophil infiltration into the subamnionic space. RESULTS:The prevalence of histologic chorioamnionitis was significantly higher in PPROM (59.3 %, 16/27) than in PTL (20.5%, 8/39). IL-6, CRP, and lipid peroxide levels in the umbilical venous plasma of histologic chorioamnionitis were significantly higher than those without histologic chorioamnionitis. IL-6, CRP, and lipid peroxide levels in the umbilical venous plasma of PTL with histologic chorioamnionitis were significantly higher than those of PTL without histologic chorioamnionitis. CRP levels in the umbilical venous plasma of PPROM with histologic chorioamnionitis were significantly higher than those of PPROM without histologic chorioamnionitis. Three suspected neonatal sepsis patients have increased IL-6 and lipid peroxide levels in the umbilical venous plasma compared with patients without neonatal sepsis. IL-6 levels in the umbilical venous plasma of histologic funisitis were significantly higher than those without funisitis. CONCLUSION:Preterm birth with chorioamnionitis is associated with an increased level of IL-6, CRP, and lipid peroxide in umbilical venous blood. Preterm birth with chorioamnionitis may have an effect on perinatal outcome.

Molecular Biological Differences Between Younger and Older Breast Cancer Patients Based on the Status of a Microsatellite Marker

PURPOSE: Younger women exhibit more aggressive pathological features of breast cancer than older women, based on previous studies. We wished to evaluate any molecular biological differences in breast cancer between younger and older women by determining the status of a microsatellite marker. METHODS: Microsatellite instability (MSI) and loss of heterozygosity (LOH) were investigated in paired tumour and normal tissue DNA from 32 younger (age less than 40 years old) and 32 older (age more than 50 years old) breast cancer patients with 12 simple repeated primer sets. RESULTS: MSI was observed at a single locus in 5 (15.6%) of the younger patients. In older patients, MSI was observed at a single locus in 5 (15.6%) and at multiple loci in 1 (3.1%) of the older patients. LOH was noted at a single locus in 7 (21.8%) and at multiple loci in 22 (68.7%) of 32 younger patients. In older patients, LOH was noted at a single locus in 9 (28.1%) and at multiple loci in 15 (46.9%). The greatest frequency of LOH was at loci UT5320 (37.5%), D8S321 (34.4%), D9S242 (31.3%), and D19S394 (31.3%) in younger patients and at loci L17686 (34.4%) and D19S394 (28.1%) in older patients. LOHs at D19S394 and L17686 were highly identified in both age groups. LOHs at D9S242 and D8S321 were significantly higher in the carcinomas of younger women (P=0.013, P=0.016, respectively). The LOH status was unrelated to clinical stage, nodal status, tumour size, histological grade or estrogen receptor (ER) status. A LOH at D8S321 was associated with tumor size (P=0.048) and a LOH at UT5320 was associated with histological grade (P=0.012) and ER status (P=0.018). CONCLUSION: These results indicate that the pattern of chromosomal alterations are not exactly the same, especially at loci D9S242 and D8S321, in the carcinomas of the two age groups and suggest that the molecular pathogenesis of the carcinomas is not similar.

Optimization and Limitation of Calcium Ionophore to Generate DCs from Acute Myeloid Leukemic Cells / Journal of the Korean Cancer Association, 대한암학회지

PURPOSE: Calcium ionophore (CI) is used to generate dendritic cells (DCs) from progenitor cells, monocytes, or leukemic cells. The aim of this study was to determine the optimal dose of CI and the appropriate length of cell culture required for acute myeloid leukemia (AML) cells and to evaluate the limitations associated with CI. MATERIALS AND METHODS: To generate leukemic DCs, leukemic cells (4 x 10(6) cells) from six AML patients were cultured with various concentrations of CI and/or IL-4 for 1, 2 or 3 days. RESULTS: Potent leukemic DCs were successfully generated from all AML patients, with an average number of 1.2 x 10(6) cells produced in the presence of CI (270 ng/ml) for 2 days. Several surface molecules were clearly upregulated in AML cells supplemented with CI and IL-4, but not CD11c. Leukemic DCs cultured with CI had a higher allogeneic T cell stimulatory capacity than untreated AML cells, but the addition of IL-4 did not augment the MLR activity of these cells. AML cells cultured with CI in the presence or absence of IL-4 showed increased levels of apoptosis in comparison to primary cultures of AML cells. CONCLUSION: Although CI appears to be advantageous in terms of time and cost effectiveness, the results of the present study suggest that the marked induction of apoptosis by CI limits its application to the generation of DCs from AML cells.

Induction of Myeloma Cell Line-specific Cytotoxic T Lymphocytes using Monocyte-derived Dendritic Cells Pulsed with Myeloma Cell Line Lysates

BACKGROUND: In multiple myeloma (MM), the idiotype (ID) determinant of the paraprotein has been used for immunotherapy using dendritic cells (DCs). However, ID-specific immune responses showed limited clinical responses after the Id vaccination. Therefore, an alternative approach using DCs pulsed with other tumor antigens is required. METHODS: We investigated the possibility of immunotherapy for MM using myeloma cell line-specific cytotoxic T lymphocytes (CTLs), that were stimulated in vitro by monocyte-derived DCs pulsed with the myeloma cell line ysates. CD14+ cells isolated from the peripheral blood of HLA-A0201+ healthy donors were cultured in the presence of GM-CSF and IL-4. On day 6, the immature DCs were pulsed with the myeloma cell line lysates (IM-9: HLA0201+ and ARH-77: HLA0201+), and then maturation of DCs was induced by the addition of TNF- alpha for 2 days. CTL lines were generated by a 2 time stimulation with DCs to the autologous CD3+ T cells. RESULTS: DCs pulsed with myeloma cell lysates showed the production of IL-12p70, but less than that of unpulsed DCs. CTLs lines stimulated with the DCs pulsing, for the myeloma cell line lysates, showed potent cytotoxic activities against autologous target cells, but not against HLA-A2-cell lines (RPMI-8226). Mature DCs pulsed with the myeloma cell line lysates showed a higher stimulatory capacity for autologous CTL when compared with mature non-pulsed DCs. CONCLUSION: These results suggest that DCs pulsed with the myeloma cell line lysates can generate potent myeloma cell line-specific CTLs for the myeloma cell-based immunotherapeutic approach in MM.

Selective Expansion of Natural Killer Cells from Peripheral Blood Mononuclear Cells by K562 Cell Line and IL-2

BACKGROUND: Several attempts have been made to expand human NK cells from peripheral blood mononuclear cells (PBMCs). This study examined the selective expansion of NK cells using interleukin 2 (IL-2) plus the K562 cell line, the expression of the NK cell receptors, and the cytotoxic activity. METHODS: The PBMCs from seven healthy volunteers were cultured in a medium containing the IL-2 plus the K562 cell line for 14 days. The expression of the activating and inhibitory receptors on the resting NK cells and the 72 hr-expanded NK cells were analyzed. A flow cytometric cytotoxic assay was used to determined the killing activity of the non-expanded NK cells and the 7 day-expanded NK cells against the K562 target cells. RESULTS: The NK cells from PBMCs expanded 4.5-fold after 7 days, and contained 56.5% CD3-CD56+ cells. The IL-2 or IL-2 plus K562 increased the expression levels of CD158b (MFI, mean florescence intensity), CD158e1/e2 (MFI), and NKp44 (MFI), while it decreased the expression levels of NKp30 (%), CD16 (MFI), and 2B4 (MFI). The non-expanded NK cells lysed 9.0% and 27.6% of the K562 target cells in the 1 : 1 and 5 : 1 effector and target ratio, respectively, and the 7-day expanded NK cells lysed 36.9% and 57.2% of the K562 target cells, respectively. CONCLUSION: The selective expansion of CD3-CD56+ NK cells occurred only during 7 days of culture. IL-2 or IL-2 plus the K562 cells altered the expression of various activating and inhibitory receptors of NK cells, and the cytotoxicity of the expanded NK cells was higher than in the non-expanded cells.

In Vitro Induction of Carcinoembryonic Antigen (CEA) Specific Cytotoxic T Lymphocytes Using Dendritic Cells Pulsed with CEA Peptide

PURPOSE: Dendritic cells (DCs) are the most potent antigen- presenting cells for initiating the T cell immune response in vivo. Recent studies have shown that active immunotherapy with tumor antigen pulsed DC tumor antigen specific cytotoxic T lymphocyte (CTL) response. The aim of this study was to establish clinically compatible procedures for generating human DCs and to determine if the CEA peptide- pulsed DCs can activate the CEA specific CTL responses in vitro. METHODS: DCs were generated from the peripheral blood monocytes (PBMCs) of HLA A2+ healthy donors using GM-CSF and IL-4. Phenotypic analysis was performed using flow cytometry with FITC- or PE-conjugated Abs against CD1a, CD14, CD80, HLA-DR, CD83 and CD86. The immature DCs were pulsed with a CEA peptide (HLA A2 epitope, [YLSGANLNL]) and the tumor lysates isolated from HLA A2+ CEA positive cell line, NCI-H498, and were incubated with the autologous PBMCs in order to generate an antigen specific CTLs in vitro. After three rounds of stimulation, the presence of a CEA-specific CTL response was determined using a CEA positive cell line as the specific targets with the standard 51Cr release assay, the ELISPOT assay, and the flow cytometry using CEA peptide-MHC tetramer. RESULTS: The DCs obtained after 6 days of culture expressed high levels of CD1a, HLA-DR, and CD80, which corresponded to the immature DC phenotype. The 51Cr- release assay showed that DCs pulsed with the CEA peptide or the lysates of the CEA-positive NCI-H498 cell line could stimulate the CEA-specific CTL responses. The CTL response to DCs pulsed with the CEA peptide was also generated using the DCs pulsed with the CEA peptide. In the ELISPOT assay, the number of CEA peptide-specific, INF-gamma-secreting spots were increased in the CTLs generated by DCs pulsed with the CEA pepide and the tumor lysates. In the peptide-MHC tetramer assay, the CD8+ T cells with the receptors specific to CEA-peptide were increased by stimulation with the DCs pulsed with the CEA peptide and the tumor lysates. CONCLUSION: These findings show that the CEA peptide pulsed DCs can generate CEA specific CTL responses and antigen bearing DCs can be used as the target cells for a cytotoxicity assay. This study provides the foundations for DC-based cancer immunotherapy for CEA expressing solid tumors.

Isolated Intracranial Granulocytic Sarcoma as a Relapse Following Unrelated Bone Marrow Transplantation for Myelodysplastic Syndrome in a 1 Year-Old Infant / 소아과

Isolated relapse of myeloid leukemia as a granulocytic sarcoma(GS) following allogeneic bone marrow transplantation(BMT) is very rare manifestation, and usually associated with a poor prognosis. We report a case of isolated intracranial GS in an infant with myelodysplastic syndrome(MDS) following unrelated BMT. A 7 month-old girl was diagnosed with refractory anemia with excess blasts (RAEB). During observation for a couple months several GS developed in the scalp and blast counts in BM increased. Induction chemotherapy resulted in partial remission of BM but GS disappeared. Four months after diagnosis, an unrelated BMT was undertaken. Engraftment was uneventful. On D+160, an intracranial GS of 6.5 cm in size developed. A craniotomy and tumor removal was done. There was no evidence of relapse in BM with complete chimerism. Reinduction chemotherapy using IDA-FLAG resulted in profound neutropenia with pneumonia. She succumbed to respiratory failure despite leukocyte recovery. The optimal management for isolated relapse as GS following BMT should be established.

Pulmonary Oxalosis Caused by Aspergillus Niger Infection / 결핵

The Aspergillus species produces metabolic products that play a significant role in the destructive processes in the lung. We experie nced a case of chronic necrotizing pulmonary aspergillosis caused by an Aspergillus niger infection, which contained numerous calcium oxalate crystals in the necrotic lung tissue. A 46-year-old man, who had a history of pulmonary tuberculosis, presented with high fever, intermittent hemoptysis and pulmonary infiltrations with a cavity indicated by the chest radiograph. Despite being treated with several antibiotics and anti-tuberculosis regimens, the high fever continued. The sputum cultures yielded A. niger repeatedly, and intravenous amphotericin B was then introduced. The pathological specimen obtained by a transbron chial lung biopsy revealed numerous calcium oxalate crystals in a background of acute inflam matory exudates with no identification of the organism. Intravenous amphotericin B was con tinued at a total dose of 1600 mg, and at that time he was afebrile, although the intermittent hemoptysis continued. On the 63rd hospital day, a massive hemoptysis (about 800 mL) developed, which could not be controlled despite embolizing the left bronchial artery. He died of respiratory failure the next day. It is believed that the oxalic acid produced by A. niger was the main cause of the patient's pulmonary injury and the ensuing massive hemoptysis.

Clonal Analysis of Neurofibroma by PCR Amplification of HUMARA Gene

BACKGROUND: While neurofibromas have generally been regarded as polyclonal hyperplastic lesions, it remains unclear whether the tumor is a true neoplasm or a hyperplastic lesion. METHODS: Determination of clonality by X chromosome inactivation pattern was investigated in twenty-one cases of neurofibroma employing enzyme digestion and PCR of the HUMARA gene. The histological, immunohistochemical, and ultrastructural characteristics of the tumors were also examined. RESULTS: Immunohistochemically, most of the tumor cells showed vimentin and S-100 protein positivity. Axons were demonstrated by neurofilament protein positivity and were seen mainly at the periphery and rarely in the central portion of the tumor. Ultrastructurally, the tumors were composed of a variety of cell types: perineurial cells, Schwann cells, fibroblasts, and axons. X chromosome inactivation analysis was completed on thirteen out of fifteen cases in which DNA was successfully extracted. Of thirteen neurofibromas that were heterozygous at the HUMARA loci, eleven showed a polyclonal pattern. The remaining two cases were considered as indeterminate for clonality because of unequal band intensity and failure to obtain the normal control DNA. CONCLUSION: The results from this study suggest that neurofibromas are polyclonal in origin and might be a neoplastic lesion comprising non-neoplastic cells among constituent components.