Laboratory tools for oligo-secretory myeloma diagnosis: capillary electrophoresis versus free light chain assay.

BACKGROUND: To investigate how capillary electrophoresis (CE) works in oligo-secretory myeloma (OSM), we report a case here of OSM using multiple diagnostic methods including gel electrophoresis (GE), CE, and free light chain assay (sFLC). Also, we provide a brief review of laboratory methods to compare their diagnostic utilities in OSM. METHODS: A 72 year-old Korean male suffering from low back pain during the past 6 months was transferred to the department of neurosurgery in order to evaluate abnormal findings in an imaging study, suggesting plasma cell myeloma (PCM) with multiple bone metastasis. CE showed no suspicious M-component; however, it showed increased Kappa components and skewing Kappa/Lambda ratio (K/L). Bone marrow examination revealed plasma cells observed up to 70%, which were compatible with sFLC results. RESULTS: Based on these results, the diagnosis turned out to be OSM with multiple bone metastases. Thereafter, the patient started the first cycle of chemotherapy accompanied by palliative radiation therapy. CONCLUSIONS: In our case, sFLC showed abnormal Kappa and K/L results from both serum and urine specimen. Therefore, it seems to be more sensitive and appropriate than both GE and CE to diagnose OSM.

Balance between SIRT1 and DBC1 expression is lost in breast cancer.

SIRT1 (silent mating-type information regulation 2 homologue 1)-mediated cellular resistance to various stresses is negatively regulated by deleted in breast cancer 1 (DBC1), which was originally reported to be deleted in breast cancer. However, the suggested functions of SIRT1 as a potential tumor promoter and of DBC1 as a potential tumor suppressor have been challenged by observations of their respective down- and up-regulation in various cancers. The aim of the present study was to simultaneously evaluate the expression levels of SIRT1 and DBC1 in the normal and tumor breast tissues from 28 breast cancer patients and to determine correlations with clinicopathological variables. SIRT1 and DBC1 expression was higher in tumor tissues than in matched normal tissues at the protein level, but not at the transcriptional level. Overexpression of SIRT1 and DBC1 in tumor tissue was correlated with favorable and unfavorable clinicopathological factors, suggesting their pleiotropic functions as a potential tumor promoter and tumor suppressor in tumorigenesis. Interestingly, although the overall expression of SIRT1 and DBC1 increased in tumor breast tissues, the correlation between SIRT1 and DBC1 expression was weaker in tumor tissue than in normal tissue. This suggests that the negative regulation of SIRT1 by DBC1 may retard tumorigenesis in breast tissue. Therefore, the correlation between SIRT1 and DBC1 is a potential prognostic indicator in breast cancer.

The association between genetic polymorphisms in CYP19 and breast cancer risk in Korean women.

Aromatase encoding by the CYP19 gene catalyzes the conversion of androgens to estrogens. In order to determine if polymorphisms of the CYP19 gene are associated with breast cancer risk, we analyzed the frequency of tetranucleotide (TTTA) tandem repeats and a 3-bp insertion (I)/deletion (D) polymorphism in intron 4 of the CYP19 gene in genomic DNA from 70 Korean breast cancer patients and 102 age-matched, healthy women. The 3-bp deletion allele was found more frequently in the breast cancer group than in the control group (p=0.001). Logistic regression analysis of the CYP 19 insertion/deletion (I/D) genotype showed a strong association between ID polymorphisms and breast cancer. The frequency of DD and ID alleles was significantly increased in the breast cancer group (DD genotype p=0.004, OR=12.81; and ID genotype p=0.005, OR=2.62). However, there were no differences in the genotype distributions of the (TTTA)n polymorphism of CYP19 between breast cancer patients and healthy controls. A positive association was noted between TTTA polymorphisms with 10 or more repeats and ER-negative tumors, as well as between lower repeat polymorphisms and ER-positive tumors (p=0.019). With respect to TTTA polymorphisms, we confirmed that the expression of aromatase in ER-positive MCF7 cells with 7-3 and 11 allele heterozygosity was significantly higher than in ER-negative MDA-MB231 cells with 11 allele homozygosity. These results suggest that 3-bp I/D polymorphisms of the CYP19 gene may be associated with breast cancer and that the (TTTA)n repeat genotype would be useful in selecting candidates for tamoxifen therapy, as well as predicting breast cancer risk in Korean women.

Aneurysmal appearance of medium-sized bronchi: a peripheral manifestation of endobronchial tuberculosis.

OBJECTIVE: The purpose of this study is to show the importance of aneurysmal appearance of medium-sized bronchi observed at CT in the diagnosis of tuberculosis (TB). CONCLUSION: Aneurysmal appearance of medium-sized bronchi is a CT feature of tuberculosis. In TB-endemic areas, the aneurysmal appearance of medium-sized bronchi at CT may suggest the diagnosis of TB.

The C1772T genetic polymorphism in human HIF-1alpha gene associates with expression of HIF-1alpha protein in breast cancer.

Hypoxia-inducible factor 1 (HIF-1) is an important genetic component involved in the cellular response to hypoxia. HIF-1 is also linked to the regulation of tumor development and growth. In previous studies, the C1772T (P582S) or the G1790A (A588T) polymorphisms of the HIF-1alpha gene have been identified in renal cell carcinoma, head and neck and esophageal squamous cell carcinomas as well as colorectal and prostate cancers. In our study, we investigated whether polymorphisms of the HIF-1alpha gene may account for the expression patterns of HIF-1alpha protein and impact of clinical progression in breast cancer. We also examined the impact of prognosis of HIF-1alpha gene polymorphism and protein expression in the prediction of biological behavior. We performed polymerase chain reaction and direct sequencing to detect polymorphisms in exon 12 of HIF-1alpha from 90 breast cancer patients and 102 healthy controls. The expression of HIF-1alpha was measured in paraffin-embedded specimens from patients by immunohistochemistry. We associated its expression with known prognostic factors. The frequency of the T allele for C1772T in breast cancer patients and healthy controls was 5.6 vs. 4.4%, whereas, the frequency of the A allele for G1790A was 1.7 vs. 4.4%. HIF-1alpha was overexpressed in 56.7% (51 of 90) of the patients. Its overexpression associated with the T1772 polymorphic allele (p=0.04). Elevated levels of HIF-1alpha protein were found in cases of breast cancer with lymph node metastasis (p=0.041), high histological grade (p=0.001) and increased Ki-67 index (p=0.031). These results suggest the potential use of C1772T (P582S) polymorphism and expression analysis in providing a new prognostic factor for unfavorable disease outcomes and may help for clinical decision-making in the treatment of breast cancer patients.

Expression of EGFR and follicular dendritic markers in lymphoid follicles from patients with Castleman's disease.

This study investigated the useful morphologic and immunophenotypic findings for the diagnosis of Castleman's disease (CD). We focused on the distribution and expression of follicular dendritic cells (FDC) in lymphoid follicles from patients with CD. Eleven CD cases of the hyaline vascular (HV) variant and six cases of the plasma cell (PC) variant were studied using tissue microarray and paraffin resistant monoclonal antibodies CD21, CD35, and EGFR, a new novel marker of FDC, as well as an antibody against human herpes virus 8 (HHV8). Epstein-Barr virus (EBV) was detected by means of in situ hybridization with a fluorescein isothiocyanate-labeled EBV-encoded RNA (EBER) specific oligonucleotide. The FDC network of the PC variant (n=4) was similar to that seen in normal or reactive germinal centers. In contrast, all HV variants and 2 cases of the PC variant were either expanded, disrupted, or exhibited multiple tight collections of FDC both in germinal centers and in mantle zone lymphocytes. The expanded mantle zone lymphocytes were CD20+, Bcl2+, PAX5+, and MUM1- with less number of CD3+ T cells admixed. Other features of the HV variant included follicular regression and vascular ingrowth of the germinal centers, whereas features of the PC variant were follicular hyperplasia and interfollicular plasmacytosis. In addition, EBV infection was positive in three CD cases, and one case had co-expression of HHV8 and EBV infection. Taken together, we found immunophenotypic differences of mantle zone lymphocytes and FDC network patterns of lymphoid follicles in CD. Thus, we conclude that these differences are relevant for the differential diagnosis of the two histopathologic variants of CD.

CCAR2 negatively regulates IL-8 production in cervical cancer cells.

Cell cycle and apoptosis regulator 2 (CCAR2) is a multifaceted protein that controls diverse cellular functions; however, its function in cancer is unclear. To better understand its potential role in cancer, we examined gene expression patterns regulated by CCAR2 in cervical cancer cells. Cytokine and chemokine production by CCAR2-deficient cells increased under oxidative conditions. In particular, H2O2-treated CCAR2-depleted cells showed a significant increase in interleukin-8 (IL-8) production, indicating a negative regulation of IL-8 by CCAR2. Upregulation of IL-8 expression in CCAR2-deficient cells occurred via activation of transcription factor AP-1. The negative correlation between CCAR2 and IL-8 expression was confirmed by examining mRNA and protein levels in tissues from cervical cancer patients. Furthermore, CCAR2-regulated IL-8 expression is associated with a shorter survival of cervical cancer patients. Overall, the data suggest that CCAR2 plays a critical role in controlling both the cancer secretome and cancer progression.

A rapid, simple, and accurate plaque assay for human respiratory syncytial virus (HRSV).

Plaque assays of human respiratory syncytial virus (HRSV) are time-consuming, requiring 4 to 7 days for plaque formation and several hours for dye staining. Here, we describe a simple method by which RSV plaques can be visualized and counted with the naked eye only 2 days after infection of HEp-2 cells. In this assay, the infected cells are stained with monoclonal antibodies and the plaques are developed using diaminobenzidine (DAB). We tested the accuracy of this new plaque assay by comparing the results obtained on days 1, 2, 3, 4, 5, and 6 post-infection. The whole procedure is significantly simpler than the traditional method, with an immunostaining process of around 1.5h. Our method is rapid, accurate, and simple; thus, it has the potential to significantly contribute to studies related to RSV disease.

Comparison of Unsatisfactory Samples from Conventional Smear versus Liquid-Based Cytology in Uterine Cervical Cancer Screening Test.

BACKGROUND: Cervical cytology for uterine cervical cancer screening has transitioned from conventional smear (CS) to liquid-based cytology (LBC), which has many advantages. The aim of this study was to compare the proportion of unsatisfactory specimens from CS versus LBC at multiple institutions including general hospitals and commercial laboratories. METHODS: Each participating institution provided a minimum of 500 Papanicolaou (Pap) test results for analysis. Pap tests were classified according to the participating institution (commercial laboratory or general hospital) and the processing method (CS, ThinPrep, SurePath, or CellPrep). The causes of unsatisfactory results were classified as technical problems, scant cellularity, or complete obscuring factors. RESULTS: A total of 38,956 Pap test results from eight general hospitals and three commercial laboratories were analyzed. The mean unsatisfactory rate of LBC was significantly lower than that of CS (1.26% and 3.31%, p = .018). In the LBC method, samples from general hospitals had lower unsatisfactory rates than those from commercial laboratories (0.65% vs 2.89%, p = .006). The reasons for unsatisfactory results were heterogeneous in CS. On the other hand, 66.2% of unsatisfactory results in LBC were due to the scant cellularity. CONCLUSIONS: Unsatisfactory rate of cervical cancer screening test results varies according to the institution and the processing method. LBC has a significantly lower unsatisfactory rate than CS.

Inhibition of Carbonyl Reductase 1 Safely Improves the Efficacy of Doxorubicin in Breast Cancer Treatment.

AIMS: Doxorubicin (DOX) is a chemotherapeutic drug that is used to treat many cancers, but its use is limited by cardiotoxic side effect. Carbonyl reductase 1 (CBR1) is an NADPH-dependent oxidoreductase that reduces DOX to doxorubicinol (DOXOL), a less potent derivative that is responsible for DOX cardiotoxicity. Thus, we aimed to demonstrate that inhibition of CBR1 enhances the chemotherapeutic efficacy of DOX and attenuates cardiotoxicity. RESULTS: Pharmacological or genetic inhibition of CBR1 improved the anticancer effects of DOX in preclinical models of breast cancer. RNA interference or chemical inhibition of CBR1 improved the anticancer effect of DOX in breast cancer. Moreover, CBR1 overexpression enabled breast cancer cells to obtain chemotherapeutic resistance to DOX treatment. Intriguingly, inhibition of CBR1 decreased DOX-induced cardiotoxicity in animal model. Innovation and Conclusions: Inhibition of CBR1 increases chemotherapeutic efficacy of DOX and reduces cardiotoxicity by blocking DOX reduction to DOXOL. Therefore, we offer preclinical proof-of-concept for a combination strategy to safely leverage the efficacy of doxorubicin by blunting its cardiotoxic effects that limit use of this cytotoxic agent used widely in the oncology clinic. Antioxid. Redox Signal. 26, 70-83.

Pseudoaneurysm at the Distal Posterior Inferior Cerebellar Artery After Blunt Head Trauma: A Case Report and Review of the Literature.

BACKGROUND: Traumatic pseudoaneurysm of the distal posterior inferior cerebellar artery (PICA) is extremely rare. We report our experience of a case of pseudoaneurysm that developed on the PICA after blunt trauma to the head. CASE DESCRIPTION: A 55-year-old woman was transferred to our emergency department presenting with a semicomatose mental status after falling to the floor from a standing position. Computed tomography showed a small intracerebral hemorrhage and subarachnoid hemorrhage in the cerebellum. Cerebral angiography revealed no causative lesion. After 3 days, the woman was nearly mentally alert. One month later, follow-up angiography showed a small, newly developed saccular aneurysm at the distal PICA. The patient underwent surgical treatment via a midline suboccipital approach. The aneurysm was surrounded by a subacute-stage subdural hematoma and protruded into the cortex. The aneurysm was coagulated and resected. On pathologic examination, pseudoaneurysm was diagnosed without infectious inflammation. No surgery-related morbidity occurred. CONCLUSIONS: Given that development of traumatic pseudoaneurysm usually is delayed, follow-up radiologic examination is required, especially for patients with severe blunt trauma.

Microneedle delivery of trivalent influenza vaccine to the skin induces long-term cross-protection.

A painless self-immunization method with effective and broad cross-protection is urgently needed to prevent infections against newly emerging influenza viruses. In this study, we investigated the cross-protection efficacy of trivalent influenza vaccine containing inactivated A/PR/8/34 (H1N1), A/Hong Kong/68 (H3N2) and B/Lee/40 after skin vaccination using microneedle patches coated with this vaccine. Microneedle vaccination of mice in the skin provided 100% protection against lethal challenges with heterologous pandemic strain influenza A/California/04/09, heterogeneous A/Philippines/2/82 and B/Victoria/287 viruses 8 months after boost immunization. Cross-reactive serum IgG antibody responses against heterologous influenza viruses A/California/04/09, A/Philippines/2/82 and B/Victoria/287 were induced at high levels. Hemagglutination inhibition titers were also maintained at high levels against these heterogeneous viruses. Microneedle vaccination induced substantial levels of cross-reactive IgG antibody responses in the lung and cellular immune responses, as well as cross-reactive antibody-secreting plasma cells in the spleen. Viral loads in the lung were significantly (p < 0.05) reduced. All mice survived after viral challenges. These results indicate that skin vaccination with trivalent vaccine using a microneedle array could provide protection against seasonal epidemic or new pandemic strain of influenza viruses.

Polarized CD163+ tumor-associated macrophages are associated with increased angiogenesis and CXCL12 expression in gastric cancer.

PURPOSE: Tumor-associated macrophages (TAMs) play a significant role in tumor progression and angiogenesis. However, the prognostic value of TAMs in different histologic locations of gastric cancer (GC) is still unknown. We evaluated the distribution of TAMs in different histologic locations to investigate its importance in predicting prognosis and the relationship with angiogenesis and CXCL12 expression in GC. METHODS AND MATERIALS: The distribution of TAMs and microvessel density (MVD) in 113 GC samples were evaluated by immunohistochemical staining of CD163 and CD105, respectively. The extent of TAM distribution in the tumor was categorized into three groups: infiltrated TAMs in the tumor nest (TN), tumor stroma (TS) and invasive tumor margin (TM). The expression of CXCL12 in GC were evaluated by immunohistochemical staining of tissues from 88 GC samples. RESULTS: The increased CD163+ TAMs in TS and TM were closely correlated with tumor size, depth of invasion, TNM stage, lymph node metastasis, and lymphovascular invasion. TAMs in TN was not related with any clinicopathologic characteristics except histologic differentiation. The high infiltration of CD163+ TAMs in TS and TM were significantly correlated with poor overall survival. Regardless of location, CD163+ TAMs were significantly correlated with increased MVD. CXCL12 expression was significantly associated with increased CD163+ TAMs in TS and TM. CONCLUSIONS: TAMs in different histologic locations in GC were related to distinct aspects of tumor progression. CD163+ TAMs in TS and TM are associated with tumor progression and CXCL12 expression in GC. TAMs may be involved in tumor progression through the angiogenesis.

Single nodular opacity of granulomatous pneumocystis jirovecii pneumonia in an asymptomatic lymphoma patient.

The radiologic findings of a single nodule from Pneumocystis jirovecii pneumonia (PJP) have been rarely reported. We described a case of granulomatous PJP manifesting as a solitary pulmonary nodule with a halo sign in a 69-year-old woman with diffuse large B cell lymphoma during chemotherapy. The radiologic appearance of the patient suggested an infectious lesion such as angioinvasive pulmonary aspergillosis or lymphoma involvement of the lung; however, clinical manifestations were not compatible with the diseases. The nodule was confirmed as granulomatous PJP by video-assisted thoracoscopic surgery biopsy.

Autophagy is related to the hedgehog signaling pathway in human gastric adenocarcinoma: prognostic significance of Beclin-1 and Gli2 expression in human gastric adenocarcinoma.

Beclin-1 induces autophagy, which is known to be involved in many physiopathological processes such as cell development, aging, stress response, immune response and cancer. Several studies showed that Beclin-1 expression is associated with several prognostic factors of gastric carcinomas. Recently, the connection between autophagy and the hedgehog (HH) signaling pathway has been studied. Here, we investigated the relationship between the autophagy and hedgehog (HH) signaling pathways in gastric adenocarcinoma. We evaluated Beclin-1 and Gli2 expression in 108 gastric adenocarcinoma tissues via immunohistochemical analysis, using a tissue microarray, in relation to survival and other prognostic factors. Our results show that increased Beclin-1 expression is correlated with favorable clinicopathological variables including histologic grade, tumor size, primary tumor (T) stage, lymph node metastasis, lymphatic invasion, neural invasion, and tumor recurrence. Furthermore, increased Gli-2 expression was correlated with several favorable clinicopathological variables including primary tumor (T) stage, lymphatic invasion, and tumor recurrence. Increased Beclin-1 expression was significantly correlated with increased Gli2. Univariate analyses for disease-free survival and overall survival revealed that the higher Beclin-1 and Gli2 expression group had a more favorable prognosis compared with the lower Beclin-1 and Gli2 expression group. Our results suggest that progressively increased Beclin-1 and Gli2 expression contributes to the inhibition of tumor growth and metastasis in gastric adenocarcinoma and Beclin-1 acts as a tumor suppressor by regulating the HH signaling pathway through Gli2 expression in gastric adenocarcinoma.