Naoko Inokuchi

Impact of miR-155 and miR-126 as novel biomarkers on the assessment of disease progression and prognosis in adult T-cell leukemia

OBJECTIVE Micro RNAs (miRNAs) provide new insight in the development of cancer, but little is known about their clinical relevance as biomarkers in the assessment of diagnosis, classification, progression and prognosis of various cancers. To explore a potential novel biomarker, we examined the cellular and plasma miRNA profiles in adult T-cell leukemia (ATL) characterized by diverse clinical features. METHODS AND RESULTS Using CD4-positive cells isolated from 2 non-infected healthy individuals, 3 chronic ATL patients and 3 acute ATL patients, cellular miRNAs were profiled by microarray. The microarray screened 5 miRNAs namely miR-155, let-7g, miR-126, miR-130a and let-7b because of the large difference in their expression in diseased vs. that of healthy controls. The expression levels of before 5 miRNAs re-quantified by reverse transcription quantifiable polymerase chain reaction (RT-qPCR) were not always accordant in cells and plasma. The high and low plasma levels of miR-155 and miR-126 changed with ATL stage. CONCLUSION The present study revealed that there is a quantitative discrepancy between cellular and plasma miRNAs. The elevation of plasma miR-155 and the reduction in miR-126 correlated with poor prognosis, indicating their usefulness as a novel biomarker for the assessment of disease stage.

Minilaparotomy May Be Independently Associated with Reduction in Inflammatory Responses after Resection for Colorectal Cancer

Objectives: A minilaparotomy approach (skin incision less than 7 cm) to resection of colon cancer is technically feasible, but objective data supporting its benefit are scarce. The aim of this study was to clarify whether minilaparotomy is independently associated with a reduction in the acute inflammatory response after resection of colorectal cancer. Design: Thirty-one patients who underwent surgical resection of colorectal cancer using minilaparotomy or conventional laparotomy were included in this nonrandomized prospective study. Inflammatory responses were evaluated with serum interleukin-6 (IL-6) and C-reactive protein (CRP) levels. Results: In both the minilaparotomy and conventional laparotomy groups, serum IL-6 and CRP levels significantly increased 24 h after the operation (1POD) compared to preoperative levels (p < 0.0001 and p < 0.0001, respectively). Median serum levels of IL-6 and CRP in the minilaparotomy group were significantly lower at 1POD versus the conventional group (p = 0.0066 and p = 0.0033, respectively). Multivariate analyses showed that a smaller increase in serum IL-6 or CRP levels at 1POD [less than 75th percentile (112.9 or 10.6 mg/ml, respectively)] was independently related to only minilaparotomy. Conclusions: These data in this nonrandomized trial suggest that minilaparotomy may be independently associated with reduced inflammatory responses in colorectal cancer resection.

Trends in HTLV‐1 prevalence and incidence of adult T‐cell leukemia/lymphoma in Nagasaki, Japan

Most previous studies aimed at estimating the number of human T‐cell leukemia virus type‐1 (HTLV‐1) carriers in endemic areas have been based on seroprevalence rates in blood donors; however, this may result in underestimation because of the healthy donor effect. People who have health problem do not donate blood. In the present study, the number of HTLV‐1 carriers in Nagasaki City was estimated based on the seroprevalence rates in a hospital‐based population from Nagasaki University Hospital. In accordance with previous reports, seroprevalence of HTLV‐1 was higher in females, and year of birth‐specific seroprevalence showed a significant annual decline in both genders (P for trend: <0.0001). The estimated number of HTLV‐1 carriers in Nagasaki City was 36,983. The incidence of adult T‐cell leukemia/lymphoma (ATLL) among HTLV‐1 carriers was estimated using data from the Nagasaki Prefectural Cancer Registry. The estimated annual incidence of ATLL was 61 per 100,000 HTLV‐1 carriers, and the crude lifetime risk of the development was 7.29% for males and 3.78% for females. There is a large pool of HTLV‐1 carriers aged over 70 years, and a continuing development of cases of ATLL among the elderly is therefore expected. J. Med. Virol. 82:668–674, 2010.

Relationship between whole-blood interferon-gamma production and extent of radiographic disease in patients with pulmonary tuberculosis.

The aim of this study was to determine whether whole-blood interferon-gamma (IFN-gamma) production correlates with the radiographic extent of pulmonary tuberculosis before treatment. The subjects were 40 human immunodeficiency virus-negative patients with pulmonary tuberculosis and 36 healthy volunteers. The concentrations of IFN-gamma in whole blood stimulated with Mycobactrium tuberculosis purified protein derivatives (tuberculous PPD) and phytohemagglutinin (PHA) were evaluated. PHA-stimulated IFN-gamma (PHA-IFN-gamma) was lower in the patients than in healthy volunteers (p < 0.05), and inversely correlated with the disease extent (p < 0.01). Tuberculous PPD-stimulated IFN-gamma as a percentage of PHA response (tuberculous PPD-IFN-gamma/PHA-IFN-gamma) was higher in the patients than in healthy volunteers (p < 0.05). However, tuberculous PPD-IFN-gamma/PHA-IFN-gamma did not correlate with the disease extent. Our results indicate that the tuberculous PPD-IFN-gamma/PHA-IFN-gamma may be useful for the diagnosis of tuberculosis but not for evaluating the disease severity, and suggest that PHA-IFN-gamma could be considered as a marker of disease severity.

Serum cytochrome c to indicate the extent of ongoing tumor cell death

Despite the significant implication of apoptosis in tumorigenesis, there is no biomarker to assess the extent of ongoing apoptosis in vivo for hematological malignancies. We investigated the potential of serum cytochrome c (cyto‐c) as a biomarker for apoptosis. Cyto‐c and lactate dehydrogenase (LD) were released into the culture medium from apoptotic cells induced by tumor necrosis factor‐related apoptosis‐inducing ligand in a time‐dependent manner in vitro, with different kinetic patterns. Only one‐third of 153 patients with hematological malignancies showed high levels of serum cyto‐c (>20 ng/ml). Although serum cyto‐c level was roughly correlated to serum LD activity, their different kinetic patterns from serial measurements indicated that serum cyto‐c rather than LD is a more sensitive indicator for tracking changes of tumor status. Furthermore, serum cyto‐c level stratified patients with acute adult T‐cell leukemia into favorable and unfavorable subgroups with 5‐year survival rates of 67%vs. 11%. In conclusion, serum cyto‐c may provide a fast real‐time biomarker for tracking changes of tumor status involved in apoptotic cell death, but lacking disease or cell‐type specificity.

Can the use of intraoperative intact parathyroid hormone monitoring be abandoned in patients with hyperparathyroidism

BACKGROUND Ultrasound (US) and technetium-99m sestamibi scintigraphy (MIBI) are used to determine the localization of abnormal glands in cases of primary hyperparathyroidism (PHPT). Intraoperative intact parathyroid hormone (iPTH) monitoring is a reliable examination used to cure PHPT. The aim was to assess the necessity of intraoperative iPTH monitoring. METHODS Sixty patients were examined using preoperative MIBI and US. iPTH was measured at 3 time points: (1) at the start of surgery; (2) 10 minutes after gland resection; and (3) more than 60 minutes after surgery. We defined a decreased iPTH level as an iPTH measured 10 minutes after resection that was less than 50% of the preoperative level. RESULTS The iPTH of 55 patients with concordant lesions decreased to within the normal range more than 60 minutes after surgery. CONCLUSIONS It is not necessary to monitor intraoperative iPTH when single concordant lesions are preoperatively identified on both MIBI and US.

RELEVANCE OF MOLECULAR TESTS FOR HTLV-1 INFECTION AS CONFIRMATORY TESTS AFTER THE FIRST SERO-SCREENING

The diagnosis of human T-cell leukemia virus type-1 (HTLV-1) infection has been widely examined by serologics. In the first screening tests, serological false negative and positive samples have been reduced thanks to advances in assay techniques that apply new emission agents and sensors. On the other hand, western blot (WB) remains problematic. For example, WB analysis yields many samples equivalent to antibody positive ones. To reduce the need for WB, an alternative testing strategy is required to detect HTLV-1 infection. Polymerase chain reaction (PCR) for the HTLV-1 provirus has recently been recommended for a final diagnosis of infection. However, although PCR is thought to be one element, the validation of detection performance for HTLV-1 infection between serological and molecular testing is not always clear. Thus, this study aimed to evaluate the accuracy and test the validity of an improved methodology for serological detection of HTLV-infection, as well as that of PCR. In conclusion, the high values of kappa-statistics are expected to deliver high quality in chemiluminescent enzyme immunoassay (or chemiluminescent immunoassay), while the problems with WB assays remain to be elucidated. As an alternative to WB, a combination of real-time qPCR and nested PCR is proposed as a suitable confirmatory test.

Screening for genetic heterogeneity in the interferon sensitivity determining region of the hepatitis C virus genome by polymerase chain reaction with melting curve analysis.

Abstract Background: Although mutations in the interferon (IFN) sensitivity determining region (ISDR) of hepatitis C virus (HCV) have been reported to be useful as a predictive viral factor for IFN therapy in patients infected with HCV-1b, such laboratory research has not been favorably translated into the clinic. To promote such translation, we attempted the establishment of a rapid and simple polymerase chain reaction (PCR) combined with melting curve analysis (MCA) to screen for mutations in the ISDR and for the monitoring of HCV quasispecies. Methods: A PCR-MCA protocol was established using in-house primers and hybridization probes designed according to the results of direct sequencing of 34 HCV-1b samples. Then, the performance of PCR-MCA was verified by comparing with mutation profiles obtained by direct sequencing and sequencing after cloning. Results: The MCA assay revealed that melting temperature (Tm) was inversely correlated with the number of nucleotide (nt) and amino acid substitutions in the ISDR deduced on the basis of the results of direct sequencing. A boundary Tm of 58.0°C allowed us to discriminate HCV genomes into two groups: one with a Tm >58.0°C had no or a low number of nt substitutions, while the other genomes with a Tm <58.0°C had a high number of nt substitutions, corresponding to wild-type in the former and mutant-type in the latter in respect of a clinical setting for IFN therapy. Moreover, this MCA assay provided precise discrimination of Tm between clones, reflecting the degree of the genetic complexity of HCV genomes. Conclusions: This study indicates that the MCA assay is useful to rapidly and simply screen the mutational status of the ISDR of HCV, as well as in using the ISDR as one of the targets for discriminating the genetic complexity of HCV genomes. The MCA assay could also be applicable as a convenient and useful screen of the genetic heterogeneity of clones relating to HCV quasispecies. Clin Chem Lab Med 2008;46:966–73.