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.

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.

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.

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.

First Japanese patient treated with parathyroid hormone peptide immunization for refractory hypercalcemia caused by metastatic parathyroid carcinoma.

Patients with unresectable parathyroid carcinoma develop severe hypercalcemia, bone fractures and renal failure, and become unresponsive to conventional treatments. It has been shown that successful induction of anti-parathyroid hormone (PTH) antibodies, using PTH peptide fragments for immunisation, normalized serum levels of calcium as well as improved clinical symptoms. Here, we report our experience of PTH immunization in a Japanese female suffering from refractory hypercalcemia and renal failure caused by unresectable metastatic parathyroid carcinoma. Upon immunization, there were apparent clinical responses including reduction of serum levels of Ca along with anti-PTH antibodies induction. Therefore, we concluded that PTH immunization was an effective treatment against hypercalcemia caused by metastatic parathyroid carcinomas that are unresponsive to conventional treatments.

Elevation of serum KL-6 glycoprotein or surfactant protein-D in adult T-cell leukemia with distinct pulmonary complications.

Patients with hematological malignancies frequently suffer from lung diseases as a complication. However, it is difficult to discriminate leukemic invasion into the lung from infectious pulmonary complications. The serum level of Krebs von den Lungen-6 (KL-6), which is a mucin-like glycoprotein, is increased in more than 70% of patients with interstitial pneumonia. Surfactant protein-D (SP-D) is produced mainly in the lung by alveolar type II and bronchiolar epithelial cells and is a useful serum marker for interstitial pneumonia. We therefore measured the levels of KL-6 and SP-D in sera from 128 patients (76 males and 52 females, mean age: 59 years) with hematological malignancies, including adult T-cell leukemia (ATL). Overall, the increase in KL-6 or SP-D, above each cut-off value (500 U/ml for KL-6 and 110 ng/ml for SP-D), was detected in 11 patients (8.6%) or 10 patients (7.8%), respectively. In contrast, among 67 ATL patients, 15 patients had high serum levels of KL-6 and/or SP-D; both were elevated in 2 patients, only KL-6 was elevated in 6 patients and only SP-D was elevated in 7 patients. Thus, serum KL-6 and SP-D appear to be elevated in a mutually exclusive manner in ATL. Indeed, high serum levels of KL-6 were closely related to the stage of ATL, while the serum SP-D was elevated in ATL patients with pulmonary infection. In conclusion, the combined measurement of KL-6 and SP-D in ATL may become a useful means to discriminate leukemic pulmonary lesions from infectious pulmonary complications.

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

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 degrees C allowed us to discriminate HCV genomes into two groups: one with a Tm >58.0 degrees C had no or a low number of nt substitutions, while the other genomes with a Tm <58.0 degrees 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.

Non-specific activities against ruthenium crosslinker as a new cause of assay interference in an electrochemilluminescent immunoassay.

Clinical assays are very important for the diagnosis and management of clinical disorders. Each assay system consists of a specific method to detect and/or quantify a substance of interest in the clinical specimen. However, clinical assays can be unfavorably influenced by non-specific activities concomitantly present in the specimen, which may mislead clinical decisions. Thus, it is very important to know how each assay works, and how and when the assay is non-specifically influenced. Here, we report three cases shown clinical data of thyroid function influenced by new type of assay interference.

Increased serum levels of interleukin-6 in malnourished patients with colorectal cancer.

Peri-operative serum levels of interleukin-6 (IL-6) were determined in 62 patients who underwent resection of colorectal cancer to clarify the relationship between nutritional status and IL-6 response. Patients were divided into two groups based on creatinine height index: malnourished group (n=13) and normally nourished group (n=49). The preoperative median serum level of IL-6 in the malnourished group was significantly higher than in the normally nourished group (P=0.041). The postoperative median serum level of IL-6 in the malnourished group also tended to be higher. In conclusion, the peri-operative IL-6 response may be activated in malnourished colorectal cancer patients.

The relationship between circulating interleukin-6 and carcinoembryonic antigen in patients with colorectal cancer.

PURPOSE: The purpose of this study was to clarify whether or not preoperative serum levels of interleukin-6 (IL-6) correlate with well-established prognostic variables (serum level of carcinoembryonic antigen (CEA), stage, histological grade, lymphatic and venous invasion) in colorectal cancer patients. PATIENTS AND METHODS: Serum levels of IL-6 and CEA were determined in 62 patients who underwent resection of colorectal cancer. The patients were divided into two groups based on a selected cut-off value: high and low IL-6 and CEA groups. RESULTS: The median serum level of IL-6 in colorectal cancer patients was significantly higher than the median level in normal controls (p = 0.0014). Multivariate logistic regression analysis showed that a high serum level of CEA is an independent predictor for a high serum level of IL-6 (odds ratio, 4.10; p = 0.046). CONCLUSION: A high serum level of IL-6 is significantly associated with a high serum level of CEA in preoperative colorectal cancer patients.

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.