Toll-like receptor 4 promoter polymorphisms in a familial mediterranean fever patient with asymptomatic bacteriuria.

A periodic fever, due to inherited inflammatory disorders, can be misdiagnosed as a common infection, when a possible pathogen is detected from a patient. TLR4 SNPs that are responsible for asymptomatic bacteriuria might disturb the pathophysiology of familial Mediterranean fever without MEFV mutations.

Ectopic expression of band 3 anion transport protein in colorectal cancer revealed in an autoimmune hemolytic anemia patient.

Cancer patients occasionally have anemia with high mean corpuscular volume in addition to iron deficiency anemia. Secondary autoimmune hemolytic anemia (AIHA) following cancer is also observed with low frequency. To date, no causal mechanisms for these disease states have been reported. Here, we present the case of an 80-year-old woman with AIHA that was resistant to prednisolone. Further examinations revealed primary adenocarcinoma of the sigmoid colon and primary squamous cell carcinoma in the right lung. After resections of these tumors, her anemia partially improved until a colon cancer-derived metastatic tumor was detected in the left lung. Immunoprecipitation of erythrocyte membrane proteins with an autoantibody followed by mass spectrometry/Western blotting identified band 3 as the target of the autoantibody. Immunohistochemical analysis revealed ectopic expression of band 3 in the colon adenocarcinoma. To our knowledge, this is the first report that identifies the cause in a case of anemia without bleeding in a cancer patient and that defines a mechanism underlying secondary AIHA following cancer progression.

Mean platelet component and mean platelet volume as useful screening markers for myelodysplastic syndrome.

BACKGROUND: Hematologic disorders, including myelodysplastic syndrome (MDS), are difficult to identify in routine hematologic examinations using automated hematology analyzers. However, the practical uses of mean platelet component and mean platelet volume (MPV) measured by these analyzers as screening markers for MDS, remain unclear. METHODS: Mean platelet component and MPV values were measured in the peripheral blood of patients with MDS, aplastic anemia, idiopathic thrombocytopenic purpura, myeloproliferative neoplasms, and in healthy controls using an automated hematologic analyzer. Cutoff values for discriminating between the MDS group and healthy controls were determined by recursive partitioning analysis. RESULTS: Mean platelet component was significantly lower in MDS patients compared with controls, while MPV was significantly higher. Combined cutoff values for MDS diagnosis of <25.3 g/dL for mean platelet component and >10.0 fL for MPV showed a specificity and positive predictive value of 99.9% and 99.1%, respectively. These cutoff values also differentiated between MDS and diagnoses of aplastic anemia, idiopathic thrombocytopenic purpura, and myeloproliferative neoplasms. CONCLUSION: Mean platelet component and MPV may, thus, be useful and convenient screening markers for MDS.

[Companion diagnostics in the era of personalized medicine--chairmen's introductory remarks].

Personalized medicine is a medical model that proposes the customization of treatment for individual patients. In this model, diagnostic tests are essential for selecting safer and more efficacious treatments. The term "companion diagnostics" has been used to describe these tests, whereby molecular assays that measure the levels of proteins or specific gene mutations are used to provide a specific therapy for an individual by stratifying the disease status, selecting the proper medication, and tailoring dosages. Examples of companion diagnostics in the field of cancer medicine for molecular targeted therapy include tests for the ALK-fusion gene in non-small cell lung cancer and expression of CCR4 in adult T-cell leukemia. For breast cancer, the expression of HER2 protein is evaluated by immunohistochemistry (IHC), and gene amplification of HER2 is tested by fluorescence in situ hybridization (FISH); both tests consist of pre-analysis, analysis, and post-analysis processes that require quality control to ensure the reliability of the results. This symposium includes: 1) future aspects of companion diagnostics addressing many of the problems that must be overcome, 2) companion diagnostics using FISH focusing on HER2 amplification and ALK alteration, 3) newly developed diagnostic tests using tumor specimens and cell-free DNA in serum, and 4) CCR4 expression detected by IHC and flow cytometry.

Quantitative mass barcode-like image of nicotine in single longitudinally sliced hair sections from long-term smokers by matrix-assisted laser desorption time-of-flight mass spectrometry imaging.

The matrix-assisted laser desorption/ionization (MALDI) imaging mass spectrometric technique (IMS) offered a new breakthrough perspective in the analysis of drug abuse in forensic science; however, it only produced barcode-like images, semi-quantitative analysis. In order to develop intermittent monitoring by this IMS for forensic and medical sciences, it is important to quantitatively measure the contents of longitudinally sliced hair sections. We developed quantitative imaging mass spectrometry (QIMS) of nicotine (NC) in longitudinally sliced hairs by MALDI-IMS with the selected reaction monitoring mode using a labeled NC ((13)C3-NC) standard for the serially chronological monitoring and traceability of NC intake in heavy smokers. The calibration curve of NC/(13)C3-NC was virtually a linear equation at ranges from 1 to 50 ng/mL, the slope was 0.020, and the intercept was almost 0.023 and the R(2) was 0.9965. The limit of quantitation of NC was calculated as 1.6 ng/mg hair (an average weight of the hair would be assumed 0.06 mg/cm) by QIMS. Moreover, NC concentrations in two separate heavy smokers (n = 3) were 8.5 ± 1.2 and 34.5 ± 2.8 ng/mg hair, respectively, and covariations were ∼10% using a single hair. Quantitative mass barcode-like image of sliced section of hair allowed for the quantitative assessment of NC concentrations in long-term smokers similar to drugs and medicines during drug histories.

Significance of the vascular concentration of angiotensin II-receptor blockers on the mechanism of lowering blood pressure in spontaneously hypertensive rats.

To clarify the hypotensive mechanism of angiotensin II receptor-blockers (ARBs), drug concentrations in plasma and vascular tissues were measured using matrix-assisted laser desorption ionization time-of-flight mass spectrometry and imaging mass spectrometry. In spontaneously hypertensive rats, systolic blood pressure (SBP) was measured 2 and 24 h after administration of candesartan cilexetil (0.3, 1, or 3 mg/kg) or azilsartan (0.3, 1, or 3 mg/kg). SBP was similarly lowered 2 h after administration of azilsartan or candesartan cilexetil, but it was significantly lower in the azilsartan-treated group than in the candesartan cilexetil-treated group at 24 h. Angiotensin II-induced vascular contractions were similarly attenuated 2 h after administration of these drugs, and the contractions were significantly lower in the azilsartan-treated group at 24 h. Although plasma concentration was significantly lower in the azilsartan-treated group at 24 h, vascular concentration of azilsartan was significantly greater than that of candesartan. Significant correlations between SBP and vascular concentrations were observed both at 2 and 24 h, while no significant correlation was observed between plasma and vascular concentrations. In conclusion, the mechanism of ARB-induced hypotension is likely to depend on vascular concentrations rather than plasma concentrations.

Topologies of amyloidogenic proteins in Congo red-positive sliced sections of formalin-fixed paraffin embedded tissues by MALDI-MS imaging coupled with on-tissue tryptic digestion.

OBJECTIVES: Matrix-assisted laser desorption time-of flight ionization (MALDI)-imaging MS (IMS) with MSMS analysis using on-tissue tryptic digests is a powerful tool for identification of disease-related proteins in formalin-fixed paraffin-embedded (FFPE) tissue sections. We applied this novel IMS technique, not only to identify tryptic peptides of deposited amyloidogenic proteins but also to clarify topologies of these proteins in amyloidosis tissue sections. METHODS: Sequence determinations of tryptic peptides derived from amyloidogenic proteins were performed using MALDI-MSMS analysis directly from Congo red positive regions in tissue sections with/without procedure for retrieval of epitopes before on-tissue digestion. RESULTS: Tryptic peptides, m/z=1073.5 and 1924.3 were identified with the sequences, from 48th to 56th and 1st to 19th positions of Ig lambda V-III region, respectively. Other peptides, m/z=1365.5 and 1523.5 were with the sequences, from 22nd to 34th and 36th to 48th positions of TTR, respectively. Heat-map images of all four tryptic peptides were overlapped with Congo red positive regions. Immunohistochemistry of FFPE tissue sections was confirmed to only react with anti-λ chain antibody in a case of AL-type amyloidosis or anti-TTR antibody in two cases of TTR-type amyloidosis. CONCLUSION: IMS with MSMS analysis using on-tissue tryptic digestion enables us not only to identify amyloidogenic molecule in a sliced tissue section but also to play a complementary role with the conventional pathological examination.

The midline electroneurography method for facial palsy reflects total nerve degeneration.

CONCLUSION: The midline electroneurography (ENoG) method might reflect total facial nerve degeneration. OBJECTIVE: We compared ENoG values in patients with facial palsy using two different methods, the midline method and five electroneurogram recordings, to reveal whether the ENoG value obtained with the midline method reflects total facial nerve degeneration. METHODS: Forty patients with facial palsy were enrolled. Compound muscle action potentials (CMAPs) were recorded using the midline method, in which the anode was placed on the mental protuberance and the cathode was placed on the philtrum. Additionally, five electroneurogram recordings were obtained by placing the anode on the skin of the parietal region and five cathodes on the skin over five facial muscles (frontalis, orbicularis oculi, nasalis, orbicularis oris, and depressor anguli oris muscles). ENoG values recorded using the two methods were compared. RESULTS: The ENoG values of the five facial muscles did not differ from those obtained using the midline method. The total ENoG value calculated by summing five CMAPs from five facial muscles, which is considered to reflect total facial nerve degeneration, was not significantly different from that using midline methods; moreover, a strong positive correlation coefficient (r = 0.87) was found between them.

Quantification of Candesartan in Mouse Plasma by MALDI-TOFMS and in Tissue Sections by MALDI-Imaging Using the Stable-Isotope Dilution Technique.

To determine the contents of candesartan in mouse plasma, and blood vessel and kidney sliced sections and also better understand its pharmacokinetics, we applied matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS) and MALDI-imaging mass spectrometry (IMS) with the selected reaction monitoring (SRM) mode using a labeled-internal standard. The results of fundamental examinations showed that the slope of the resulting curves of candesartan in the plasma from the equation was 0.91 and the y-intercept was 0.02. Both intra- and inter-day accuracies (n=10) and the precision of candesartan in the plasma by MALDI-TOFMS with the SRM mode were in the range of 3.4 to 17.3% and 93.2%, respectively. The detection limit of candesartan in spiked plasma was 0.2 nmol/L. IMS analysis enabled us to clarify distinct spacial time-distribution images in sliced mouse blood vessel and kidney sections although it still needed to improve a protocol of quantification. Typical pharmacokinetic patterns of candesartan were obtained in the plasma and sliced kidney sections, but those in the blood vessel sections gradually increased 24 h after administration. MALDI-TOFMS and IMS with the SRM mode are powerful tools to identify the spacial distribution and traceability of candesartan in sliced blood vessel and tissue sections as well as in the plasma.

[Acquisition of accreditation ISO 15189 and view of the future--case of a private college].

We received ISO 15189 accreditation (International Organization for Standardization 15189) in 2009. Several effects from the subjective points are shown below, 1. The reliability of the test results has improved. 2. Skill or knowledge level of staff has improved. 3. Internal recognition has improved. 4. Procedures to prevent accidents have been enforced. 5. Management system has improved. 6. Effective management tool for the manager. In addition to high costs for qualification and maintenance, extensive human resources are required to prepare documents. We need to make further efforts to aim at better cost-effectiveness.

[Clinical applications of MALDI imaging using sliced sections of formalin-fixed paraffin-embedded tissues and longitudinal sliced hairs].

MALDI-imaging MS (IMS) with MSMS analysis is a new powerful tool for the identification of not only disease-related proteins in formalin-fixed paraffin-embedded (FFPE) tissue sections but also protein/peptides/drugs/medicine in fresh-frozen tissues. IMS is used to reveal the mass profiles and spatial distribution of proteins in tissue sections and/or digested peptides derived from deposited protein in pathologic organs and then MSMS analysis identifies the amino acid sequence of the detected proteins in the tissue section. Moreover, on-tissue digestion combined with the MALDI-IM-TOF-IMS approach allows a proteomics "bottom-up" strategy with clinical samples, especially perioperative isolated tissues and FFPE tissues conserved for a long time in a clinical sample bank. The mass barcode-like image (MBI) on a longitudinal sliced hair by IMS is used in the selected reaction monitoring mode for serially chronological monitoring and traceability every few hours after drug and medicine intake. The advances of quantitative MBI for sliced sections of hair allow a new universal standardized assessment of drugs and medicines throughout the drug history.

[In a disaster, what should medical technologists at a university hospital do? What do they have?].

"Disaster medicine" involves "cases that cannot be supported by the normal medical service system of the hospital because many injuries have occurred by an explosion/chemical pollution/radioactive pollution or a pandemic caused deliberately as well as natural disasters". In "disaster medicine", university hospitals should become the base for advanced medical services and wide area transportation. Also, the clinical laboratory/medical technologists of the university hospital should offer high quality laboratory analysis. On the other hand, one of the advantages of a university hospital is an experimental medicine laboratory (integrated universities also have a Department of Science and Department of Engineering). Development of testing equipment to cover all the functions necessary for disaster medicine may be a possibility. A system to conduct simple testing is expected at the actual location of the emergency, and an identification system must be established. "Emergency medicine" and "disaster medicine" have different aspects, but the essence is the same, and medical technologists must have knowledge/techniques to report and interpret results quickly. Because a university hospital is the core of logistical support, daily training is important.