An automated rapid detection system using the quenching probe method for detecting interleukin 28B and inosine triphosphatase single nucleotide polymorphisms in chronic hepatitis C.

Single nucleotide polymorphisms (SNPs) in the interleukin 28B gene (IL28B) are good pretreatment predictors of anti-hepatitis C virus (HCV) therapy with interferon. SNPs of the inosine triphosphatase (ITPA) gene are associated with reduced haemoglobin levels during treatment with ribavirin. The i-densy™ (Arkray, Inc.), which is based on the quenching probe (QP) method, automatically detects target genes in blood samples by fluorescence quenching within 100 min. Using a QP and primer set, a gene amplification response is generated that can quickly and easily detect a specific gene's arrangement by fluorometry. The present study was conducted to compare the utility of i-densy (QP method) with that of conventional direct sequencing (DS) for detecting SNPs in the IL28B and ITPA genes in chronic hepatitis C patients. Between June 2011 and January 2012, 73 consecutive patients underwent genotyping of IL28B, and 54 patients underwent genotyping of ITPA. All of the patients were seropositive for HCV-RNA. The IL28B and ITPA genotypes were tested for bi-allelic polymorphisms in rs8099917 (T/T, T/G and G/G; minor allele, G) and rs1127354 (C/C, C/A and A/A; minor allele, A), respectively. The results obtained with the QP method were identical to those obtained with the conventional DS method. The frequency of the IL28B genotypes TT, GT and GG were 74%, 24.7% and 1.4%, respectively, and those of the ITPA genotypes CC, AC and AA were 68.5%, 29.6% and 1.9%, respectively. These results indicate that the i-densy using the QP method can automatically, quickly and easily identify genotypes of IL28B and ITPA.

Hemodynamic assessment of celiaco-mesenteric anastomosis in patients with pancreaticoduodenal artery aneurysm concomitant with celiac artery occlusion using flow-sensitive four-dimensional magnetic resonance imaging.

OBJECTIVES: Many pancreaticoduodenal artery (PDA) aneurysms are associated with celiac artery (CA) stenosis. The pathogenesis of PDA aneurysm may be associated with hemodynamic changes due to CA stenosis/occlusion. The aim of this study was to assess the hemodynamic changes of celiaco-mesenteric anastomosis in patients with PDA aneurysms concomitant with CA occlusion using four-dimensional flow-sensitive magnetic resonance imaging (4D-Flow). METHODS: 4D-Flow was performed preoperatively on five patients. Seven age- and sex-matched individuals were used as controls. Hemodynamic parameters such as flow volume and maximum flow velocity in PDAs, gastroduodenal arteries, common hepatic arteries, and superior mesenteric arteries were compared between both groups. Wall shear stress (WSS) and oscillatory shear index (OSI) were mapped in both groups. RESULTS: In the patient group, 4D-Flow identified retrograde flow of both gastroduodenal arteries and common hepatic arteries. Heterogeneous distribution patterns of both WSS and OSI were identified across the entire PDA in the patient group. OSI mapping showed multiple regions with extremely high OSI values (OSI > 0.3) in all patients. All PDA aneurysms, which were surgically resected, were atherosclerotic. CONCLUSIONS: 4D-Flow identified hemodynamic changes in celiaco-mesenteric arteries in patients with PDA aneurysms with concomitant CA occlusion. These hemodynamic changes may be associated with PDA aneurysm formation.

Assessment of wastewater-irrigated soil containing heavy metals and establishment of specific biomarkers.

Irrigation with treated wastewater (TWW) is a vital alternative for arid and semi-arid lands but it poses pollution-risk to soil, vegetation and groundwater. Therefore, in the present study, in vitro bioassays were used to evaluate the adverse effects of TWW and irrigated-soil extract sample, on mammalian cells, with respect to heavy metal--Ni, Cd, Pb, Fe, Al-content. The heat shock protein (HSP) 47, E-screen, and transepithelial electrical resistance (TEER) assays served to investigate the stress response of treated-HSP47-transfected Chinese hamster ovary (CHO) cells, the estrogenic activity of the samples in MCF-7 breast cancer cells, and the barrier function (BF) of Caco-2 cells. Furthermore, proteomics analyses were performed to shed light on involved mechanisms and to establish pollution biomarkers. Results showed that the TWW elicited a stress response on HSP cells from 0.1% concentration while soil extract samples exhibited a stress at 1%. TWW induced an estrogenic activity at 10%; up-regulating cell proliferation and tumor-related proteins. Soil extract triggered the enhanced expression of HSP70 family proteins as survival mechanisms against their cytotoxicity toward MCF-7 cells. Moreover, depending on the concentration, 1% of soil extract from 20 cm depth (T20) resulted in a disruption of BF in Caco-2 cells involving cell metabolism, protein synthesis and tumor marker proteins, whereas, 5% of T20 induced the expression of BF-related proteins associated to heat shock, oxidative stress, cell proliferation and glycolytic metabolic pathway. These biological techniques were found to be extremely useful to evaluate the impact of wastewater reuse and to establish specific biomarkers that are common proteins for humans, other mammals and plants. Future studies should focus on exposure quantifications.

Olive leaf tea may have hematological health benefit over green tea.

Olive leaf extracts are rich in several polyphenols having potential health benefits. We conducted the current parallel-group randomized controlled trial to compare the effects of long-term consumption of olive leaf tea (OLT) and green tea (GT) on hematological parameters in 31 female volunteers aged between 40 and 70 years of old. We found that RBC count, hemoglobin, and hematocrit were increased significantly in the OLT group than those of in the GT group at 6 and 12 weeks of intervention. Within-group comparison showed that hematocrit was significantly increased in the OLT group at 6 weeks of intervention, whereas RBC count and serum iron was significantly decreased in the GT group at 12 weeks of intervention. This is the first clinical study reporting the beneficial effects of continuous intake of OLT on hematological parameters. This observation is supported by our previous in vitro study reporting the differentiation-inducing effect of certain olive leaf components on human hematopoietic stem cells. However, further investigations in larger cohorts with a careful consideration of target population are required to confirm the preventive effect of OLT against anemia and other red cell disorders.

Inhibitory effect of various Tunisian olive oils on chemical mediator release and cytokine production by basophilic cells.

Tunisian olive oils have been traditionally used as a medicinal food for chronic inflammation. To investigate the antiallergic effect of virgin olive oil samples from five principal olive varieties grown in various regions of Tunisia, we used the type I allergy reaction model using rat basophilic leukemia (RBL-2H3) cells and different dilutions of olive oil samples to determine beta-hexosaminidase release inhibition at two different response stages. Results showed that the Sayali olive oil significantly inhibited beta-hexosaminidase release by the IgE antibody-sensitized, BSA antigen-stimulated RBL-2H3 cells at the antibody-antigen binding stage. The result of our experiment shows that the anti-allergic effect of olive oil at this binding stage may be dependent on their flavone content. The Zarrazi olive oil significantly inhibited beta-hexosaminidase release at the antigen-receptor binding stage. Moreover, we investigated the effect of olive oil samples on histamine release and production of cytokines by activated human basophilic (KU812) cells. Different dilutions of Sayali olive oil dose-dependently inhibited the production of tumor necrosis factor-alpha (TNF-alpha) and interleukin-4 (IL-4), and different dilutions of Zarrazi olive oil dose-dependently inhibited histamine release and IL-4 production by calcium ionophore A23187 plus phorbol 12-myristate 13-acetate (PMA)-stimulated KU812 cells.

Quantitative Analysis of Conebeam CT for Delineating Stents in Stent-Assisted Coil Embolization.

BACKGROUND AND PURPOSE: Innovative techniques and device-related advances have improved the outcomes of neuroendovascular treatment. 3D imaging has previously used 2 × 2 binning, but 1 × 1 binning has recently been made available. The aim of this study was to evaluate the quantitative ability of conebeam CT for stent delineation and to investigate its effectiveness in the clinical environment. MATERIALS AND METHODS: Four acquisition groups of 3D MIP images acquired using conebeam CT with varying conditions (acquisition time, 10 or 20 seconds and binning, 1 × 1 or 2 × 2) were compared. Two methods of analysis were performed, a phantom study and an analysis of 28 randomly selected patients. The phantom study assessed the contrast-to-noise ratio and full width at half maximum values in conebeam CT images of intracranial stent struts. In the clinical subjects, we assessed contrast-to-noise ratio, full width at half maximum, and dose-area product. RESULTS: In the phantom study, the contrast-to-noise ratio was not considerably different between 10- and 20-second acquisition times at equivalent binning settings. Additionally, the contrast-to-noise ratio at equivalent acquisition times did not differ considerably by binning setting. For the full width at half maximum results, equivalent acquisition times differed significantly by binning setting. In the clinical analyses, the 10-second/1 × 1 group (versus 20 second/2 × 2) showed a higher contrast-to-noise ratio (P < .05) and a dose-area product reduced by approximately 70% (P < .05), but the difference in full width at half maximum was not significant (P = .20). CONCLUSIONS: For stent-assisted coil embolization, quantitative assessment of conebeam CT showed that 10 second/1 × 1 was equivalent to 20 second/2 × 2 for imaging deployed intracranial stents. Furthermore, the 10-second/1 × 1 settings resulted in a much smaller DAP.

A Multiple-Cell Microenvironment in a 3-Dimensional System Enhances Direct Cellular Reprogramming Into Hepatic Organoids.

OBJECTIVES: The difficulty in proliferation and availability and the rapid loss functions of primary human hepatocytes highlight the need to develop an alternative, preferably renewable source of human induced hepatocytes in regenerative medicine. Liver organoids generated on a multiple-cell microenvironment in a 3-dimensional (3D) system can provide a highly efficient solution to this issue. METHODS: Human hepatocytes were induced from fibroblasts by the lentiviral expression of FOXA3, HNF1A, and HNF4A. Together with these induced hepatocytes, human umbilical vein endothelial cells and mesenchymal stem cells in a 3D system were used to produce liver organoids. Liver-related gene and protein expression of liver organoids and induced hepatocytes were tested using a 2-dimensional (2D) system. RESULTS: Liver organoids notably increased the expression of hepatic transcription factors, marker genes, transporter genes, and liver metabolism enzyme genes, while it decreased the specific gene expression of fibroblasts. Liver organoids expressed comparable liver-specific proteins, such as ALB, AAT, and HNF4A in the 3D system. CONCLUSION: Direct reprogramming in multiple-cell microenvironments in 3D systems is more controllable and efficient than cell reprogramming in 2D systems. Liver organoids have the potential for use in disease modeling, pharmaceutical applications, and cellular transplantation.

Visualization of hemodynamics in a silicon aneurysm model using time-resolved, 3D, phase-contrast MRI.

We performed time-resolved 3D phase-contrast MR imaging by using a 1.5T MR scanner to visualize hemodynamics in a silicon vascular model with a middle cerebral aneurysm. We ran an aqueous solution of glycerol as a flowing fluid with a pulsatile pump. Time-resolved images of 3D streamlines and 2D velocity vector fields clearly demonstrated that the aneurysm had 3D complex vortex flows within it during systolic phase. This technique provided us with time-resolved 3D hemodynamic information about the intracranial aneurysm.

Expressions of galectin-3, glutathione S-transferase A2 and peroxiredoxin-1 by nonylphenol-incubated Caco-2 cells and reduction in transepithelial electrical resistance by nonylphenol.

Nonylphenol, an estrogenic xenobiotic widely used in the manufacture of plastics and detergents, has been found in drinking water and may therefore enter the body through the oral route. Thus, intestinal cells lining the alimentary tract serve as the body's first line of defense against this compound. In this study, the effects of nonylphenol on the human intestinal cell line Caco-2 were determined using transepithelial electrical resistance (TEER) measurement and proteomics. Results show that 10 microM nonylphenol can disrupt the tight-junction permeability of Caco-2 cells in approximately 15 min. Incubating the cells with 1 or 10 microM nonylphenol for 6 days resulted in the enhanced expressions of galectin-3 (approximately 4-fold vs. control with 1 microM; 2-fold with 10 microM), glutathione S-transferase A2 (approximately 8-fold with 1 microM; 5-fold with 10 microM) and peroxiredoxin-1 (approximately 6-fold with 1 microM; 4-fold with 10 microM). These expressions may represent a possible consortium of mechanisms by which the cells protect themselves against nonylphenol-induced stresses. To the best of our knowledge, this is the first study on the effects of nonylphenol on Caco-2 cells.

Change in ascorbate radical production in an irradiated experimental tumor with increased tumor size.

We have reported that ascorbate radical (Asc.-) could serve as an indicator of the amount of hydroxyl radical and superoxide produced by irradiation in vivo. Using this method, we investigated the relationship between tumor size and Asc.- production after irradiation (10 Gy) and between tumor size and the radical-scavenging ability of WR-2721 (300 mg/kg). Asc.- was measured in normal muscle and SCC-VII tumors transplanted into mice (n = 6). In tumors, the increase in Asc.- significantly decreased with increasing tumor size (r = -0.483; P < 0.05). The increase in Asc.- production after irradiation was more inhibited by WR-2721 in normal muscle tissue than in tumor tissue at various sizes. In tumors, the increase in Asc.- was less inhibited by WR-2721 with increasing tumor size. These results demonstrate that the increase in radical production after irradiation and drug distribution decreased with increasing tumor size and that WR-2721 has excellent differential protection. This method is expected to measure changes in the amounts of local hydroxyl radical and superoxide modified by a change of tumor environment or drug administration.

Gastrointestinal visualization during xenon-133 ventilation study in a patient with tracheoesophageal fistula.

A 37-yr-old female with H-type tracheoesophageal fistula demonstrated visualization of the gastrointestinal tract during a ventilation study using 133Xe gas. Gastrointestinal visualization by 133Xe on a ventilation study may prove to be a useful screening procedure for this condition in appropriate patients.

Enhancement of thermal damage in murine tumors by hydralazine-induced modification of blood flow and oxygen tension.

We investigated changes in blood flow in normal muscle and in SCC-VII tumors treated by hyperthermia combined with hydralazine, to evaluate the enhancement of thermal tumor damage by hydralazine. We studied SCC-VII tumor-bearing C3H/He mice. Hydralazine was administered by intraperitoneal injection, and tumors were heated by a water bath. We measured blood flow using the laser Doppler method, and oxygen tension using polarography. The response of tumors to therapy was assessed using a growth delay analysis. In tumors, blood flow and O-2 tension significantly decreased with increasing doses of hydralazine. Compared to tumors treated by hydralazine alone or by hyperthermia alone, tumor blood flow was significantly decreased in the group treated by hyperthermia with hydralazine. In tumors treated by hyperthermia with hydralazine, blood flow was significantly decreased with increasing Hyd doses, heat durations, and temperatures. In normal muscle, no decrease in blood flow was induced by hyperthermia, hydralazine, or their combination. In tumors treated by hyperthermia (43 degrees C, 20 min) with hydralazine, a maximum additional growth delay was observed. Our results suggest that a decrease in tumor blood flow caused by hydralazine plays an important role in enhancement of the hyperthermic antitumor effect.

Radiosensitization with a 3-nitrotriazole (ak-2123).

We investigated to clarify the mechanism of action of AK-2123. V79 cells were used, and the effect of the substance was estimated with different end-points viz. induction of DNA breaks, micronuclei and clonogenic survival after hypoxic irradiation with different X-ray doses. After treatment with 0.1 mmol dm(-3) of the substance, sensitization by a factor of 1.8, 1.1 and 1.4 respectively, was calculated. By determining the enhancement ratio after treatment with the drug at 7-8 different concentrations varying between 0.001 and 50 mmol dm(-3), the concentration of the substance for half of the maximum sensitization (K-value) was estimated to be 40 mu mol dm(-3) (DNA breaks), 1200 mu mol dm(-3) (micronuclei) and 2000 mu mol dm(-3) (survival); the maximum sensitizer enhancement ratio was 2.4 (DNA breaks), 2.3 (micronuclei) and 2.4 (survival). No cytotoxic effects were found when the cells were treated with the drug in different concentrations for 30 minutes. However, after longer periods the compound was found to be cytotoxic both under aerobic and hypoxic conditions, in a concentration and time dependent manner. A post-irradiation enhancement of the sensitivity was noted when the drug was left in the medium for 24 hours after hypoxic as well as aerobic radiation exposure.

Dissociation of writing processes: functional magnetic resonance imaging during writing of Japanese ideographic characters.

Dissociation between copying letters and writing to dictation has been reported in the clinical neuropsychological literature. Functional magnetic resonance imaging (fMRI) was conducted in normal volunteers to detect the neurofunctional differences between 'copying Kanji', the Japanese ideographic characters, and 'writing Kanji corresponding to phonological information'. Four tasks were conducted: the copying-Kanji task, the writing-Kanji-corresponding-to-phonogram task, the Kanji-grapheme-puzzle task, and the control task. The right superior parietal lobule was extensively activated during the copying-Kanji task (a model of the copying letters process) and the Kanji-grapheme-puzzle task. These observations suggested that this area was involved in referring the visual stimuli closely related to the ongoing handwriting movements. On the other hand, Broca's area, which is crucial for language production, was extensively activated during the writing-Kanji-corresponding-to-phonogram task (a model of the writing-to-dictation process). The Kanji-grapheme-puzzle task activated the bilateral border portions between the inferior parietal lobule and the occipital lobe, the left premotor area, and the bilateral supplementary motor area (SMA). Since the Kanji-grapheme-puzzle task involved manipulospatial characteristics, these results suggested cooperation between visuospatial and motor executive functions, which may be extensively utilized in demanding visual language processing. The neurofunctional difference between 'copying Kanji' and 'writing Kanji corresponding to phonogram' was efficiently demonstrated by this fMRI experiment.

The effects of listening comprehension of various genres of literature on response in the linguistic area: an fMRI study.

Using fMRI at a static magnetic field strength of 1.5T, we investigated how comprehension and humor of sentences would correlate to activation of the language areas in listening comprehension of a native language. Sentences with a high comprehension score augmented activation in the left inferior parietal lobule and posterior part of the left superior temporal gyrus, which may be related to semantic processing. Sentences with a high humor score induced activation in Broca's area, which may be associated with syntactic processing and auditory working memory. Furthermore, sentences with a high humor factor and/or a low comprehension score activated the middle frontal gyrus, which may be attributed to auditory working memory.

Involvement of motor cortices in retrieval of kanji studied by functional MRI.

Functional magnetic resonance imaging was successfully used to study the activation of the motor cortices during retrieval of Japanese ideogram, kanji. The subjects performed kanji completion tasks to generate a kanji in response to an element which is always written first. In most of the subjects, the contralateral premotor cortex, the presupplementary motor area (pre-SMA) and the bilateral intraparietal sulcus were activated during retrieval of kanji without actual writing nor intentional mental writing. Activation associated with actual writing was shown in the contralateral primary sensorimotor cortex and the SMA proper. These results suggested that retrieval of kanji would share the neural basis of motor representation with writing of kanji except for regions directly working for motor output.

Ideographic characters call for extra processing to correspond with phonemes.

Cortical areas used in the copying of Japanese ideographic characters and syllabic characters were studied using functional magnetic resonance imaging in healthy volunteers. Complexity of characters was controlled to illustrate differences resulting from character to sound conversion differences between the ideographic and syllabic characters. Statistical comparisons indicated extensive activation in the fusiform gyrus, posterior portions around the intraparietal sulcus and in the conjunction area of BA 6, 9 and 44 (which is assumed to be Exner's area) during the copying of ideographic characters. These findings suggested that indirectness between ideographic characters and their pronunciation demands extra processing such as semantic mediation and intensive grapheme processing in comparison with syllabic characters.

Proton magnetic resonance spectroscopy of lenticular nuclei in simple schizophrenia.

1. The lenticula nuclei have been suggested to be the site of structural and functional abnormalities in schizophrenia. 2. Recently, several studies involving proton magnetic resonance spectroscopy (1H MRS) showed that the ratio of N-acetyl-aspartate (NAA) to choline-containing compounds (Cho) was significantly reduced in the basal ganglia region in patients with schizophrenia. 3. Simple schizophrenia is characterized by social withdrawal and affective flattening, but not by prominent catatonic, hebephrenic or paranoid features. 4. We studied, using 1H MRS, the lenticula nuclei of 10 patients with simple schizophrenia, and 10 age- and sex-matched healthy controls. 5. No differences between the patients and the controls were found in any of the measured ratios, i.e. Cho/Cr, NAA/Cr and NAA/Cho. 6. Our results suggest the normal viability of neuronal cells, as found on quantification of NAA, Cr and Cho, in the lenticular nuclei of patients with simple schizophrenia. 7. The pathophysiology of simple schizophrenia may be different from those of other types of schizophrenia.

MR angiography of aneurysm models of various shapes and neck sizes.

PURPOSE: To investigate the signal intensity of lateral and terminal saccular aneurysm models with differing neck sizes using three-dimensional time-of-flight (TOF) MR angiography with various imaging parameters. METHODS: The study included four lateral and four terminal saccular aneurysm models with pulsatile flow. The height and fundus diameter were 10 mm; the neck diameters were 2.5 mm, 5 mm, 7.5 mm, and 10 mm, respectively. Each aneurysm model was examined with fast imaging with steady-state precession MR sequences with parameters of 20-140/7 (repetition time/echo time) and flip angles of 10 degrees to 30 degrees. Signal intensity was measured and compared among the models. RESULTS: Three-dimensional TOF MR angiography with the shorter repetition time and/or larger flip angle showed weaker signal intensity in the aneurysm models. Stronger signal intensity was obtained in the terminal saccular aneurysm models and/or the models with a wider neck than in the lateral saccular aneurysm models and/or the models with a narrower neck. In some aneurysm models, longer repetition times produced greater signal intensity than that of background brain models, but not in aneurysms with narrow necks. CONCLUSION: Noncontrast 3-D TOF MR angiography delineated terminal saccular aneurysms and/or aneurysms with wider necks and did not delineate lateral saccular aneurysms and/or aneurysms with narrower necks. Longer repetition times are recommended to allow the spins flowing into the aneurysms to recover.