Imaging discrepancies between magnetic resonance imaging and brain perfusion single-photon emission computed tomography in the diagnosis of Alzheimer's disease, and verification with amyloid positron emission tomography.

BACKGROUND: In the diagnosis of Alzheimer's disease (AD), discrepancies are often observed between magnetic resonance imaging (MRI) and brain perfusion single-photon emission computed tomography (SPECT) findings. MRI, brain perfusion SPECT, and amyloid positron emission tomography (PET) findings were compared in patients with mild cognitive impairment or early AD to clarify the discrepancies between imaging modalities. METHODS: Several imaging markers were investigated, including the cortical average standardized uptake value ratio on amyloid PET, the Z-score of a voxel-based specific regional analysis system for AD on MRI, periventricular hyperintensity grade, deep white matter hyperintense signal grade, number of microbleeds, and three indicators of the easy Z-score imaging system for a specific SPECT volume-of-interest analysis. Based on the results of the regional analysis and the three indicators, we classified patients into four groups and then compared the results of amyloid PET, periventricular hyperintensity grade, deep white matter hyperintense signal grade, and the numbers of microbleeds among the groups. RESULTS: The amyloid deposition was the highest in the group that presented typical AD findings on both the regional analysis and the three indicators. The two groups that showed an imaging discrepancy between the regional analysis and the three indicators demonstrated intermediate amyloid deposition findings compared with the typical and atypical groups. The patients who showed hippocampal atrophy on the regional analysis and atypical AD findings using the three indicators were approximately 60% amyloid-negative. The mean periventricular hyperintensity grade was highest in the typical group. CONCLUSIONS: Patients showing discrepancies between MRI and SPECT demonstrated intermediate amyloid deposition findings compared with patients who showed typical or atypical findings. Strong white matter signal abnormalities on MRI in patients who presented typical AD findings provided further evidence for the involvement of vascular factors in AD.

A Randomized, Double-Blind, Placebo-Controlled Study to Evaluate the Efficacy of Teprenone in Patients with Alzheimer's Disease.

BACKGROUND: Teprenone (geranylgeranylacetone), an anti-ulcer agent, has been reported to inhibit amyloid-ß increase, senile plaque formation, and neuronal degeneration, and improve memory in mouse models of Alzheimer's disease (AD). OBJECTIVE: We conducted a randomized, double-blind, placebo-controlled study to ascertain teprenone's therapeutic ability for AD. METHODS: Patients with mild to moderate AD, with a Mini-Mental State Examination (MMSE) score of 13 to 26, were randomly allocated into two groups depending on the administered drug: donepezil +  placebo (placebo group) and donepezil + teprenone (teprenone group). The primary and secondary endpoints included changes in scores of the Japanese version of the AD Assessment Scale-cognitive subscale (ADAS-J cog) and other evaluations, respectively, including MMSE scores, during a 12-month period after the first administration. RESULTS: Forty-two and thirty-seven patients were allocated to the teprenone and placebo groups, respectively. ADAS-J cog score changes were not different between groups (placebo, 0.6±0.8; teprenone, 0.4±0.8; p = 0.861). However, MMSE scores significantly improved in the teprenone group (placebo, - 1.2±0.5; teprenone, 0.2±0.5; p = 0.044). Subgroup analysis considering the severity of medial temporal area atrophy revealed that this improvement by teprenone was significant in patients with mild (p = 0.013) but not with severe atrophy (p = 0.611). Adverse events were observed in 17.8 and 10.4% of patients in the placebo and teprenone group, respectively. CONCLUSION: Teprenone may be effective for AD when administered before atrophy progression in the medial temporal areas. TRIAL REGISTRATION: UMIN ID: UMIN000016843.

Use of ANGPTL2 mRNA levels in formalin-fixed paraffin-embedded tissues as a biomarker to diagnose gastric cancer and to evaluate the extent of vascular invasion.

With the recent advances in medical technologies, gastric cancer can often be removed with minimally invasive surgical techniques when identified early. Surgery must remove all gastric cancer, since residual cancerous tissue may lead to recurrence. Resected cancerous tissues are pathologically evaluated to determine whether all cancerous areas have been removed, but such assessments are rarely straightforward, and cancer markers could inform such pathological evaluations of cancer. An ideal marker would be identifiable in formalin-fixed paraffin-embedded (FFPE) tumor tissue. The first objective of the present study was to compare levels of angiopoietin-like protein 2 (ANGPTL2) in cancerous and noncancerous areas of FFPE tissues to determine whether ANGPTL2 is a marker relevant to the pathological diagnosis of cancer. The second objective was to evaluate whether ANGPTL2 mRNA is useful as a marker of the extent of vascular invasion of gastric cancer. Out of the 15 patients studied, 12 had a higher ANGPTL2 mRNA levels in cancerous areas compared with noncancerous areas. This finding indicated that ANGPTL2 mRNA is useful as a biomarker for identifying cancerous areas in FFPE tissues, at least for male patients. Spearman's rank correlation analysis showed a significant correlation between the ANGPTL2 mRNA level and the degree of vascular invasion of cancer (r=0.66; P=0.01). In receiver operating characteristic curve analysis of the association between the ANGPTL2 mRNA level and the degree of vascular invasion, the area under the curve was 0.92 (95% confidence interval, 0.78-1.00; P=0.01), indicating a significant association. The present study demonstrates that ANGPTL2 mRNA in FFPE tissues is a potential biomarker that informs the pathological diagnosis of gastric cancer and that ANGPTL2 mRNA may be predictive of vascular invasion, which is an indicator of metastasis in gastric cancer.

A novel isolation method for cancer prognostic factors via the p53 pathway by a combination of in vitro and in silico analyses.

Identifying new therapeutic target genes affecting the survival of patients with cancer is crucial for the development of new cancer therapies. Here, we developed a novel technology combining in vitro short hairpin RNA (shRNA) library screening and in silico analysis of the tumor transcriptome to identify prognostic factors via the p53 tumor-suppressor pathway. For initial screening, we screened 5,000 genes through selection of shRNAs in p53 wild-type tumor cells that altered sensitivity to the p53 activator actinomycin D (ActD) to identify p53 regulatory genes; shRNAs targeting 322 genes were obtained. Among these 322 genes, seven were prognostic factor candidates whose high expression increased ActD sensitivity while prolonging the survival period in patients with the p53 wild-type genotype. Conversely, we identified 33 genes as prognostic factor candidates among ActD-resistant genes related to a shortened survival period only in p53 wild-type tumors. These 40 genes had biological functions such as apoptosis, drug response, cell cycle checkpoint, and cell proliferation. The 40 genes selected by this method contained many known genes related to the p53 pathway and prognosis in patients with cancer. In summary, we developed an efficient screening method to identify p53-dependent prognostic factors with in vitro experimental data and database analysis.

Autoclavable physically-crosslinked chitosan cryogel as a wound dressing.

Moist wounds were known to heal more rapidly than dry wounds. Hydrogel wound dressings were suitable for the moist wound healing because of their hyperhydrous structure. Chitosan was a strong candidate as a base material for hydrogel wound dressings because the polymer had excellent biological properties that promoted wound healing. We previously developed physically-crosslinked chitosan cryogels, which were prepared solely by freeze-thawing of a chitosan-gluconic acid conjugate (CG) aqueous solution, for wound treatment. The CG cryogels were disinfected by immersing in 70% ethanol before applying to wounds in our previous study. In the present study, we examined the influence of autoclave sterilization (121°C, 20 min) on the characteristics of CG cryogel because complete sterilization was one of the fundamental requirements for medical devices. We found that optimum value of gluconic acid content of CG, defined as the number of the incorporated gluconic acid units per 100 glucosamine units of chitosan, was 11 for autoclaving. An increased crosslinking level of CG cryogel on autoclaving enhanced resistance of the gels to enzymatic degradation. Furthermore, the autoclaved CG cryogels retained favorable biological properties of the pre-autoclaved CG cryogels in that they showed the same hemostatic activity and efficacy in repairing full-thickness skin wounds as the pre-autoclaved CG cryogels. These results showed the great potential of autoclavable CG cryogels as a practical wound dressing.

Angiopoietin-like Protein 2 is a Useful Biomarker for Pancreatic Cancer that is Associated with Type 2 Diabetes Mellitus and Inflammation.

Pancreatic cancer is one of the tumors with the worst prognosis, with the 5-year survival rate reported to be 6%. The number of patients suffering from pancreatic cancer in recent years has continued to increase dramatically. Carbohydrate antigen 19-9 is an established biomarker of pancreatic cancer, but it does not have sufficient ability to detect pancreatic cancer at an early stage. We focused on angiopoietin-like protein 2 (ANGPTL2), which has been reported to be related to chronic inflammation and Type 2 diabetes mellitus. In this study, whether ANGPTL2 can detect early pancreatic cancer was evaluated. It was found that the concentration of serum ANGPTL2 was significantly higher in pancreatic cancer patients and tumor stage 0-I patients than in healthy individuals (5.84 ± 1.82 ng/mL vs 3.61 ± 0.64 ng/mL; P < 0.001) (5.68 ± 0.79 ng/mL vs 3.61 ± 0.64 ng/mL; P = 0.010). In addition, the diagnostic capability of serum ANGPTL2 levels for pancreatic cancer was evaluated using receiver operating characteristic (ROC) curve analysis. The area under the ROC curve (AUC) for ANGPTL2 was 0.906 (95% confidence interval (CI): 0.815-0.997; P < 0.001). To identify the risk factors for pancreatic cancer, multivariate regression models were used. Ten factors were included, and increasing age (odds ratio (OR), 1.318, 95% CI, 1.058-1.642; P = 0.014) and high ANGPTL2 levels (OR, 22.219, 95% CI, 1.962-251.659, P = 0.012) were found to be independent risk factors for pancreatic cancer, with ANGPTL2 having the strongest relationship. In addition, serum ANGPTL2 levels were strongly correlated with inflammatory markers, with blood sugar levels showing the strongest correlation with serum ANGPTL2 levels. In conclusion, this study suggested that an elevated serum ANGPTL2 level has the potential to be a biomarker capable of early detection of pancreatic cancer, and it was correlated with inflammation of the pancreas and the risk of developing diabetes mellitus.

Combined assessment of serum folate and hemoglobin as biomarkers of brain amyloid ß accumulation.

A relationship between Alzheimer's disease (AD) and folate has been reported. Amyloid positron emission tomography (PET) is currently one of the most reliable biomarkers for AD. We investigated the correlation between serum folate levels and amyloid imaging to clarify whether serum folate could be a biomarker for AD. We also examined the usefulness of a combined assessment of serum folate levels and red blood cell hemoglobin content. Apolipoprotein E (APOE) gene polymorphisms were also assessed. Serum folate levels and hemoglobin content were evaluated by receiver operating characteristic analysis for their diagnostic capability as AD biomarkers relating to brain amyloid ß accumulation. The area under the ROC curve (AUC) for serum folate was 0.136 (95% confidence interval [CI]: 0.000-0.312; p = 0.016). The AUC for hemoglobin content was 0.848 (95% CI: 0.661-1.000; p = 0.021). Therefore, the folate deficiency with low folate levels or the non-anaemia with high hemoglobin content levels were found to have a high probability of also testing positive for amyloid. Furthermore, eight patients were found to be folate deficiency and non-anaemia, those who were consist of 7 amyloid positive patients (87.5%), and only one of the amyloid negative patients (12.5%). These results suggest that a deficiency of serum folate and high hemoglobin levels may reflect an increased risk of amyloid ß accumulation in the brain. Additionally, we demonstrated that these biomarkers could enhance the effectiveness of APOE as an AD biomarker. This study reveals that the combined assessment of serum folate levels and red blood cell hemoglobin content may be a useful biomarker for amyloid ß accumulation in the brain. We also found that the combination of serum folate levels and hemoglobin content is a more specific and sensitive blood biomarker for AD than APOE or folate alone. These findings may be used to support clinical diagnosis of AD using a simple blood test.

Vascular-like network prepared using hollow hydrogel microfibers.

One major challenge in the field of tissue engineering was the creation of volumetric tissues and organs in vitro. To achieve this goal, the development of a three-dimensional vascular-like network that extended throughout the tissue-engineered construct was essential to supply sufficient oxygen and nutrients to all of the cells in the constructs. For sufficient oxygenation and nutrition of the tissue-engineered constructs, the distance between each microvessel-like channel in the network should ideally be within 100-200 µm. In addition, the medium or blood should be perfused through the microchannels as soon as possible after the seeding of cells into the templates (scaffolds) of the constructs. In the present study, we proposed a novel technique for fabricating an engineered vascular-like network that satisfied these two requirements. The network comprised assembled hollow alginate hydrogel microfibers with mammalian cells enclosed in the gel portions. We controlled the distance between each flow microchannel (hollow core portions and interspace of the microfibers) to be within 150 µm by using microfibers with a gel thickness of approximately 50 µm. Furthermore, we confirmed that medium could be perfused into the flow channels quickly (within 10 min) after immobilization of the cells in the assembly. A human hepatoblastoma cell line (HepG2) proliferated in the gel portions of the microfibers and maintained their specific function during perfusion culture for 7 days. These results showed that the novel vascular-like networks fabricated here had the potential to allow the creation of volumetric tissues in vitro.

Angiopoietin-like protein 2 is a potential biomarker for gastric cancer.

Globally, gastric cancer is one of the most common types of cancer and is the second leading cause of cancer­induced mortality. Early detection of gastric cancer is able to contribute to a reduction of its mortality. For early detection, more specific and sensitive biomarkers than the classic biomarkers, including carcinoembryonic antigen, carbohydrate antigen 19­9 and C­reactive protein, are required. The present study focused on the evaluation of the potential of angiopoietin­like protein 2 (ANGPTL2) as a novel biomarker for gastric cancer. The expression levels of ANGPTL2 in undifferentiated and differentiated gastric cancer cell lines (HGC­27 and MKN7, respectively) were therefore investigated. Additionally, ANGPTL2 levels in the serum of gastric cancer patients were compared with those of healthy individuals to evaluate the possibility of the protein as a predictive biomarker for gastric cancer. It was established that the expression levels of ANGPTL2 mRNA and protein were higher in undifferentiated HGC­27 cells than those in differentiated MKN7 cells. In a patient study, it was indicated that the levels of ANGPTL2 in the serum of gastric cancer patients were higher than those in healthy controls. The diagnostic performance of ANGPTL2 was assessed by constructing a receiver operating characteristic (ROC) curve and was evaluated by calculating the area under each ROC curve (AUC). For the discrimination of patients with gastric cancer from healthy individuals, the AUC for ANGPTL2 was 0.774 (P=0.005) (95% confidence interval, 0.615­0.933). These results suggested that ANGPTL2 was a potential biomarker for gastric cancer.

Angiopoietin-like protein 2 as a potential biomarker for colorectal cancer.

Colorectal cancer (CRC) is the third most common malignancy worldwide. Disease progression leads to its spread to other organs, such as the liver, and is associated with higher mortality rates. Early CRC detection is therefore crucial for maximizing the chances of complete cure. The measurement of serum-based tumor biomarkers has shown great potential for the detection of CRC. In this study, we investigated the feasibility of using angiopoietin-like protein 2 (ANGPTL2) as a candidate biomarker for CRC. We first investigated ANGPTL2 expression in 7 CRC cell lines, among which Colo320, NCC-CoCK-115P, Caco-2 and Colo205 exhibited comparatively high ANGPTL2 expression. The serum levels of ANGPTL2 in CRC patients (3.45±1.30 ng/ml) were higher compared with those in healthy controls (2.74±0.64 ng/ml) (P<0.05). A receiver operating characteristic analysis demonstrated that the diagnostic performance of ANGPTL2 was marginally lower compared with that of the established biomarker C-reactive protein, but higher compared with that of carbohydrate antigen 19-9. These results suggested that the simultaneous measurement of ANGPTL2, along with previously established serum biomarkers, may increase the likelihood of early detection of CRC.