DNA hypomethylator phenotype reprograms glutamatergic network in receptor tyrosine kinase gene-mutated glioblastoma.

DNA methylation is crucial for chromatin structure and gene expression and its aberrancies, including the global "hypomethylator phenotype", are associated with cancer. Here we show that an underlying mechanism for this phenotype in the large proportion of the highly lethal brain tumor glioblastoma (GBM) carrying receptor tyrosine kinase gene mutations, involves the mechanistic target of rapamycin complex 2 (mTORC2), that is critical for growth factor signaling. In this scenario, mTORC2 suppresses the expression of the de novo DNA methyltransferase (DNMT3A) thereby inducing genome-wide DNA hypomethylation. Mechanistically, mTORC2 facilitates a redistribution of EZH2 histone methyltransferase into the promoter region of DNMT3A, and epigenetically represses the expression of DNA methyltransferase. Integrated analyses in both orthotopic mouse models and clinical GBM samples indicate that the DNA hypomethylator phenotype consistently reprograms a glutamate metabolism network, eventually driving GBM cell invasion and survival. These results nominate mTORC2 as a novel regulator of DNA hypomethylation in cancer and an exploitable target against cancer-promoting epigenetics.

Dendrimer-based micelles with highly potent targeting to sites of active bone turnover for the treatment of bone metastasis.

Bone-drug targeting therapies using nanoparticles based on targeting ligands remain challenging due to their uptake clearance at non-target sites such as the liver, kidney, and spleen. Furthermore, the distribution sites of nanoparticles in bones have not been fully investigated, thus halting the development of more effective bone metastasis treatment strategies. In this study, we developed nanoparticles self-assembled from cholesterol-terminated, polyethylene glycol-conjugated, aspartic acid (Asp)-modified polyamidoamine dendrimer (Asp-PAMAM-Micelles) with targeting to active bone turnover sites associated with bone metastasis pathogenesis. On analysis through whole-body single photon emission computed tomography/computed tomography (SPECT/CT) imaging, 111In-Asp-PAMAM-Micelles showed high specificity to active bone turnover sites (especially the joints in the lower limbs, shoulder, and pelvis) after intravenous injection in mice. The lower limb bone uptake clearance for 111In-Asp-PAMAM-Micelles encapsulating paclitaxel (PTX) was 3.5-fold higher than that for 111In-unmodified PAMAM-Micelles (PTX). 3H-PTX encapsulated Asp-PAMAM-Micelles effectively accumulated in the lower limb bones in a similar manner as the 111In-Asp-PAMAM-Micelles (PTX). In a bone metastatic tumor mouse model, the tumor growth in the lower limb bones was significantly inhibited by injection of Asp-PAMAM-Micelles (PTX) compared to unmodified PAMAM-Micelles (PTX). Our results demonstrate that Asp-PAMAM-Micelles are sophisticated drug delivery systems for highly potent targeting to active bone turnover sites.

Acute Effects of Whole-Body Vibration Training on Endothelial Function and Cardiovascular Response in Elderly Patients with Cardiovascular Disease.

The aim of this single-arm pilot study was to determine the effects of whole-body vibration training (WBVT) on endothelial function in elderly patients with cardiovascular diseases, as well as its safety. A total of 20 elderly patients with stable cardiovascular diseases underwent WBVT, which consisted of five static resistance training exercises (squats, wide stance squats, toe-stands, squats + band, and front lunges). The parameters of WBVT included vertical vibrations, 30 Hz frequency, and a 3-mm peak-to-peak amplitude. Each vibration session lasted 30 seconds, with 120 seconds of rest between sessions. Before and after WBVT, the reactive hyperemia peripheral arterial tonometry index (RH-PAT index) and transcutaneous oxygen pressure (tcPO2) were recorded as a measure of endothelial function and peripheral blood circulation. Systolic blood pressure, diastolic blood pressure, heart rate, and arterial oxygen saturation of pulse oximetry (SpO2) were measured at each rest interval as well as before and after WBVT. All patients completed our WBVT protocol without adverse events. The RH-PAT index significantly increased following WBVT (1.42 to 2.06, P < 0.001). There were no significant changes in heart rate (P = 0.777), systolic blood pressure (P = 0.183), diastolic blood pressure (P = 0.925), or SpO2 (P = 0.248) during WBVT. In conclusion, we demonstrated the acute effects of WBVT on endothelial function, with no reports of adverse events. These findings support the need for further randomized controlled studies to investigate the long-term effects of WBVT.

Characterization of novel bovine papillomavirus type 12 (BPV-12) causing epithelial papilloma.

Bovine papillomavirus type 12 (BPV-12, putative type BAA1) was detected in epithelial papilloma located on the tongue of an infected cow. Then, the whole genome was sequenced, and phylogenetic analysis illustrated that it should be classified as a member of the genus Xipapillomavirus. The viral genome is 7197 base pairs in length and contains five early ORFs (E1, E2, E4, E7 and E8), three late ORFs (L1, L2 and L3), and a long control region that possesses replication regulatory elements. Meanwhile, mRNA of each gene was detected in the papilloma sample. The papilloma was identified as epithelial papilloma by histological and immunohistochemical examination. Based on the genome information and pathological properties, BAA1 was designated as BPV-12 in this study.

Association between serum lipoprotein (a) level and progression of non-proliferative diabetic retinopathy in Type 2 diabetes.

PURPOSE: To investigate independent risk factors related to the progression of non-proliferative diabetic retinopathy (NPDR) for Japanese Type 2 diabetic patients. METHODS: One hundred and six patients with NPDR were followed up for 2 years. Diabetic retinopathy (DR) was determined by colour fundus photography. Multivariate logistic regression analysis was performed to assess variables independently associated with the progression of NPDR. Serum concentrations of novel risk factors for atherosclerotic vascular disease, including lipoprotein (a) [Lp(a)] and fibrinogen, were measured. RESULTS: Thirty-three patients (31%) had progressed by two scale steps or more in 2 years. The progression of NPDR was significantly associated with HbA(1c) [odds ratio (OR) 2.12; 95% confidence interval (CI) 1.14-4.87], systolic blood pressure (OR 1.72; 95% CI 1.14-2.91), Lp(a) (OR 2.70; 95% CI 1.09-5.12) and fibrinogen (OR 1.68; 95% CI 1.03-3.08). Multivariate logistic regression analysis showed that HbA(1c) (OR 1.74; 95% CI 1.12-3.21) and Lp(a) level (OR 1.90; 95% CI 1.06-4.33) were significant and independent predictors of the progression of NPDR. CONCLUSION: These data suggest that serum Lp(a) level is an independent risk factor for the progression of NPDR in Type 2 diabetes patients. We recommend that further prospective validation of our findings be undertaken to confirm these observations.

Glutamic acid decarboxylase autoantibody prevalence and association with HLA genotype in patients with younger-onset type 1 diabetes and proliferative diabetic retinopathy.

OBJECTIVE: To investigate the relationship of HLA status and autoantibodies to glutamic acid decarboxylase (GAD) in proliferative diabetic retinopathy (PDR) to assess the role of autoimmunity and genetic markers in retinopathy. DESIGN: Retrospective, nonrandomized, comparative study. PARTICIPANTS: Patients who had suffered from type 1 diabetes for >10 years and who had been first diagnosed as diabetic under 30 years of age were studied. They were classified into 3 groups: 20 patients with diabetes and PDR (PDR group), 22 patients who had diabetes and severe nonproliferative diabetic retinopathy (SNPDR group), and 25 patients who had diabetes with no diabetic retinopathy (non-DR group). METHODS: Blood was collected, and the relationship between HLA status and GAD autoantibody positivity in diabetic retinopathy was investigated in a cross-sectional study. MAIN OUTCOME MEASURES: Human leukocyte antigen status and GAD autoantibody positivity. RESULTS: The highest positive rate of GAD autoantibody was 56.0% in the non-DR group, followed by the SNPDR group (40.1%) and the PDR group (15.0%). The frequencies of the HLA-DQ4 and -DR4/-DQ4 haplotypes were significantly higher in the PDR group (75.0% and 65%, respectively) than in the SNPDR group (40.9% and 31.8%) or the non-DR group (40.0% and 28.0%) (P = 0.035 and P = 0.026, respectively). The prevalence of GAD antibodies was lower in patients with the HLA-DR4 and HLA-DQ4 alleles and -DR4/-DQ4 haplotype frequencies in the PDR group (P = 0.018, P = 0.0088, and P = 0.0031, respectively). CONCLUSIONS: We found that the existence of GAD antibodies is inversely related and HLA status is directly related to the stage or severity of retinopathy.

Quantitative measurement of retinal thickness in patients with diabetic macular edema is useful for evaluation of therapeutic agents.

The effect of lisinopril (an angiotensin-converting enzyme inhibitor) on diabetic macular edema (DME) was investigated by quantitative measurement of macular thickness. In a nonrandomized clinical trial, 19 normotensive type 2 diabetic patients with DME prospectively received oral lisinopril therapy for 2 months. Another 10 normotensive type 2 diabetic patients with similar DME were prospectively followed for two months without treatment. Central macular thickness was measured with a retinal thickness analyzer (RTA). In the lisinopril group, visual acuity improved by two lines or more in two out of 19 eyes (11%), was unchanged in 15 eyes (78%), and deteriorated by two lines or more in two eyes (11%). The mean central macular thickness was significantly reduced after 2 months of treatment (381.3 +/- 121.1 microm) compared with that before administration (475.2 +/- 171.0 microm, P = 0.0093). In the control group, central macular thickness was not significantly decreased after 2 months (458.5 +/- 113.7 microm, P = 0.2178) compared with the baseline value (464.7 +/- 152.2). Fluorescein angiography showed that macular leakage was decreased in 10 patients from the lisinopril group (53%) and was unchanged in nine patients (47%). There was a significant difference of central macular thickness between the patients with and without improvement of macular leakage (P = 0.0040). Lisinopril therapy may reduce macular thickness in patients with DME, as shown by this quantitative study. In addition, quantitative measurement of retinal thickness is useful when evaluating therapeutic agents for DME.

Diabetic retinopathy with repeated amaurosis fugax caused by orthostatic hypotension.

PURPOSE: To evaluate the blood flow velocity in the ophthalmic artery using color Doppler imaging in two patients who had diabetic retinopathy with amaurosis fugax caused by orthostatic hypotension. DESIGN: Interventional case reports. METHODS: A 40-year-old woman (Case 1) and a 65-year-old man (Case 2) had diabetic retinopathy and amaurosis fugax caused by orthostatic hypotension. We evaluated the blood flow velocity in the ophthalmic artery using color Doppler imaging in the two patients. SETTING: Department of Ophthalmology, Diabetes Center Tokyo Women's Medical University, Tokyo, Japan. RESULTS: For Case 1, the maximum systolic blood flow velocity in the ophthalmic artery (V max) was 10 cm/sec in the supine position. In a seated position, visual acuity was decreased from 20/20 to light perception as V max fell to 1 cm/sec. For Case 2, the V max was 14 cm/sec in the supine position, but it was too low to measure in a seated position. CONCLUSIONS: The decrease of V max by orthostatic hypotension may cause amaurosis fugax in patients with diabetic retinopathy.

Questionnaire survey on periodic ocular examination in Japanese diabetic patients.

PURPOSE: To identify the reasons why diabetic patients do not undergo periodic examination. DESIGN: Cohort study. METHODS: A questionnaire survey of 1,333 Japanese type 2 diabetic patients was conducted at Tokyo Women's Medical University. Performance of ocular examinations was investigated over 5 years and the reasons for not undergoing examination were analyzed. RESULTS: Periodic ocular examination was performed in 69.5% of patients, but not in 30.5%. The questionnaire survey showed that physicians usually explained the risk of ocular complications and recommended periodic examination. More than 98% of the patients were aware of diabetic eye disease. Multivariate logistic regression analysis showed the main reason for skipping examination as a reason was that patients believed they had no diabetic retinopathy. CONCLUSIONS: Patients should be informed about the necessity of receiving periodic ocular examination even if they do not yet have retinopathy.

Relationship between human leukocyte antigen status and proliferative diabetic retinopathy in patients with younger-onset type 1 diabetes mellitus.

PURPOSE: To investigate the relationship between human leukocyte antigen (HLA) status and proliferative diabetic retinopathy (PDR) to assess the genetic basis of PDR in younger type 1 diabetic patients. DESIGN: Retrospective, nonrandomized, comparative trial. METHOD: Patients who were diagnosed with type 1 diabetes under 30 years of age (range, 13-28) and had a disease duration of more than 10 years were studied. The patients were divided into two groups: 30 patients who had undergone vitreous surgery younger than 40 years old (the PDR group) and 50 patients without retinopathy (the non-DR group). The duration of diabetes and the level of glycemic control were matched between the two groups. The control group consisted of 50 healthy patients. HLA-A, -B, -C, -DR, and -DQ typing of blood samples was done using the standard microcytotoxicity method. RESULTS: The frequency of HLA-B62, Cw4, and DQ4 was significantly higher in the PDR group than in the non-DR group (P =.0020, P =.048, and P =.0026, respectively). The Cw4-DR4-DQ4 haplotype frequency was significantly higher in the PDR group than in the control group and the non-DR group (P =.0059). CONCLUSIONS: Our results suggests that HLA-B62, Cw4, and DQ4 may be useful for predicting the prognosis of retinopathy in patients with younger-onset type 1 diabetes.

[Attitude survey of diabetic patients visiting the department of internal medicine as outpatients].

PURPOSE: To investigate the awareness and understanding of ophthalmologic complications by diabetic patients and assess the problems. SUBJECTS AND METHODS: In August 1999, a questionnaire survey was done of 3,613 diabetic outpatients attending the Department of Internal Medicine at Tokyo Women's Medical University Diabetes Center. RESULTS: Patients who understood diabetic retinopathy as ophthalmological complications were 54.4% and those to whom physicians explained the complications were 33.8%. Patients who were recommended to receive an eye examination by physicians were 66.4% and those who continued to attend the eye clinic were 61.8%. Patients who had ophthalmological complications explained by physicians were 74.2%. CONCLUSION: Few patients understood diabetic retinopathy as a diabetic ophthalmological complication. To encourage the awareness and understanding concerning diabetic retinopathy, it is important to form a tight relationship between physicians and ophthalmologists and to improve the system for educating patients.

Relation of diabetic macular edema to cytokines and posterior vitreous detachment.

PURPOSE: To investigate whether angiotensin II (AII) or vascular endothelial growth factor (VEGF) is related to diabetic macular edema (DME) in patients with and without posterior vitreous detachment (PVD). DESIGN: A case-control study. METHODS: Vitreous fluid samples were obtained at vitreoretinal surgery from 28 eyes of 28 DME patients without PVD, 8 eyes of 8 DME patients with PVD, 14 eyes of 14 nondiabetic patients, and 8 eyes of diabetic patients without retinopathy. The VEGF levels in vitreous fluid and plasma were determined by enzyme-linked immunosorbent assay, while AII levels were measured by radioimmunoassay. RESULTS: The vitreous levels of AII and VEGF were significantly higher in DME patients with or without PVD than in nondiabetic patients or diabetic patients without retinopathy (without PVD: P < .0061, P < .0001, P = .0261, and P < .0001; with PVD: P < .0012, P < .0001, P = .0473, and P < .0001, respectively). There was no significant difference in the vitreous levels of AII or VEGF between patients with or without PVD (P = .4948 and P = .6642, respectively). The vitreous level of AII significantly correlated with that of VEGF in DME patients without PVD (P = .576) or with PVD (P = .488). AII and VEGF levels in vitreous fluid were significantly higher than the respective plasma levels. CONCLUSIONS: We found that the vitreous levels of AII and VEGF were elevated in DME patients irrespective of the status of PVD. Angiotensin II and VEGF may be induced in the eyes and be related to the pathogenesis of DME.

Prediction of macular edema exacerbation after phacoemulsification in patients with nonproliferative diabetic retinopathy.

PURPOSE: To ascertain whether the aqueous humor levels of vascular endothelial growth factor (VEGF), interleukin-6 (IL-6), and protein can predict the postoperative exacerbation of macular edema in patients with nonproliferative diabetic retinopathy (NPDR) after phacoemulsification surgery for cataract. SETTING: Department of Ophthalmology, Diabetes Center, Tokyo Women's Medical University, Tokyo, Japan. METHODS: This prospective study included 104 consecutive patients (104 eyes) with NPDR who had cataract surgery. The concentrations of VEGF and IL-6 in aqueous humor specimens obtained during cataract surgery were measured by enzyme-linked immunosorbent assay. Patients were followed for 6 months to assess the postoperative exacerbation of macular edema. RESULTS: Ninety patients (87%) achieved a visual acuity of 20/40 or better. Exacerbation of macular edema was seen in 30 eyes (29%) after 6 months. Hypertension and the aqueous levels of VEGF, IL-6, and protein were significantly correlated with the exacerbation of macular edema (odds ratio 1.16, 1.33, 1.27, and 1.28, respectively). In contrast, there was no correlation between the exacerbation of macular edema and other systemic factors. Multivariate logistic regression analysis showed that the aqueous VEGF level increase of 100 pg/mL increased the macular edema after phacoemulsification surgery (odds ratio 1.53). CONCLUSIONS: A high VEGF level in the aqueous humor predicted a significant risk for the postoperative exacerbation of macular edema. A model was developed to predict the risk exacerbation.

Plasma level of interleukin-6 is an indicator for predicting diabetic macular edema.

PURPOSE: To find a predictor in the pathogenesis of macular edema, we investigated ocular and systemic risk factors. METHODS: One hundred and fifty-nine patients with mild diabetic retinopathy who showed one or more soft exudates were recruited. We selected the possible predictors on the basis of relevant factors, including concentration of vascular endothelial growth factor, interleukin-6 (IL-6), transforming growth factor (TGF)-beta(1), tumor necrosis factor (TNF)-alpha, and lipoprotein(a) in plasma, and serum level of von Willebrand factor and thrombomodulin. RESULTS: Macular edema was not detected in 94 eyes; focal macular edema was detected in 46 eyes; diffuse macular edema was detected in 18 eyes; and cystoid macular edema was present in 1 eye. The plasma level of IL-6 concentration and the state of the posterior vitreous detachment (PVD) correlated significantly with the severity of macular edema (odds ratios = 3.68, 1.70, respectively). Other risk factors were not significantly associated with macular edema. We estimated the probability of macular edema according to the IL-6 level in plasma and the state of the PVD, and were able to predict the probability of macular edema. CONCLUSION: The results of the present study indicate that IL-6 concentration in plasma and the state of the PVD can be predictors of macular edema.