Twenty Years of Anti-Vascular Endothelial Growth Factor Therapeutics in Neovascular Age-Related Macular Degeneration Treatment.

Neovascular age-related macular degeneration (nAMD) is the primary disastrous retinal disease that leads to blindness in the elderly population. In the early 2000s, nAMD resulted in irreversible vision loss and blindness with no available treatment options. However, there have been breakthrough advances in the drug development of anti-angiogenic biological agents over the last two decades. The primary target molecule for treating nAMD is the vascular endothelial growth factor (VEGF), and there are currently several anti-VEGF drugs such as bevacizumab, ranibizumab, and aflibercept, which have made nAMD more manageable than before, thus preventing vision loss. Nevertheless, it should be noted that these anti-VEGF drugs for nAMD treatment are not effective in more than half of the patients, and even those who initially gain visual improvements lose their vision over time, along with potential deterioration in the geography of atrophy. As a result, there have been continuous endeavors to improve anti-VEGF agents through better efficacy, fewer doses, expanded intervals, and additional targets. This review describes past and current anti-VEGF therapeutics used to treat nAMD and outlines future directions to improve the effectiveness and safety of anti-VEGF agents.

Glucose-Thymidine Ratio as a Metabolism Index Using 18F-FDG and 18F-FLT PET Uptake as a Potential Imaging Biomarker for Evaluating Immune Checkpoint Inhibitor Therapy.

Immune checkpoint inhibitors (ICIs) are widely used in cancer immunotherapy, requiring effective methods for response monitoring. This study evaluated changes in 18F-2-fluoro-2-deoxy-D-glucose (FDG) and 18F-fluorothymidine (FLT) uptake by tumors following ICI treatment as potential imaging biomarkers in mice. Tumor uptakes of 18F-FDG and 18F-FLT were measured and compared between the ICI treatment and control groups. A combined imaging index of glucose-thymidine uptake ratio (GTR) was defined and compared between groups. In the ICI treatment group, tumor growth was effectively inhibited, and higher proportions of immune cells were observed. In the early phase, 18F-FDG uptake was higher in the treatment group, whereas 18F-FLT uptake was not different. There was no difference in 18F-FDG uptake between the two groups in the late phase. However, 18F-FLT uptake of the control group was markedly increased compared with the ICI treatment group. GTR was consistently higher in the ICI treatment group in the early and late phases. After ICI treatment, changes in tumor cell proliferation were observed with 18F-FLT, whereas 18F-FDG showed altered metabolism in both tumor and immune cells. A combination of 18F-FLT and 18F-FDG PET, such as GTR, is expected to serve as a potentially effective imaging biomarker for monitoring ICI treatment.

Oral bioavailability and pharmacokinetics of esculetin following intravenous and oral administration in rats.

Esculetin is the main active ingredient isolated from Artemisia montana (Nakai) Pamp. and Euphorbia lathyris L. The present study investigated the oral bioavailability and pharmacokinetics of esculetin in rats, following intravenous and oral administration.Twenty Sprague-Dawley rats were randomly assigned to receive 10 mg/kg of esculetin either by the intravenous or oral route. Plasma concentrations of esculetin were measured using liquid chromatography-tandem mass spectrometry. Pharmacokinetic parameters were estimated using non-compartmental analysis as well as a compartmental modelling approach using WinNonlinTM and ADAPT 5 software, respectively.According to non-compartmental analysis, the mean oral bioavailability of esculetin was 19%. Mean ± standard deviation values of esculetin half-life, steady-state volume of distribution and clearance, following intravenous dosing, were 2.08 ± 0.46 h, 1.81 ± 0.52 L/kg and 1.27 ± 0.26 L/h/kg, respectively. As indicated by compartmental modelling, a two-compartment pharmacokinetic model with first-order absorption and elimination rate constants of 0.98 ± 0.18 h-1 and 2.47 ± 0.28 h-1, respectively, sufficiently described the plasma concentration-time curve of esculetin.Improving our understanding of the pharmacokinetic properties of esculetin could help with future development of herbal medicine products with appropriate bioactivity.

Identification of Lymphatic and Hematogenous Routes of Rapidly Labeled Radioactive and Fluorescent Exosomes through Highly Sensitive Multimodal Imaging.

There has been considerable interest in the clinical use of exosomes as delivery vehicles for treatments as well as for promising diagnostic biomarkers, but the physiological distribution of exosomes must be further elucidated to validate their efficacy and safety. Here, we aimed to develop novel methods to monitor exosome biodistribution in vivo using positron emission tomography (PET) and optical imaging. Exosomes were isolated from cultured mouse breast cancer cells and labeled for PET and optical imaging. In mice, radiolabeled and fluorescently labeled exosomes were injected both via lymphatic and hematogenous metastatic routes. PET and fluorescence images were obtained and quantified. Radioactivity and fluorescence intensity of ex vivo organs were measured. PET signals from exosomes in the lymphatic metastatic route were observed in the draining sentinel lymph nodes. Immunohistochemistry revealed greater exosome uptake in brachial and axillary versus inguinal lymph nodes. Following administration through the hematogenous metastasis pathway, accumulation of exosomes was clearly observed in the lungs, liver, and spleen. Exosomes from tumor cells were successfully labeled with 64Cu (or 68Ga) and fluorescence and were visualized via PET and optical imaging, suggesting that this simultaneous and rapid labeling method could provide valuable information for further exosome translational research and clinical applications.

Evaluation of high nutrient diets on litter performance of heat-stressed lactating sows.

OBJECTIVE: The present study investigated the litter performance of multiparous sows fed 3% and 6% densified diets at farrowing to weaning during summer with mean maximum room temperature of 30.5°C. METHODS: A total of 60 crossbred multiparous sows were allotted to one of three treatments based on body weight according to a completely randomized design. Three different nutrient levels based on NRC were applied as standard diet (ST; metabolizable energy, 3,300 kcal/kg), high nutrient level 1 (HE1; ST+3% higher energy and 16.59% protein) and high nutrient level 2 (HE2; ST+6% higher energy and 17.04% protein). RESULTS: There was no variation in the body weight change. However, backfat thickness change tended to reduce in HE1 in comparison to ST treatment. Dietary treatments had no effects on feed intake, daily energy intake and weaning-to-estrus interval in lactating sows. Litter size, litter weight at weaning and average daily gain of piglets were significantly greater in sows in HE1 compared with ST, however, no difference was observed between HE2 and ST. Increasing the nutrient levels had no effects on the blood urea nitrogen, glucose, triglyceride, and creatinine at post-farrowing and weaning time. The concentration of follicle stimulating hormone, cortisol and insulin were not affected by dietary treatments either in post-farrowing or weaning time. The concentration of blood luteinizing hormone of sows in ST treatment was numerically less than sows in HE2 treatment at weaning. Milk and colostrum compositions such as protein, fat and lactose were not affected by the treatments. CONCLUSION: An energy level of 3,400 kcal/kg (14.23 MJ/kg) with 166 g/kg crude protein is suggested as the optimal level of dietary nutrients for heat stressed lactating sows with significant beneficial effects on litter size.

A new fluorescence/PET probe for targeting intracellular human telomerase reverse transcriptase (hTERT) using Tat peptide-conjugated IgM.

Despite an increasing need for methods to visualize intracellular proteins in vivo, the majority of antibody-based imaging methods available can only detect membrane proteins. The human telomerase reverse transcriptase (hTERT) is an intracellular target of great interest because of its high expression in several types of cancer. In this study, we developed a new probe for hTERT using the Tat peptide. An hTERT antibody (IgG or IgM) was conjugated with the Tat peptide, a fluorescence dye and (64)Cu. HT29 (hTERT+) and U2OS (hTERT-) were used to visualize the intracellular hTERT. The hTERT was detected by RT-PCR and western blot. Fluorescence signals for hTERT were obtained by confocal microscopy, live cell imaging, and analyzed by Tissue-FAXS. In nude mice, tumors were visualized using the fluorescence imaging devices Maestro™ and PETBOX. In RT-PCR and western blot, the expression of hTERT was detected in HT29 cells, but not in U2OS cells. Fluorescence signals were clearly observed in HT29 cells and in U2OS cells after 1 h of treatment, but signals were only detected in HT29 cells after 24 h. Confocal microscopy showed that 9.65% of U2OS and 78.54% of HT29 cells had positive hTERT signals. 3D animation images showed that the probe could target intranuclear hTERT in the nucleus. In mice models, fluorescence and PET imaging showed that hTERT in HT29 tumors could be efficiently visualized. In summary, we developed a new method to visualize intracellular and intranuclear proteins both in vitro and in vivo.

Superficial Echogenic Lesions Detected on Neonatal Cranial Sonography: Possible Indicators of Severe Birth Injury.

OBJECTIVES: The purpose of this study was to evaluate the characteristics and importance of superficial echogenic lesions around cranial sutures on neonatal cranial sonography. METHODS: We retrospectively reviewed the clinical records and neuroimaging studies of 40 neonates who had superficial echogenic lesions around sutures on neonatal cranial sonography. Magnetic resonance imaging (n = 18) and computed tomography (n = 2) were performed within 2 weeks after sonography. We correlated sonographic findings with computed tomographic and magnetic resonance imaging findings and analyzed them. We also evaluated the associated lesions, neurologic signs, and follow-up changes. RESULTS: Sonographically, the superficial echogenic lesions involved both sulci and perisulcal parenchyma in 39 neonates and were located in the frontal and parietal areas around the sagittal suture in 38 neonates. Magnetic resonance imaging revealed a pattern of hypoxic ischemic encephalopathy in 9 neonates, birth trauma in 3 neonates, a mixed pattern of hypoxic ischemic encephalopathy and trauma in 3 neonates, nonspecific single infarctions in 2 neonates, and lack of a defined lesion in 1 neonate. The associated lesions were subdural hemorrhage (n = 12), epidural hematoma (n = 4), germinal matrix hemorrhage (n = 3), intraventricular hemorrhage (n = 2), and periventricular leukomalacia (n = 1). All epidural hematomas were associated with scalp hematoma, and 2 patients had skull fractures. One neonate with epidural hematoma associated with a hypoxic ischemic encephalopathy pattern showed mild spasticity in both ankles until 16 months. CONCLUSIONS: Superficial echogenic lesions detected around cranial sutures on neonatal sonography may be an indicator of more serious intracranial lesions such as more extensive hypoxic ischemic encephalopathy and intracranial hematomas, including epidural hematoma.

Oligonol suppresses lipid accumulation and improves insulin resistance in a palmitate-induced in HepG2 hepatocytes as a cellular steatosis model.

BACKGROUND: Oligonol is a low molecular weight form of polyphenol polymers derived from lychee fruits. Several studies suggest that Oligonol has an anti-obesity effect. Since obesity is tightly associated with insulin resistance, we investigated a possible remission effect of Oligonol on lipid accumulation and insulin resistance in human hepatic HepG2 cells. METHODS: HepG2 cells were treated with palmitate for 24 h to induce cellular hepatic steatosis and insulin resistance. The cells were then treated with Oligonol at subtoxic concentrations and examined for lipid metabolism, cytokine production, and insulin signaling using quantitative RT-PCR and western blot analysis. RESULTS: Oligonol treatment reversed the palmitate-induced intracellular lipid accumulation, down regulated the expression of lipogenic genes, and up-regulated genes for fatty acid degradation. Oligonol restored insulin sensitivity, as was determined by the phosphorylation states of IRS-1. Oligonol also inhibited STAT3-SOCS3 signaling and increased AMPK phosphorylation in HepG2 cells. CONCLUSION: Oligonol treatment improved palmitate-induced cellular steatosis and insulin resistance in HepG2 cells with concomitant reduction of inflammatory cytokines and decrease in STAT3-SOCS3 and AMPK-mTOR pathways. Oligonol may have beneficial effects in lipid metabolism and insulin resistance in the liver.

Relationship between apoptosis imaging and radioiodine therapy in tumor cells with different sodium iodide symporter gene expression.

The therapeutic efficacy of radioiodine (¹³¹I) therapy has been reported to be variable among cancer patients and even between metastatic regions in the same patients. Because the expression level of sodium iodide symporter (NIS) cannot reflect the efficacy of therapy, other strategies are required to predict the precise therapeutic effect of ¹³¹I therapy. In this research, we investigated the correlation between iodine (I) uptake, apoptosis imaging, and therapeutic efficacy. Two HT29 cell lines, cytomegalovirus (CMV)-NIS (or NIS+++) and TERT-NIS (or NIS+), were established by retroviral transfection. I uptake was estimated by I-uptake assay and gamma camera imaging. Apoptosis was evaluated by confocal microscopy and a Maestro fluorescence imaging system (CRi Inc., Woburn, MA) using ApoFlamma (BioACTs, Seoul, Korea), a fluorescent dye-conjugated apoptosis-targeting peptide 1 (ApoPep-1). Therapeutic efficacy was determined by tumor size. The CMV-NIS showed higher I uptake and ApoFlamma signals than TERT-NIS. In xenograft models, CMV-NIS also showed high 99m technetium signals and ApoFlamma signals. Tumor reduction had a stronger correlation with apoptosis imaging signals than with gamma camera imaging signals, which reflect I uptake. Higher NIS-expressing tumors showed increased apoptosis and I uptake, resulting in a significant tumor reduction. Moreover, tumor reduction showed a strong correlation with ApoFlamma imaging compared to I-uptake imaging.

Optimal extraction conditions and quantification of lignan phytoestrogens in cereal grains using targeted LC-MS/MS.

Lignans are phytoestrogens found in various forms such as glycosides, ester-linked oligomers, and aglycones in a variety of foods, including soy products, legumes, grains, nuts, vegetables, and fruits. This study aimed to optimize the extraction of lignans from cereal grains using response surface methodology (RSM). Lignans, including secoisolariciresinol (Seco), matairesinol (Mat), pinoresinol (Pin), lariciresinol (Lar), and syringaresinol (Syr), were quantified using high-performance liquid chromatography-tandem mass spectrometry. A Box-Behnken design was employed to determine the optimal values for three extraction parameters: temperature (X1: 20°C-60°C), methanol concentration (X2: 60%-100%), and extraction time (X3: 30-90 min). The highest lignan contents were obtained at X1 = 44.24°C, X2 = 84.64%, and X3 = 53.63 min. To apply these experimental conditions to the actual experiment, the optimal conditions were slightly adjusted to X1 = 40°C, X2 = 80%, and X3 = 60 min. The predicted results closely matched the experimental results obtained using the modified optimal extraction conditions. The highest lignan content found in barley sprouts (85.930 µg/100 g), however, most grains exhibited relatively low concentrations of lignans. These findings provide valuable insights into the lignan content of grains and contribute to the generation of reliable data in this field.

Comparison of the Antioxidant Potency of Four Triterpenes of Centella asiatica against Oxidative Stress.

We comparatively evaluated the antioxidant properties of key triterpenes from Centella asiatica, including asiatic acid (AA), asiaticoside, madecassic acid, and madecassoside, in several cell types, including skin fibroblasts, macrophages, hepatocytes, and endothelial cells, under conditions promoting oxidative stress. AA conferred the highest viability on Hs68 cells exposed to ultraviolet B (UVB) irradiation. Triterpene pretreatment attenuated the UVB-induced generation of reactive oxygen species (ROS) and malondialdehyde (MDA), as well as the UVB-induced depletion of glutathione (GSH) in skin fibroblasts. AA most potently inhibited UVB-induced MMP generation, resulting in increased intracellular collagen levels. Pretreatment with triterpenes, particularly AA, significantly improved cell viability and attenuated TBHP-induced levels of ROS, alanine aminotransferase, and aspartate aminotransferase in HepG2 cells. Triterpenes attenuated ROS levels and reduced MDA and GSH expression in EA.hy926 cells. In RAW264.7 macrophages, production of nitric oxide, tumor necrosis factor-α, and interleukin-6 (indicators of LPS-induced oxidative damage) was significantly reduced by treatment with any of the triterpenes. Statistical analyses of triterpene biological activities using principal component analysis and hierarchical clustering revealed that AA exerted the greatest overall influence and showed remarkable activity in Hs68 and HepG2 cells.

Profiles of volatile sulfur compounds in various vegetables consumed in Korea using HS-SPME-GC/MS technique.

Volatile sulfur compounds (VSCs) are not only important for their therapeutic potential but also significantly influence the flavor profiles of agricultural products. VSCs exhibit various chemical structures due to their stability and volatility, and they may form or be altered as a result of enzymatic and chemical reactions during storage and cooking. This study has focused on profiles of VSCs in 58 different vegetable samples by using HS-SPME-GC/MS technique and chemometric analyses. The validation was carried out using cabbage juice as a vegetable matrix for VSCs analysis, showing satisfactory repeatability (RSD 8.07% ~ 9.45%), reproducibility (RSD 4.22% ~ 7.71%), accuracy and specificity. The established method was utilized on various vegetables, revealing that 21 VSCs such as sulfides, disulfides, trisulfides, isothiocyanates, sulfhydryls, and thiophenes were successfully identified and quantified. These compounds were found in a range of vegetables including Allium species, Cruciferae, Capsicum species, green leafy vegetables, and mushrooms. In particular, isocyanate and allyl groups were abundant in Cruciferae and Allium vegetables, respectively. Cooking conditions were shown to reduce the levels of certain sulfur compounds such as dimethyl sulfide and dimethyl trisulfide in vegetables like broccoli and cabbage, suggesting that heat treatment can lead to the volatilization and reduction of these compounds. The present study provides reliable insights into the compositions of VSCs in various vegetables and examines the changes induced by different cooking methods.

Protective Effects of Cereal Grain Extracts on Alcohol-Induced Hepatocyte Damage.

This study investigated the protective effects of cereal grains on alcohol-induced hepatocyte damage. Cereal grains were extracted with methanol, and their radical scavenging properties and total phenolic contents were examined. Black rice extract exhibited the highest total polyphenol content and radical scavenging capacity. Treatment with sorghum extract increased the viability of cells exposed to alcohol by up to 81.6%. All cereal grain extracts decreased reactive oxygen species and malondialdehyde production and glutathione depletion in HepG2 cells exposed to ethanol. In particular, black rice and sorghum extracts exhibited greater antioxidant effects than other cereal grains. Treatment with black rice extract increased the levels of alanine aminotransferase and aspartate aminotransferase of alcohol-exposed cells to control levels. Overall, black rice extract showed a greater protective effect compared with other cereal grains against alcohol exposure in HepG2 cells and could improve alcohol-induced liver problems.

Hydrolyzed cow colostrum extract (BCFM) inhibits alpha-MSH-induced melanogenesis in B16F1 cells via regulation of the MC1R-cAMP signaling pathway.

Cow colostrum is the first milk produced after birth and is a rich natural source of nutrients, immunoglobulins, peptides, and growth factors. The bioconversion of milk and whey changes the immobilization and biochemical characterization. However, the cellular mechanism and the anti-melanin synthesis effects of hydrolyzed cow colostrum extract (BCFM) in alpha-MSH-induced B16F1 cells have not been examined. In this study, we investigated the anti-melanogenesis mechanism by examining the effects of BCFM in alpha-MSH-induced B16F1 cells. Cells were treated with BCFM in the presence or absence of alpha-MSH and co-cultured for 24, 48, and 72 h. The treatment of B16F1 cells with alpha-MSH resulted in the darkening of the color of the cells and induction of melanin synthesis. In addition, the expression levels of MC1R and cAMP, as well as phosphorylation levels of CREB and PKA, were increased by alpha-MSH treatment. However, concomitant treatment with BCFM resulted in a significant decrease in these factors and phosphorylated MITF. At the same time, the expressive amount of TRP-1 and tyrosinase was also decreased in B16F1 cells. These results demonstrate the potential of BCFM for the prevention of melanogenesis progression via the regulation of the MC1R-cAMP signaling pathway in alpha-MSH-induced B16F1 cells. The administration of BCFM suppressed the expression of TRP-1 and/or tyrosinase by regulating the CREB/MITF signaling pathways in the B16F1 cells. We propose that hydrolyzed cow colostrum extract (BCFM) is suitable for use as a novel active agent for skin whitening or pharmaceutical applications.

Hypoglycemic and hypolipidemic effects of unsaponifiable matter from okra seed in diabetic rats.

BACKGROUND/OBJECTIVES: Okra seed is a rich source of various nutritional and bioactive constituents, but its mechanism of action is still unclear. The aim of this study was to evaluated the effects on glucose uptake and serum lipid profiles of unsaponifiable matter (USM) from okra seed in adipocytes and diabetic animal models. MATERIALS/METHODS: USM was prepared from okra seed powder by saponification. The contents of phytosterols and vitamin E in USM were measured. 3T3-L1 preadipocytes were cultured for 6 days with different concentrations of USM (0-200 µg/mL). The diabetic rats were administered with or without USM for 5 wk. RESULTS: In the USM, the contents of phytosterols and vitamin E were 394.13 mg/g USM and 31.16 mg/g USM, respectively. USM showed no cytotoxicity and led to an approximately 1.4-fold increase in glucose uptake in 3T3-L1 adipocytes. The treatment of USM also increased the expressions of peroxisome proliferator-activated receptor-γ and glucose transporter-4 in a dose-dependent manner in adipocytes. The body weight change was not significantly different in all diabetic rats. However, blood glucose and the weights of liver and adipose tissues were significantly reduced compared to those in the control diabetic rats. Treatment with USM decreased the levels of triglycerides, total cholesterol, and low-density lipoprotein cholesterol compared to the control group. The USM group also showed significantly decreased atherogenic indices and cardiac risk factors. CONCLUSION: These results suggest that USM from okra seed improves the hypoglycemic and hypolipidemic effects in diabetic rats, and provides valuable information for improving the functional properties of okra seed.

Technological Tools and Artificial Intelligence in Estrus Detection of Sows-A Comprehensive Review.

In animal farming, timely estrus detection and prediction of the best moment for insemination is crucial. Traditional sow estrus detection depends on the expertise of a farm attendant which can be inconsistent, time-consuming, and labor-intensive. Attempts and trials in developing and implementing technological tools to detect estrus have been explored by researchers. The objective of this review is to assess the automatic methods of estrus recognition in operation for sows and point out their strong and weak points to assist in developing new and improved detection systems. Real-time methods using body and vulvar temperature, posture recognition, and activity measurements show higher precision. Incorporating artificial intelligence with multiple estrus-related parameters is expected to enhance accuracy. Further development of new systems relies mostly upon the improved algorithm and accurate data provided. Future systems should be designed to minimize the misclassification rate, so better detection is achieved.

MyD88-5 links mitochondria, microtubules, and JNK3 in neurons and regulates neuronal survival.

The innate immune system relies on evolutionally conserved Toll-like receptors (TLRs) to recognize diverse microbial molecular structures. Most TLRs depend on a family of adaptor proteins termed MyD88s to transduce their signals. Critical roles of MyD88-1-4 in host defense were demonstrated by defective immune responses in knockout mice. In contrast, the sites of expression and functions of vertebrate MyD88-5 have remained elusive. We show that MyD88-5 is distinct from other MyD88s in that MyD88-5 is preferentially expressed in neurons, colocalizes in part with mitochondria and JNK3, and regulates neuronal death. We prepared MyD88-5/GFP transgenic mice via a bacterial artificial chromosome to preserve its endogenous expression pattern. MyD88-5/GFP was detected chiefly in the brain, where it associated with punctate structures within neurons and copurified in part with mitochondria. In vitro, MyD88-5 co-immunoprecipitated with JNK3 and recruited JNK3 from cytosol to mitochondria. Hippocampal neurons from MyD88-5-deficient mice were protected from death after deprivation of oxygen and glucose. In contrast, MyD88-5-null macrophages behaved like wild-type cells in their response to microbial products. Thus, MyD88-5 appears unique among MyD88s in functioning to mediate stress-induced neuronal toxicity.

Quantitative positron emission tomography imaging of angiogenesis in rats with forelimb ischemia using (68)Ga-NOTA-c(RGDyK).

Gallium-68-labeled 1,4,7-triazacyclononane-1,4,7-triacetic acid (NOTA)-cyclic Arg-Gly-Asp-D-Tyr-Lys (c(RGDyK)) was developed for αvß3 targeting, and is a promising agent for imaging of cancer and disorders related to angiogenesis. In this study, we performed kinetic analysis of (68)Ga-NOTA-c(RGDyK) in rats with surgically induced forelimb ischemia, and immunohistochemical analysis was also performed to assess αvß3 immuno-staining level. Animal models were created by excision of the left brachial vessels, and a sham operation was performed on the right brachial region under 2 % isoflurane anesthesia. Using an animal positron emission tomography/computed tomography (PET/CT) scanner, a list mode PET scan (120 min) was started with the injection of (68)Ga-NOTA-c(RGDyK) via the tail vein at 3, 5 and 7 days after ischemic surgery. Volumes of interest were drawn on the left ventricle, sham operation, control, and ischemic regions. Compartmental and two graphical analyses (Logan and RE plots) were performed for kinetic parameter estimation. The immunohistochemical analysis was also performed after the last PET scan, and cell components were scored on a six point scale for quantification of immuno-staining level (0-negative to 5-very high). A 3-compartment model with reversible binding best described the tissue time-activity curves. The distribution volume of the ischemic region was significantly higher than that of the sham operation (P < 10(-6)) and control region (P < 10(-9)). Both the Logan and RE plots showed high correlation with compartmental analysis (R(2) = 0.96 and 0.95 for Logan and RE, respectively). The temporal changes in distribution volume and binding potential were not significant. The immuno-staining level of the ischemic region was significantly higher than that of sham operation (P < 10(-4)) and control region (P < 10(-8)). Kinetic modeling studies with dynamic (68)Ga-NOTA-c(RGDyK) PET scan are feasible based on an image-derived input function in a rat ischemia model. The kinetic modeling analysis performed in this study will be useful for the quantitative evaluation of (68)Ga-NOTA-c(RGDyK) binding to αvß3 in angiogenic tissues.

Betaine protects against rotenone-induced neurotoxicity in PC12 cells.

Rotenone is an inhibitor of mitochondrial complex I-induced neurotoxicity in PC12 cells and has been widely studied to elucidate the pathogenesis of Parkinson's disease. We investigated the neuroprotective effects of betaine on rotenone-induced neurotoxicity in PC12 cells. Betaine inhibited rotenone-induced apoptosis in a dose-dependent manner, with cell viability increasing from 50 % with rotenone treatment alone to 71 % with rotenone plus 100-µM betaine treatment. Flow cytometric analysis demonstrated cell death in the rotenone-treated cells to be over 50 %; the number of live cells increased with betaine pretreatment. Betaine pretreatment of PC12 cells attenuated rotenone-mediated mitochondrial dysfunction, including nuclear fragmentation, ATP depletion, mitochondrial membrane depolarization, caspase-3/7 activation, and reactive oxygen species production. Western blots demonstrated activation of caspase-3 and caspase-9, and their increased expression levels in rotenone-treated cells; betaine decreased caspase-3 and caspase-9 expression levels and suppressed their activation. Together, these results suggest that betaine may serve as a neuroprotective agent in the treatment of neurodegenerative diseases.

Downregulated lipid metabolism in differentiated murine adipocytes by procyanidins from defatted grape seed meal.

Recent reports have proposed possible anti-obesity mechanisms for antioxidants involving increased energy expenditure, pre-adipocyte differentiation and proliferation, decreased lipogenesis and increased lipolysis, and fat oxidation. The aim of this study was to examine and to confirm the anti-obesity effect of the oligomeric and polymeric procyanidin fractions from defatted grape seeds. The lipid metabolism-related mRNA level in the mouse preadipocytes, 3T3-L1 cells, was determined to evaluate the anti-obesity effect of the phenolic fractions from a grape seed meal. Lipid accumulation was reduced by 19% of the control level by the procyanidin fraction originating from the grape seed meal. Emerging from the effect of the treatment on HSL and LPL mRNA expression, lipolytic enzyme activity was not involved in the anti-obesity effects of CPE and FPP from the defatted grape seed meal. We tested and confirmed in this study the effect of the biological activities of oligomeric and polymeric procyanidins from the defatted grape seed meal. It is suggested from the results of this brief study that further studies would be desirable to focus on the anti-obesity effect of the purified extracts of a defatted grape seed meal.