In vitro evaluation of PET radiotracers for imaging synaptic density, the acetylcholine transporter, AMPA-tarp-γ8 and muscarinic M4 receptors in Alzheimer's disease.

Several therapeutics and biomarkers that target Alzheimer's disease (AD) are under development. Our clinical positron emission tomography (PET) research programs are interested in six radiopharmaceuticals to image patients with AD and related dementias, specifically [11C]UCB-J and [18F]SynVesT-1 for synaptic vesicle glycoprotein 2A as a marker of synaptic density, two vesicular acetylcholine transporter PET radiotracers: [18F]FEOBV and [18F]VAT, as well as the transmembrane AMPA receptor regulatory protein (TARP)-γ8 tracer, [18F]JNJ-64511070, and the muscarinic acetylcholine receptor (mAChR) M4 tracer [11C]MK-6884. The goal of this study was to compare all six radiotracers (labeled with tritium or 18F) by measuring their density variability in pathologically diagnosed cases of AD, mild cognitive impairment (MCI) and normal healthy volunteer (NHV) human brains, using thin-section in vitro autoradiography (ARG). Region of interest analysis was used to quantify radioligand binding density and determine whether the radioligands provide a signal-to-noise ratio optimal for showing changes in binding. Our preliminary study confirmed that all six radiotracers show specific binding in MCI and AD. An expected decrease in their respective target density in human AD hippocampus tissues compared to NHV was observed with [3H]UCB-J, [3H]SynVesT-1, [3H]JNJ-64511070, and [3H]MK-6884. This preliminary study will be used to guide human PET imaging of SV2A, TARP-γ8 and the mAChR M4 subtype for imaging in AD and related dementias.

Performance of modified first-stage French Vertical Flow Constructed Wetlands under extreme operational conditions.

Operation conditions considerably affect the removal efficiency of wastewater treatment systems, and yet we still lack data on how these systems function under extreme dilution rates and climatic conditions at high altitudes. Here, we applied two modified First-Stage French Vertical Flow Constructed Wetlands (FS-FVFCWs) for sewage treatment in Northern Tropical Andes. Specifically, within 18 months, we conducted a pilot-scale experiment at two hydraulic loading rates (HLRs) of 0.94 and 0.56 m d-1, representing 2.5 and 1.5 times the recommended design values, with two different feeding/resting periods to investigate the impact of HLRs and operational strategy on system performance. We found that chemical oxygen demand (COD) and total suspended solids (TSS) removal was satisfactory, with average values of 53 ± 18 and 69 ± 16%, respectively. Moreover, reducing HLRs resulted in higher removal efficiency for COD, from 46 ± 15 to 64 ± 15%, but had no impact on TSS removal, with 3 days of feeding and 6 days of resting. For an equal time of feeding and resting, COD and TSS removals were not affected by the modified HLR. These findings suggest that high HLRs can be applied to FS-FVFCW without compromising the system operation and obtaining satisfactory results, leading to opportunities to reduce areas and costs.

PET Imaging of Fructose Metabolism in a Rodent Model of Neuroinflammation with 6-[18F]fluoro-6-deoxy-D-fructose.

Fluorine-18 labeled 6-fluoro-6-deoxy-D-fructose (6-[18F]FDF) targets the fructose-preferred facilitative hexose transporter GLUT5, which is expressed predominantly in brain microglia and activated in response to inflammatory stimuli. We hypothesize that 6-[18F]FDF will specifically image microglia following neuroinflammatory insult. 6-[18F]FDF and, for comparison, [18F]FDG were evaluated in unilateral intra-striatal lipopolysaccharide (LPS)-injected male and female rats (50 µg/animal) by longitudinal dynamic PET imaging in vivo. In LPS-injected rats, increased accumulation of 6-[18F]FDF was observed at 48 h post-LPS injection, with plateaued uptake (60-120 min) that was significantly higher in the ipsilateral vs. contralateral striatum (0.985 ± 0.047 and 0.819 ± 0.033 SUV, respectively; p = 0.002, n = 4M/3F). The ipsilateral-contralateral difference in striatal 6-[18F]FDF uptake expressed as binding potential (BPSRTM) peaked at 48 h (0.19 ± 0.11) and was significantly decreased at one and two weeks. In contrast, increased [18F]FDG uptake in the ipsilateral striatum was highest at one week post-LPS injection (BPSRTM = 0.25 ± 0.06, n = 4M). Iba-1 and GFAP immunohistochemistry confirmed LPS-induced activation of microglia and astrocytes, respectively, in ipsilateral striatum. This proof-of-concept study revealed an early response of 6-[18F]FDF to neuroinflammatory stimuli in rat brain. 6-[18F]FDF represents a potential PET radiotracer for imaging microglial GLUT5 density in brain with applications in neuroinflammatory and neurodegenerative diseases.

PET imaging of glycogen synthase kinase-3 in pancreatic cancer xenograft mouse models.

Glycogen synthase kinase-3 (GSK-3) contributes to tumorigenesis in pancreatic cancer by modulating cell proliferation and survival. This study evaluated the lead GSK-3 targeted PET radiotracers for neuro-PET imaging, [11C]PF-367 and [11C]OCM-44, in pancreatic cancer xenograft mice. Immunohistochemistry showed that GSK-3α and GSK-3ß were overexpressed in PANC-1 xenografts. In autoradiography studies, higher specific binding was observed for [3H]PF-367 compared to [3H]OCM-44 when co-incubated with unlabeled PF-367 (59.2±1.8% vs 22.6±3.75%, respectively). Co-incubation of [11C]OCM-44 with OCM-44 did not improve the specific binding (25.5±30.2%). In dynamic PET imaging of PANC-1 xenograft mouse models, tumors were not visualized with [11C]PF-367 but were well visualized with [11C]OCM-44. Time-activity curves revealed no difference in accumulation in PANC-1 tumor tissue compared to muscle tissue in [11C]PF-367 baseline studies, while a significant difference was observed for [11C]OCM-44 with a tumor-to-muscle ratio of 1.6. Tumor radioactivity accumulation following injection with [11C]OCM-44 was not displaced by pre-treatment with unlabeled PF-367. Radiometabolite analysis showed that intact [11C]PF-367 accounted for 7.5% of tumor radioactivity, with >30% in plasma, at 40 min post-injection of the radiotracer, and that intact [11C]OCM-44 accounted for 20% of tumor radioactivity, with >60% in plasma. [11C]OCM-44 is superior to [11C]PF-367 for detecting lesions in preclinical mouse models of pancreatic cancer, however, both radiotracers undergo rapid metabolism in vivo. GSK-3 PET radiotracers with improved in vivo stability are needed for clinical translation. To our knowledge this work represents the first PET imaging study of GSK-3 in oncology.

Repurposing [11C]MC1 for PET Imaging of Cyclooxygenase-2 in Colorectal Cancer Xenograft Mouse Models.

PURPOSE: Cyclooxygenase-2 (COX-2) is a target for inflammation and colorectal cancer (CRC). This study evaluated the COX-2 neuro-PET radiopharmaceutical, [11C]MC1, in CRC xenograft mice. PROCEDURES: [11C]MC1 was evaluated in ICRscid mice with HT-29 and HCT-116 CRC xenografts, with high and low COX-2 expression, respectively, by immunohistochemistry, cellular uptake, dynamic PET/MR imaging, ex vivo biodistribution, and radiometabolite analysis. RESULTS: HT-29 xenografts were well visualized with [11C]MC1 using PET/MR. Time-activity curves revealed steady tumor radioactivity accumulation in HT-29 xenografts that plateaued from 40 to 60 min (3.07 ± 0.65 %ID/g) and was significantly reduced by pre-treatment with MC1 or celecoxib (1.62 ± 0.29 and 1.18 ± 0.21 %ID/g, respectively, p = 0.045 and p = 0.005). Radiometabolite analysis showed that [11C]MC1 accounted for >90 % of tumor radioactivity, with <10 % in plasma, at 40 min post-injection of the radiotracer. CONCLUSIONS: [11C]MC1 is a promising PET imaging agent for COX-2 in CRC and translation for cancer research should be considered.

Visual Signaling in the Semi-Fossorial Lizard Pholidobolus montium (Gymnophthalmidae).

It has been suggested that gymnophthalmids, like most semi-fossorial lacertoids, rely more in chemical cues to communicate, in comparison to other groups, like Iguanids, on which communication is mostly based on visual signaling. We present the first description of visual signaling in the Andean lizard Pholidobolusmontium (Gymnophthalmidae) and a complete ethogram based on ex situ observations (34 different types of behaviors including positions and simple movements). Through the design of conspecific stimulus experiments, we were able to recognize leg-waving as a visual signal, as it is only displayed in presence of conspecifics or in presence of a mirror and was one of first and most frequent displays in this context. We also detected other visual displays like neck-arching and tail-undulation which may also be relevant as visual signals. Based on our results, we propose that visual signaling is also possible in semi-fossorial lizards; however, further studies regarding chemical signal recognition and color detection are required to confirm our hypothesis.

A new giant anole (Squamata: Iguanidae: Dactyloinae) from southwestern Ecuador.

We describe a new species of Anolis lizard from the Pacific slopes of the Andes of southwestern Ecuador at elevations between 3721,000 m. The new species belongs to the Dactyloa clade and may be distinguished from other Anolis by size, external anatomy, mitochondrial DNA divergence, and dewlap color. Based on phylogenetic analyses of mitochondrial and nuclear DNA sequence data, we found that the new species is sister to A. fraseri in a clade composed primarily of large Dactyloid species. The new species is known from a protected area in southern Ecuador, Buenaventura Reserve, which suggests that at least some its populations are well protected.

Assessment of a Multidisciplinary Intervention in Patients With BMI ≥35 kg/m2 and Recently Diagnosed Type 2 Diabetes.

CONTEXT: Patients with a body mass index (BMI) ≥35 kg/m2 have lower benefits with intensive treatments and metabolic control goals are more difficult to reach. OBJECTIVE: Evaluate the effect of a comprehensive care program in patients with a BMI ≥35 kg/m2. DESIGN: Prospective cohort study. SETTING: Comprehensive Care Center in a National Institute of Health. PATIENTS: Patients with type 2 diabetes, ≤5 years of diagnosis, without disabling complications, nonsmokers, and a BMI <45 kg/m2. INTERVENTIONS: Exercise and nutritional interventions are modified for patients with a higher BMI to achieve metabolic control. MAIN OUTCOME MEASURES: Main outcome is achievement of treatment goals defined as HbA1c <7%, low-density lipoprotein cholesterol (LDL-c) <100 mg/dL, and blood pressure <130/80 mm Hg. Secondary measures were the percentage of patients achieving three metabolic goals. RESULTS: A total of 587 patients with annual evaluation were included. Mean age was 55.3 ± 9.5 years, 56.6% women, time since diagnosis of 1 year (0 to 5). BMI <35 kg/m2 group included 521 patients and BMI ≥35 kg/m2 group included 66 patients. In the BMI ≥35 kg/m2 group, we observed a greater decrease in weight and fat mass at 3 months compared with <35 kg/m2 group, but the HbA1c, LDL-c, or blood pressure goals were similar at 3 months and 1 year between groups. CONCLUSIONS: Comprehensive interventions are equally effective in patients with recently diagnosed type 2 diabetes with BMI ≥35 kg/m2 compared with patients with a lower BMI.

Mating patterns and post-mating isolation in three cryptic species of the Engystomops petersi species complex.

Determining the extent of reproductive isolation in cryptic species with dynamic geographic ranges can yield important insights into the processes that generate and maintain genetic divergence in the absence of severe geographic barriers. We studied mating patterns, propensity to hybridize in nature and subsequent fertilization rates, as well as survival and development of hybrid F1 offspring for three nominal species of the Engystomops petersi species complex in Yasuní National Park, Ecuador. We found at least two species in four out of six locations sampled, and 14.3% of the wild pairs genotyped were mixed-species (heterospecific) crosses. We also found reduced fertilization rates in hybrid crosses between E. petersi females and E. "magnus" males, and between E. "magnus" females and E. "selva" males but not in the reciprocal crosses, suggesting asymmetric reproductive isolation for these species. Larval development times decreased in F1 hybrid crosses compared to same species (conspecific) crosses, but we did not find significant reduction in larval survival or early metamorph survival. Our results show evidence of post-mating isolation for at least two hybrid crosses of the cryptic species we studied. The general decrease in fertilization rates in heterospecific crosses suggests that sexual selection and reinforcement might have not only contributed to the pattern of call variation and behavioral isolation we see between species today, but they may also contribute to further signal divergence and behavioral evolution, especially in locations where hybridization is common and fertilization success is diminished.