Nonopioid Actions of U50,488 Enantiomers Contribute to Their Peripheral Cutaneous Antinociceptive Effects

ABSTRACT The ability of arylacetamide -opioid receptor agonists (-ORAs) to block sodium channels by a nonopioid mechanism has been previously documented. The present experiments were undertaken to test whether two enantiomers of the aryla

Transcript expression of vesicular glutamate transporters in lumbar dorsal root ganglia and the spinal cord of mice – Effects of peripheral axotomy or hindpaw inflammation

Using specific riboprobes, we characterized the expression of vesicular glutamate transporter (VGLUT)1–VGLUT3 transcripts in lumbar 4–5 (L4–5) dorsal root ganglions (DRGs) and the thoracolumbar to lumbosacral spinal cord in male BALB/c mice after a 1- or 3-day hindpaw inflammation, or a 7-day sciatic nerve axotomy. Sham animals were also included. In sham and contralateral L4–5 DRGs of injured mice, VGLUT1-, VGLUT2- and VGLUT3 mRNAs were expressed in ∼45%, ∼69% or ∼17% of neuron profiles (NPs), respectively. VGLUT1 was expressed in large and medium-sized NPs, VGLUT2 in NPs of all sizes, and VGLUT3 in small and medium-sized NPs. In the spinal cord, VGLUT1 was restricted to a number of NPs at thoracolumbar and lumbar segments, in what appears to be the dorsal nucleus of Clarke, and in mid laminae III–IV. In contrast, VGLUT2 was present in numerous NPs at all analyzed spinal segments, except the lateral aspects of the ventral horns, especially at the lumbar enlargement, where it was virtually absent. VGLUT3 was detected in a discrete number of NPs in laminae III–IV of the dorsal horn. Axotomy resulted in a moderate decrease in the number of DRG NPs expressing VGLUT3, whereas VGLUT1 and VGLUT2 were unaffected. Likewise, the percentage of NPs expressing VGLUT transcripts remained unaltered after hindpaw inflammation, both in DRGs and the spinal cord. Altogether, these results confirm previous descriptions on VGLUTs expression in adult mice DRGs, with the exception of VGLUT1, whose protein expression was detected in a lower percentage of mouse DRG NPs. A detailed account on the location of neurons expressing VGLUTs transcripts in the adult mouse spinal cord is also presented. Finally, the lack of change in the number of neurons expressing VGLUT1 and VGLUT2 transcripts after axotomy, as compared to data on protein expression, suggests translational rather than transcriptional regulation of VGLUTs after injury.Fil: Malet, Mariana. Universidad Austral. Facultad de Ciencias Biomédicas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Vieytes, C. A.. Universidad Austral. Facultad de Ciencias Biomédicas; ArgentinaFil: Lundgren, K. H.. University of Cincinnati; Estados UnidosFil: Seal, R. P.. University of Pittsburgh; Estados UnidosFil: Tomasella, María Eugenia. Universidad Austral. Facultad de Ciencias Biomédicas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Seroogy, K. B.. University of Cincinnati; Estados UnidosFil: Hökfelt, T.. Karolinska Huddinge Hospital. Karolinska Institutet; SueciaFil: Gebhart, G. F.. University of Pittsburgh; Estados UnidosFil: Brumovsky, Pablo Rodolfo. Universidad Austral. Facultad de Ciencias Biomédicas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. University of Pittsburgh; Estados Unido

Luminal hypertonicity and acidity modulate colorectal afferents and induce persistent visceral hypersensitivity

Carbohydrate malabsorption such as in lactose intolerance or enteric infection causes symptoms that include abdominal pain. Because this digestive disorder increases intracolonic osmolarity and acidity by accumulation of undigested carbohydrates and fermented products, we tested whether these two factors (hypertonicity and acidity) would modulate colorectal afferents in association with colorectal nociception and hypersensitivity. In mouse colorectum-pelvic nerve preparations in vitro, afferent activities were monitored after application of acidic hypertonic saline (AHS; pH 6.0, 800 mosM). In other experiments, AHS was instilled intracolonically to mice and behavioral responses to colorectal distension (CRD) measured. Application of AHS in vitro excited 80% of serosal and 42% of mechanically-insensitive colorectal afferents (MIAs), sensitizing a proportion of MIAs to become mechanically sensitive and reversibly inhibiting stretch-sensitive afferents. Acute intracolonic AHS significantly increased expression of the neuronal activation marker pERK in colon sensory neurons and augmented noxious CRD-induced behavioral responses. After three consecutive daily intracolonic AHS treatments, mice were hypersensitive to CRD 4-15 days after the first treatment. In complementary single fiber recordings in vitro, the proportion of serosal class afferents increased at day 4; the proportion of MIAs decreased, and muscular class stretch-sensitive afferents were sensitized at days 11-15 in mice receiving AHS. These results indicate that luminal hypertonicity and acidity, two outcomes of carbohydrate malabsorption, can induce colorectal hypersensitivity to distension by altering the excitability and relative proportions of colorectal afferents, suggesting the potential involvement of these factors in the development of abdominal pain.Fil: La, Jun Ho. University of Pittsburgh; Estados UnidosFil: Feng, Bin. University of Pittsburgh; Estados UnidosFil: Schwartz, Erica S.. University of Pittsburgh; Estados UnidosFil: Brumovsky, Pablo Rodolfo. Universidad Austral. Facultad de Ciencias Biomédicas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Gebhart, G. F.. University of Pittsburgh; Estados Unido

Performance Analysis of Throughput Efficient Switch-over between FSO and mmW Links

Free Space Optics (FSO) links provide usage of high bandwidth and the flexibility of wireless communication links. However, weather patterns like fog and heavy snow fall limit the availability of FSO. Another technology providing similar properties regarding offered data rates and flexibility of setup is Millimeter Wave Technology (mmW), operating at several tens of GHz. In this case, heavy rain limits mmW link availability. A combination of both technologies had been proved to be very effective to achieve very high availability. Different hybrid architectures of these two links and switch-over techniques had been proposed in the recent years. All of these techniques require redundant transmission on either both transmission links or waste bandwidth of backup link when main FSO link is operational. In this paper, a switch-over between these technologies is proposed, to maintain high availability without the loss of transmission bandwidth. The performance of this switch-over has been simulated for more than one year measured availability data for hybrid network of mmW link and FSO link. The switch over behavior has also been simulated for fog, rain and snow events. It has been shown that the availability with switch-over reaches the redundant link availability but switchover can save more than 90% redundant transmission and increase the hybrid network throughput significantly

Expression of vesicular glutamate transporters in sensory and autonomic neurons innervating the mouse urinary bladder

Purpose: Vesicular glutamate transporters (VGLUTs), essential for loading glutamate into synaptic vesicles, are present in various neuronal systems. However, the expression of VGLUTs in neurons innervating the urinary bladder has not yet been analyzed. Here, we study the presence of VGLUTs type-1, -2 and -3 (VGLUT1, VGLUT2 and VGLUT3, respectively) in mouse urinary bladder neurons. Materials and Methods: Expression of VGLUT1, VGLUT2 and calcitonin gene-related peptide (CGRP) was analyzed by immunohistochemistry in retrogradely labeled primary afferent and autonomic neurons of BALB/C mice after injecting Fast Blue in the urinary bladder wall. To study VGLUT3, retrograde tracing of the urinary bladder was performed in transgenic mice where VGLUT3 is identified by detection of enhanced green fluorescent protein (EGFP). Results: Most urinary bladder DRG neurons expressed VGLUT2. A smaller percentage of neurons also expressed VGLUT1 or VGLUT3. Coexpression with CGRP was only observed for VGLUT2. Occasional VGLUT2-immunoreactive (IR) neurons were seen in the major pelvic ganglion (MPG). Abundant VGLUT2-IR nerves were detected in the urinary bladder dome, trigone and also the urethra; VGLUT1-IR nerves were discretely present. Conclusions: We present novel data on the expression of VGLUTs in sensory and autonomic neurons innervating the mouse urinary bladder. The frequent association of VGLUT2 and CGRP in sensory neurons suggests interactions between glutamatergic and peptidergic neurotransmissions, potentially influencing commonly perceived sensations in the urinary bladder, such as discomfort and pain.Fil: Brumovsky, Pablo Rodolfo. Universidad Austral. Facultad de Ciencias Biomédicas. Laboratorio de Investigaciones Biomédicas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. University of Pittsburgh. Department of Anesthesiology. Pittsburgh Center for Pain Research; Estados UnidosFil: Seal, Rebecca P.. University of Pittsburgh. Department of Anesthesiology. Pittsburgh Center for Pain Research; Estados UnidosFil: Lundgren, Kerstin H.. University of Cincinnati. Department of Neurology; Estados UnidosFil: Seroogy, Kim B.. University of Cincinnati. Department of Neurology; Estados UnidosFil: Watanabe, Masahiko. Hokkaido University School of Medicine. Department of Anatomy; JapónFil: Gebhart, G. F.. University of Pittsburgh. Department of Anesthesiology. Pittsburgh Center for Pain Research; Estados Unido

Collecting Service-Based Maintainability Metrics from RESTful API Descriptions: Static Analysis and Threshold Derivation

While many maintainability metrics have been explicitly designed for service-based systems, tool-supported approaches to automatically collect these metrics are lacking. Especially in the context of microservices, decentralization and technological heterogeneity may pose challenges for static analysis. We therefore propose the modular and extensible RAMA approach (RESTful API Metric Analyzer) to calculate such metrics from machine-readable interface descriptions of RESTful services. We also provide prototypical tool support, the RAMA CLI, which currently parses the formats OpenAPI, RAML, and WADL and calculates 10 structural service-based metrics proposed in scientific literature. To make RAMA measurement results more actionable, we additionally designed a repeatable benchmark for quartile-based threshold ranges (green, yellow, orange, red). In an exemplary run, we derived thresholds for all RAMA CLI metrics from the interface descriptions of 1,737 publicly available RESTful APIs. Researchers and practitioners can use RAMA to evaluate the maintainability of RESTful services or to support the empirical evaluation of new service interface metrics.Comment: Accepted at CSE/QUDOS workshop (collocated with ECSA 2020

Interpretable Subgroup Discovery in Treatment Effect Estimation with Application to Opioid Prescribing Guidelines

The dearth of prescribing guidelines for physicians is one key driver of the current opioid epidemic in the United States. In this work, we analyze medical and pharmaceutical claims data to draw insights on characteristics of patients who are more prone to adverse outcomes after an initial synthetic opioid prescription. Toward this end, we propose a generative model that allows discovery from observational data of subgroups that demonstrate an enhanced or diminished causal effect due to treatment. Our approach models these sub-populations as a mixture distribution, using sparsity to enhance interpretability, while jointly learning nonlinear predictors of the potential outcomes to better adjust for confounding. The approach leads to human-interpretable insights on discovered subgroups, improving the practical utility for decision suppor

Trastuzumab and pertuzumab without chemotherapy in early-stage HER2+ breast cancer: a plain language summary of the PHERGain study

This is a summary of a publication about the PHERGain study, which was published in The Lancet Oncology in May 2021. The study includes 376 women with a type of breast cancer called HER2-positive breast cancer that can be removed by surgery. In the study, researchers wanted to learn if participants could be treated with two medicines called trastuzumab and pertuzumab without the need for chemotherapy. To identify HER2-positive tumors with more sensitivity to anti-HER2 therapies, the researchers used a type of imaging called a FDG-PET scan to check how well the treatments were working.Participants took a treatment before surgery, consisting of either chemotherapy (docetaxel and carboplatin) plus trastuzumab and pertuzumab (group A) or trastuzumab and pertuzumab alone (plus hormone therapy if the tumor was hormone receptor-positive; group B). After two cycles of treatment, participants underwent a FDG-PET scan. Participants assigned to group A completed 6 cycles of treatment regardless of 18F-FDG-PET results. Participants in group B continued the same treatment until surgery if their FDG-PET scan showed the treatment was working. While participants who did not show a response started treatment with chemotherapy in addition to trastuzumab and pertuzumab. All participants then had surgery.The results revealed that, of the participants in group B who showed a response using FDG-PET scan, 37.9% achieved a disappearance of all invasive cancer in the breast and axillary lymph nodes. This rate appears to be higher than those reported in previous studies evaluating the same treatment. These participants also had less side effects and improved overall quality of life compared with participants taking chemotherapy plus trastuzumab and pertuzumab.Early monitoring of how well participants respond to treatment by FDG-PET scan seems to identify participants with operable HER2-positive breast cancer who were more likely to benefit from trastuzumab and pertuzumab without the need to have chemotherapy. The PHERGain study is still ongoing and results on long-term survival are expected to be released in 2023. Clinical Trial Registration: NCT03161353 (ClinicalTrials.gov)

Expression of vesicular glutamate transporters type 1 and 2 in sensory and autonomic neurons innervating the mouse colorectum

Vesicular glutamate transporters (VGLUTs) have been extensively studied in various neuronal systems, but their expression in visceral sensory and autonomic neurons remains to be analyzed in detail. Here we studied VGLUTs type 1 and 2 (VGLUT(1) and VGLUT(2) , respectively) in neurons innervating the mouse colorectum. Lumbosacral and thoracolumbar dorsal root ganglion (DRG), lumbar sympathetic chain (LSC), and major pelvic ganglion (MPG) neurons innervating the colorectum of BALB/C mice were retrogradely traced with Fast Blue, dissected, and processed for immunohistochemistry. Tissue from additional naïve mice was included. Previously characterized antibodies against VGLUT(1) , VGLUT(2) , and calcitonin gene-related peptide (CGRP) were used. Riboprobe in situ hybridization, using probes against VGLUT(1) and VGLUT(2) , was also performed. Most colorectal DRG neurons expressed VGLUT(2) and often colocalized with CGRP. A smaller percentage of neurons expressed VGLUT(1) . VGLUT(2) -immunoreactive (IR) neurons in the MPG were rare. Abundant VGLUT(2) -IR nerves were detected in all layers of the colorectum; VGLUT(1) -IR nerves were sparse. A subpopulation of myenteric plexus neurons expressed VGLUT2 protein and mRNA, but VGLUT1 mRNA was undetectable. In conclusion, we show 1) that most colorectal DRG neurons express VGLUT(2) , and to a lesser extent, VGLUT(1) ; 2) abundance of VGLUT2-IR fibers innervating colorectum; and 3) a subpopulation of myenteric plexus neurons expressing VGLUT(2). Altogether, our data suggests a role for VGLUT(2) in colorectal glutamatergic neurotransmission, potentially influencing colorectal sensitivity and motility.Fil: Brumovsky, Pablo Rodolfo. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Univeristy of Pittsburgh. School of Medicine; Estados Unidos. Universidad Austral; ArgentinaFil: Robinson, David R.. Univeristy of Pittsburgh. School of Medicine; Estados Unidos. University of Pittsburgh at Johnstown; Estados Unidos. University of Pittsburgh; Estados UnidosFil: La, Jun Ho. Univeristy of Pittsburgh. School of Medicine; Estados UnidosFil: Seroogy, Kim B.. No especifíca;Fil: Lundgren, Kerstin H.. No especifíca;Fil: Albers, Kathryn M.. Univeristy of Pittsburgh. School of Medicine; Estados UnidosFil: Kiyatkin, Michael E.. Univeristy of Pittsburgh. School of Medicine; Estados UnidosFil: Seal, Rebecca P.. No especifíca;Fil: Edwards, Robert H.. No especifíca;Fil: Watanabe, Masahiko. No especifíca;Fil: Hökfelt, Tomas. Karolinska Huddinge Hospital. Karolinska Institutet; SueciaFil: Gebhart, G. F.. Univeristy of Pittsburgh. School of Medicine; Estados Unido