bluuebunny/crossref_metadata_2025_split huggingface dataset:
Influence of spice and grape pomace extracts on migraine‐related indicators in vitro
Authors: Slavin Margaret , Bourguignon Julia , Jackson Kyle , Orciga Michael‐Angelo 
Published: June 2021

Calcitonin gene‐related peptide (CGRP) is well‐recognized as a pivotal contributor to the inflammatory process in migraine. Limited evidence suggests that diet may impact circulating levels of CGRP. The objective of this study was to investigate the effects of ginger, turmeric and grape pomace extracts on two cellular models of migraine: CGRP secretion from neuroendocrine CA77 cells, and calcium uptake by nerve growth factor‐differentiated PC12 cells.Dried, ground samples were passed through a 40‐mesh sieve prior to extraction in solvents chosen for their efficiency: grape pomace (50% acetone), ginger root (methanol), and turmeric (acetone). Treatment doses, in 0.1% DMSO vehicle solvent, were based on cell viability results via methylene blue cytoxicity assay. CA77 cells were incubated with treatment for 1 h, then ELISA was used to quantify CGRP. Calcium influx was measured in PC12 cells via the GCaMP5 genetically encoded fluorescent calcium indicator, after 10 min pre‐treatment with extracts, upon stimulation with 50 mM KCl.Results showed a statistically significant decrease in CGRP secretion following treatment with high doses of ginger (0.2 mg ginger equivalent/mL) and grape pomace (1.0 mg pomace equivalent/mL) extracts. Relative to vehicle control, CGRP secretion was decreased by 22% and 87%, respectively. A non‐statistical trend of lower calcium influx was observed in PC12 cells treated with ginger extracts. Solubility of turmeric extracts may have precluded detection of an effect. These results suggest that grape pomace and ginger extracts may have anti‐inflammatory propensity in migraine, which future studies may elucidate.
Carnitine Palmitoyltransferase 2 (CPT2) Inhibition in Rats And Dogs Resembles Inborn Human CPT2 Deficiency
Authors: DE VERA MUDRY MARIA CRISTINA , FESTAG MATTHIAS , HOFLACK JEAN‐CHRISTOPHE , CRAMERI FLAVIO 
Published: June 2021

IntroductionCarnitine palmitoyltransferases (CPT1 and 2) regulate the rate of fatty acid oxidation through intake of long‐chain fatty acids (LCFA) into mitochondria. Inhibition of CPT can reduce gluconeogenesis by decreasing LCFA and acetyl‐CoA in the liver. CPT inhibitors are therefore proposed as a potential therapeutic approach to reduce hyperglycemia in diabetes. Inborn human CPT deficiency is associated with multiorgan toxicity such as muscle weakness, rhabdomyolyis, myoglobinuria, tissue accumulation of lipids and acylcarnitines, and associated degeneration and mitochondrial damage.MethodsTo assess the safety of therapeutic CPT inhibition, a selective inhibitor of CPT2 was given daily by oral gavage to Wistar rats for 14 days, or to Beagle dogs in a single ascending‐dose phase followed by a 7‐day fixed‐dose phase.ResultsClinical signs of CPT2 inhibition consisted of hypoactivity, salivation, emesis (dogs), and body weight loss. At high doses, rats were moribund or died after overnight fasting. Laboratory parameters, gross and histopathology, and transcriptomic profiles (rat liver) indicated liver, kidney, and muscle toxicity. Pale liver, kidney, and heart correlated with increased lipid deposits. Hepatocellular apoptosis, skeletal muscle atrophy and myodegeneration were also present. Electron microscopy of rat liver and muscle revealed mitochondrial damage.ConclusionOn‐target toxicity of a CPT2 inhibitor in the liver, kidney, muscle, and heart in rats and dogs recapitulates and serves as a model of inherited human CPT2 deficiency.
Mouse Cerebellar Neuronal Nitric Oxide Synthase (nNOS) mediates Nicotine ‐‐THC Functional Antagonism (FA) and Cross‐Tolerance
Authors: Dar M Saeed 
Published: June 2021

Despite cannabinoid and nicotine co‐abuse, no study has examined co‐infused intracerebellar (ICB) THC and nicotine effect on cerebellar ataxia (CA).Aims(i) evaluate ICB THC + nicotine effect on CA; (ii) obtain evidence for nNOS's role in THC‐induced CA, THC – nicotine FA and cross‐tolerance.MethodsRotorod evaluated CA in male CD‐1 mice. All drugs [THC 20μg; nicotine 5ng; nNOS inhibitor (nNOSI; 1,2,5 ng), N’‐nitroguanidine tristrifluracetate] were given by ICB infusion.ResultsIncreased THC‐induced CA by nNOSI indicated nNOS mediation. nNOSI opposed nicotine‐induced attenuation of CA by THC. The antagonism was abolished by 5 ng nNOSI. The response curves of nNOSI + nicotine + THC and aCSF + THC treatments overlapped, further evidence of nNOS mediation. We have previously observed robust cross‐tolerance between nicotine and THC. Results of ongoing chronic study involving nNOSI and nicotine infusion<em style=”mso‐bidi‐font‐style: normal;”> (once daily x 5 days followed 16 h later by acute THC) may further support nNOS role.DiscussionNicotine activates NOS [present in cerebellar neurons except Purkinje cells (PCs)] via release of glutamate. Alleviation of CA is likely due to: increased release of glutamate ►enhanced glutamate transmission ►diffusion of NO into PCs► activation of GC and cGMP production ►low PC firing► removal of CA. Overall, the results support a role of cerebellar nNOS in the nicotine ‐‐THC FA and cross‐tolerance.
Central Amygdala PACAP in the Behavioral Stress Response
Authors: Seiglie Mariel , Iemolo Attilio , Blasio Angelo , Cottone Pietro , Sabino Valentina 
Published: June 2021

Anxiety‐related disorders are the most common forms of mental disorders; characterized by feelings of excessive worry in the absence of specific external stimuli, they are accompanied by physical, affective and behavioral symptoms.Pituitary adenylate cyclase‐activating polypeptide (PACAP) is a 38‐amino acid peptide located in the hypothalamus and in various extra‐hypothalamic regions including the central nucleus of the amygdala (CeA). PACAP and PAC1 receptor (PAC1R) have been proposed to play a key role in mediating the behavioral and endocrine responses to stress; however, few studies have examined the involvement of the extrahypothalamic PACAP system in the modulation of stress. Our aim was to elucidate the role of the PACAP/PAC1 system of the CeA in the context of anxiety.We first assessed the effects of PACAP microinfused into the CeA of male rats in the elevated plus maze (EPM) and the acoustic startle response (ASR), behavioral tests sensitive to states of anxiety and fear. PACAP was also given into the basolateral amygdala (BlA) as a control brain region. We then used restraint stress to determine whether the endogenous PACAP‐PAC1 system is recruited by acute stressors.Infusion of PACAP into the CeA significantly reduced percent open arm time in the EPM and increased plasma corticosterone levels, while BlA microinfusion had no effect. Infusion of PACAP into the CeA also increased acoustic startle reactivity. Finally, we observed that an acute, 1h exposure to restraint produced a rapid elevation of PACAP and PAC1R expression in the CeA, but not the BlA.These data prove an anxiogenic role for the PACAP/PAC1 system of the CeA and suggest that hyperactivity of this system may underlie the anxiogenic effects of stress.
Pharmacological treatment(?) of Sleep Problems in a Swedish Pediatric Setting
Authors: Holmbäck Ulf , Forslund Anders 
Published: June 2021

AimThe purpose of this study is to investigate the clinical use of sleep aids, specifically melatonin for sleep disorders in children and adolescents in treated at various facilities in Uppsala County, SwedenMethodA cross sectional study was conducted using a short 9 question web‐based survey. The survey contained questions about sleep medications in general and some specific questions about melatonin/Circadin®. The questionnaire was then sent to doctors in various clinics in Uppsala County, Sweden.ResultsPreliminary results shows that the majority of the doctors prescribe some sort of sleep aid, where Circadin® was the most commonly prescribed sleep aid; with melatonin as a close second (Melatonin not an OTC‐drug in Sweden). Sleep aid was prescribed to children of all ages; and most often in conjunction to various neuropsychiatric disorders. The average dose was 2‐3 mg melatonin /Circadin® per day, with some prescribing up to 6 or 9 mg per day.ConclusionThese preliminary results indicate that the doctors prescribe Circadin® to a larger degree. This despite that Circadin®'s current indication is for people >55 years old. Moreover, high doses of melatonin are prescribed, most probably leading to next‐day drowsiness and sub‐optimal functioning of the children. Doctors need to be informed about proper choice of drug and dose for the pediatric patient
Nicotine Increases Codeine Analgesia Through the Induction of Brain CYP2D Central Activation of Codeine to Morphine
Authors: McMillan Douglas , Tyndale Rachel 
Published: June 2021

Cytochrome P450 2D (CYP2D) metabolizes codeine to morphine, a step required for codeine analgesia. However lower morphine permeability across the blood‐brain barrier, as well as exportation of morphine, suggest that after codeine the initial morphine in the brain, and resulting analgesia, may be due to brain CYP2D activation. Higher brain, but not liver, CYP2D6 is found in smokers, and 7‐day nicotine treatment induces rat brain, but not liver, CYP2D. We hypothesized that inducing rat‐brain CYP2D would increase the CNS CYP2D‐mediated activation of codeine to morphine resulting in increased analgesia, despite first‐pass metabolism, which could be blocked by inhibition of brain CYP2D. Codeine (20mg/kg ip) analgesia was tested: compared to pretreatment with vehicle, 7‐day nicotine (1mg/kg sc, brain CYP2D inducer) increased analgesia (1.59, AUC0‐30min fold change vs. vehicle; p<0.001), propranolol (20 μg in 4 μl icv, brain CYP2D inhibitor) decreased analgesia (0.56; p<0.05), and co‐pretreatment was similar to vehicle (1.23; p>0.1). 7‐day nicotine increased and propranolol decreased brain, but not plasma, morphine levels; analgesia correlated with brain (p<0.02), but not plasma (p>0.4), morphine levels at 15 min after codeine. Pretreatments (alone or together) had no effect on baseline nociception or morphine (3.5mg/kg ip) analgesia. Thus, variation in brain CYP2D activity may contribute to clinically relevant differences in individual response to centrally acting CYP2D substrates. Individuals exposed to nicotine may have increased brain metabolism contributing to altered drug efficacy and abuse liability.Funding: CIHR and the CAMH Foundation
Fumaric Acid Esters Attenuate Secondary Degeneration and Promote Functional Recovery Following Experimental Spinal Cord Injury
Authors: Esposito Emanuela , Paterniti Irene , Cordaro Marika , Cuzzocrea Salvatore 
Published: June 2021

Spinal cord injury (SCI) causes a significant physical, emotional, social and economic burden. The main clinical hallmark of SCI is the permanent loss of motor, sensory and autonomic functions below the level of injury that are in part due to the ensuing inflammatory response. Fumaric acid esters (FAEs) display immunomodulatory properties and may also exert neuroprotective effects, as shown in vitro as well as in experimental models of multiple sclerosis.The aim of the present study was to evaluate the potential beneficial effects of FAE in a mouse model of traumatic spinal cord injury. SCI was induced in mice by spinal cord compression, and dimethyl fumarate (DMF) and monomethyl fumarate (MF) (both at 30 mg/kg) were orally administered to the mice 1 and 6 h after SCI, and once daily thereafter for 10 d. Motor function (Basso Mouse Scale) improved gradually in mice treated with DMF. However, those treated with DMF exhibited a significantly more rapid and sustained recovery of motor function. FAEs also significantly reduced the severity of inflammation (proinflammatory cytokines, apoptosis, and neuroptrophic factors) that characterized the secondary effects of SCI. Again, the effects of DMF were superior to those of MF for several parameters. These results showed marked protective effects of DMF in an animal model of SCI, significantly enhancing recovery of motor function, possibly by reducing the secondary inflammation and tissue injury that characterize this model. In summary, DMF may constitute a promising target for future SCI therapies.
Spinal Cord Injury in Rat – Therapeutic Effect of Pentadecapeptide BPC 157
Authors: Perovic Darko , Buljat Gojko , Kolenc Danijela , Drmic Domagoj , Seiwerth Sven , Sikiric Predrag 
Published: June 2021

BackgroundPreviously pentadecapeptide BPC 157 exhibited a strong healing effect on brain traumas and peripheral nerve injuries (Regul Pept. 2010;160(1‐3):26‐32; Regul Pept. 2010;160(1‐3):33‐41). Aim. To verify influence of BPC 157 on rat spinal cord injury. Materials and methods. Deeply anaesthetized Wistar Albino male rats, 350 g b.w., were subjected to surgery (all experimental procedures were approved by the local Ethics Committee): after laminectomy on level L2‐L3 (correspond to sacrocaudal spinal cord) 60 second compression with neurosurgical piston (60‐66 g) on exposed dural sac. One single medication (IP) (saline 5ml/kg; pentadekapeptide BPC 157 200µg/kg; 2 µg/kg) was 10 min post‐injury and sacrifice was at 1, 2, 4, 12, 26 and 52 weeks. Tail motor function (1 paralysis; 5 normal) and spasticity (1 normal; 5 maximum) was scored daily. Histology samples of spinal cord were taken from lesion site, and tail samples 10 mm distal from base. Results. Tail motor function. BPC 157 rats have significantly higher motor score in all investigated time. Tail spasticity. BPC 157 rats have significantly less spasticity starting from 4 weeks to the end of experiments. Histology of spinal cord. From 2 weeks to the end of follow up BPC 157 rats have less grey matter edema and less cyst and axonal necrosis in white matter. Histology of rat tail. All treatment groups show increased number of all and small axons starting from 4 weeks but without statistically difference between groups. Conclusions. Given after sacrocaudal spinal cord injury, BPC 157 recovered rat tail function with single treatment. Specifically, BPC 157 decreased secondary injury of spinal cord, and consequently, improved neurologic function recovery. Supported by Grant 108‐1083570‐3635, Croatia.
Nanoparticle‐Bound Cytolytic Peptide Prodrugs for Cancer Therapy
Authors: Jallouk Andrew , Palekar Rohun , Pan Hua , Schlesinger Paul , Wickline Samuel 
Published: June 2021

ObjectiveTo develop a nanoparticle platform which promotes the delivery of cytolytic peptide prodrugs to tumors and their activation by locally produced proteases.Introduction: Melittin is a peptide toxin derived from bee venom which has been proposed as a cancer therapy due to its cytolytic activity. Yet, its therapeutic applications have been limited by its nonspecific cytotoxicity and rapid clearance from the circulation. We have previously developed a prodrug form of melittin which is activated upon cleavage by matrix metalloproteinase 9 (MMP‐9), a protease closely linked with tumor growth and metastasis. Here, we present a platform for delivery of this prodrug based on a perfluorocarbon nanoparticle system designed to enhance intratumoral accumulation while permitting prodrug activation by local MMP‐9 activity.Methods and Results: Incubation of the melittin prodrug with perfluorocarbon nanoparticles (NPs) resulted in stable loading of 10,250 peptides per NP with no significant change in NP size. Addition of 20 nM MMP‐9 led to activation of 57.8% of NP‐bound prodrug within 3 hours. B16F10 melanoma cell viability was virtually eliminated following incubation with 50 μM free or NP‐bound prodrug in the presence of MMP‐9 (p < 0.001 compared to control). In a mouse model of B16F10 lung metastasis, NP accumulation in lung tumors was demonstrated by both fluorescence microscopy (Figure A; NPs in red) and quantitative 19F magnetic resonance spectroscopy (Figure B). Gelatin zymography confirmed production of active MMP‐9 by these tumors.ConclusionPerfluorocarbon nanoparticles accumulate within tumors and permit activation of bound prodrugs, making them promising delivery vehicles for cytolytic peptide agents.Research Support: NIH R01 HL073646
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Identifying the Structure‐Function Activity of Human Cytosolic Sulfotransferase (SULT) 1C4
Authors: Guidry Amber , Falany Charles 
Published: June 2021

Human cytosolic sulfotransferases (hSULTs) are phase II conjugation enzymes that play a critical role in the metabolism of drugs and hormones, the bioactivation of carcinogens, and the detoxification of xenobiotics. SULTs transfer a sulfuryl‐moiety from the obligate donor PAPS (3′‐phosphoadenosine 5′‐phosphosulfate) to a suitable hydroxyl or amine group of the substrate. While some of the SULTs have been studied extensively, others, such as the SULT 1C family, remain poorly characterized. The objective of this study is to determine the role of hSULT1C4 in drug metabolism by identifying novel substrates and defining the substrate binding, selectivity, and activity of the SULT1C4 enzyme using molecular modeling and kinetic analysis. SULT1C4 was bacterially expressed and purified for in vitro kinetic analysis. Steady‐state kinetic parameters for 1‐naphthol were analyzed using four concentrations of 1‐naphthol (0.5, 1, 2.5, and 7.5 uM) and four concentrations of PAPS (0.5, 1.5, 5, 15 uM). The Km of 1‐naphthol was approximately 1 uM, whereas the data for PAPS suggests the existence of two Kms (1 uM and 12 uM). To determine if these two Kms are due to SULT1C4's dimeric protein state, two different dimerization domain mutants, SULT1C4‐K272A and SULT1C4‐V277E, have been generated. Native page gel analysis has shown that these mutations prevent dimerization resulting in monomeric SULT1C4 proteins. Our data suggests that the dimeric state of SULT1C4 influences PAPS’ ability to bind to the enzyme.Funding sources: NIH ES022606 and GM038953
Helicobacter heilmannii sensu lato‐induced MALT Lymphoma Formation in Lacrimal Gland as well as Fundic Gland in C57BL/6 mouse: Effect of Aprepitant
Authors: Nakamura Masahiko , Matsui Hidenori , Takahashi Tetsufumi 
Published: June 2021

As to the pathogenesis of the Sjogren syndrome, Helicobacter pylori has been suggested to have some relation mostly from the epidemiological studies, while few experimental studies have clarified this point.We have established a gastric low‐grade MALT lymphoma model in C57BL/6 mouse by the infection of Helicobacter heilmannii sensu lato obtained from cynomolgus monkey. After the long term infection, we have detected the accumulation of lymphocytes in the lacrimal glands similar to the gastric MALT lymphoma. The characteristics of this lesion were investigated as well as the effect of aprepitant, an NK‐1R antagonist on this lesion.Materials and MethodsThe characteristics of the lymphocyte accumulation were investigated by the histochemical analysis using lymphocyte marker, CD4, B220, CD10, epithelial cell marker cytokeratin, mucus marker MUC7 and aquaporin‐4 and 5. The localization of the autonomic nerves was also clarified by the choline acetyltransferase, substance P and calcitonin gene‐related peptide.ResultsBy the histochemical analysis, the lymphoepithelial lesion was detected in the lymphocyte aggregation and the germinal center was formed in the center of the lesion, showing its characteristics as a mucosa‐associated lymphoid tissue (MALT) lymphoma. The lesion was also richly innervated by the autonomic nerves. Intraperitoneal administration of aprepitant induced the significant reduction in the lesion size.ConclusionHelicobacter heilmannii sensu lato infection to the C57BL/6 mouse induced the formation of the MALT lymphoma in the mouse lacrimal gland. This model could be one of the animal models of the Sjogren syndrome.
Time to Reach Steady State Resting Energy Expenditure in Healthy Adults Using Indirect Calorimetry
Authors: Popp Collin , Tisch Jocelyn , Bridges William , Traylor Dan , Najm Nadia , Jesch Elliot 
Published: June 2021

BackgroundResting energy expenditure (REE) measures the amount of calories needed for the body to maintain basic function over a 24‐hour period. Traditionally, REE protocols through indirect calorimetry take 45 minutes to complete. Generally, the first 10 to 20 minutes of data collected are excluded to allow the participants to reach steady state.ObjectiveThe purpose of this study was to determine a more precise time to reach a resting steady state.MethodsFifty healthy, adult male and female participants volunteered for this study. Each participant completed indirect calorimetry after a 12‐hour overnight fast. Participants were instructed to lay supine and awake under a plastic canopy for approximately 30‐minute, while oxygen consumption (VO2) and carbon dioxide production (VCO2) measures were collected. These values were used to determine steady state REE (kcals•day‐1) and respiratory quotient (RQ). Statistical analysis for the response of variables to time was modeled with a logistics decay function. Data are reported as mean ± SD with an alpha level of 0.05.ResultsPredicted time to steady state REE in kcals•day‐1 and in kcals•day‐1•kg‐1 was reached at 15.09 ± 0.1 minutes and 15.10 ± 0.1 minutes, respectively. There was no significant change in REE thereafter. In addition, respiratory quotient values from the first 15‐minutes (0.91) to the last 15‐minutes (0.86) were significantly different (p < 0.001).ConclusionIndirect calorimetry to measure steady state resting energy expenditure should exclude the first 15‐minutes in a young, healthy population.
Involvement of HCN channels in diaphragmatic lymphatic pacemaking
Authors: Negrini Daniela , Marcozzi Cristiana , Solari Eleonora , Moriondo Andrea , Bossi Elena , Cinquetti Raffaella , Reguzzoni Marcella 
Published: June 2021

Pacemaking in spontaneously contracting lymphatics is thought to depend upon: a) small transient depolarisations sustained by an inward current through calcium‐dependent chloride channels, seen in mesenteric lymphatic vessels of guinea pigs, and, b) a current, observed only in bovine and sheep lymphatics, resembling cardiac If current. This study aimed at performing, a molecular, functional, and immuno‐hystological study to verify the potential involvement of hyperpolarization cyclic nucleotide gated (HCN) channels in pacing of intrinsic pumping activity. Experiments were performed in conformance with the FASEB Statement of Principles for the use of animals in research and education. After deep anesthesia, rat diaphragmatic specimens containing spontaneously contracting FITC‐dextran‐filled lymphatics were excised. Whole‐tissue perfusion with Ivabradine (300 mM) or CsCl (10 mM) caused a significant reduction in contraction frequency. CsCl also caused vasorelaxation and an increase in stroke amplitude. HCN channels were immunolocalised in the lymphatic vessel wall and this was further confirmed by RT‐PCR analysis of mRNA transcripts from the contracting vessels. Therefore, our results provide for the first time the molecular, functional, and immuno‐hystological evidences that HCN channels are responsible for pacemaking of lymphatic smooth muscle, setting the lymphatic vessel wall contractility.Supported by the FAR2014 Research fund by the University of Insubria