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Abstract Background In experimental models of fetal alcohol spectrum disorder (FASD), cerebellar hypoplasia and hypofoliation are associated with insulin and insulin-like growth factor (IGF) resistance with impaired signaling through pathways that mediate growth, survival, plasticity, metabolism, and neurotransmitter function. To more directly assess the roles of impaired insulin and IGF signaling during brain development, we administered intracerebroventricular (ICV) injections of si-RNA targeting the insulin receptor, (InR), IGF-1 receptor (IGF-1R), or IGF-2R into postnatal day 2 (P2) Long Evans rat pups and examined the sustained effects on cerebellar function, structure, and neurotransmitter-related gene expression (P20). Results Rotarod tests on P20 demonstrated significant impairments in motor function, and histological studies revealed pronounced cerebellar hypotrophy, hypoplasia, and hypofoliation in si-InR, si-IGF-1R, and si-IGF-2R treated rats. Quantitative RT-PCR analysis showed that si-InR, and to a lesser extent si-IGF-2R, broadly inhibited expression of insulin and IGF-2 polypeptides, and insulin, IGF-1, and IGF-2 receptors in the brain. ELISA studies showed that si-InR increased cerebellar levels of tau, phospho-tau and β-actin, and inhibited GAPDH. In addition, si-InR, si-IGF-1R, and si-IGF-2R inhibited expression of choline acetyltransferase, which mediates motor function. Although the ICV si-RNA treatments generally spared the neurotrophin and neurotrophin receptor expression, si-InR and si-IGF-1R inhibited NT3, while si-IGF-1R suppressed BDNF. Conclusions early postnatal inhibition of brain InR expression, and to lesser extents, IGF-R, causes structural and functional abnormalities that resemble effects of FASD. The findings suggest that major abnormalities in brains with FASD are mediated by impairments in insulin/IGF signaling. Potential therapeutic strategies to reduce the long-term impact of prenatal alcohol exposure may include treatment with agents that restore brain insulin and IGF responsiveness.

Abstract Background Aspartyl-(Asparaginyl)-β-Hydroxylase (AAH) is a hydroxylating enzyme that promotes cell motility by enhancing Notch-Jagged-HES-1 signaling. Ethanol impaired cerebellar neuron migration during development is associated with reduced expression of AAH. Methods To further characterize the role of AAH in relation to cerebellar development, structure, and function, we utilized an in vivo model of early postnatal (P2) intracerebro-ventricular gene delivery to silence AAH with small interfering RNA (siAAH), or over-express it with recombinant plasmid DNA (pAAH). On P20, we assessed cerebellar motor function by rotarod testing. Cerebella harvested on P21 were used to measure AAH, genes/proteins that mediate AAH's downstream signaling, i.e. Notch-1, Jagged-1, and HES-1, and immunoreactivity corresponding to neuronal and glial elements. Results The findings demonstrated that: 1) siAAH transfection impaired motor performance and blunted cerebellar foliation, and decreased expression of neuronal and glial specific genes; 2) pAAH transfection enhanced motor performance and increased expression of neuronal and glial cytoskeletal proteins; and 3) alterations in AAH expression produced similar shifts in Notch-1, Jagged-1, and HES-1 protein or gene expression. Conclusions The results support our hypothesis that AAH is an important mediator of cerebellar development and function, and link AAH expression to Notch signaling pathways in the developing brain.

Unemployment and underemployment have consistently been shown to be higher in autistic adults relative to non-autistic adults. This may be due, in part, to a lack of workplace accommodations being made for autistic people. One factor that may contribute to employment inequalities in autistic people is differences in attitudes towards interpersonal touch. This study acts as a preliminary investigation into whether employed autistic and non-autistic participants differ in their attitudes towards touch in the workplace, and in their loneliness and wellbeing. The current dataset was drawn from a larger online survey (the Touch Test) designed to explore attitudes and experiences towards touch. We found that employed autistic participants had more negative attitudes to general, social and workplace touch relative to non-autistic participants. Autistic participants also experienced greater loneliness and reduced wellbeing. Attachment-related anxiety was the only significant predictor of wellbeing in employed autistic adults. However, attachment-related anxiety, general attitudes to touch and the role of touch in the workplace predicted wellbeing in employed non-autistic adults. With regards to loneliness, general attitudes to touch and the role of touch in the workplace predicted loneliness in autistic participants. We also replicated the finding that a greater proportion of autistic participants were unemployed relative to non-autistic participants. Collectively, this research highlights the importance of considering touch in research investigating employment, and its impact on loneliness and wellbeing, in autistic participants., (© 2022. The Author(s).)

The coronavirus disease 2019 (COVID-19) outbreak emerged at the end of 2019 and quickly spread around the world. Measures to counter COVID-19, including social distancing and lockdowns, created an unusual situation that had the potential to impact a variety of behaviours, including sleep, which is crucial for health and well-being. Data were obtained through an online survey. The total sample comprised 19,482 participants from the UK. Participants were asked several questions regarding sleep quality and quantity. Each participant completed the questionnaires once during a data collection period spanning January 20 to March 31, 2020. Data provided by different participants during different weeks (spanning time-periods just before COVID-19 was identified in the UK and during the early weeks following its arrival) were compared using analysis of variance tests and regressions. Regression analyses controlling for age, sex and ethnicity revealed significant associations of small magnitude between date of survey completion and sleep quality, sleep latency, number of awakenings and composite score of poor sleep quality. These analyses also indicated small increases in eveningness tendency as the study progressed. There was no change in sleep duration or time spent awake at night. The COVID-19 outbreak did not appear to impact negatively sleep in a substantial manner during the early stages in the UK. The small increases in sleep quality variables (except for time spent awake at night and sleep duration) and eveningness are nonetheless of interest. Further research is needed to understand how best to provide support to those most in need of a good night's sleep during this unprecedented time., (© 2021 The Authors. Journal of Sleep Research published by John Wiley & Sons Ltd on behalf of European Sleep Research Society.)

[Bis(pyridine)iodine(I)] + complexes offer controlled access to halonium ions under mild conditions. The reactivity of such stabilized halonium ions is primarily determined by their three-center, four-electron [N-I-N] + halogen bond. We studied the importance of chelation, strain, steric hindrance and electrostatic interaction for the structure and reactivity of halogen bonded halonium ions by acquiring their 15 N NMR coordination shifts and measuring their iodenium release rates, and interpreted the data with the support of DFT computations. A bidentate ligand stabilizes the [N-I-N] + halogen bond, decreasing the halenium transfer rate. Strain weakens the bond and accordingly increases the release rate. Remote modifications in the backbone do not influence the stability as long as the effect is entirely steric. Incorporating an electron-rich moiety close by the [N-I-N] + motif increases the iodenium release rate. The analysis of the iodine(I) transfer mechanism highlights the impact of secondary interactions, and may provide a handle on the induction of stereoselectivity in electrophilic halogenations., (© 2021 The Authors. Chemistry - A European Journal published by Wiley-VCH GmbH.)

The epidemic of opioid and other drug use and related arrests are a growing public health crisis in the United States. The national prevalence of pregnant women with opioid use disorder (OUD) has increased dramatically from 1.5 per 1,000 delivery hospitalizations in 1999 to 6.5 in 2014. The combination of these factors has led to an increased frequency of pregnant women with OUDs in the correctional health care system. This protocol provides evidence-based treatment recommendations including the initiation of methadone and buprenorphine in the inpatient or jail setting. It also explores many of the nuances around caring for this vulnerable patient population and discusses ways in which the medical and correctional health care teams can efficiently collaborate to improve patient outcomes.

Touch is our most interpersonal sense, and so it stands to reason that we represent not only our own bodily experiences, but also those felt by others. This review will summarise brain and behavioural research on vicarious tactile perception (mirror touch). Specifically, we will focus on vicarious touch across the lifespan in typical and atypical groups, and will identify the knowledge gaps that are in urgent need of filling by examining what is known about how individuals differ within and between typical and atypical groups.

Mirror-touch synaesthesia is a condition where observing touch to another's body induces a subjective tactile sensation on the synaesthetes body. The present study explores which characteristics of the inducing stimulus modulate the synaesthetic touch experience. Fourteen mirror-touch synaesthetes watched videos depicting a touch event while indicating (i) whether the video induced a tactile sensation, (ii) on which side of their body they felt this sensation and (iii) the intensity of the experienced sensation. Results indicate that the synaesthetes experience stronger tactile sensations when observing touch to real bodies, whereas observing touch to dummy bodies, pictures of bodies and disconnected dummy body parts elicited weaker sensations. These results suggest that mirror-touch synaesthesia is not entirely bottom-up driven, but top-down information, such as knowledge about real and dummy body parts, also modulate the intensity of the experience., (Copyright © 2011 Elsevier Inc. All rights reserved.)

Background: Aspartyl-(Asparaginyl)-β-Hydroxylase (AAH) is a hydroxylating enzyme that promotes cell motility by enhancing Notch-Jagged-HES-1 signaling. Ethanol impaired cerebellar neuron migration during development is associated with reduced expression of AAH., Methods: To further characterize the role of AAH in relation to cerebellar development, structure, and function, we utilized an in vivo model of early postnatal (P2) intracerebro-ventricular gene delivery to silence AAH with small interfering RNA (siAAH), or over-express it with recombinant plasmid DNA (pAAH). On P20, we assessed cerebellar motor function by rotarod testing. Cerebella harvested on P21 were used to measure AAH, genes/proteins that mediate AAH's downstream signaling, i.e. Notch-1, Jagged-1, and HES-1, and immunoreactivity corresponding to neuronal and glial elements., Results: The findings demonstrated that: 1) siAAH transfection impaired motor performance and blunted cerebellar foliation, and decreased expression of neuronal and glial specific genes; 2) pAAH transfection enhanced motor performance and increased expression of neuronal and glial cytoskeletal proteins; and 3) alterations in AAH expression produced similar shifts in Notch-1, Jagged-1, and HES-1 protein or gene expression., Conclusions: The results support our hypothesis that AAH is an important mediator of cerebellar development and function, and link AAH expression to Notch signaling pathways in the developing brain.

Glioblastomas are high-grade, malignant CNS neoplasms that are nearly always fatal within 12 months of diagnosis. Immunotherapy using proinflammatory cytokines such as IL-2 or IL-12 may prolong survival with glioblastoma. Thymosin-alpha1 (Talpha1) is a thymic hormone and immunemodulator that increase IL-2 production and T-cell proliferation. We examined potential therapeutic effects of Talpha1 in experimental in vivo glioblastoma, and characterized Talpha1's anti-tumor effects in vitro. Rar 9L cells (10(4)) were implanted into the right frontal lobe of adult Long Evans rats that were subsequently treated with vehicle, BCNU, Talpha1, or Talpha1+BCNU from postoperative day 6. Talpha1+BCNU significantly lowered tumor burdens, and increased cure rates. In vitro experiments demonstrated that Talpha1 had no direct effect on viability or mitochondrial function, and instead, it increased expression of pro-apoptosis genes, including FasL, FasR and TNFalpha-R1 (65.89%, 44.08%, and 22.18%, resp.), and increased 9L cell sensitivity to oxidative stress. Moreover, Talpha1 enhanced 9L cell sensitivity to both Granzyme B- and BCNU-mediated killing. The findings suggest that Talpha1 enhances BCNUmediated eradication of glioblastoma in vivo, and that Talpha1 mediates its effects by activating pro-apoptosis mechanisms, rendering neoplastic cells more sensitive to oxidative stress and immune-mediated killing by Granzyme B and chemotherapeutic agents.

We have previously demonstrated that protein kinase C (PKC)- alpha expression is significantly elevated in failing human left ventricle, with immunostaining showing increased PKC- alpha localization at the intercalated disks of cardiomyocytes. In the present study we sought to determine, in the failing heart, if PKC- alpha interacted with connexin-43 (Cx-43) both spatially and functionally, and to compare the association of PKC- alpha/Cx-43 with that of PKC- epsilon, a PKC isozyme that does not significantly increase in failing hearts. The possibility of a PKC- alpha or PKC- epsilon/Cx-43 association in non-failing hearts was also investigated. Co-immunoprecipitation of PKC- alpha or PKC- epsilon and Cx-43 in non-failing and failing left ventricle was achieved using antibodies to PKC- alpha or Cx-43. Confocal microscopy confirmed that PKC- alpha distribution within the cardiomyocyte included co-localization with connexin-43 in both failing and non-failing myocardium. In a similar manner, confocal imaging of PKC- epsilon showed cardiomyocyte distribution in both cytosol and membrane, and colocalization of PKC- epsilon with Cx-43. Recombinant PKC- alpha or - epsilon increased PKC activity significantly above endogenous levels in the co-immunoprecipitated Cx-43 complexes (P<0.05). However, phosphorylation of purified human Cx-43 (isolated from failing human left ventricle) by recombinant PKC- alpha or PKC- epsilon resulted in only PKC- epsilon mediated Cx-43 phosphorylation. Thus, in the human heart PKC- alpha, PKC- epsilon, and Cx-43 appear to form a closely associated complex. Whereas only PKC- epsilon directly phosphorylates Cx-43, both PKC isoforms result in increased phosphorylation within the Cx-43 co-immunoprecipitated complex.

Background: Increased expression of Ca2+-sensitive protein kinase C (PKC) isoforms may be important markers of heart failure. Our aim was to determine the relative expression of PKC-beta1, -beta2, and -alpha in failed and nonfailed myocardium., Methods and Results: Explanted hearts of patients in whom dilated cardiomyopathy or ischemic cardiomyopathy was diagnosed were examined for PKC isoform content by Western blot, immunohistochemistry, enzymatic activity, and in situ hybridization and compared with nonfailed left ventricle. Quantitative immunoblotting revealed significant increases of >40% in PKC-beta1 (P<0.05) and -beta2 (P<0.04) membrane expression in failed hearts compared with nonfailed; PKC-alpha expression was significantly elevated by 70% in membrane fractions (P<0.03). PKC-epsilon expression was not significantly changed. In failed left ventricle, PKC-beta1 and -beta2 immunostaining was intense throughout myocytes, compared with slight, scattered staining in nonfailed myocytes. PKC-alpha immunostaining was also more evident in cardiomyocytes from failed hearts with staining primarily localized to intercalated disks. In situ hybridization revealed increased PKC-beta1 and -beta2 mRNA expression in cardiomyocytes of failed heart tissue. PKC activity was significantly increased in membrane fractions from failed hearts compared with nonfailed (1021+/-189 versus 261+/-89 pmol. mg-1. min-1, P<0.01). LY333531, a selective PKC-beta inhibitor, significantly decreased PKC activity in membrane fractions from failed hearts by 209 pmol. min-1. mg-1 (versus 42.5 pmol. min-1. mg-1 in nonfailed, P<0.04), indicating a greater contribution of PKC-beta to total PKC activity in failed hearts., Conclusions: In failed human heart, PKC-beta1 and -beta2 expression and contribution to total PKC activity are significantly increased. This may signal a role for Ca2+-sensitive PKC isoforms in cardiac mechanisms involved in heart failure.

Ethinyl estradiol (EE2), administered orally to ovariectomized (ovex) rats, has been shown to prevent loss of bone mineral density and to decrease serum cholesterol levels. Radioligand binding studies with the dihydropyridine (DHP) [3H]PN200-110 were undertaken to characterize calcium (Ca++) channels in cardiac and aortic tissues from ovex rats treated for 35 days with EE2 (0.1 mg/kg day p.o.) or vehicle, and from vehicle-treated sham-operated controls (sham). Cardiac tissues from EE2-treated rats displayed significant increases in the density (Bmax) of high-affinity DHP binding sites (505 +/- 46 fmol/mg) compared with vehicle-treated ovex rats (303 +/- 35 fmol/mg); DHP Bmax values from EE2-treated cardiac tissues were not significantly different from vehicle-treated shams (385 +/- 76 fmol/mg). Cardiac Ca++ efflux channels from sarcoplasmic reticulum were assessed with [3H]ryanodine. [3H]Ryanodine Bmax values were not affected by EE2 treatment. However, [3H]ryanodine Kd values in preparations from EE2-treated rats were significantly decreased (10 +/- 2 nM) compared with ovex rats (35 +/- 11 nM) and were similar to values in sham rats (8 +/- 2 nM). Cardiac beta adrenoceptors were not affected by EE2 treatment, which thus confirmed the selective regulation of DHP receptors by EE2. Aortic preparations from EE2-treated rats exhibited significant increases in DHP receptors (125 +/- 37 fmol/mg) compared with both vehicle-treated ovex rats (32 +/- 3 fmol/mg) and vehicle-treated shams (24 +/- 9 fmol/mg). There were no differences in the binding affinity (Kd) of [3H]PN200-110 for cardiac or aortic sites among the three groups. To ascertain if EE2-mediated increases in Ca++ channel density and ryanodine binding affinity affected in vivo responses to agonists that use extracellular and intracellular Ca++ stores, responses to BAY k 8644 and norepinephrine were examined in pithed rats from the same three groups. No significant differences in hemodynamic responses occurred among the three groups to BAY k 8644 or norepinephrine. Thus, in female ovex rats, prolonged treatment with EE2 resulted in increased density of cardiac and aortic calcium channels which did not translate into increased calcium-mediated inotropic, rate or pressor responses. Conversely, EE2 treatment in ovex rats prevented the decrease in cardiac [3H]ryanodine binding affinity evident in vehicle-treated ovex rats. These data suggest that EE2 treatment in normotensive ovex rats resulted in modulation of both the L-type and sarcoplasmic reticulum Ca++ channels, and these alterations maintained cardiovascular homeostasis.

An 125I-iodinated ryanodine analog, modified by attaching an iodo-Cbz-beta-alanyl group to the C10eq hydroxy of ryanodine (iodo-carbobenzyloxy-beta-alanyl-ryanodine), binds to cardiac sarcoplasmic reticulum Ca2+ release channels with equal affinity as [3H]ryanodine. In the present study, both iodo-Cbz-beta-alanyl-ryanodine and ryanodine bound to canine cardiac microsomal membrane preparations in a Ca2+ dependent manner. At 10 microM free Ca2+ doxorubicin increased specific binding of both ligands, with doxorubicin concentrations of 4.06 +/- 0.44 and 6.22 +/- 1.31 microM inducing 50% maximal enhancement of binding for ryanodine and iodo-Cbz-beta-alanyl-ryanodine, respectively. Effects of ryanodine and iodo-Cbz-beta-alanyl-ryanodine +/- doxorubicin in vitro on cardiac sarcoplasmic reticulum Ca2+ release were compared indirectly by determining Ca2+ accumulation in cardiac microsomal vesicles loaded with 45Ca2+. In the absence of oxalate, neither ryanodine nor iodo-Cbz-beta-alanyl-ryanodine (10 microM) decreased net Ca2+ uptake, whereas doxorubicin reduced Ca2+ accumulation 20 +/- 2%. In the presence of oxalate and 0.4 microM free Ca2+ ("low"), both ryanodine and iodo-Cbz-beta-alanyl-ryanodine modestly decreased (by 19% and 17% at 10 nM, respectively) maximum Ca2+ accumulation. Increasing concentrations of ryanodine (100 nM-100 microM) and iodo-Cbz-beta-alanyl-ryanodine (100 nM-30 microM) had no greater effect, but 100 microM iodo-Cbz-beta-alanyl- ryanodine decreased net Ca2+ uptake 57 +/- 3%. Doxorubicin (30 microM) alone reduced Ca2+ uptake 36%; its effects with 1 nM-10 microM ryanodine or 1 nM-100 microM iodo-Cbz-beta-alanyl-ryanodine were additive.(ABSTRACT TRUNCATED AT 250 WORDS)

Thromboxane A2 (TXA2) and prostaglandin H2 (PGH2) induce platelet aggregation and are potent vasoconstrictors, and they have been implicated in coronary vasospasm and myocardial infarction. The TXA2 mimetic [1S-(1 alpha, 2 beta (5Z), 3 alpha (1E,3S*), 4 alpha)]-7-[3-(3-hydroxy-4-(4'- iodophenoxy)-1-butenyl)-7-oxabicyclo-[2.2.1]heptan-2-yl]-5-h eptenoic acid (IBOP) was used to characterize binding to microsomal membrane preparations from saline-perfused guinea pig atria (GPA) and ventricles (GPV). [125I]IBOP bound to GPA and GPV in a protein-dependent and saturable manner, although total binding was two-fold greater and non-specific binding was proportionately less in GPA compared to GPV. Analysis of equilibrium binding data indicated one class of binding sites in both GPA and GPV with Kd values of 333 +/- 117 and 645 +/- 187 pM, respectively, which were in close agreement with kinetically determined Kd values of 226 and 882 pM, respectively. Bmax values of GPA and GPV of 57 +/- 5.6 and 24 +/- 4.3 fmol/mg protein were significantly different (P < 0.01). Ki values (from IC50s) were determined for various TXA2/PGH2 analogues and prostaglandins in competition binding assays with [125I]IBOP. The rank order for ability to inhibit binding in GPA was U46619 = SQ29548 > I-PTA-OH > PGF2 alpha = PGE2. In GPV, the rank order was U46619 = SQ29548 > PGF2 alpha = I-PTA-OH = PGE2. [125I]IBOP binding to GPA and GPV was completely displaced by the TXA2/PGH2 agonist U46619 and by the TXA2/PGH2 antagonist SQ29548.(ABSTRACT TRUNCATED AT 250 WORDS)

Cardiovascular effects of S-dobutamine were compared with effects of vehicle and other catecholamines in dogs during and after 3 days of approximately 90% ligation of the left anterior descending coronary artery (LAD). Twenty-four hours after LAD ligation, dogs infused with S-dobutamine (2.5 micrograms/kg/min intravenously, i.v.) maintained systolic blood pressure (SBP 149 +/- 6 mm Hg), diastolic blood pressure (DBP 100 +/- 6 mm Hg), and aortic dP/dt60 (2.8 +/- 0.2 s-1), with no significant changes from preligation values. In comparison, saline-treated dogs showed decreases in arterial BP and contractility: SBP 121 +/- 4 mm Hg; DBP 85 +/- 3 mm Hg; and aortic dP/dt60 was 1.9 +/- 0.1 s-1. S-Dobutamine-infused dogs had a heart rate (HR) of 148 +/- 5 beats/min with 44 +/- 14 beats/min premature ventricular contractions (PVCs), whereas dogs infused with saline, R-dobutamine, dopamine, norepinephrine (NE), or isoproterenol (ISO) all displayed a significantly greater number of PVCs at 24 h. Myocardial necrosis was limited by S-dobutamine treatment (2.5 micrograms/kg/min i.v. for 54 h). As demonstrated by histologic examination, S-dobutamine ameliorated the effects of ischemia as compared with vehicle, R-dobutamine, dopamine, hexamethonium, NE, or ISO. Myocardial tissue electrolytes, quantified 72 h after LAD ligation, were maintained by S-dobutamine-infused dogs in all sections of left ventricle (LV); but in saline-treated dogs, Ca2+ increased eightfold, Na+ increased twofold, and both K+ and Mg2+ decreased 50% in tissue "at risk" as compared with tissues "not at risk." Coronary nutrient blood flow (CNBF) to myocardial capillary vessels was calculated by radiolabeled microspheres 2 h after LAD ligation. As compared with CNBF in untreated hearts, endocardial CNBF in hearts receiving S-dobutamine (5 micrograms/kg/min i.v.) increased from 26 +/- 8 to 49 +/- 15 ml/min/100 g in tissue at risk, from 102 +/- 26 to 217 +/- 50 in "border zone," and from 133 +/- 13 to 215 +/- 41 in tissue not at risk. CNBF values in animals receiving vehicle infusion were not significantly different from CNBF values measured after ligation only. The S-enantiomer of dobutamine, infused in dogs for 54 h after coronary artery ligation, maintained cardiac performance, electrolyte balance, and myocardial cellular viability and reduced incidences of arrhythmias through its ability to increase CNBF without increasing HR.

The synthesis of a novel radioiodinated ryanodine-O10eq-N-acylamino acylate along with biological data are reported. The affinity of the iodinated product, 7, was comparable to ryanodine, 7.97 nM and 6.47 nM, respectively. Conversion of the non-radioactive iodinated ryanodine analog to the [125I] isotope was accomplished by conversion of 7 to the trimethyltin derivative followed by [125I] exchange using chloramine-T in organic solvent. The radioiodinated ryanodine analog, 9, bound to cardiac membrane preparations in a protein dependent and saturable manner indicating that this analog may represent a useful new tool for the study of ryanodine receptors and that modifications about the C-10 hydroxy group of ryanodine may be carried out without loss in biological activity.

The selective contractile effects of s-dobutamine were studied in vitro in selected canine arteries and vein preparations; propranolol was included to block potential beta-mediated vasodilation. These in vitro data were expanded by quantifying the in vivo effects of s-dobutamine on venous blood return and redistribution of regional nutrient blood flow (NBF) and non-nutrient blood flow (non-NBF) in anesthetized dogs. In in vitro studies with isolated canine arteries and veins, s-dobutamine exhibited vein-selective constriction. At maximally efficacious concentrations of agonist, contractions of carotid, coronary, and femoral arteries in response to s-dobutamine were only 7, 25 and 45% as great as those elicited by norepinephrine (NE). Similarly, in jugular vein, s-dobutamine-mediated contractions were 55% as great as those obtained in response to NE. Coronary and femoral arteries precontracted with NE were relaxed in a dose-related manner by increasing concentrations of s-dobutamine. Effects of NE and s-dobutamine on venous blood return (VR) were compared in dogs. s-Dobutamine increased VR by 49 +/- 10 ml, whereas NE increased VR by 14 +/- 6 ml during 5-min infusion. s-Dobutamine significantly increased coronary NBF in left ventricular (LV) endocardium from 115 +/- 10 to 194 +/- 13 and 263 +/- 9 ml/min/100 g at doses of 10 and 20 micrograms/kg/min, respectively. In addition, LV epicardium flow was increased from 87 +/- 8 to 189 +/- 15 and 262 +/- 11 ml/min/100 g at 10 and 20 micrograms/kg/min, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

Tibenelast (LY186655), 5,6,-diethoxybenzo(b)thiophene-2-carboxylic acid, sodium salt, is an orally active anti-anaphylactic compound in guinea pigs, and has been shown to prevent bronchospasm in moderately severe asthmatic patients. Pharmacological studies with tibenelast demonstrated that it is a selective phosphodiesterase (PDE) inhibitor in that it is moderately active against the lung and stomach enzyme while being a very weak inhibitor of the heart enzyme. The compound was without cardiovascular effects at anti-anaphylactic doses. In contrast to theophylline, tibenelast did not have a direct inotropic effect in the cat papillary muscle system. The concentration that inhibited 50% of the enzymatic activity (IC50) for tibenelast was 20- to 30-fold lower for neutrophil PDE than for PDE of other tissues. It was 100 times more potent than aminophylline in inhibiting superoxide generation from platelet-activating factor (PAF)-primed polymorphonuclear leukocytes (PMNL) challenged with chemotactic factor, N-formyl-methionyl-leucyl-phenylalanine. However, tibenelast was less effective in the tumor necrosis factor-primed system, and did not inhibit superoxide generation during phagocytosis or when other soluble stimuli, such as phorbo-12-myristate-13-acetate or the calcium ionophore A23187, were used. Furthermore, tibenelast did not inhibit enzymes involved in arachidonic acid metabolism. These results suggest that tibenelast probably inhibits superoxide release from PMNL via a selective inhibition on PDE.

Effects of prolonged isoproterenol infusion upon the density of cardiac calcium channels, calcium-mediated contractile responses, and the ability of forskolin to enhance tension development and cyclic AMP accumulation were studied in ventricular muscle preparations from Sprague-Dawley rats. Isoproterenol infusion (400 micrograms/kg per h s.c., 4 days) significantly decreased calcium channel density (Bmax) in cardiac microsomal membranes as quantified by a 32% decrease in specific [3H]nitrendipine binding sites; binding affinity (KD) was unchanged. A 57% decrease of beta-adrenoceptors confirmed homologous down regulation. To examine functional effects of decreased [3H]nitrendipine binding sites, responses to calcium, BAY K8644 and nifedipine were determined in isolated right ventricular strips. Significant decreases in basal developed tension were observed in muscles from isoproterenol-infused rats. However, concentration-dependent increases in contractility in response to CaCl2 or BAY K8644 were comparable, and the negative inotropic effect of nifedipine was unchanged. Whereas isoproterenol infusion was associated with significantly decreased basal cardiac cyclic AMP concentrations, exposure of ventricular strips from either vehicle- or isoproterenol-infused rats to 10 microM forskolin resulted in comparable increases in cyclic AMP and in developed tension. Cumulative, submaximal concentrations of forskolin also produced similar increases in contractility with maximum responses in ventricular strips from vehicle-infused animals attained at 4.4 microM forskolin. Higher concentrations resulted in automaticity. By contrast, ventricle from isoproterenol-infused animals responded to 14.4 microM forskolin with maximal increases in force of contraction.

A variety of lipoxygenase products such as 12- and 15-hydroxyeicosatetraenoic acid (12- and 15-HETE) inhibit thromboxane A2 (TXA2) mimetic induced human platelet aggregation in a stereoselective manner. The mechanism of this inhibition remains unclear. To determine if this inhibition is due to a receptor level interaction of the lipoxygenase products at the thromboxane A2/prostaglandin H2 (TXA2/PGH2) receptor, radioligand binding studies were performed using a new [125I]-labelled thromboxane mimetic [125I]BOP. The mono-HETES 5(S), 12(R), 12(S) and 15(S) inhibited binding of the radioligand to the TXA2/PGH2 receptor in washed human platelets with IC50 values of greater than 25, 0.73, 2.06 and 2.0 microM respectively. LTB4 and its positional isomer 5(S), 12(S)-diHETE were less potent with IC50 values greater than 10 microM for LTB4 and 9.38 microM for 5(S), 12(S)-diHETE. Thus, stereoselective inhibition of the binding of the radioligand was demonstrated between 12(R)- and 12(S)-HETE. These lipoxygenase products also inhibited IBOP (10nM) induced platelet aggregation in a concentration dependent fashion with a similar rank order of potency as that obtained in the competition binding assay. These results suggest that, at least in part, the platelet inhibitory properties of these HETEs may be mediated through their interaction at the TXA2/PGH2 receptor.

Compound LY195115 is a novel cardiotonic with both inotropic and vasodilator activities. In cat papillary muscles, LY195115 increased contractility in a concentration-dependent manner; its actions were not blocked by either prazosin or propranolol. An intravenous dose of 7.0 micrograms/kg LY195115 resulted in a 50% increase in contractility in anesthetized dogs; comparable inotropic responses were observed in anesthetized cats receiving 10 micrograms/kg i.v. These doses of LY195115 increased heart rates of both dogs and cats by less than 10%. Oral administration of 25 micrograms/kg to conscious dogs was associated with a selective inotropic response that was maximal at 3 h and maintained in excess of 23 h. This effect was not accompanied by gross behavioral changes or emesis. The hemodynamic profile of LY195115 was evaluated in anesthetized beagle dogs. A 60-min infusion of 1.0 microgram/kg/min LY195115 followed by a 5-min infusion of 10 micrograms/kg/min resulted in dose-dependent increases in contractility (LV dP/dt60) and heart rate; doses that increased LV dP/dt60 by 50% increased heart rate by less than 10%. Doses of greater than 5.0 micrograms/kg decreased left ventricular end-diastolic pressure and systemic vascular resistance; mean arterial blood pressure and cardiac output were unchanged. Estimated myocardial oxygen consumption (heart rate times either systolic or mean arterial blood pressure) was not altered by doses as high as 110 micrograms/kg. This balance of inotropic/vasodilator activities may provide a means of improving cardiac function while maintaining myocardial oxygen supply/demand.

Isoproterenol increased contractility in isolated cat papillary muscles 2-fold with an EC50 of 6.3 X 10(-8) M. Nifedipine (3 X 10(-7) M) reduced contractility in control muscles by 43%; however, inotropic state was restored by isoproterenol with a comparable EC50 of 5 X 10(-8) M. To test the hypothesis that this effect might result from cAMP-dependent phosphorylation of a Ca2+ channel-associated protein, [3H]nitrendipine binding was used to probe the high-affinity 1,4-dihydropyridine site in either phosphorylated or dephosphorylated sarcolemmal vesicles. Kd and Bmax values for binding to phosphorylated sarcolemmal vesicles (0.14 +/- 0.027 nM and 479 +/- 62 fmol/mg protein, respectively) were not significantly different from control values P greater than 0.4). Similarly, dephosphorylation of sarcolemmal vesicles did not alter binding parameters. These data demonstrate that phosphorylation of sarcolemmal vesicles neither alters the binding affinity for [3H]nitrendipine nor promotes an interconversion of dihydropyridine-binding sites from high to low affinity or vice versa. While phosphorylation may regulate the slow Ca2+ channel, this is not reflected as changes in [3H]nitrendipine-binding parameters determined in vitro. Furthermore, the cyclic AMP-dependent phosphorylation state of sarcolemmal proteins does not appear to account for wide variations (more than 100-fold) between Kd values from binding studies and IC50 values determined in pharmacological investigations.