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Key points We investigated the role played by lactate and hydrogen in evoking the exercise pressor reflex (EPR) in decerebrated rats whose hindlimb muscles were either freely perfused or ischaemic. Production of lactate and hydrogen by the contracting hindlimb muscles was manipulated by knocking out the myophosphorylase gene (pygm). In knockout rats (pygm−/−; n = 13) or wild‐type rats (pygm+/+; n = 13), the EPR was evoked by isometrically contracting the triceps surae muscles. Blood pressure, tension, blood flow, renal sympathetic nerve activity and blood lactate concentrations were measured. Intramuscular metabolites and pH changes induced by the contractions were quantified by 31P‐magnetic resonance spectroscopy (n = 5). In a subset of pygm−/− rats (n = 5), contractions were evoked with prior infusion of lactate (pH 6.0) in an attempt to restore the effect of lactate and hydrogen ions. Contraction of freely perfused muscles increased blood lactate and decreased muscle pH in pygm+/+ rats only. Despite these differences, the reflex pressor and sympathetic responses to freely perfused contraction did not differ between groups (P = 0.992). During ischaemia, contraction increased muscle lactate and hydrogen ion production in pygm+/+ rats (P < 0.0134), whereas it had no effect in pygm−/− rats (P > 0.783). Likewise, ischaemia exaggerated the reflex pressor, and sympathetic responses to contraction in pygm+/+ but not in pygm−/− rats. This exaggeration was restored when a solution of lactate (pH 6.0) was infused prior to the contraction in pygm−/− rats. We conclude that lactate and hydrogen accumulation in contracting myocytes play a key role in evoking the metabolic component of the EPR during ischaemic but not during freely perfused contractions. Conflicting results exist about the role played by lactate and hydrogen ions in evoking the exercise pressor reflex. Using CRISP‐Cas9, we rendered the myophosphorylase gene non‐functional to block the production of lactate and hydrogen ions. The exercise pressor reflex was evoked in decerebrated rats by statically contracting the triceps surae muscles with or without muscle ischaemia. Static contraction elevated the concentration of lactate and hydrogen ions in pygm+/+ but not in pygm−/− rats. Despite these differences, the exercise pressor reflex was not different between groups. Acute muscle ischaemia exaggerated the concentration of lactate and hydrogen ions in pygm+/+ but not in pygm−/− rats. Likewise, acute muscle ischaemia exaggerated the exercise pressor reflex in pygm+/+ but not in pygm−/− rats. We conclude that lactate and hydrogen play a key role in evoking the exercise pressor reflex during ischaemic but not during freely perfused contractions. [ABSTRACT FROM AUTHOR]

The role played by the transient receptor potential vanilloid 1 (TRPV1) channel on the thin fibre afferents evoking the exercise pressor reflex is controversial. To shed light on this controversy, we compared the exercise pressor reflex between newly developed TRPV1+/+, TRPV1+/− and TRPV1−/− rats. Carotid arterial injection of capsaicin (0.5 μg), evoked significant pressor responses in TRPV1+/+ and TRPV1+/− rats, but not in TRPV1−/− rats. In acutely isolated dorsal root ganglion neurons innervating the gastrocnemius muscles, capsaicin evoked inward currents in neurons isolated from TRPV1+/+ and TRPV1+/− rats but not in neurons isolated from TRPV1−/− rats. The reflex was evoked by stimulating the tibial nerve in decerebrated rats whose femoral artery was either freely perfused or occluded. We found no difference between the reflex in the three groups of rats regardless of the patency of the femoral artery. For example, the peak pressor responses to contraction in TRPV1+/+, TRPV1+/− and TRPV1−/− rats with patent femoral arteries averaged 17.1 ± 7.2, 18.9 ± 12.4 and 18.4 ± 8.6 mmHg, respectively. Stimulation of the tibial nerve after paralysis with pancuronium had no effect on arterial pressure, findings which indicated that the pressor responses to contraction were not caused by electrical stimulation of afferent tibial nerve axons. We also found that expression levels of acid‐sensing ion channel 1 and endoperoxide 4 receptor in the L4 and 5 dorsal root ganglia were not upregulated in the TRPV1−/− rats. We conclude that TRPV1 is not needed to evoke the exercise pressor reflex in rats whose contracting muscles have either a patent or an occluded arterial blood supply. Key points: A reflex arising in contracting skeletal muscle contributes to the increases in arterial blood pressure, cardiac output and breathing evoked by exercise.The sensory arm of the reflex comprises both mechanoreceptors and metaboreceptors, of which the latter signals that blood flow to exercising muscle is not meeting its metabolic demand.The nature of the channel on the metaboreceptor sensing a mismatch between supply and demand is controversial; some believe that it is the transient receptor potential vanilloid 1 (TRPV1) channel.Using genetically engineered rats in which the TRPV1 channel is rendered non‐functional, we have shown that it is not needed to evoke the metaboreflex. [ABSTRACT FROM AUTHOR]

Tubal ligation keeps the fimbriated end of the fallopian tube intact while interrupting the conduit for sperm and egg between the uterus and ovary. Tubal ligation is associated with an approximately 20% decreased risk of high-grade serous ovarian cancers, which mounting evidence suggests arise from the distal fallopian tube epithelium. We postulated that biological changes at the epithelial cellular level of the distal fallopian tube may account for the surgical procedure’s observed risk reduction. We compared the histology, presence of epithelial progenitors (basally located CD44-positive cells), and degree of epithelial proliferation (Ki67-positive cells) of distal fallopian tube from 10 patients with previous tubal ligation and 10 age-matched patients with uncut fallopian tubes. A significantly reduced population of proliferating epithelial progenitors (basally located CD44/Ki67 dual-positive cells) was detected in the tubal ligated specimens (P = .0002). To functionally assess the effect of tubal ligation, a murine model was utilized to compare the growth capacity of distal fallopian tube epithelial cells isolated from either ligated or sham-operated tubal epithelia. Murine fallopian tube epithelial cells isolated after tubal ligation showed a significantly reduced capacity to grow organoids in culture compared to sham-operated controls (P = .002). The findings of this study show that tubal ligation is associated with a reduced presence and decreased proliferation of progenitor cells in the distal fallopian tube epithelium. These compositional and functional changes suggest that tubal ligation induces quiescence of distal fallopian tube epithelial cells. [ABSTRACT FROM AUTHOR]

In preclinical rodent models, spinal cord injury (SCI) manifests as gastric vagal afferent dysfunction both acutely and chronically. However, the mechanism that underlies this dysfunction remains unknown. In the current study, we examined the effect of SCI on gastric nodose ganglia (NG) neuron excitability and on voltage-gated Na+ (Na V) channels expression and function in rats after an acute (i.e. 3-days) and chronic (i.e. 3-weeks) period. Rats randomly received either T3-SCI or sham control surgery 3-days or 3-weeks prior to experimentation as well as injections of 3% DiI solution into the stomach to identify gastric NG neurons. Single cell qRT-PCR was performed on acutely dissociated DiI-labeled NG neurons to measure Na V 1.7, Na V 1.8 and Na V 1.9 expression levels. The results indicate that all 3 channel subtypes decreased. Current- and voltage-clamp whole-cell patch-clamp recordings were performed on acutely dissociated DiI-labeled NG neurons to measure active and passive properties of C- and A-fibers as well as the biophysical characteristics of Na V 1.8 channels in gastric NG neurons. Acute and chronic SCI did not demonstrate deleterious effects on either passive properties of dissociated gastric NG neurons or biophysical properties of Na V 1.8. These findings suggest that although Na V gene expression levels change following SCI, Na V 1.8 function is not altered. The disruption throughout the entirety of the vagal afferent neuron has yet to be investigated. • SCI decreased the gene expression of Na V 1.7, Na V 1.8 and Na V 1.9 channels of rat gastric nodose ganglion (NG) neurons. • Downregulation of Na V channels within gastric vagal afferents may explain the reduction in vagal afferent sensitivity after SCI. • SCI did not impair either Na V 1.8 biophysical properties or the passive membrane properties of gastric NG neurons. • Further studies are necessary to identify the mechanisms responsible for diminished gastric vagal afferent signaling in vivo. [ABSTRACT FROM AUTHOR]

Abstract: The direct impact of solar activity on climate has been widely studied through Total Solar Irradiance (TSI). Biological processes also impact climate and are deeply affected by TSI. Marine phytoplankton emissions into the atmosphere have been proposed to change cloud albedo through cloud formation. In this work, we use wavelet analysis to investigate the decadal relation between high-latitude concentrations of methane sulphonic acid, a product of seawater algae, and TSI. We found that some of the methane sulphonic acid main periodicities coincide with periods of solar activity periods. [Copyright &y& Elsevier]

Abstract: The sphenopalatine ganglion (SPG) is the main parasympathetic ganglion that is involved in regulating cerebral vascular tone and gland secretion. SPG neurons have been implicated in some types of migraine headaches but their precise role has yet to be determined. In addition, very little information is available regarding ion channel modulation by neurotransmitters that are involved in the parasympathetic drive of SPG neurons. In this study, acute isolation of adult rat SPG neurons was developed in order to begin the electrophysiological characterization of this ganglion. Under our dissociation conditions, the average number of neurons obtained per ganglion was greater than 1200. Immunofluorescence imaging results showed positive labeling with acethylcholinesterase (AChE), confirming the parasympathetic nature of SPG neurons. On the other hand, weak tyrosine hydroxylase immunostaining was observed in these neurons. Whole-cell patch-clamp recordings revealed that most of the Ca2+ current is carried by N-type (53%) and SNX-482 resistant R-type (30%) Ca2+ channels. In addition, Ca2+ currents were inhibited in a voltage-dependent manner following exposure to oxotremorine-M (Oxo-M), norepinephrine and ATP via muscarinic acetylcholine receptor 2 (M2 AChR) subtype, adrenergic and P2Y purinergic receptors, respectively. The peptides VIP and angiotensin II failed to modulate Ca2+ currents, suggesting that these receptors are not present on the SPG soma or do not couple to Ca2+ channels. In summary, our data suggest that the Ca2+ current inhibition mediated by Oxo-M, NE and ATP in adult rat SPG neurons plays an integral part in maintaining parasympathetic control of cranial functions. [Copyright &y& Elsevier]

A catalog containing an unprecedented amount of historical data in the southeastern part of Mexico covering almost four centuries (1502–1899) is used to construct a drought time series. The catalog records information of agricultural disasters and includes events associated with hydrometeorological phenomena or hazards whose effects were mainly felt in the agricultural sector, such as droughts. An analysis of the historical series of droughts in southeastern Mexico for the period 1502–1899 is performed. The highest drought frequency occurred around the years 1650, 1782, and 1884; no droughts were reported around 1540, between 1630 and 1640, along the largest time lapse of 1672–1714, and between 1740 and 1760. From 1760 until the end of the period of study droughts definitively occur more often than they did from ∼1550 to 1760. In addition, most droughts lasted for 1–2 yr. Analyzing the frequencies of the drought time series it is found that the most conspicuous cycles are ∼3–4 and 7 yr, although cycles of ∼12, 20, 43, and 70 yr are also evident. The relation between droughts and El Niño events indicates that 38% of droughts are associated with El Niño. Sea surface temperature changes, the Southern Oscillation index, and solar activity leave their signals in the southeastern part of Mexico, with the signs in Oaxaca clearer than in the Yucatan Peninsula. However, the dominance of some phenomena over others depends on the time scales considered. [ABSTRACT FROM AUTHOR]

Introduction: Because of the strong correlation between the blood concentration of circulating resistin and the illness severity of septic patients, resistin has been proposed as a mediator of sepsis pathophysiology. In vitro data indicate that human resistin directly impairs neutrophil migration and intracellular bacterial killing, although the significance of these findings in vivo remain unclear. Objective: The objectives of the present study were: (1) to validate the expression of human resistin in a clinically relevant, murine model of surgical sepsis, (2) to assess how sepsis-induced changes in resistin correlate with markers of infection and organ dysfunction, and (3) to investigate whether the expression of human resistin alters immune function or disease outcomes in vivo. Methods: 107 male, C57BL/6 mice transgenic for the human resistin gene and its promoter elements (Retn+/−/−, or Retn+) were generated on a Retn−/− (mouse resistin knockout, or Rko) background. Outcomes were compared between age-matched transgenic and knockout mice. Acute sepsis was defined as the initial 24 h following cecal ligation and puncture (CLP). Physiologic and laboratory parameters correlating to the human Sequential Organ Failure Assessment (SOFA) Score were measured in mice, and innate immune cell number/function in the blood and peritoneal cavity were assessed. Results: CLP significantly increased circulating levels of human resistin. The severity of sepsis-induced leukopenia was comparable between Retn+ and Rko mice. Resistin was associated with increased production of neutrophil reactive oxygen species, a decrease in circulating neutrophils at 6 h and an increase in peritoneal Ly6Chi monocytes at 6 h and 24 h post-sepsis. However, intraperitoneal bacterial growth, organ dysfunction and mouse survival did not differ with resistin production in septic mice. Significance: Ex vivo resistin-induced impairment of neutrophil function do not appear to translate to increased sepsis severity or poorer outcomes in vivo following CLP. [ABSTRACT FROM AUTHOR]

Background: Silent inflammatory bowel disease (IBD) is a condition in which individuals with the active disease experience minor to no pain. Voltage‐gated Na+ (NaV) channels expressed in sensory neurons play a major role in pain perception. Previously, we reported that a NaV1.8 genetic polymorphism (A1073V, rs6795970) was more common in a cohort of silent IBD patients. The expression of this variant (1073V) in rat sympathetic neurons activated at more depolarized potentials when compared to the more common variant (1073A). In this study, we investigated whether expression of either NaV1.8 variant in rat sensory neurons would exhibit different biophysical characteristics than previously observed in sympathetic neurons. Methods: Endogenous NaV1.8 channels were first silenced in DRG neurons and then either 1073A or 1073V human NaV1.8 cDNA constructs were transfected. NaV1.8 currents were recorded with the whole‐cell patch‐clamp technique. Key Results: The results indicate that 1073A and 1073V NaV1.8 channels exhibited similar activation values. However, the slope factor (k) for activation determined for this same group of neurons decreased by 5 mV, suggesting an increase in voltage sensitivity. Comparison of inactivation parameters indicated that 1073V channels were shifted to more depolarized potentials than 1073A‐expressing neurons, imparting a proexcitatory characteristic. Conclusions and Inferences: These findings differ from previous observations in other expression models and underscore the challenges with heterologous expression systems. Therefore, the use of human sensory neurons derived from induced pluripotent stem cells may help address these inconsistencies and better determine the effect of the polymorphism present in IBD patients. [ABSTRACT FROM AUTHOR]

Key points We determined the role played by the transient receptor potential canonical 6 (TRPC6) channel in evoking the mechanical component of the exercise pressor reflex in male decerebrated Sprague–Dawley rats. TRPC6 channels were identified by quadruple‐labelled (DiI, TRPC6, neurofilament‐200 and peripherin) immunohistochemistry in dorsal root ganglion (DRG) cells innervating the triceps surae muscles (n = 12). The exercise pressor reflex was evoked by statically contracting the triceps surae muscles before and after injection of the TRPC6 antagonist BI‐749327 (n = 11; 12 μg kg−1) or SAR7334 (n = 11; 7 μg kg−1) or the TRPC6 positive modulator C20 (n = 11; 18 μg kg−1). Similar experiments were conducted while the muscles were passively stretched (n = 8–12), a manoeuvre that isolated the mechanical component of the reflex. Blood pressure, tension, renal sympathetic nerve activity (RSNA) and blood flow were recorded. Of the DRG cells innervating the triceps surae muscles, 85% stained positive for the TRPC6 antigen, and 45% of those cells co‐expressed neurofilament‐200. Both TRPC6 antagonists decreased the reflex pressor responses to static contraction (−32 to −42%; P < 0.05) and to passive stretch (−35 to −52%; P < 0.05), whereas C20 increased these responses (55–65%; P < 0.05). In addition, BI‐749327 decreased the peak and integrated RSNA responses to both static contraction (−39 to −43%; P < 0.05) and passive stretch (−56 to −62%; P < 0.05), whereas C20 increased the RSNA to passive stretch only. The onset latency of the decrease or increase in RSNA occurred within 2 s of the onset of the manoeuvres (P < 0.05). Collectively, our results show that TRPC6 plays a key role in evoking the mechanical component of the exercise pressor reflex. The exercise pressor reflex plays a key role in the sympathetic and haemodynamic responses to exercise. This reflex is composed of two components, namely the mechanoreflex and the metaboreflex. The receptors responsible for evoking the mechanoreflex are poorly documented. A good candidate for this function is the transient receptor potential canonical 6 (TRPC6) channel, which is activated by mechanical stimuli and expressed in dorsal root ganglia of rats. Using two TRPC6 antagonists and one positive modulator, we investigated the role played by TRPC6 in evoking the mechanoreflex in decerebrated rats. Blocking TRPC6 decreased the renal sympathetic and the pressor responses to both contraction and stretch, the latter being a manoeuvre that isolates the mechanoreflex. In contrast, the positive modulator increased the pressor reflex to contraction and stretch, in addition to the sympathetic response to stretch. Our results provide strong support for a role played by the TRPC6 channel in evoking the mechanoreflex. [ABSTRACT FROM AUTHOR]

The exercise pressor reflex is initiated by the contraction-induced activation of group III and IV muscle afferents. The reflex is manifested by increases in arterial blood pressure and cardiac output, which, in turn, are generated by increases in the sympathetic outflow to the heart and vasculature and decreases in the vagal outflow to the heart. In previous experiments, we used a pharmacological approach to assess the role played by the acid-sensing ion channel 3 (ASIC3) on group III and IV afferents in evoking the exercise pressor reflex. In the present experiments, we used an alternative approach, namely functional knockout (KO) of the ASIC3 gene, to confirm and extend our previous finding that pharmacological blockade of the ASIC3 had only a small impact on the expression of the exercise pressor reflex when the arterial supply to the contracting hindlimb muscles of rats was patent. Using this alternative approach, we compared the magnitude of the exercise pressor reflex evoked in ASIC3 KO rats with that evoked in their wild-type (WT) counterparts. We found both WT and ASIC3 KO rats displayed similar pressor responses to static contraction (WT, n = 10, +12 ± 2 mmHg; KO, n = 9, +11 ± 2 mmHg) and calcaneal tendon stretch (WT, n = 9, +13 ± 2 mmHg; KO, n = 7, +11 ± 2 mmHg). Likewise, both WT and ASIC3 KO displayed similar pressor responses to intra-arterial injection of 12 mM lactic acid (WT, n = 9, +14 ± 3 mmHg; KO, n = 8, +18 ± 5 mmHg), 24 mM lactic acid (WT, n = 9,+24 ± 2 mmHg; KO, n = 8, +20 ± 5 mmHg), capsaicin (WT, n = 9,+27 ± 5 mmHg; KO, n = 10, +29 ± 5 mmHg), and diprotonated phosphate (H2PO4- ; WT, n = 6,+22 ± 3 mmHg; KO, n = 6, +32 ± 6 mmHg). We conclude that redundant receptors are responsible for evoking the pressor reflexes arising from group III and IV afferents. [ABSTRACT FROM AUTHOR]

The nodose ganglion (NG) is the main parasympathetic ganglion conveying sensory signals to the CNS from numerous visceral organs including digestive signals such as gastric distension or the release the gastrointestinal peptides. The response characteristics of NG neurons to ATP and ADP and pharmacological interrogation of purinergic receptor subtypes have been previously investigated but often in NG cells of undetermined visceral origin. In this study, we confirmed the presence of P2X3 and P2Y1 receptors and characterized P2X and P2Y responses in gastric-innervating NG neurons. Application of ATP-evoked large inward currents and cytosolic Ca2+ increases in gastric-innervating NG neurons. Despite the expression of P2Y1 receptors, ADP elicited only minor modulation of voltage-gated Ca2+ channels. Considering the sensitivity of NG neurons to comorbidities associated with disease or neural injury, purinergic modulation of gastric NG neurons in disease- or injury-states is worthy of further investigation. [ABSTRACT FROM AUTHOR]

The impact of rare and damaging variants in genes associated with platelet function in large-vessel ischemic stroke (LVIS) remains unknown. The aim of this study was to investigate the contribution of some of these variants to the genetic susceptibility to LVIS in Polish patients using a deep re-sequencing of 54 selected genes, coding for proteins associated with altered platelet function. Targeted pooled re-sequencing (Illumina HiSeq 2500) was performed on genomic DNA of 500 cases (patients with history of clinically proven diagnosis of LVIS) and 500 age-, smoking status-, and sex-matched controls (no history of any type of stroke), and from the same population as patients with LVIS. After quality control and prioritization based on allele frequency and damaging probability, individual genotyping of all deleterious rare variants was performed in patients from the original cohort, and stratified to concomitant cardiac conditions differing between the study and stroke groups. We demonstrated a statistically significant increase in the number of rare and potentially damaging variants in some of the investigated genes in the LVIS pool (an increase in the genomic variants burden). Furthermore, we identified an association between LVIS and 6 rare functional and damaging variants in the Kv7.1 potassium channel gene (KCNQ1). The predicted functional properties (partial loss-of function) for the three most damaging variants in KCNQ1 coding locus were further confirmed in vitro by analyzing the membrane potential changes in cell lines co-transfected heterogeneously with human muscarinic type 1 receptor and wild-type or mutated KCNQ1 cDNA constructs using fluorescence imaging plate reader. The study demonstrated an increased rare variants burden for 54 genes associated with platelet function, and identified a putative role for rare damaging variants in the KCNQ1 gene on LVIS susceptibility in the Polish population. [ABSTRACT FROM AUTHOR]

All currently employed pharmaceutical formulations of hydroxychloroquine (HCQ) sulfate are a racemate, consisting of equal parts mixture of two stereoisomers: R(−)HCQ and S(+)HCQ sulfates. The aims of the current study were first, to obtain and characterize pure HCQ enantiomers. The separation and purification of free base HCQ enantiomers from the racemate form were performed using semi‐preparative chiral high‐performance liquid chromatography. Second, we compared the pharmacological properties of both optical isomers and racemic mixture on the intracellular Ca2+ oscillations employing an in vitro model of human cardiomyocytes derived from induced pluripotent stem cells (iPSCs). The results of the pharmacological investigations indicate that the racemic and pure stereoisomer forms of HCQ sulfate exhibited a dose‐dependent inhibition of spontaneous Ca2+ oscillations (as measured from their frequency and Ca2+ peak widths) in cardiomyocytes 5–45 min following exposure. In addition, the concentration‐response relationships for all three compounds indicated that the rank order of potency (IC50) was R(−)HCQ >racemic HCQ >S(+)HCQ for the frequency of the Ca2+ oscillations and width of Ca2+ peaks for all time points examined. These studies indicate that both R(−) and S(+) stereoisomers exhibit differing pharmacological actions on hiPSC cardiomyocytes, with the former effecting a greater potency on cell Ca2+ handling. [ABSTRACT FROM AUTHOR]

The contribution of low-frequency and damaging genetic variants associated with platelet function to ischemic stroke (IS) susceptibility remains unknown. We employed a deep re-sequencing approach in Polish patients in order to investigate the contribution of rare variants (minor allele frequency, MAF < 1%) to the IS genetic susceptibility in this population. The genes selected for re-sequencing consisted of 26 genes coding for proteins associated with the surface membrane of platelets. Targeted pooled re-sequencing (Illumina HiSeq 2500) was performed on genomic DNA of 500 cases (patients with history of clinically proven diagnosis of large-vessel IS) and 500 controls. After quality control and prioritization based on allele frequency and damaging probability, follow-up individual genotyping of deleterious rare variants was performed in patients from the original cohort. Gene-based analyses identified an association between IS and 6 rare functional and damaging variants in the purinergic genes (P2RY1 and P2RY12 locus). The predicted properties of the most damaging rare variants in P2RY1 and P2RY12 were confirmed by using mouse fibroblast cell cultures transfected with plasmid constructs containing cDNA of mutated variants (FLIPR on FlexStation3). This study identified a putative role for rare variants in P2RY1 and P2RY12 genes involved in platelet reactivity on large-vessel IS susceptibility in a Polish population. [ABSTRACT FROM AUTHOR]

The nociceptin/orphanin FQ peptide (NOP) opioid receptors regulate neurotransmitter release via inhibition of voltage-gated Ca 2+ channels (Ca V 2.2) in sympathetic and sensory neurons. Stimulation of NOP receptors by its endogenous agonist, nociception (Noc), leads to membrane-delimited, voltage-dependent (VD) block of Ca V 2.2 channel currents mediated by Gβγ protein subunits. Previously we reported that the pertussis toxin-sensitive Gαi1 and Gβ2/β4 isoforms mediate the functional coupling of NOP opioid receptors with Ca V channels in rat stellate ganglion (SG) sympathetic neurons. In the present report we extended our studies by identifying the Gγ subunit that forms the heterotrimer within this signaling pathway. Small interference RNA (or siRNA) was employed to silence the expression of the natively expressed Gγ subunits. Initial PCR assays indicated that SG neurons expressed seven Gγ subunits. Silencing Gγ3 subunits did not alter signaling between NOP receptors and Ca 2+ channels. However, after Gγ7 isoforms were silenced, the Noc-mediated inhibition of Ca V channels was significantly decreased when compared to SG neurons transfected with scrambled siRNA. We observed that Gγ10 and Gγ11 mRNA levels increased 2.5- and 2.7-fold, respectively, after Gγ7 subunits were silenced. However, this compensatory increase in mRNA expression did not appear to fully rescue the NOP receptor coupling efficiency. Additionally, both Gγ2 and Gγ5 levels increased 50 and 75%, respectively, while Gγ3 and Gγ4 expression levels remained relatively unchanged. Taken together, our findings suggest that the Gαi1/Gβ2(β4)/Gγ7 heterotrimeric G protein complex determines the NOP receptor-mediated modulation of Ca V channels in SG neurons. [ABSTRACT FROM AUTHOR]

Amiloride has been shown to inhibit acid-sensing ion channels (ASICs), which contribute to ischemia-related muscle pain during exercise. The purpose of this study was to determine if a single oral dose of amiloride would improve exercise tolerance and attenuate blood pressure during blood-flow-restricted (BFR) exercise in healthy adults. Ten subjects (4 females) performed isometric plantar flexion exercise with BFR (30% maximal voluntary contraction) after ingesting either a 10-mg dose of amiloride or a volume-matched placebo (random order). Time to failure, time-tension index (TTI), and perceived pain (visual analog scale) were compared between the amiloride and placebo trials. Mean blood pressure, heart rate, blood pressure index (BPI), and BPI normalized to TTI (BPInorm) were also compared between trials using both time-matched (TM50 and TM100) and effort-matched (T50 and T100) comparisons. Time to failure (+69.4 ± 63.2 s, P < 0.01) and TTI (+1,441 ± 633 kg·s, P = 0.02) were both significantly increased in the amiloride trial compared with placebo, despite no increase in pain (+0.4 ± 1.7 cm, P = 0.46). In contrast, amiloride had no significant influence on the mean blood pressure or heart rate responses, nor were there any significant differences in BPI or BPInorm between trials when matched for time (all P ≥ 0.13). When matched for effort, BPI was significantly greater in the amiloride trial (+5,300 ± 1,798 mmHg·s, P = 0.01), likely owing to an increase in total exercise duration. In conclusion, a 10-mg oral dose of amiloride appears to significantly improve the tolerance to BFR exercise in healthy adults without influencing blood pressure responses. [ABSTRACT FROM AUTHOR]

Objectives: To assess the reversibility of acute kidney injury-induced neutrophil dysfunction and to identify involved mechanisms.Design: Controlled laboratory experiment and prospective observational clinical study.Setting: University laboratory and hospital.Subjects: C57BL/6 wild-type mice.Patients: Patients with septic shock with or without acute kidney injury.Interventions: Murine acute kidney injury was induced by intraperitoneal injections of folic acid (nephrotoxic acute kidney injury) or by IM injections of glycerol (rhabdomyolysis-induced acute kidney injury). After 24 hours, we incubated isolated neutrophils for 3 hours in normal mouse serum or minimum essential medium buffer. We further studied the effects of plasma samples from 13 patients with septic shock (with or without severe acute kidney injury) on neutrophilic-differentiated NB4 cells.Measurements and Main Results: Experimental acute kidney injury significantly inhibited neutrophil migration and intracellular actin polymerization. Plasma levels of resistin, a proinflammatory cytokine and uremic toxin, were significantly elevated during both forms of acute kidney injury. Incubation in serum or minimum essential medium buffer restored normal neutrophil function. Resistin by itself was able to induce acute kidney injury-like neutrophil dysfunction in vitro. Plasma resistin was significantly higher in patients with septic shock with acute kidney injury compared with patients with septic shock alone. Compared with plasma from patients with septic shock, plasma from patients with septic shock and acute kidney injury inhibited neutrophilic-differentiated NB4 cell migration. Even after 4 days of renal replacement therapy, plasma from patients with septic shock plus acute kidney injury still showed elevated resistin levels and inhibited neutrophilic-differentiated NB4 cell migration. Resistin inhibited neutrophilic-differentiated NB4 cell migration and intracellular actin polymerization at concentrations seen during acute kidney injury, but not at normal physiologic concentrations.Conclusions: Acute kidney injury-induced neutrophil dysfunction is reversible in vitro. However, standard renal replacement therapy does not correct this defect in patients with septic shock and acute kidney injury. Resistin is greatly elevated during acute kidney injury, even with ongoing renal replacement therapy, and is sufficient to cause acute kidney injury-like neutrophil dysfunction by itself. [ABSTRACT FROM AUTHOR]

00974.2015.--Mechanical and metabolic stimuli arising from contracting muscles evoke the exercise pressor reflex. This reflex is greater in a rat model of simulated peripheral arterial disease in which a femoral artery is chronically ligated than it is in rats with freely perfused femoral arteries. The role played by the mechanically sensitive component of the exaggerated exercise pressor reflex in ligated rats is unknown. We tested the hypothesis that the mechano-gated channel inhibitor GsMTx4, a relatively selective inhibitor of mechano-gated Piezo channels, reduces the exercise pressor reflex in decerebrate rats with ligated femoral arteries. Injection of 10 μg of GsMTx4 into the arterial supply of the hindlimb reduced the pressor response to Achilles tendon stretch (a purely mechanical stimulus) but had no effect on the pressor responses to intra-arterial injection of α,β-methylene ATP or lactic acid (purely metabolic stimuli). Moreover, injection of 10 μg of GsMTx4 into the arterial supply of the hindlimb reduced both the integrated pressor area (control 535 ± 21, GsMTx4 218 ± 24 mmHg·s; P < 0.01), peak pressor (control 29 ± 2, GsMTx4 14 ± 3 mmHg; P < 0.01), and renal sympathetic nerve responses to electrically induced intermittent hindlimb muscle contraction (a mixed mechanical and metabolic stimulus). The reduction of the integrated pressor area during contraction caused by GsMTx4 was greater in rats with ligated femoral arteries than it was in rats with freely perfused femoral arteries. We conclude that the mechanically sensitive component of the reflex contributes to the exaggerated exercise pressor reflex during intermittent hindlimb muscle contractions in rats with ligated femoral arteries. [ABSTRACT FROM AUTHOR]

The mechanisms by which G proteins modulate voltage-gated Ca2+ channel currents (CaV), particularly CaV2.2 and CaV2.3, are voltage dependent (VD) or voltage independent (VI). VD pathways are typically mediated by Gαi/o and GαS subfamilies. On the other hand, VI inhibition modulation is coupled to the Gαq subfamily and signaling pathways downstream of phospholipase C stimulation. In most studies, this latter pathway has been shown to be linked to Gαq and/or Gα11 protein subunits. However, there are no studies that have examined whether natively expressed Gα14 subunits (Gαq subfamily member) couple G protein-coupled receptors (GPCR) with CaV2.2 channels. We report that Gα14 subunits functionally couple the substance P (SP)/neurokinin-1 (NK-1) receptor pathway to CaV2.2 channels in acutely dissociated rat celiac-superior mesenteric ganglion (CSMG) neurons. Exposure of CSMG neurons to SP blocked the CaV2.2 currents in a predominantly VD manner that was pertussis toxin and cholera toxin resistant, as well as Gαq/11 independent. However, silencing Gα14 subunits significantly attenuated the SP-mediated Ca2+ current block. In another set of experiments, exposure of CSMG neurons to SP led to the inhibition of KCNQ K+ M-currents. The SP-mediated M-current block was significantly reduced in neurons transfected with Gα14 small-interference RNA. Finally, overexpression of the GTP-bound Gαq/11 binding protein RGS2 did not alter the block of M-currents by SP but significantly abolished the oxotremorine methiodide-mediated M-current inhibition. Taken together, these results provide evidence of a new Gα14-coupled signaling pathway that modulates CaV2.2 and M-currents via SP-stimulated NK-1 receptors in CSMG neurons. [ABSTRACT FROM AUTHOR]

Key points Mechanical and metabolic stimuli from contracting muscles evoke reflex increases in blood pressure, heart rate and sympathetic nerve activity. Little is known, however, about the nature of the mechano-gated channels on the thin fibre muscle afferents that contribute to evoke this reflex, termed the exercise pressor reflex., We determined the effect of GsMTx4, an inhibitor of mechano-gated Piezo channels, on the exercise pressor reflex evoked by intermittent contraction of the triceps surae muscles in decerebrated, unanaesthetized rats., GsMTx4 reduced the pressor, cardioaccelerator and renal sympathetic nerve responses to intermittent contraction but did not reduce the pressor responses to femoral arterial injection of compounds that stimulate the metabolically-sensitive thin fibre muscle afferents., Expression levels of Piezo2 channels were greater than Piezo1 channels in rat dorsal root ganglia., Our findings suggest that mechanically-sensitive Piezo proteins contribute to the generation of the mechanical component of the exercise pressor reflex in rats., Abstract Mechanical and metabolic stimuli within contracting skeletal muscles evoke reflex autonomic and cardiovascular adjustments. In cats and rats, gadolinium has been used to investigate the role played by the mechanical component of this reflex, termed the exercise pressor reflex. Gadolinium, however, has poor selectivity for mechano-gated channels and exerts multiple off-target effects. We tested the hypothesis that GsMTX4, a more selective mechano-gated channel inhibitor than gadolinium and a particularly potent inhibitor of mechano-gated Piezo channels, reduced the exercise pressor reflex in decerebrate rats. Injection of 10 μg of GsMTx4 into the arterial supply of the hindlimb reduced the peak pressor (control: 24 ± 5, GsMTx4: 12 ± 5 mmHg, P < 0.01), cardioaccelerator and renal sympathetic nerve responses to tendon stretch, a purely mechanical stimulus, but had no effect on the pressor responses to intra-arterial injection of α,β-methylene ATP or lactic acid. Moreover, injection of 10 μg of GsMTx4 into the arterial supply of the hindlimb reduced the peak pressor (control: 24 ± 2, GsMTx4: 14 ± 3 mmHg, P < 0.01), cardioaccelerator and renal sympathetic nerve responses to electrically-induced intermittent hindlimb muscle contractions. By contrast, injection of 10 μg of GsMTx4 into the jugular vein had no effect on the pressor, cardioaccelerator, or renal sympathetic nerve responses to contraction. Quantitative RT-PCR and western blot analyses indicated that both Piezo1 and Piezo2 channel isoforms were natively expressed in rat dorsal root ganglia tissue. We conclude that GsMTx4 reduced the exercise pressor reflex in decerebrate rats and that the reduction was attributable, at least in part, to its effect on mechano-gated Piezo channels. [ABSTRACT FROM AUTHOR]

Introduction and hypothesis: We sought to develop a Spanish translation of the Female Genitourinary Pain Index (GUPI) and to validate this instrument in US Latina women. Methods: Translation back-translation was performed to create the initial Spanish version. Bilingual women with pelvic and/or genitourinary pain were recruited from clinical sites and social media. Participants reported demographics and completed the Female GUPI in both English and Spanish. Agreement was assessed for each item, subscale and total score. Additionally, we performed cognitive debriefing interviews to further test face validity. A consensus group of bilingual physicians and healthcare personnel utilized comments from the interviews to create a final Spanish version. Results: Thirty-four participants completed the questionnaire. Their average age was 33 years, 80% reported attending some college, and 20% reported an undergraduate degree or higher. Most were born in mainland USA (57%) or Mexico (27%). Agreement for the pain, urinary and quality of life subscales between the English and Spanish versions of the measure were excellent (0.91, 0.89 and 0.92, respectively) with 0.96 agreement for the measure as a whole. Despite favorable psychometrics, preferences for alternate wording were reported over 50 times. Based on that feedback, a consensus group was formed, which recommended changes to 13 of the 15 items, 3 of which required complete rewriting. Conclusions: The Spanish Female GUPI is strongly correlated with the English original; however, participants reported the language was overly complex. Translation and validation should include review of the measure and feedback by the target audience for optimal clarity and readability. [ABSTRACT FROM AUTHOR]

Key point Sympathetic stellate ganglion (SG) neurons that innervate cardiac muscle play an important role in regulating heart rate and contractility., In this study, we examined which Gβ protein subunit couples Ca2+ channels to opioid and adrenergic receptors expressed in SG neurons., We show that Gβ2 and Gβ4 are important signalling elements that maintain the transduction pathway of both receptor subtypes and Ca2+ channels, and how their expression levels are differentially compensated., Our results also indicate that Gβ1 subunits do not appear to play a role in either signalling pathway., The elucidation of specific G proteins that couple Ca2+ channels with opioid and adrenergic receptors will help us better understand how these receptors regulate synaptic transmission of SG neurons that innervate cardiac muscle., Abstract Cardiac function is regulated in part by the sympathetic branch of the autonomic nervous system via the stellate ganglion (SG) neurons. Neurotransmitters, such as noradrenaline (NA), and neuropeptides, including nociceptin (Noc), influence the excitability of SG neurons by modulating Ca2+ channel function following activation of the adrenergic and nociceptin/orphanin FQ peptide (NOP) opioid receptors, respectively. The regulation of Ca2+ channels is mediated by Gβγ, but the specific Gβ subunit that modulates the channels is not known. In the present study, small interference RNA (siRNA) was employed to silence the natively expressed Gβ proteins in rat SG tissue and to examine the coupling specificity of adrenergic and NOP opioid receptors to Ca2+ channels employing the whole-cell variant of the patch-clamp technique. Western blotting analysis showed that Gβ1, Gβ2 and Gβ4 are natively expressed. The knockdown of Gβ2 or Gβ4 led to a significant decrease of the NA- and Noc-mediated Ca2+ current inhibition, while Gβ1 silencing was without effect. However, sustaining low levels of Gβ2 resulted in an increased expression of Gβ4 and a concomitant compensation of both adrenergic and opioid signalling pathways modulating Ca2+ channels. Conversely, Gβ4-directed siRNA was not accompanied with a compensation of the signalling pathway. Finally, the combined silencing of Gβ2 and Gβ4 prevented any additional compensatory mechanisms. Overall, our studies suggest that in SG neurons, Gβ2 and Gβ4 normally maintain the coupling of Ca2+ channels with the receptors, with the latter subtype responsible for maintaining the integrity of both pathways. [ABSTRACT FROM AUTHOR]

Using unprecedented catalogues of past severe drought data for the Yucatan Peninsula between 1502 and 1900 coming from historical written documentation, we identified five conspicuous time lapses with no droughts between 1577-1647, 1662-1724, 1728-1764, 1774-1799 and 1855-1880, as well as time epochs with most frequent droughts between 1800 and 1850. Moreover, the most prominent periodicity of the historical drought time series was that of ∼40 years. Using the Palmer Drought Severity Index for the Yucatan Peninsula for the period 1921-1987 we found prominent negative phases between ∼1942-1946 and 1949-1952, 1923-1924, 1928-1929, 1935-1936, 1962-1963, 1971-1972 and 1986-1987. Two prominent periodicities clearly appear at ∼5 and 10 years. Most modern and historical severe droughts lasted 1 year, and share a quasidecadal frequency. Also, in the first 66 years of the twentieth century the frequency of occurrence of severe drought has been lower compared with the nineteenth century. Some of the major effects and impacts of the most severe droughts in the Yucatan region are examined. We also studied the relation between historical and modern droughts and several large scale climate phenomena represented by the Atlantic Multidecadal Oscillation (AMO) and the Southern Oscillation Index (SOI). Our results indicate that historical droughts and the cold phase of the AMO coincide, while the influence of the SOI is less clear. The strongest coherence between historical droughts and AMO occurred at periodicities of ∼40 years. For modern droughts the coherence of a drought indicator (the Palmer Drought Severity Index) is similar with AMO and SOI, although it seems more sustained with the AMO. They are strongest at ∼10 years and very clearly with the AMO cold phase. Concerning the solar activity proxies and historical droughts, the coherence with a record of beryllium isotope Be10, which is a good proxy of cosmic rays, is higher than with Total Solar Irradiance. We notice that the strongest coherence between historical droughts and Be10 occurs at periods ∼60-64 years. When studying modern droughts and solar activity, frequencies of ∼8 years appear, and the coherences are similar for both sunspots and cosmic rays. Comparing natural terrestrial and solar phenomena, we found that the most sustained and strongest modulation of historical drought occurrence is at ∼60-64 years and is between the historical drought series and the solar proxy Be10. For modern droughts we notice that the coherence is similar among AMO, SOI and the solar indices. We can conclude that the sea surface temperatures (AMO) and solar activity leave their signal in terms of severe droughts in the Maya lands, however in the long term, the influence of the SOI on this type of phenomenon is less clear. [ABSTRACT FROM AUTHOR]

NaV1.8 channels play a crucial role in regulating the action potential in nociceptive neurons. A single nucleotide polymorphism in the human NaV1.8 gene SCN10A, A1073V (rs6795970, G>A), has been linked to the diminution of mechanical pain sensation as well as cardiac conduction abnormalities. Furthermore, studies have suggested that this polymorphism may result in a "loss-of-function" phenotype. In the present study, we performed genomic analysis of A1073V polymorphism presence in a cohort of patients undergoing sigmoid colectomy who provided information regarding perioperative pain and analgesic use. Homozygous carriers reported significantly reduced severity in postoperative abdominal pain compared with heterozygous and wild-type carriers. Homozygotes also trended toward using less analgesic/opiates during the postoperative period. We also heterologously expressed the wild-type and A1073V variant in rat superior cervical ganglion neurons. Electrophysiological testing demonstrated that the mutant NaV1.8 channels activated at more depolarized potentials compared with wild-type channels. Our study revealed that postoperative abdominal pain is diminished in homozygous carriers of A1073V and that this is likely due to reduced transmission of action potentials in nociceptive neurons. Our findings reinforce the importance of NaV1.8 and the A1073V polymorphism to pain perception. This information could be used to develop new predictive tools to optimize patient pain experience and analgesic use in the perioperative setting. NEW & NOTEWORTHY We present evidence that in a cohort of patients undergoing sigmoid colectomy, those homozygous for the NaV1.8 polymorphism (rs6795970) reported significantly lower abdominal pain scores than individuals with the homozygous wild-type or heterozygous genotype. In vitro electrophysiological recordings also suggest that the mutant NaV1.8 channel activates at more depolarizing potentials than the wild-type Na+ channel, characteristic of hypoactivity. This is the first report linking the rs6795970 mutation with postoperative abdominal pain in humans. [ABSTRACT FROM AUTHOR]

Background: Abdominal pain is a frequent and persistent problem in the most common gastrointestinal disorders, including irritable bowel syndrome and inflammatory bowel disease. Pain adversely impacts quality of life, incurs significant healthcare expenditures, and remains a challenging issue to manage with few safe therapeutic options currently available. It is imperative that new methods are developed for identifying and treating this symptom. A variety of peripherally active neuroendocrine signaling elements have the capability to influence gastrointestinal pain perception. A large and growing body of evidence suggests that voltage‐gated sodium channels (VGSCs) play a critical role in the development and modulation of nociceptive signaling associated with the gut. Several VGSC isoforms demonstrate significant promise as potential targets for improved diagnosis and treatment of gut‐based disorders associated with hyper‐ and hyposensitivity to abdominal pain. Purpose: In this article, we critically review key investigations that have evaluated the potential role that VGSCs play in visceral nociception and discuss recent advances related to this topic. Specifically, we discuss the following: (a) what is known about the structure and basic function of VGSCs, (b) the role that each VGSC plays in gut nociception, particularly as it relates to human physiology, and (c) potential diagnostic and therapeutic uses of VGSCs to manage disorders associated with chronic abdominal pain. Voltage‐gated sodium channels (VGSCs) are critical for the transmission of nociceptive signals that are interpreted as pain, including from the gastrointestinal (GI) tract. Here, we review the basic function and pathophysiology of VGSCs in the context of gastrointestinal pain perception. [ABSTRACT FROM AUTHOR]

The single nucleotide polymorphism of the μ-opioid receptor, OPRM1 A118G, has been associated with greater drug and alcohol use, increased sensitivity to pain, and reduced sensitivity to the antinociceptive effects of opiates. In the present studies, we employed a ‘humanized’ mouse model containing the wild-type (118AA) or variant (118GG) allele to examine behavior in a model of heroin-induced devaluation of an otherwise palatable saccharin cue when repeated saccharin-heroin pairings occurred every 24 h (Experiment 1) or every 48 h (Experiment 2). The results showed that, while both the 118AA and 118GG mice demonstrated robust avoidance of the heroin-paired saccharin cue following daily taste-drug pairings, only the 118AA mice suppressed intake of the heroin-paired saccharin cue when 48 h elapsed between each taste-drug pairing. Humanized 118GG mice, then, defend their intake of the sweet cue despite saccharin-heroin pairings and this effect is illuminated by the use of spaced, rather than massed, trials. Given that this pattern of strain difference is not evident with saccharin-cocaine pairings (Freet et al., 2015), reduced avoidance of the heroin-paired saccharin cue by the 118GG mice may be due to an interaction between the opiate and the subjects’ drive for the sweet or, alternatively, to differential downstream sensitivity to the aversive kappa mediated properties of the drug. These alternative hypotheses are addressed. [ABSTRACT FROM AUTHOR]

Objective: Sepsis is a condition associated with a dysregulated inflammatory response to infection with significant morbidity. Recent advances have elucidated the vital role that the short chain fatty acid glycoprotein receptor 43 (FFA2/GPR43) plays in inflammatory and immunomodulatory pathways. We hypothesized that elevated whole blood GPR43 RNA expression would be associated with increased 30-day survival in patients admitted with sepsis. Patients (n = 93) admitted to the intensive care unit with the diagnosis of sepsis underwent quantitative real time PCR within 48 h of intensive care unit admission. Clinical and demographical parameters were retrospectively extracted from the chart and compared to quantitative measurements of GPR43 RNA expression. Results: Utilizing logistic regression, we found that the odds of mortality decreased for every one-unit increase in GPR43 RNA expression for patients that survived to 30 days [OR = 0.71; 95% CI (0.50, 0.99) p = 0.049]. Using linear regression, we determined that the increase in whole blood GPR43 expression was not associated with whole blood white cell count [r = 0.04; 95% CI (-0.16, 0.24); p = 0.70] or body mass index [r = - 0.07; 95% CI (- 0.23, 0.18); p = 0.81]. We conclude that the GPR43 receptor plays an integral role in survival during and after sepsis. [ABSTRACT FROM AUTHOR]

Key points Chronic limb ischaemia, characterized by inflammatory mediator release and a low extracellular pH, leads to acid-sensing ion channel (ASIC) activation and reflexively increases mean arterial pressure; endomorphin release is also increased under inflammatory conditions., We examined the modulation of ASIC currents by endomorphins in sensory neurons from rats with freely perfused and ligated femoral arteries: peripheral artery disease (PAD) model., Endomorphins potentiated sustained ASIC currents in both groups of dorsal root ganglion neurons, independent of mu opioid receptor stimulation or G protein activation., Intra-arterial administration of lactic acid (to simulate exercising muscle and evoke a pressor reflex), endomorphin-2 and naloxone resulted in a significantly greater pressor response than lactic acid alone, while administration of APETx2 inhibited endomorphin's enhancing effect in both groups., These results suggest a novel role for endomorphins in modulating ASIC function to effect lactic acid-mediated reflex increase in arterial pressure in patients with PAD., Abstract Chronic muscle ischaemia leads to accumulation of lactic acid and other inflammatory mediators with a subsequent drop in interstitial pH. Acid-sensing ion channels (ASICs), expressed in thin muscle afferents, sense the decrease in pH and evoke a pressor reflex known to increase mean arterial pressure. The naturally occurring endomorphins are also released by primary afferents under ischaemic conditions. We examined whether high affinity mu opioid receptor (MOR) agonists, endomorphin-1 (E-1) and -2 (E-2), modulate ASIC currents and the lactic acid-mediated pressor reflex. In rat dorsal root ganglion (DRG) neurons, exposure to E-2 in acidic solutions significantly potentiated ASIC currents when compared to acidic solutions alone. The potentiation was significantly greater in DRG neurons isolated from rats whose femoral arteries were ligated for 72 h. Sustained ASIC current potentiation was also observed in neurons pretreated with pertussis toxin, an uncoupler of G proteins and MOR. The endomorphin-mediated potentiation was a result of a leftward shift of the activation curve to higher pH values and a slight shift of the inactivation curve to lower pH values. Intra-arterial co-administration of lactic acid and E-2 led to a significantly greater pressor reflex than lactic acid alone in the presence of naloxone. Finally, E-2 effects were inhibited by pretreatment with the ASIC3 blocker APETx2 and enhanced by pretreatment with the ASIC1a blocker psalmotoxin-1. These findings have uncovered a novel role of endomorphins by which the opioids can enhance the lactic acid-mediated reflex increase in arterial pressure that is MOR stimulation-independent and APETx2-sensitive. [ABSTRACT FROM AUTHOR]

R2656 --> 560.3 --> Background: During exercise, blood pressure increases via augmented central command and stimulation of mechano‐ and metabosensitive muscle afferents (i.e., the exercise pressor reflex). Recent evidence has reported a strong role of acid sensing ion channels (ASIC's) in mediating this reflex, as well as exercise induced muscle pain. Therefore, it is logical to suspect that inhibiting ASIC's would mitigate the exaggerated blood pressure responses and exercise related muscle pain observed in individuals with peripheral artery disease (PAD). Purpose:. We tested the hypothesis that a single 10mg dose of amiloride (a K+‐sparing diuretic known to inhibit ASIC's) would attenuate the pressor response to blood flow restricted (BFR) plantar flexion exercise in young healthy adults. Methods: Six healthy subjects (5 male, 26 ± 2 y/o, 180.8 ± 12.1 cm, 79.7 ± 12.9 Kg) completed three separate visits. Each visit included two bouts of isometric plantar flexion, both performed until failure at 30% of the predetermined maximal voluntary isometric contraction. During one bout, blood flow to the limb was restricted via a pressurized cuff placed around the proximal portion of the lower leg (BFR); the other bout was performed under free flow conditions. In the first visit, subjects performed these two exercise bouts (in random order) under normal, non‐treated conditions. In the second and third visits, subjects performed these exercise bouts (same order as in visit 1) ~3 hrs after oral ingestion of 10mg of amiloride, or ingestion of a placebo (random order). Blood pressure and heart rate were continuously recorded throughout exercise, and time to fatigue and muscle pain (on a 12cm visual analog scale) was recorded at the end of each exercise bout. Only data from the placebo and amiloride conditions are presented here. Results: The addition of BFR reduced total exercise time by ~40.8% (t=2.99, p=0.03), but had no significant effect on peak muscle pain (p>0.10). Due to the preliminary nature of these data, no significant main effects of treatment (amiloride vs. placebo) were observed for free flow or BFR exercise (all p>0.05). However, we did observe a net decrease in the average heart rate response (+10.7 ± 3.0 bpm vs. +11.3 ± 3.7 bpm, ηp2=0.539, p>0.47), the total blood pressure response (5477.5 ± 373.7 mmHg*sec vs. 5841.3 ± 518.9 mmHg*sec, ηp2=0.250, p>0.30), the total time‐tension index (595.9 ± 406.5 Kg*sec vs. 691.6 ± 394.2 Kg*sec, ηp2=0.211, p>0.30), and the blood pressure to time‐tension index relationship (9.09 ± 4.40 mmHg/Kg vs. 11.38 ± 6.30 mmHg/Kg, ηp2=0.08, p>0.50) in the amiloride condition compared to the placebo condition during BFR exercise. Conclusions: While these preliminary findings do not indicate significant effects of treatment, these observations serve as strong pilot data to support the continued investigation of amiloride as a method of attenuating the pressor response to ischemic exercise. [ABSTRACT FROM AUTHOR]

Septic shock, the most severe complication associated with sepsis, is manifested by tissue hypoperfusion due, in part, to cardiovascular and autonomic dysfunction. In many cases, the splanchnic circulation becomes vasoplegic. The celiac-superior mesenteric ganglion (CSMG) sympathetic neurons provide the main autonomic input to these vessels. We used the cecal ligation puncture (CLP) model, which closely mimics the hemodynamic and metabolic disturbances observed in septic patients, to examine the properties and modulation of Ca2+ channels by G protein-coupled receptors in acutely dissociated rat CSMG neurons. Voltage-clamp studies 48 hr post-sepsis revealed that the Ca2+ current density in CMSG neurons from septic rats was significantly lower than those isolated from sham control rats. This reduction coincided with a significant increase in membrane surface area and a negligible increase in Ca2+ current amplitude. Possible explanations for these findings include either cell swelling or neurite outgrowth enhancement of CSMG neurons from septic rats. Additionally, a significant rightward shift of the concentration-response relationship for the norepinephrine (NE)-mediated Ca2+ current inhibition was observed in CSMG neurons from septic rats. Testing for the presence of opioid receptor subtypes in CSMG neurons, showed that mu opioid receptors were present in ~70% of CSMG, while NOP opioid receptors were found in all CSMG neurons tested. The pharmacological profile for both opioid receptor subtypes was not significantly affected by sepsis. Further, the Ca2+ current modulation by propionate, an agonist for the free fatty acid receptors GPR41 and GPR43, was not altered by sepsis. Overall, our findings suggest that CSMG function is affected by sepsis via changes in cell size and α2-adrenergic receptor-mediated Ca2+ channel modulation. [ABSTRACT FROM AUTHOR]

Several studies have shown that human carriers of the single nucleotide polymorphism of the μ-opioid receptor, OPRM1 A118G, exhibit greater drug and alcohol use, increased sensitivity to pain, and reduced sensitivity to the antinociceptive effects of opiates. In the present study, we employed a ‘humanized’ mouse model containing the wild-type (118AA) or variant (118GG) allele to examine behavior in our model of drug-induced suppression of a natural reward cue and to compare the morphine pharmacological profile in acutely isolated sensory neurons. Compared with 118AA mice, our results demonstrate that homozygous 118GG mice exhibit greater avoidance of the cocaine-paired saccharin cue, a behavior linked to an aversive withdrawal-like state. Electrophysiological recordings confirmed the reduced modulation of Ca 2 + channels by morphine in trigeminal ganglion (TG) neurons from 118GG mice compared to the 118AA control cells. However, repeated cocaine exposure in 118GG mice led to a leftward shift of the morphine concentration–response relationship when compared with 118GG control mice, while a rightward shift was observed in 118AA mice. These results suggest that cocaine exposure of mice carrying the 118G allele leads to a heightened sensitivity of the reward system and a blunted modulation of Ca 2 + channels by morphine in sensory neurons. [ABSTRACT FROM AUTHOR]

The exercise pressor reflex, a crucial component of the cardiovascular response under physiological and pathophysiological states, is activated via metabolic and mechanical mediators that originate from contracting muscles and stimulate group III and IV afferents. We reported previously that stimulation of mu opioid receptors (MOR), expressed in both afferents, led to a significant attenuation of the reflex in rats whose femoral arteries had been occluded for 72 h. The present study examined the effect of arterial occlusion on the signaling components involved in the opioid-mediated modulation of Ca2+ channels in rat dorsal root ganglion neurons innervating the triceps surae muscles. We focused on neurons that were transfected with cDNA coding for enhanced green fluorescent protein whose expression is driven by the voltagegated Na+ channel 1.8 (NaV1.8) promoter region, a channel expressed primarily in nociceptive neurons. With the use of a small interference RNA approach, our results show that the pertussis toxin-sensitive Gαi3 subunit couples MOR with Ca2+ channels. We observed a significant leftward shift of the MOR agonist [D-Ala2-N-Me-Phe4- Glycol5]-enkephalin concentration-response relationship in neurons isolated from rats with occluded arteries compared with those that were perfused freely. Femoral occlusion did not affect Ca2+ channel density or the fraction of the main Ca2+ channel subtype. Furthermore, Western blotting analysis indicated that the leftward shift did not result from either increased Gαi3 or MOR expression. Finally, all neurons from both groups exhibited an inward current following exposure of the transient potential receptor vanilloid 1 (TRPV1) agonist, 8-methyl-N-vanillyl-6-nonenamide. These findings suggest that sensory neurons mediating the exercise pressor reflex express NaV1.8 and TRPV1 channels, and femoral occlusion alters the MOR pharmacological profile. [ABSTRACT FROM AUTHOR]

In rats with ligated femoral arteries, the exercise pressor reflex is exaggerated, an effect that is attenuated by stimulation of peripheral μ-opioid receptors on group IV metabosensitive afferents. In contrast, δ-opioid receptors are expressed mostly on group III mechanosensitive afferents, a finding that prompted us to determine whether stimulation of these opioid receptors could also attenuate the exaggerated exercise pressor reflex in "ligated" rats. We found femoral arterial injection of [D-Pen2,D-Pen5]enkephalin (DPDPE; 1.0 μg), a δ-opioid agonist, significantly attenuated the pressor and cardioaccelerator components of the exercise pressor reflex evoked by hindlimb muscle contraction in both rats with ligated and patent femoral arteries. DPDPE significantly decreased the pressor responses to muscle mechanoreflex activation, evoked by tendon stretch, in ligated rats only. DPDPE (1.0 μg) had no effect in either group on the pressor and cardioaccelerator responses to capsaicin (0.2 μg), which primarily stimulates group IV afferents. DPDPE (1.0 μg) had no effect on the pressor and cardioaccelerator responses to lactic acid (24 mM), which stimulates group III and IV afferents, in rats with patent femoral arteries but significantly decreased the pressor response in ligated rats. Western blots revealed the amount of protein comprising the δ-opioid receptor was greater in dorsal root ganglia innervating hindlimbs with ligated femoral arteries than in dorsal root ganglia innervating hindlimbs with patent femoral arteries. Our findings support the hypothesis that stimulation of δ-opioid receptors on group III afferents attenuated the exercise pressor reflex. [ABSTRACT FROM AUTHOR]

Abstract: Background: The endogenous opioid peptide, nociception (Noc), contributes to the regulation of systemic blood pressure and regional blood flow. Recent clinical and animal studies have reported that Noc and its receptor (nociceptin/orphanin FQ [NOP]) are involved in inflammation and sepsis. The purpose of the present study was to examine the modulation of Ca2+ channels by Noc in acutely isolated stellate ganglion (SG) neurons from control and septic rats. Materials and methods: Sepsis was induced in male Sprague–Dawley rats via cecal ligation and puncture. SG neurons were isolated 24 and 72 h after sepsis induction. Thereafter, the concentration–response relationships for the Noc-stimulated NOP receptor Ca2+ current inhibition were determined using the whole-cell patch clamp technique. In addition, the Noc precursor (prepronociceptin [PNOC]) and NOP receptor messenger RNA (mRNA) levels were determined by quantitative real-time polymerase chain reaction, and PNOC protein levels were measured by Western blot analysis. Results: Comparison of the Noc concentration–response relationships in SG neurons from control and septic rats 24 h after sepsis revealed similar potency and efficacy. Moreover, 72 h after sepsis, neurons from control and septic rats exhibited an increased potency compared with both groups at the 24-h time point—an effect that was more pronounced in neurons from septic rats. PNOC mRNA levels were significantly greater in SG neurons isolated from septic rats compared with control neurons, but NOP receptor mRNA levels remained unchanged during the 72-h period. Conclusions: Our study demonstrates the cecal ligation and puncture model–induced temporal upregulation of components within the NOP receptor signaling pathway in rat sympathetic neurons. As SG neurons provide the main sympathetic input to the heart, an increased Noc release and potency during sepsis may compromise cardiovascular function. [Copyright &y& Elsevier]

Cardiovascular adjustments to exercise are partially mediated by group III/IV (small to medium) muscle afferents comprising the exercise pressor reflex (EPR). However, this reflex can be inappropriately activated in disease states (e.g. peripheral vascular disease) leading to increased risk of myocardial infarction. Here we investigate the voltage-dependent calcium (CaV) channels expressed in small to medium muscle afferent neurons as a first step towards determining their potential role in controlling the EPR. Using specific blockers and 5 mM Ba2+ as the charge carrier, we found the major calcium channel types to be CaV.2 (N-type) > CaV.1 (P/Q-type) > CaV1.2 (L-type). Surprisingly, the CaV2.3 channel (R-type) blocker SNX482 was without effect. However, R-type currents are more prominent when recorded in Ca2+ (Liang and Elmslie 2001). We reexamined the channel types using 10 mM Ca2+ as the charge carrier, but results were similar to those in Ba2+. SNX482 was without effect even though ~27% of the current was blocker-insensitive. Using multiple methods, we demonstrate that CaV2.3 channels are functionally expressed in muscle afferent neurons. Finally, ATP is an important modulator of the EPR and we examined the effect on CaV currents. ATP reduced CaV current primarily via G protein βγ-mediated inhibition of CaV2.2 channels. We conclude that small to medium muscle afferent neurons primarily express CaV2.2 > CaV2.1 ≥ CaV2.3 > CaV1.2 channels. As with chronic pain, CaV2.2 channel blockers may be useful in controlling inappropriate activation of the EPR. [ABSTRACT FROM AUTHOR]

Key points In decerebrated rats, the exercise pressor reflex arising from a hindlimb whose femoral artery was occluded for 72 h was significantly higher than that arising from a hindlimb whose femoral artery was freely perfused., Blockade of endoperoxide 4 receptors, but not blockade of endoperoxide 3 receptors, prevented the exaggerated exercise pressor reflex in rats with ligated femoral arteries., Blockade of endoperoxide 3 or 4 receptors in rats with freely perfused femoral arteries had no effect on the exercise pressor reflex., Western immunoblots showed that ligation of the femoral artery for 72 h increased the endoperoxide 4 receptor protein in the L4 and L5 dorsal root ganglia over their freely perfused counterparts by 24% ( P < 0.05)., Abstract Ligating the femoral artery for 72 h in decerebrated rats exaggerates the exercise pressor reflex. The sensory arm of this reflex is comprised of group III and IV afferents, which can be either sensitized or stimulated by PGE2. In vitro studies showed that endoperoxide (EP) 3 and 4 receptors were responsible for the PGE2-induced sensitization of rat dorsal root ganglion cells. This in vitro finding prompted us to test the hypothesis that blockade of EP3 and/or EP4 receptors attenuated the exaggerated exercise pressor reflex in rats with ligated femoral arteries. We measured the cardiovascular responses to static hindlimb contraction or tendon stretch before and after femoral arterial injection of L798106 (an EP3 antagonist) or L161982 (an EP4 antagonist). The pressor and cardioaccelerator responses to either contraction or tendon stretch were not attenuated by L798106 in either the ligated or freely perfused rats. Likewise in five rats whose hindlimb muscles were freely perfused, the pressor and cardioaccelerator responses to either contraction or tendon stretch were not attenuated by L161982. In the six ligated rats, however, the pressor response to contraction was attenuated by L161982, averaging 37 ± 3 mmHg before, 18 ± 2 mmHg afterward ( P < 0.05). Western blotting analysis revealed that ligation of the femoral artery for 72 h increased the EP4 receptor protein in the L4 and L5 dorsal root ganglia over their freely perfused counterparts by 24% ( P < 0.05). We conclude that EP4 receptors, but not EP3 receptors, play an important role in the exaggerated exercise pressor reflex found in rats with ligated femoral arteries. [ABSTRACT FROM AUTHOR]

Highlights: [•] A803467 (1mg) reduced pressor responses to stimulation of group III & IV afferents. [•] A803467 (1mg) reduced muscle spindle activity to stretch and succinylcholine. [•] The utility of A803467 in vivo is challenged as a selective antagonist for NaV 1.8. [Copyright &y& Elsevier]

Summary: Background: Haemophilus influenzae type b (Hib) conjugate vaccine reduces the risk of pneumonia in infants. Objective: To determine the effect of Hib conjugate vaccine (HibCV) on the prevention of pneumonia as a complication among infants hospitalized for bronchiolitis. Methods: This record-based case–control study was conducted at The Children''s Hospital “Dr. Ovidio Aliaga U” in La Paz, Bolivia during 2003 and 2004. Cases were infants hospitalized for bronchiolitis under 1 year of age who developed radiological pneumonia during hospitalization. Controls were patients who had good clinical progress without the use of antibiotics. Pneumonia was defined by alveolar consolidation on chest X-ray that justified the use of antibiotics. Results: Eighty patients were studied (16 cases and 64 controls). Their median age was 4.5 months. Demographic and clinical features were similar in both groups, except for a higher proportion of vomiting (56.3% vs. 28.1%; p <0.05) in the case group. The percentage of unvaccinated infants was significantly higher in cases (68.8% vs. 26.6%; p <0.05) and the length of hospital stay longer (8.5±5.4 vs. 3.1±2.2 days; p <0.05). There was a strong association between unvaccinated infants and the occurrence of pneumonia as a complication (odds ratio 6.1, 95% confidence interval 1.8–20.1; p <0.01). Conclusions: Unvaccinated infants admitted for bronchiolitis have a higher risk of radiologically confirmed pneumonia. Larger studies are needed to validate these results and reconsider the burden of Hib infection among infants in less developed countries. [Copyright &y& Elsevier]

Background: Cold storage in any of the commonly used preservation solutions is not always adequate for donation after cardiac death (DCD) liver grafts due to prolonged warm ischemic time. In this study, we used a third-generation perfluorocarbon (PFC), Oxycyte, for DCD liver graft preservation in a rat model. Materials and Methods: Twenty-eight rats (14 in each group) were used. Thirty minutes after cardiopulmonary arrest, livers were harvested and flushed with a cold and pre-oxygenated solution of either University of Wisconsin (UW) or UW + 20% PFC. After 8 h of cold preservation in either of the investigated solutions, liver graft specimens were analyzed for evidence of ischemic injury. Hemotoxylin and eosin staining (H and E), as well as immunohistochemical analysis with anti-cleaved caspase 3 antibody, was performed. Levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in the preservation solution were analyzed at 1 and 8 h during preservation. Results: In the PFC group, the degree of cell congestion, vacuolization and necrosis were all significantly less than in the UW group (P = 0.002–0.004). The number of cells with a positive cleaved caspase 3 antibody reaction was reduced by about 50% in comparison with the UW group (P < 0.006). The AST level in the PFC group was significantly less than in the UW group after 8 h of preservation (P < 0.048). Conclusion: The addition of PFC to UW solution significantly decreases the degree of histologic damage in rat DCD liver grafts. This preservation strategy can be potentially helpful for organ preservation after prolonged warm ischemia. [Copyright &y& Elsevier]

Background: The most common functional single nucleotide polymorphism of the human OPRM1 gene, A118G, has been shown to be associated with interindividual differences in opioid analgesic requirements, particularly with morphine, in patients with acute postoperative pain. The purpose of this study was to examine whether this polymorphism would modulate the morphine and fentanyl pharmacological profile of sensory neurons isolated from a humanized mouse model homozygous for either the 118A or 118G allele.Methods: The coupling of wild-type and mutant μ opioid receptors to voltage-gated Ca channels after exposure to either ligand was examined by employing the whole cell variant of the patch-clamp technique in acutely dissociated trigeminal ganglion neurons. Morphine-mediated antinociception was measured in mice carrying either the 118AA or 118GG allele.Results: The biophysical parameters (cell size, current density, and peak current amplitude potential) measured from both groups of sensory neurons were not significantly different. In 118GG neurons, morphine was approximately fivefold less potent and 26% less efficacious than that observed in 118AA neurons. On the other hand, the potency and efficacy of fentanyl were similar for both groups of neurons. Morphine-mediated analgesia in 118GG mice was significantly reduced compared with the 118AA mice.Conclusions: This study provides evidence to suggest that the diminished clinical effect observed with morphine in 118G carriers results from an alteration of the receptor's pharmacology in sensory neurons. In addition, the impaired analgesic response with morphine may explain why carriers of this receptor variant have an increased susceptibility to become addicted to opioids. [ABSTRACT FROM AUTHOR]

Abstract: The 20th century surface air temperature (SAT) records of China from various sources are analyzed using data which include the recently released Twentieth Century Reanalysis Project dataset. Two key features of the Chinese records are confirmed: (1) significant 1920s and 1940s warming in the temperature records, and (2) evidence for a persistent multidecadal modulation of the Chinese surface temperature records in co-variations with both incoming solar radiation at the top of the atmosphere as well as the modulated solar radiation reaching ground surface. New evidence is presented for this Sun–climate link for the instrumental record from 1880 to 2002. Additionally, two non-local physical aspects of solar radiation-induced modulation of the Chinese SAT record are documented and discussed. Teleconnections that provide a persistent and systematic modulation of the temperature response of the Tibetan Plateau and/or the tropospheric air column above the Eurasian continent (e.g., 30°N–70°N; 0°–120°E) are described. These teleconnections may originate from the solar irradiance-Arctic–North Atlantic overturning circulation mechanism proposed by . Also considered is the modulation of large-scale land–sea thermal contrasts both in terms of meridional and zonal gradients between the subtropical western Pacific and mid-latitude North Pacific and the continental landmass of China. The Circum-global teleconnection (CGT) pattern of summer circulation of provides a physical framework for study of the Sun–climate connection over East Asia. Our results highlight the importance of solar radiation reaching the ground and the concomitant importance of changes in atmospheric transparency or cloudiness or both in motivating a true physical explanation of any Sun–climate connection. We conclude that ground surface solar radiation is an important modulating factor for Chinese SAT changes on multidecadal to centennial timescales. Therefore, a comprehensive view of local and remote factors of climate change in China must take account of this as well as other natural and anthropogenic forcings. [Copyright &y& Elsevier]

Trypanosoma cruzi kinetoplastid membrane protein-11 (KMP-11) is able to induce protective immunity in mice. In humans, T-cell immunity during Chagas disease has been documented using parasite antigens allowing the identification of specific CD8+ T cells. However, little is known about the CD4+ T-cell response during the evolution of the disease. In this paper, the induction of a natural CD4+ T-cell response against the KMP-11 protein in T. cruzi infected humans was studied to assess whether this parasite-derived protein could be processed, presented and detected as a major histocompatibility complex class II restricted epitope. The results show that helper T cells from 5 out of 13 chagasic patients specifically produced interferon-γ after exposure to the KMP-11 antigen, whereas healthy donors and non-chagasic cardiopathic patients did not respond. This is the first description of T. cruzi KMP-11 protein recognition by CD4+ T cells in chronic chagasic patients.Immunology and Cell Biology (2009) 87, 149–153; doi:10.1038/icb.2008.76; published online 28 October 2008 [ABSTRACT FROM AUTHOR]

A catalog containing an unprecedented amount of historical data in central Mexico, covering almost six centuries (1450–1900), is used. This is a catalog of agricultural disasters that includes events associated with hydrometeorological phenomena, or hazards, whose effects were mainly felt in the agricultural sector, such as droughts. An analysis of the historical series of droughts in central Mexico for the period of 1450–1900 is performed. Periods of frequent drought centered at the years 1483, 1533, 1571, 1601, 1650, 1691, 1730, 1783, 1818, and 1860 have been identified. In particular, droughts in Mexico City and northwest Mexico that were identified through poor tree-ring growth are included in the frequent drought periods obtained in this work. Moreover, it was found that droughts occurred in El Niño years mainly for events of very strong and strong strengths, at a significant level. Also, most droughts lasted for 1 or 2 yr. Last, by analyzing the periodicities of the drought time series it was found that those that are the most conspicuous are the quasi-bidecadal frequencies of 18.9 and 21 yr. [ABSTRACT FROM AUTHOR]

Stellate ganglion (SG) neurons provide the main sympathetic innervation to the heart and help to regulate cardiac function. The purpose of this study was to determine if ultrasound imaging could be employed to retrograde label rat SG neurons innervating the heart without employing thoracotomy. In addition, electrophysiological experiments were performed to characterize the modulation of Ca2+ channels by neurotransmitters in unlabeled and dye-labeled SG neurons. Fluorescence imaging of actutely isolated cells revealed that dye uptake was successful within five days following injection of dye in the cardiac muscle. Whole-cell voltage-clamp recordings revealed that the majority of the Ca2+ current was carried by N-type Ca2+ channels. Finally, fluorescence dye uptake did not appear to affect the modulation of Ca2+ currents following exposure of SG neurons to norepinephrine, adenosine and neurokinin A. These results demonstrate that ultrasound imaging-guided percutaneous injection can be effectively employed to retrograde label neurons innervating the heart. [Copyright &y& Elsevier]

• ER and ZD methods provide phasor estimates removing the exponential DC offset. • The proposals outperform the DFT behaviour in both steady and dynamic states. • The proposals prevent the oscillatory behaviour in the impedance estimation. • The proposals improve the operating time under fault and oscillatory conditions. This paper introduces the eigensystem realization (ER)-based identification algorithm and the Z-domain method (ZDM) to improve the impedance estimation of transmission lines under fault conditions, which is achieved by removing the exponential decaying DC offset from current signals in distance relays. Both methods effectively work employing a one-cycle rectangular window; specifically, the Z-domain method works in less than a fifth of one-cycle window, ensuring timely, accurate, and prompt estimates. To confirm the effectiveness and performance of the proposed algorithms, they are evaluated under steady-state and dynamic conditions by using time-domain simulations performed in PSCAD©and DIgSILENT©PowerFactory. They also are compared with the well-known discrete Fourier transform (DFT). The attained results indicate that both the ER-based and Z-domain methods can become a powerful tool to reduce the adverse influence of exponentially decaying DC offsets in distance relays. [ABSTRACT FROM AUTHOR]

Background: Sepsis-induced immunosuppression is a key factor contributing to the morbidity and mortality of critically ill patients, and polymorphonuclear neutrophil dysfunction is believed to be a hallmark of this immunosuppression. Circulating myeloid cells produce the cytokine resistin (RETN), which has been associated with poor outcomes in sepsis/septic shock and can directly inhibit neutrophil function. We previously demonstrated that resistin caused a dose-dependent impairment in neutrophil migration, reactive oxygen species production, and bacterial clearance in neutrophil cell lines. However, the relative antimicrobial responses of other innate immune cells to Gram-positive and Gram-negative infections in the presence of elevated levels of resistin have not been evaluated. We hypothesized that resistin directly contributes to sepsis-induced immunosuppression by selectively targeting the neutrophil component of the innate cellular immune system. Thus, the goal of the present study was to compare the effect of resistin on bacterial killing using monocultures or co-cultures of monocyte and neutrophil cell lines, as well as to extend our findings to primary immune cells. Results: Our results indicate that human resistin impairs the ability of neutrophils to kill the Gram-negative bacterium Pseudomonas aeruginosa and the Gram-positive bacterium Staphylococcus aureus. In contrast, with the exception of macrophages incubated with P. aeruginosa, resistin did not affect the ability of macrophages or monocytes to kill either Gram-positive or Gram-negative organisms. Furthermore, co-incubation of neutrophils with increasing proportions of monocytes did not enhance bacterial killing. Resistin blocked bactericidal activity through partial reduction of F-actin polymerization and suppression of the oxidative burst in neutrophils. Conclusions: Our studies indicate that resistin selectively impairs neutrophil bacterial killing. These findings further support the notion that resistin can mimic cell type-dependent immunosuppressive effects. This is consistent with its putative role in the pathogenesis of bacterial sepsis. [ABSTRACT FROM AUTHOR]