Does the Presence or a High Titer of Yellow Fever Virus Antibodies Interfere with Pregnancy Outcomes in Women with Zika Virus Infection?

Zika virus (ZIKV) and yellow fever virus (YFV) originated in Africa and expanded to the Americas, where both are co-circulated. It is hypothesized that in areas of high circulation and vaccination coverage against YFV, children of pregnant women have a lower risk of microcephaly. We evaluated the presence and titers of antibodies and outcomes in women who had ZIKV infection during pregnancy. Pregnancy outcomes were classified as severe, moderate, and without any important outcome. An outcome was defined as severe if miscarriage, stillbirth, or microcephaly occurred, and moderate if low birth weight and/or preterm delivery occurred. If none of these events were identified, the pregnancy was defined as having no adverse effects. A sample of 172 pregnant women with an acute ZIKV infection confirmed during pregnancy were collected throughout 2016. About 89% (150 of 169) of them presented immunity against YFV, including 100% (09 of 09) of those who had severe outcomes, 84% (16 of 19) of those who had moderate outcomes, and 89% (125 of 141) of those who had non-outcomes. There was no difference between groups regarding the presence of anti-YFV antibodies (p = 0.65) and YFV titers (p = 0.6). We were unable to demonstrate a protective association between the presence or titers of YFV antibodies and protection against serious adverse outcomes from exposure to ZIKV in utero.

The role of APC/C in cell cycle dynamics, growth and development in cereal crops.

Cereal crops can be considered the basis of human civilization. Thus, it is not surprising that these crops are grown in larger quantities worldwide than any other food supply and provide more energy to humankind than any other provision. Additionally, attempts to harness biomass consumption continue to increase to meet human energy needs. The high pressures for energy will determine the demand for crop plants as resources for biofuel, heat, and electricity. Thus, the search for plant traits associated with genetic increases in yield is mandatory. In multicellular organisms, including plants, growth and development are driven by cell division. These processes require a sequence of intricated events that are carried out by various protein complexes and molecules that act punctually throughout the cycle. Temporal controlled degradation of key cell division proteins ensures a correct onset of the different cell cycle phases and exit from the cell division program. Considering the cell cycle, the Anaphase-Promoting Complex/Cyclosome (APC/C) is an important conserved multi-subunit ubiquitin ligase, marking targets for degradation by the 26S proteasome. Studies on plant APC/C subunits and activators, mainly in the model plant Arabidopsis, revealed that they play a pivotal role in several developmental processes during growth. However, little is known about the role of APC/C in cereal crops. Here, we discuss the current understanding of the APC/C controlling cereal crop development.

Towards the Laboratory Maintenance of Haemagogus janthinomys (Dyar, 1921), the Major Neotropical Vector of Sylvatic Yellow Fever.

Haemagogus (Haemagogus) janthinomys (Dyar, 1921), the major neotropical vector of sylvatic yellow fever virus, is notoriously difficult to maintain in captivity. It has never been reared beyond an F1 generation, and almost no experimental transmission studies have been performed with this species since the 1940s. Herein we describe installment hatching, artificial blood feeding, and forced-mating techniques that enabled us to produce small numbers of F3 generation Hg. janthinomys eggs for the first time. A total of 62.8% (1562/2486) F1 generation eggs hatched during ≤10 four-day cycles of immersion in a bamboo leaf infusion followed by partial drying. Hatching decreased to 20.1% (190/944) in the F2 generation for eggs laid by mosquitoes copulated by forced mating. More than 85% (79/92) female F2 mosquitoes fed on an artificial blood feeding system. While we were unable to maintain a laboratory colony of Hg. janthinomys past the F3 generation, our methods provide a foundation for experimental transmission studies with this species in a laboratory setting, a critical capacity in a region with hyper-endemic transmission of dengue, Zika, and chikungunya viruses, all posing a risk of spillback into a sylvatic cycle.

Protective effect of anti-erosive solutions enhanced by an aminomethacrylate copolymer.

OBJECTIVE: To investigate if an aminomethacrylate copolymer (AMC) could potentiate the anti-erosive effect of solutions containing sodium fluoride -F (225 ppm F-) and sodium fluoride associated to stannous chloride -FS (800 ppm Sn2+). METHODS: The experimental solutions (F, FS, AMC, AMC + F, AMC + FS, and deionized water-DW as negative control) were tested in the presence of acquired pellicle. Polished bovine enamel specimens (n = 13/group) were submitted to an erosion-rehardening cycle (2 h immersion in human saliva, 5 min in 0.3 % citric acid, 1 h in human saliva, 4×/day, 5 days). Treatment with the solutions was performed for 2 min, 2×/day. The rehardening (%Re) and protective (%Prot) potential of the solutions were assessed in the beginning of the experiment, and the surface loss (SL) by contact profilometry after 5 days. Additional bovine specimens (n = 5/group) were prepared to evaluate the contact angle on the treated enamel surface. The zeta potential of the dispersed hydroxyapatite (HA) crystals after the treatment with the solutions was also measured (n = 3/group). Data were statistically analyzed (α = 0.05). RESULTS: The association with AMC improved the %Re and the %Prot for W and F, but not for FS. The results of SL were: AMC + F = AMC + FS < AMC < FS < F < DW. The presence of AMC significantly reduced the contact angle on enamel surfaces. The HA presented a strong negative surface charge after the treatment with DW, F and FS, whereas after the treatment with the solutions containing AMC it became positive. CONCLUSION: AMC has potential to enhance the anti-erosive effect of fluoride solutions. CLINICAL SIGNIFICANCE: The aminomethacrylate copolymer (AMC) may be a promising agent to be added to oral care products for the prevention of erosive tooth wear.

Automated needle-sleeve based online hyphenation of solid-phase microextraction and liquid chromatography.

A novel approach for the online coupling of solid-phase microextraction (SPME) and liquid chromatography (LC) is introduced. An innovative Si@GO@ßCD coated needle-sleeve extractant device was developed and then employed in the automated online SPME-LC-UV determination of estrogen-like isoflavones from human urine samples. The extractant SPME device is easily attachable at the endpoint of an analytical syringe needle and operated by a lab-made autosampler. Fully automated online SPME-LC is accomplished by proper autosampler programming to perform the following steps: i) the analytes extraction by direct immersion of the extractant device into the stirred sample, ii) a rinsing step iii) the analytes desorption/enrichment, iv) the online transference of the extract to the LC injection valve. Besides allowing the online SPME hyphenation, this extraction modality efficiently addressed the drawbacks associated with the clogging and dispersion of graphene-based microextraction techniques performed in packed-bed and dispersive formats. The main extraction parameters and the performance of the automated online SPME-LC method developed were carefully studied. The results show a good sensitivity, reliability, and straightforward analytical strategy for the determination of organic compounds in complex samples. The detection limit of the method was 20 µg L1 for DAI and 10 µg L-1 for GEN, FOR and BIO. The intra-day RSD was below 10% and inter-day RSD was below 13%. The total analysis time was less than 17 min per sample.

ß-Cyclodextrin coupled to graphene oxide supported on aminopropyl silica as a sorbent material for determination of isoflavones.

ß-Cyclodextrin, coupled to graphene oxide supported on aminopropyl silica, was synthesized and characterized. This material was combined with microextraction by packed sorbent to act as the sample preparation step. The analytical method optimization was carried out by employing experimental design and had its figures of merit determined. The resulting linearity ranged from 1.0 to 200 µg/L for daidzein, from 2.0 to 200 µg/L for genistein, from 3.0 to 200 µg/L for formononetin, and from 2.0 to 200 µg/L for biochanin A with all R2 values above 0.993 and limit of quantification ranging from 0.5 to 1.5 µg/L. The accuracy ranged from 93.3 to 123.3%, and intraday and interday precision reported by the relative standard deviations were <16%. This work aimed to synthesize and evaluate cyclodextrins coupled to graphene-based sorbents to be used as a high sorption capacity and selective sorbent for sample preparation of complex matrices using microextraction techniques. The synthesized material kept the high absorption characteristic of graphene-based materials while maintaining the cyclodextrins' selectivity to extract the target analyte. Four isoflavones were determined in soy-based juice samples from the local market, confirming the excellent performance of the proposed method.

The Current Role of Graphene-Based Nanomaterials in the Sample Preparation Arena.

Since its discovery in 2004 by Novoselov et al., graphene has attracted increasing attention in the scientific community due to its excellent physical and chemical properties, such as thermal/mechanical resistance, electronic stability, high Young's modulus, and fast mobility of charged atoms. In addition, other remarkable characteristics support its use in analytical chemistry, especially as sorbent. For these reasons, graphene-based materials (GBMs) have been used as a promising material in sample preparation. Graphene and graphene oxide, owing to their excellent physical and chemical properties as a large surface area, good mechanical strength, thermal stability, and delocalized π-electrons, are ideal sorbents, especially for molecules containing aromatic rings. They have been used in several sample preparation techniques such as solid-phase extraction (SPE), stir bar sorptive extraction (SBSE), magnetic solid-phase extraction (MSPE), as well as in miniaturized modes as solid-phase microextraction (SPME) in their different configurations. However, the reduced size and weight of graphene sheets can limit their use since they commonly aggregate to each other, causing clogging in high-pressure extractive devices. One way to overcome it and other drawbacks consists of covalently attaching the graphene sheets to support materials (e.g., silica, polymers, and magnetically modified supports). Also, graphene-based materials can be further chemically modified to favor some interactions with specific analytes, resulting in more efficient hybrid sorbents with higher selectivity for specific chemical classes. As a result of this wide variety of graphene-based sorbents, several studies have shown the current potential of applying GBMs in different fields such as food, biological, pharmaceutical, and environmental applications. Within such a context, this review will focus on the last five years of achievements in graphene-based materials for sample preparation techniques highlighting their synthesis, chemical structure, and potential application for the extraction of target analytes in different complex matrices.

Effects of increasing nitrogen levels in Mombasa grass on pasture characteristics, chemical composition, and beef cattle performance in the humid tropics of the Amazon.

Improving beef cattle production in pasture-based systems without expanding agricultural land has been the focus of several studies over the last decades. Nitrogen fertilization is one of the available strategies with high potential to optimize cattle performance in tropical systems. This study aimed to evaluate the effects of increasing nitrogen levels (150, 300, and 450 kg ha-1 of N year-1) in Megathyrsus maximus cv. Mombasa in the humid tropics of the Amazon. The following parameters were evaluated: pasture agronomic characteristics, herbage chemical composition, and beef cattle performance. A total of 24 nine-month-old Nellore bulls with initial mean weight of 173 ± 1.95 kg were used for the performance test. The experimental design was a randomized block with three treatments. Herbage mass and leaf mass contents were increased by the highest nitrogen level (P < 0.05). Nitrogen levels elevated the forage accumulation rate, final leaf blade length, and pre-grazing canopy height(P < 0.05). The crude protein content increased at the highest N level (P < 0.05). The neutral detergent fiber and acid detergent fiber contents decreased with increasing N levels. The stocking rate and total weight gain per hectare were higher in bulls grazing pastures fertilized with 450 compared with 150 kg ha-1 of N year-1(P < 0.05). The average daily gain was influenced by N levels (P < 0.05). Nitrogen levels at 300 and 450 kg increased animal weight gain and productivity per area by increasing forage yield and improving the chemical composition of Mombasa grass.

Rostral ventromedial medulla connections in Cavia porcellus and their relation with tonic immobility defensive behavior: a biotinylated dextran amine neurotracing study.

The rostroventromedial medulla (RVM), together with the periaqueductal gray matter (PAG), constitutes the descendent antinociceptive system. Additionally, these structures mutually regulate defensive behaviors, including tonic immobility (TI) in guinea pigs. The current study was undertaken to evaluate the connections of the RVM with the PAG and the spinal cord in guinea pigs in order to provide an anatomical basis for the role played by RVM in the modulation of TI. To address this goal, five guinea pigs were treated with non-fluorescent biotinylated dextran amine (BDA) neurotracer by injection into the RVM. After four days of survival, the encephalon and spinal cord were removed from each rodent, and BDA labeling was visualized with a standard avidin-biotinylated horseradish peroxidase method through reaction with nickel-intensified peroxidase 3,3'-diaminobenzidine dihydrochloride. The microinjection of BDA into the RVM stained fibers in the ventral horn, dorsal horn and intermediate gray matter of the spinal cord. BDA-labeled fibers, terminal buttons suggesting synaptic contacts, and perikarya were found in the dorsomedial, dorsolateral, lateral and ventrolateral PAG, and neuronal somata were identified in the cuneiform nucleus. Together, the current data demonstrate neuroanatomical evidence that supports the role of the RVM in the modulation of TI defensive behavior.

Rostral ventromedial medulla modulates nociception and tonic immobility behavior through connections with the A7 catecholaminergic region.

Cholinergic stimulation of the rostral ventromedial medulla (RVM) produces antinociception and reduces the duration of tonic immobility (TI) behavior in guinea pigs. Previous studies indicated that cholinergic antinociception in the RVM is mediated through connections with the A7 catecholaminergic cell group (A7). In the current study, we tested the role of the A7 in both the antinociception and reduction of TI duration mediated by cholinergic stimulation of the RVM. In addition, we used biotinylated dextran amines (BDA) to evaluate the connections between the RVM and A7. The microinjection of the cholinergic agonist carbachol into the RVM produced antinociception and reduced TI behavior duration. These effects were blocked by prior administration of lidocaine to the A7. However, the microinjection of lidocaine into the A7 prior to saline injection into the RVM had no effect on either the nociceptive or TI responses. The microinjection of the neurotracer BDA into the RVM positively stained fibers and synaptic boutons in the A7, indicating that there are direct projections from the RVM to the A7. Taken together, our results indicate that the antinociception and reduction of TI behavior duration after cholinergic stimulation of the RVM depends on connections with the A7.

LQB-118, a pterocarpanquinone structurally related to lapachol [2-hydroxy-3-(3-methyl-2-butenyl)-1,4-naphthoquinone]: a novel class of agent with high apoptotic effect in chronic myeloid leukemia cells.

Despite the relevant therapeutic progresses obtained with imatinib, clinical resistance to this drug has emerged and reemerged after cytogenetic remission in a group of patients with chronic myeloid leukemia (CML). Therefore, novel treatment strategies are needed. In this study, we evaluated the anti-CML activity and mechanisms of action of LQB-118, a pterocarpanquinone structurally related to lapachol [2-hydroxy-3-(3-methyl-2-butenyl)-1,4-naphthoquinone]. LQB-118 treatment resulted in an important reduction of cell viability in cell lines derived from CML, both the vincristine-sensitive K562 cell line, and the resistant K562-Lucena (a cell line overexpressing P-glycoprotein). In agreement with these results, the induction of caspase-3 activation by this compound indicated that a significant rate of apoptosis was taking place. In these cell lines, apoptosis induced by LQB-118 was accompanied by a reduction of P-glycoprotein, survivin, and XIAP expression. Moreover, this effect was not restricted to cell lines as LQB-118 produced significant apoptosis rate in cells from CML patients exhibiting multifactorial drug resistance phenotype such as P-glycoprotein, MRP1 and p53 overexpression. The data suggest that LQB-118 has a potent anti-CML activity that can overcome multifactorial drug resistance mechanisms, making this compound a promising new anti-CML agent.

Changes in expression of NMDA-NR1 receptor subunits in the rostral ventromedial medulla modulate pain behaviors.

NMDA receptors have an important role in pain facilitation in rostral ventromedial medulla (RVM) and the NR1 subunit is essential for its function. Studies suggest that the NMDA receptors in RVM are critical to modulate both cutaneous and muscle hypersensitivity induced by repeated intramuscular acid injections. We propose that increased expression of the NR1 subunit in the RVM is critical for the full development of hypersensitivity. To test this we used recombinant lentiviruses to over-express the NR1 subunit in the RVM and measured nociceptive sensitivity to cutaneous and muscle stimuli. We also downregulated the expression of NR1 in the RVM and measured the hyperalgesia produced by repeated-acid injections. Increasing the expression of NR1 in the RVM reduces cutaneous and muscle withdrawal threshold, and decreasing the expression of NR1 in the RVM increases the muscle withdrawal threshold and prevents the development of hyperalgesia in an animal model of muscle pain. These results suggest that the NR1 subunits in the RVM are critical for modulating NMDA receptor function, which in turn sets the 'tone' of the nervous system's response to noxious stimuli and tissue injury.

Opioidergic and GABAergic mechanisms in the rostral ventromedial medulla modulate the nociceptive response of vocalization in guinea pigs.

Vocalization generated by the application of a noxious stimulus is an integrative response related to the affective-motivational component of pain. The rostral ventromedial medulla (RVM) plays an important role in descending pain modulation, and opiates play a major role in modulation of the antinociception mediated by the RVM. Further, it has been suggested that morphine mediates antinociception indirectly, by inhibition of tonically active GABAergic neurons. The current study evaluated the effects of the opioids and GABA agonists and antagonists in the RVM on an affective-motivational pain model. Additionally, we investigated the opioidergic-GABAergic interaction in the RVM in the vocalization response to noxious stimulation. Microinjection of either morphine (4.4nmol/0.2microl) or bicuculline (0.4nmol/0.2microl) into the RVM decreased the vocalization index, whereas application of the GABA(A) receptor agonist, muscimol (0.5nmol/0.2microl) increased the vocalization index during noxious stimulation. Furthermore, prior microinjection of either the opioid antagonist naloxone (2.7nmol/0.2microl) or muscimol (0.25nmol/0.2microl) into the RVM blocked the reduction in vocalization index induced by morphine. These observations suggest an antinociceptive and pro-nociceptive role of the opioidergic and GABAergic neurotransmitters in the RVM, respectively. Our data show that opioids have an antinociceptive effect in the RVM, while GABAergic neurotransmission is related to the facilitation of nociceptive responses. Additionally, our results indicate that the antinociceptive effect of the opioids in the RVM could be mediated by a disinhibition of tonically active GABAergic interneurons in the downstream projection neurons of the descending pain control system; indicating an interaction between the opioidergic and GABAergic pathways of pain modulation.

Cholecystokinin receptors mediate tolerance to the analgesic effect of TENS in arthritic rats.

Transcutaneous electrical nerve stimulation (TENS) is a treatment for pain that involves placement of electrical stimulation through the skin for pain relief. Previous work from our laboratory shows that repeated application of TENS produces analgesic tolerance by the fourth day and a concomitant cross-tolerance at spinal opioid receptors. Prior pharmacological studies show that blockade of cholecystokinin (CCK) receptors systemically and spinally prevents the development of analgesic tolerance to repeated doses of opioid agonists. We therefore hypothesized that systemic and intrathecal blockade of CCK receptors would prevent the development of analgesic tolerance to TENS, and cross-tolerance at spinal opioid receptors. In animals with knee joint inflammation (3% kaolin/carrageenan), high (100Hz) or low frequency (4Hz) TENS was applied daily and the mechanical withdrawal thresholds of the muscle and paw were examined. We tested thresholds before and after inflammation, and before and after TENS. Animals treated systemically, prior to TENS, with the CCK antagonist, proglumide, did not develop tolerance to repeated application of TENS on the fourth day. Spinal blockade of CCK-A or CCK-B receptors blocked the development of tolerance to high and low frequency TENS, respectively. In the same animals we show that spinal blockade of CCK-A receptors prevents cross-tolerance at spinal delta-opioid receptors that normally occurs with high frequency TENS; and blockade of CCK-B receptors prevents cross-tolerance at spinal mu-opioid receptors that normally occurs with low frequency TENS. Thus, we conclude that blockade of CCK receptors prevents the development of analgesic tolerance to repeated application of TENS in a frequency-dependent manner.

Sebaceous carcinoma of the penis.

Sebaceous carcinoma is a very aggressive malignant tumor, derived from the adnexal epithelium of sebaceous glands. Extraocular sebaceous carcinoma is a very uncommon neoplasm usually localized on the head and neck. To our knowledge, there are only 2 previously reported cases of sebaceous carcinoma on the penis. We report the clinicopathologic data on 3 additional cases of sebaceous carcinoma arising in the penis. Treatment is debatable in view of the fact that this kind of tumor has a high recurrence rate and early regional lymph node involvement. Considering these facts, we used preoperative lymphoscintigraphy, intraoperative lymph node mapping and sentinel node biopsy before performing a bilateral inguinal lymphadenectomy in 1 of 3 patients treated in our institute.

Modulation of tonic immobility in guinea pig PAG by homocysteic acid, a glutamate agonist.

Tonic immobility (TI) is an innate defensive behavior elicited by physical restriction and postural inversion, and is characterized by a profound and temporary state of motor inhibition. The participation of the periaqueductal gray matter (PAG) in TI modulation has previously been described. In addition, the excitatory amino acids (EAA) are important mediators involved in the adjustment of several defensive responses produced by PAG. In the present study, we investigated the effect of microinjection of the EAA agonist dl-homocysteic acid (DLH) and the N-methyl-d-aspartate (NMDA) receptor antagonist (MK-801) into the ventrolateral and dorsal PAG over the duration of TI in guinea pigs. Microinjection of 15 nmol/0.2 microl of DLH into the ventrolateral PAG (vlPAG) and 30 nmol/0.2 microl of DLH into the dorsal PAG (dPAG) promoted an increase and decrease in TI duration, respectively. These responses were blocked by prior microinjection of the NMDA receptor antagonist, MK-801 (3.6 nmol/0.2 microl) at the same site. Microinjection of MK-801 alone into the vlPAG and dPAG did not alter the duration of TI episodes. These results suggest that NMDA receptors are involved in the modulation of TI in both the vlPAG and dPAG. In addition, PAG excitatory amino acids modulate the TI response via columnar organization of the PAG. In this manner, the vlPAG facilitates TI modulation whereas dPAG has an inhibitory role in TI.

Role of opioidergic and GABAergic neurotransmission of the nucleus raphe magnus in the modulation of tonic immobility in guinea pigs.

Tonic immobility (TI) is an inborn defensive behavior characterized by a temporary state of profound and reversible motor inhibition elicited by some forms of physical restraint. Previous results from our laboratory have demonstrated that nucleus raphe magnus (NRM) is also a structure involved in the modulation of TI behavior, as chemical stimulation through carbachol decreases the duration of TI in guinea pigs. In view of the fact that GABAergic and opioidergic circuits participate in the regulation of neuronal activity in the NRM and since these neurotransmitters are also involved in the modulation of TI, the objective of the present study was to evaluate the role of these circuits of the NRM in the modulation of the behavioral TI response. Microinjection of morphine (4.4 nmol/0.2 microl) or bicuculline (0.4 nmol/0.2 microl) into the NRM increased the duration of TI episodes while muscimol (0.5 nmol/0.2 microl) decreased it. The effect of morphine injection into the NRM was blocked by previous microinjection of naloxone (2.7 nmol/0.2 microl). Muscimol at 0.25 nmol did not produce any change in TI duration; however, it blocked the increased response induced by morphine. Our results indicate a facilitatory role of opioidergic neurotransmission in the modulation of the TI response within the NRM, whereas GABAergic activity plays an inhibitory role. In addition, in the present study the modulation of TI in the NRM possibly occurred via an interaction between opioidergic and GABAergic systems, where the opioidergic effect might be due to inhibition of tonically active GABAergic interneurons.

Role of probucol in inhibiting intimal hyperplasia after coronary stent implantation: a randomized study.

BACKGROUND: Oxygen-free radicals can stimulate smooth muscle cell proliferation and may therefore be involved in the genesis of in-stent restenosis. Thus, treatment with probucol, a potent antioxidant agent that has been shown to reduce restenosis after balloon angioplasty, may be an effective strategy to prevent intimal hyperplasia after stenting. METHODS: In a prospective double-blind study, 59 patients submitted to coronary stent implantation were randomly assigned to treatment with either probucol (1 g/d) or placebo, starting two weeks before the procedure and continued for 6 months. The primary end point was the intimal hyperplasia volume at 6 months measured by intravascular ultrasound (IVUS) imaging. RESULTS: Of the 59 randomized patients, 54 underwent successful stent implantation, completed the follow-up period, and underwent repeat angiography, 6.1 +/- 1.1 months after the procedure. Volumetric IVUS analysis revealed similar intimal hyperplasia volumes (403 +/- 26.7 mm3 for probucol vs 44.8 +/- 28.3 mm3 for placebo) and percent volume obstruction of the lumen (30.4% +/- 14.5% for probucol versus 30.7% +/- 17.2% for placebo) in both groups. In addition, quantitative coronary angiography showed no differences in late loss (1.0 +/- 0.8 mm vs 1.1 +/- 0.8 mm), loss index (0.5 +/- 0.4 for both groups), or angiographic restenosis rates (19.4% vs 18.5%) between the probucol and placebo groups, despite the observation of significant changes in the lipid profile and in the plasma antioxidant defenses in patients receiving probucol. CONCLUSIONS: Treatment with the antioxidant probucol failed to reduce neointimal formation after coronary stent implantation as assessed by IVUS volumetric analysis.

Cholinergic modulation of tonic immobility and nociception in the NRM of guinea pig.

Tonic immobility (TI) is an inborn defensive behavior characterized by a temporary state of profound and reversible motor inhibition elicited by some forms of physical restraint. It is known that endogenous antinociceptive systems are activated during the emission of defensive behaviors including TI. The nucleus raphe magnus (NRM) is related to the modulation of nociceptive and behavioral responses. In the present study, we investigated the role of the cholinergic system of the NRM in the modulation of TI and nociception in guinea pigs. Microinjection of the cholinergic agonist carbachol (0.5 microg/0.2 microl) into the NRM promoted a reduction in the duration of TI episodes and nociception, the latter measured by the vocalization test in guinea pigs. The effect of microinjection of carbachol on TI reduction and antinociception was blocked by the previous microinjection of the cholinergic antagonist atropine (0.5 microg/0.2 microl and 1 microg/0.2 microl, respectively), demonstrating the participation of muscarinic receptors in the modulation of these responses. Microinjection of atropine per se did not interfere with the duration of TI episodes. In summary, the present results demonstrate that cholinergic stimulation of the NRM promoted analgesia and a reduction in the duration of TI in guinea pigs. These data indicate that the NRM possibly contributes to the modulation of defensive and nociceptive behavioral responses, probably by modulating the activity of neurons in the ventral and dorsal horn of the spinal cord, respectively.