RESUMO
PURPOSE: In vivo and ex vivo inhibition of ectopic activity of clinically used and newly developed sodium channel (NaV) blockers were quantified in the rat spinal nerve ligation (SNL) model using a pharmacokinetic-pharmacodynamic (PKPD) approach and correlated to in vitro NaV1.7 channel inhibition and clinical effective concentrations. METHODS: In vivo, drug exposure and inhibition of ectopic activity were assessed in anaesthetized SNL rats at two dose levels. Ex vivo, compounds were applied at increasing concentrations to dorsal root ganglias isolated from SNL rats. The inhibitory potency (IC 50 ) was estimated using PKPD analysis. In vitro IC 50 was estimated using an electrophysiology-based assay using recombinant rat and human NaV1.7 expressing HEK293 cells. RESULTS: In vivo and ex vivo inhibition of ectopic activity correlated well with the in vitro inhibition on the rat NaV1.7 channel. The estimated IC 50s for inhibition of ectopic activity in the SNL model occurred at similar unbound concentrations as clinical effective concentrations in humans. CONCLUSIONS: Inhibition of ectopic activity in the SNL model could be useful in predicting clinical effective concentrations for novel sodium channel blockers. In addition, in vitro potency could be used for screening, characterization and selection of compounds, thereby reducing the need for in vivo testing.
Assuntos
Canal de Sódio Disparado por Voltagem NAV1.7/metabolismo , Neuralgia/tratamento farmacológico , Bloqueadores dos Canais de Sódio/sangue , Bloqueadores dos Canais de Sódio/farmacologia , Nervos Espinhais/efeitos dos fármacos , Animais , Células HEK293 , Humanos , Ligadura , Masculino , Ratos , Ratos Sprague-Dawley , Nervos Espinhais/cirurgiaRESUMO
AIM: Through the use of CBCT images, many unidentified features of impacted canines can be easily resolved. The potential collision of impacted maxillary canines and adjacent teeth could lead to root resorption (RR). The aim of this study is to examine the prevalence, location and severity of RR on adjacent teeth caused by impacted maxillary canines and the association between the adjacent teeth and the features of maxillary impacted canines on CBCT. METHODS: This study examined 89 subjects with 108 maxillary impacted canines, having had no previous orthodontic treatment (mean age: 18.3 ± 4.1 years). The following impacted-canine-related parameters were analyzed on the CBCT images: location; RR levels on adjacent teeth; occlusal line and midline distances of impacted canines; and the angulations of impacted canines to the midline, lateral incisor and occlusal line. Logistic regression was used to evaluate the association between RR and the measured parameters on CBCT. RESULTS: In this study, we found that the majority of our patients with impacted maxillary canines were female (62.5%). Of the total 108 maxillary impacted canines, 60.2% resorbed the adjacent teeth of the affected quadrants. Lateral incisors were the most affected (34.3%). The mean age of subjects with RR was 16.7 ± 3.5 years. The frequency of RR was statistically significantly higher in female subjects (40.4%). Slight RR was the most frequent (30.5%) and the highest incidence noted at the apical third of the root (29.6%). Regarding the impacted maxillary canine angulation to the midline and adjacent tooth, higher values of angulation caused severe forms of RR (p < 0.05). CONCLUSION: The sensitivity of CBCT allows for the accurate diagnosis of the location and the degree of RR, alongside the angulation and distance of impacted canines to adjacent teeth. The association between the linear and angular features of the impacted maxillary canines and RR was confirmed.
RESUMO
Increasing attention has been paid to the possible link between periodontal disease and atherosclerosis over the past decade. The aim of this study is to investigate the presence of five periopathogens: Porphyromonas gingivalis (P.g.), Aggregatibacter actinomycetemcomitans (A.a.), Tannerella forsythia (T.f.), Treponema denticola (T.d.), and Prevotella intermedia (P.i.) in atheromatous plaques obtained from the carotid and coronary arteries in patients who underwent coronary artery bypass graft surgery and carotid endarterectomy. Group I (carotid arteries) consisted of 30 patients (mean age: 54.5 ± 14.8), and group II (coronary arteries) consisted of 28 patients (mean age: 63 ± 12.1). Clinical periodontal examinations consisted of plaque index, gingival index, sulcus bleeding index, and periodontal probing depth and were performed on the day of vascular surgery. The presence of periopathogens in periodontal pockets and atherosclerotic vessels was detected using polymerase chain reaction. In both subgingival plaque and atherosclerotic plaque of carotid arteries, P.g., A.a., T.f., T.d., and P.i. were detected in 26.7%, 6.7%, 66.7%, 10.0%, and 20.0%, respectively, while for coronary arteries, P.g. was detected in 39.3%, A.a. in 25%, T.f. in 46.4%, T.d. in 7.1%, and P.i. in 35.7%. The presence of five periopathogens in carotid and coronary atherosclerotic vessels showed correlation in regard to the degree of periodontal inflammation. The present study suggests the relationship between periodontal pathogenic bacteria and atherogenesis. Further studies are necessary in relation to the prevention or treatment of periodontal disease that would result in reduced mortality and morbidity associated with atherosclerosis.
Assuntos
Artérias Carótidas/microbiologia , Vasos Coronários/microbiologia , Placa Aterosclerótica/microbiologia , Adulto , Idoso , Aggregatibacter actinomycetemcomitans , Aterosclerose , Feminino , Humanos , Inflamação , Masculino , Pessoa de Meia-Idade , Doenças Periodontais/imunologia , Reação em Cadeia da Polimerase , Porphyromonas gingivalis , Prevotella intermedia , RNA Ribossômico 16S/metabolismo , Fatores de Risco , Tannerella forsythia , Treponema denticolaRESUMO
Previously we showed that neuropeptide Y (NPY), a sympathetic vasoconstrictor neurotransmitter, stimulates endothelial cell migration, proliferation, and differentiation in vitro. Here, we report on NPY's actions, receptors, and mediators in ischemic angiogenesis. In rats, hindlimb ischemia stimulates sympathetic NPY release (attenuated by lumbar sympathectomy) and upregulates NPY-Y2 (Y2) receptor and a peptidase forming Y2/Y5-selective agonist. Exogenous NPY at physiological concentrations also induces Y5 receptor, stimulates neovascularization, and restores ischemic muscle blood flow and performance. NPY-mediated ischemic angiogenesis is not prevented by a selective Y1 receptor antagonist but is reduced in Y2(-/-) mice. Nonischemic muscle vascularity is also lower in Y2(-/-) mice, whereas it is increased in NPY-overexpressing rats compared with their WT controls. Ex vivo, NPY-induced aortic sprouting is markedly reduced in Y2(-/-) aortas and spontaneous sprouting is severely impaired in NPY(-/-) mice. NPY-mediated aortic sprouting, but not cell migration/proliferation, is blocked by an antifetal liver kinase 1 antibody and abolished in mice null for eNOS. Thus, NPY mediates neurogenic ischemic angiogenesis at physiological concentrations by activating Y2/Y5 receptors and eNOS, in part due to release of VEGF. NPY's effectiveness in revascularization and restoring function of ischemic tissue suggests its therapeutic potential in ischemic conditions.
Assuntos
Isquemia/tratamento farmacológico , Músculo Esquelético/irrigação sanguínea , Neovascularização Patológica/induzido quimicamente , Neuropeptídeo Y/farmacologia , Neuropeptídeo Y/fisiologia , Animais , Dipeptidil Peptidase 4/fisiologia , Fatores de Crescimento Endotelial/fisiologia , Peptídeos e Proteínas de Sinalização Intercelular/fisiologia , Isquemia/patologia , Isquemia/fisiopatologia , Linfocinas/fisiologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Óxido Nítrico/fisiologia , Óxido Nítrico Sintase/deficiência , Óxido Nítrico Sintase/genética , Óxido Nítrico Sintase/fisiologia , Óxido Nítrico Sintase Tipo II , Óxido Nítrico Sintase Tipo III , Ratos , Ratos Sprague-Dawley , Ratos Wistar , Receptores de Neuropeptídeo Y/agonistas , Receptores de Neuropeptídeo Y/deficiência , Receptores de Neuropeptídeo Y/genética , Receptores de Neuropeptídeo Y/fisiologia , Fator A de Crescimento do Endotélio Vascular , Fatores de Crescimento do Endotélio VascularRESUMO
In anaesthetised rats, Fos-immunoreactive and NADPH-diaphorase-positive neurons in the medulla and, for comparison, in the spinal cord were studied after fatiguing stimulation of the hindlimb muscles. Following both direct muscle stimulation and L5 ventral root stimulation, fatigue-related c-fos gene expression was most prominent in the dorsal horn of the ipsilateral L2-L5 segments and within the ipsilateral nucleus tractus solitarius, the caudoventrolateral and rostroventrolateral reticular nuclei, and the intermediate reticular nucleus at levels of -14.3 and -13.8 mm, and contralaterally at -13.2 mm caudal to the bregma. The order of intensity of c-fos expression was as follows: nucleus tractus solitarius>caudoventrolateral and rostroventrolateral reticular nuclei>intermediate reticular nucleus>lateral paragigantocellular nucleus. NADPH-diaphorase reactivity was changed in the following sequence: NTS>intermediate reticular nucleus lateral paragigantocellular nucleus>rostroventrolateral reticular nucleus. Fos-immunoreactive neurons were codistributed with NADPH-diaphorase-reactive cells within the dorsomedial and ventrolateral medulla, and double-staining neurons were found in the nucleus tractus solitarius, intermediate reticular nucleus and lateral paragigantocellular nucleus. The patterns of distribution of c-fos expression and NADPH-diaphorase reactivity show that afferent signals arising from fatiguing muscles may activate spinal and medullary neurons which are involved in nociceptive and cardiovascular reflex pathways. The functional role of nitric oxide (NO) in the generation of cardiovascular and somatosensory responses in the medulla during fatigue of skeletal muscles is discussed.
Assuntos
Bulbo/metabolismo , Músculo Esquelético/fisiologia , NADPH Desidrogenase/metabolismo , Neurônios/enzimologia , Proteínas Proto-Oncogênicas c-fos/biossíntese , Animais , Barorreflexo , Pressão Sanguínea/fisiologia , Estimulação Elétrica , Frequência Cardíaca/fisiologia , Membro Posterior/fisiologia , Imuno-Histoquímica , Masculino , Bulbo/citologia , Contração Muscular/fisiologia , Fadiga Muscular/fisiologia , Músculo Esquelético/inervação , Neurônios/química , Ratos , Ratos Wistar , Medula Espinal/citologia , Medula Espinal/metabolismo , Raízes Nervosas Espinhais/fisiologiaRESUMO
It is by now well established that tetrodotoxin-resistant (TTX-R) afferent fibres from muscle in the rat exhibit a multisensitive profile, including nociception. TTX-R afferent fibres play an important role in motor control, via spinal and supraspinal loops, but their activation and function during muscle exercise and fatigue are still unknown. Therefore, the specific effect of isometric fatiguing muscle contraction on the responsiveness of TTX-R C-fibres has been investigated in this study. To quantify the TTX-R afferent input we recorded the cord dorsum potential (CDP), which is the result of the electrical fields set up within the spinal cord by the depolarisation of the interneurons located in the dorsal horn, activated by an incoming volley of TTX-R muscle afferents. The changes in TTX-R CDP size before, during and after fatiguing electrical stimulation of the gastrocnemius-soleus (GS) muscle have been taken as a measure of TTX-R C-unit activation. At the end of the fatiguing protocol, following an exponential drop in force, TTX-R CDP area decreased in the majority of trials (9/14) to 0.75 ± 0.03% (mean ± SEM) of the pre-fatigue value. Recovery to the control size of the TTX-R CDP was incomplete after 10 min. Furthermore, fatiguing trials could sensitise a fraction of the TTX-R C-fibres responding to muscle pinch. The results suggest a long-lasting activation of the TTX-R muscle afferents after fatiguing stimulation. The role of this behaviour in chronic muscle fatigue in connection with pain development is discussed. Accumulation of metabolites released into the interstitium during fatiguing stimulation might be one of the reasons underlying the C-fibres' long-lasting activation.
Assuntos
Anestésicos Locais/farmacologia , Contração Isométrica/efeitos dos fármacos , Fadiga Muscular/efeitos dos fármacos , Fadiga Muscular/fisiologia , Fibras Nervosas Amielínicas/efeitos dos fármacos , Fibras Nervosas Amielínicas/fisiologia , Tetrodotoxina/farmacologia , Vias Aferentes , Animais , Potenciais Evocados , Masculino , RatosRESUMO
Investigation and interpretation of defective motor circuitries in transgenic mice required further basic results from wild-type mice. Therefore, we investigated the lumbar motor reflex pattern in anaesthetised mice using intracellular motoneuronal recording and monosynaptic reflex testing. Thresholds and latencies in mice were similar to those in cats: thresholds for monosynaptic (group I) EPSPs were slightly above 1T (T=threshold for the lowest threshold fibres), around 1.5T for group II EPSPs and above 10T for group III EPSPs; group I EPSPs were maximal with a stimulus strength around 2T, group II EPSPs were maximal with 5-8T; latencies to the group I incoming volley were below 1ms for monosynaptic group I EPSPs, around 3ms for polysynaptic group II EPSPs and above 4ms for polysynaptic group III EPSPs. In contrast to reflex actions in the cat, monosynaptic gastrocnemius-soleus reflexes were facilitated by conditioning stimulation of the peroneal, sural and tibial nerves, i.e. by a variety of different, probably flexor reflex afferents. This facilitation persisted after high lumbar spinalisation indicating an independency to supraspinal influences. Nociceptive muscle afferents facilitated the peroneal monosynaptic reflex while nociceptive cutaneous afferents from the foot sole inhibited the ipsilateral but facilitated the contralateral peroneal reflex.
Assuntos
Vias Aferentes/fisiologia , Neurônios Motores/fisiologia , Neurônios Aferentes/fisiologia , Reflexo/fisiologia , Medula Espinal/fisiologia , Animais , Gatos , Estimulação Elétrica , Potenciais Pós-Sinápticos Excitadores/fisiologia , Pé/inervação , Região Lombossacral , CamundongosRESUMO
Responses of gastrocnemius-soleus (G-S) motoneurones to stretches of the homonymous muscles were recorded intracellularly in decerebrate cats before, during and after fatiguing stimulation (FST) of G-S muscles. Ventral roots (VR) L7 and S1 were cut, and FST was applied to VR S1, a single FST session including 4 to 5 repetitions of 12-s periods of regular 40 s(-1) stimulation. Muscle stretches consisted of several phases of slow sinusoidal shortening-lengthening cycles and intermediate constant lengths. The maximal stretch of the muscles was 8.8 mm above the rest length. Effects of FST on excitatory postsynaptic potentials (EPSPs) and spikes evoked by the muscle stretches were studied in 12 motoneurones from ten experiments. Stretch-evoked EPSPs and firing were predominantly suppressed after FST, with the exception of a post-contraction increase of the first EPSP after FST, which was most likely due to after-effects in the activity of muscle spindle afferents. The post-fatigue suppression of EPSPs and spike activity was followed by restoration within 60-100 s. Additional bouts of FST augmented the intensity of post-fatigue suppression of EPSPs, with the spike activity sometimes disappearing completely. FST itself elicited EPSPs at latencies suggesting activation of muscle spindle group Ia afferents via stimulation of beta-fibres. The suppression of the stretch-evoked responses most likely resulted from fatigue-evoked activity of group III and IV muscle afferents. Presynaptic inhibition could be one of the mechanisms involved, but homosynaptic depression in the FST-activated group Ia afferents may also have contributed.
Assuntos
Neurônios Motores/fisiologia , Fadiga Muscular/fisiologia , Músculo Esquelético/citologia , Músculo Esquelético/fisiologia , Animais , Gatos , Potenciais Evocados/fisiologia , Potenciais Pós-Sinápticos Excitadores/fisiologia , Feminino , Masculino , Fusos Musculares/fisiologia , Neurônios Aferentes/fisiologia , Reflexo de Estiramento/fisiologia , Medula Espinal/citologia , Medula Espinal/fisiologiaRESUMO
The objective of this investigation was to determine the percentage of microinvasive and invasive cancers found when CIN 3 was treated by hysterectomy. The postoperative histologic findings of all patients with CIN 3 treated by conization or hysterectomy were analyzed. In 295 patients treated by conization, 14 (4.75%) microinvasive and 11 (3.73%) invasive cancers were found. Histologic analysis of 106 hysterectomy specimens revealed microinvasive carcinoma in 11 (10.38%) and invasive carcinoma in 17 (16.04%) cases. The finding of unexpected invasive cervical cancer in hysterectomy specimens seems significantly higher compared with conization. One must be cautious to avoid hysterectomy as the primary mode of treatment for women with coincident uterine pathology, postmenopausal women, or those who do not desire further reproduction. Hysterectomy as a mode of treatment for CIN 3 should be performed only if the existence of invasive cancer was excluded with certainty.
RESUMO
In decerebrate cats, changes in the monosynaptic reflex (MSR) of gastrocnemius-soleus (G-S) motoneurones were studied after fatiguing stimulation (FST) of the G-S muscles. Monosynaptic reflexes were evoked by stimulation of Ia fibres in the G-S nerve and recorded from a filament of ventral root (VR) L7. FST (intermittent 40 s(-1) stimulation for 10-12 min) was applied to the distal part of the cut VR S1. FST reduced MSR amplitudes to 0.64 +/- 0.04 (mean +/-s.e.m.) of the prefatigue values. The suppression remained stable for approximately 25 min and then MSR amplitudes gradually returned towards the normal. To test for the involvement of presynaptic and recurrent inhibition, MSRs were conditioned by stimulation of the nerve to the posterior biceps and semitendinosus (PBSt) muscles or a filament of VR L7, respectively. The intensity of presynaptic inhibition (reduction of the normalized value of MSR amplitude during conditioning) increased from 0.19 +/- 0.02 in prefatigue to 0.44 +/- 0.04 within a 5.3-18.2 min interval after FST, followed by a recovery. In contrast, the intensity of recurrent inhibition first diminished from 0.23 +/- 0.02 in prefatigue to 0.15 +/- 0.01 within 15.6-30.1 min after FST and then gradually recovered. Both primary afferent depolarization and the intensity of antidromic discharges in primary afferents increased with the presynaptic inhibition intensity. These results demonstrate a fatigue-related suppression of Ia excitation of synergistic motoneurones, probably arising from the activation of group III and IV afferents. The effects could in part be due to increased presynaptic inhibition, while recurrent inhibition plays a minor role.
Assuntos
Fadiga Muscular/fisiologia , Músculo Esquelético/fisiologia , Reflexo Monosináptico/fisiologia , Animais , Gatos , Estado de Descerebração , Estimulação Elétrica , Feminino , Membro Posterior , Masculino , Músculo Esquelético/inervação , Fibras Nervosas/fisiologia , Inibição Neural , Terminações Pré-Sinápticas/fisiologia , Fatores de TempoRESUMO
The aim of the study was to characterize the species-related differences in the distribution of nitric oxide synthase (NOS)-containing neurons in rodents and carnivores medullary cardiovascular centers that take part in regulation of the sympathetic or parasympathetic drives. The order of the mean number of NOS-containing neurons in the dorsomedial and ventrolateral medulla (per section) in different animals was as follows: dog>cat>rat. Although the density of the positive cells in the both regions was changed in the following sequence: rat=cat>dog. Within the dorsal motor nucleus of vagus significant exceeding of the mean number and density of positive cells (preganglionic vagal neurons) in dog were found. Differences in the distribution of NO-generating neurons in the medullary cardiovascular centers and the heterogeneity in the basal level of NO release may contribute to the distinctive alterations of the hemodynamic reactions in the studied species after administration of NOS inhibitors.