RESUMO
The present study investigated the effects of 25 and 50% dietary inclusion of yellow mealworm, Tenebrio molitor (TM), larvae on the intestinal antioxidant enzymes, lipid peroxidation and immune system of rainbow trout, Oncorhynchus mykiss. Fish were fed for 90 days with a control diet (TM0) or with 2 test diets (TM25 and TM50) where 35 and 67% of the fishmeal was replaced by TM larvae meal respectively. At the end of the growth trial, humoral immune parameters were assessed in the sera of the fish and the activities of antioxidant enzymes were measured in intestine homogenates. The present results show a strong increase of the activity of the antioxidant intestinal enzymes and reduction of lipid peroxidation. Increased trypsin inhibition and faster antibacterial activity of the serum were also detected. These evidences could be due to the physiological similarities between the exoskeleton of parasites and insects. The present findings reinforce the previous results which revealed a positive effect of the dietary TM on rainbow trout growth and further confirm its adequacy for fishmeal partial replacement in aquafeed.
Assuntos
Suplementos Nutricionais , Larva , Oncorhynchus mykiss/imunologia , Tenebrio , Animais , Dieta/veterinária , Proteínas de Peixes/metabolismo , Glucosefosfato Desidrogenase/metabolismo , Glutationa Peroxidase/metabolismo , Glutationa Redutase/metabolismo , Imunidade Inata , Intestinos/enzimologia , Oncorhynchus mykiss/metabolismo , Superóxido Dismutase/metabolismoRESUMO
Chaga's disease is an important communicable neglected disease that is gaining wider attention due to its increasing incidence worldwide. Achalasia due to chagasic megaesophagus (CM), a complication of this disease, is a known-yet, poorly understood-etiological factor for esophageal squamous cell carcinoma (ESCC) development. In this study, we aimed to perform the analysis of TP53 mutations in a series of Brazilian patients with ESCC that developed in the context CM (ESCC/CM), and to compare with the TP53 mutation profile of patients with benign CM and patients with nonchagasic ESCC. Additionally, we intended to correlate the TP53 mutation results with patient's clinical pathological features. By polymerase chain reaction (PCR) followed by direct sequencing of the hotspot regions of TP53 (exon 5 to 8), we found that TP53 mutations were present in 40.6% (13/32) of the ESCC/CM group, 45% (18/40) of the nonchagasic ESCC group, and in only 3% (1/33) of the benign CM group. Missense mutations were the most common in the three groups, yet, the type and mutated exon mutation varied significantly among the groups. Clinically, the groups exhibited distinct features, with both cancer groups (ESCC and ESCC/CM) been significantly associated higher consumption of alcohol and tobacco, older age, worse Karnofsky performance status, poor outcome than the patients with benign CM. No significant association was found between TP53 mutation profile and clinical-pathological features in any of the three groups. We describe first the time the analysis of TP53 mutations in ESCC that developed in the context of CM, and the observed high frequency of mutations, suggest that TP53 also plays an important role in the tumorigenic process of this unexplored etiological condition.
Assuntos
Carcinoma de Células Escamosas/genética , Doença de Chagas/genética , Acalasia Esofágica/genética , Neoplasias Esofágicas/genética , Genes p53 , Mutação , Adulto , Idoso , Brasil , Doença de Chagas/complicações , Acalasia Esofágica/complicações , Carcinoma de Células Escamosas do Esôfago , Éxons , Feminino , Estudos de Associação Genética , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Fatores de Risco , Adulto JovemRESUMO
The immune response of European sea bass, Dicentrarchus labrax, to a natural infection by the copepod parasite Lernanthropus kroyeri was evaluated for the first time in vivo. The results clearly demonstrated the triggering of the fish immune system by the parasite. Lysozyme activity and alternative complement pathway were involved in the early action against the parasitical infection, whilst classical complement and respiratory burst (RB) activity took over in the later stages of infection. It was hypothesized that the levels of alternative and classical complement and RB stimulation indexes may determine the resistance capacity of the fish to the parasite. It is not clear how parasites can survive despite the strong immunological arsenal deployed by the fish. The continual increase of prevalence and severity of parasite infection suggested that the parasite's mechanism of evasion of the immune system was extremely successful. The contrasting decrease in the negative effects of parasites on the fish health (such as gills anaemia) suggested that an equilibrium between the parasites and their hosts was reached in chronic infection. These dynamic interactions between parasites and fish hosts were probably the main determinant of host specificity.
Assuntos
Bass/imunologia , Bass/parasitologia , Copépodes , Doenças dos Peixes/imunologia , Doenças dos Peixes/parasitologia , Interações Hospedeiro-Parasita/imunologia , Anemia/patologia , Anemia/veterinária , Animais , Atividade Bactericida do Sangue/imunologia , Proteínas do Sistema Complemento/análise , Proteínas do Sistema Complemento/imunologia , Feminino , Pesqueiros , Brânquias/parasitologia , Brânquias/patologia , Grécia , Imunidade Inata , Masculino , Muramidase/sangue , Óxido Nítrico/sangue , Explosão Respiratória/imunologiaRESUMO
Tumor necrosis factor alpha (TNFalpha), a pro-inflammatory cytokine, enhances the development of pain and hyperalgesia, although the molecular mechanisms are not well understood. This study evaluated the hypothesis that TNFalpha increases the sensitivity of rat trigeminal neurons to capsaicin via two different mechanisms triggered by either brief or sustained exposure to the cytokine. A brief (5 min) application of TNFalpha significantly sensitized capsaicin-evoked accumulation of intracellular calcium ([Ca2+]i) (226.4+/-37.7 nM vs. 167.5+/-31.3 nM) and increased capsaicin-evoked nocifensive behavior (78.3+/-9.7 vs. 30.9+/-3.6 s) as compared with vehicle pretreatment (P<0.01 for both). Sustained (30 min to 4 h) exposure of cultured neurons to TNFalpha evoked a twofold increase in mRNA transcript (P<0.05) and protein levels (P<0.01) of transient potential receptor vanilloid type 1 (TRPV1). This long-term up-regulation of TRPV1 expression by TNFalpha correlated with enhancement in capsaicin-induced calcitonin gene-related peptide release (P<0.05). Demonstration of colocalization of TNFalpha receptor subtypes I and II with TRPV1 in almost all (>90%) TRPV1 expressing neurons provides evidence consistent with a direct interaction on the same subpopulation of sensory neurons. In summary, our data demonstrate that TNFalpha directly enhances the sensitivity of rat trigeminal neurons to capsaicin via both rapid, non-genomic mechanisms as well as sustained genomic regulation in TRPV1 expression. Thus, increased sensitization and up-regulation of TRPV1 constitutes a potential mechanism by which TNFalpha mediates inflammatory hyperalgesia and pain.
Assuntos
Capsaicina/farmacologia , Neurônios/efeitos dos fármacos , Nervo Trigêmeo/citologia , Neuralgia do Trigêmeo/fisiopatologia , Fator de Necrose Tumoral alfa/farmacologia , Animais , Peptídeo Relacionado com Gene de Calcitonina/metabolismo , Cálcio/metabolismo , Células Cultivadas , Sinergismo Farmacológico , Expressão Gênica/efeitos dos fármacos , Masculino , Neurônios/citologia , Neurônios/fisiologia , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-Dawley , Receptores Tipo I de Fatores de Necrose Tumoral/genética , Receptores Tipo II do Fator de Necrose Tumoral/genética , Fármacos do Sistema Sensorial/farmacologia , Canais de Cátion TRPV/genética , Canais de Cátion TRPV/metabolismo , Tempo , Regulação para Cima/efeitos dos fármacosRESUMO
Feeding small European sea bass, Dicentrarchus labrax, for 6 weeks with Tenebrio molitor larval meal showed significant anti-inflammatory responses (ceruloplasmin, myeloperoxidase and nitric oxide). Serum bacteriolytic activity against a Gram negative bacterium was not significantly affected by dietary Tenebrio, while both lysozyme antibacterial activity and serum trypsin inhibition usually linked to the anti-parasite activity of the fish, were significantly enhanced. The latter may be due to the similarities in the composition of the exoskeleton of parasites and insects that may therefore act as an immunostimulant potentially increasing the anti-parasitic activity. The addition of exogenous proteases significantly decreased both trypsin-inhibition and serum bacteriolytic activity probably through direct inhibition of the proteins responsible for these immune functions. Further investigation involving bacterial or parasitic challenges will be necessary to assess if the effects of dietary mealworm meal on the immune system observed in the present study are translated into an improved resistance to diseases.
Assuntos
Anti-Inflamatórios/metabolismo , Bass/imunologia , Dieta , Imunidade Inata , Inflamação/imunologia , Insetos/metabolismo , Tenebrio/metabolismo , Animais , Ceruloplasmina/metabolismo , Proteínas de Peixes/metabolismo , Sistema Imunitário , Muramidase/metabolismo , Óxido Nítrico/metabolismo , Peroxidase/metabolismo , Tripsina/metabolismoRESUMO
BACKGROUND: Currently, Roux-en-Y gastric bypass (RYGB) is one of the most widely used bariatric surgeries. Banding the pouch forms a banded gastric bypass operation, an accepted and frequently used variant. Placing a silastic ring around the pouch to band the gastric bypass operation increases the restriction mechanism. However, the ubiquitous use of the banded gastric bypass remains controversial. One of the controversies is the effect of the silastic ring on patients' perception of their well being after surgery because of the frequency of vomiting. A prospective, blindly randomized, comparative trial was undertaken to resolve this controversy. METHOD: Four hundred subjects scheduled for gastric bypass surgery were randomized into two arms of the trial, 200 with a silastic ring (WR) and 200 without (NR). After 2-year follow-up, the variables associated with the scores of Bariatric Analysis and Reporting Outcome System (BAROS) were analyzed. RESULTS: The initial median weight (125 kg), BMI (47), and age (36 years) were the same in both the NR and WR groups. The median excess weight loss, weight regain, and incidence of vomiting were 71, 10.5, and 7.75%, respectively, in the NR group vs. 75.4 and 1.1, and 24.4% in the WR group. The mean QOL score was 79% in the NR group vs. 80% in the WR group. CONCLUSION: After 2-year follow-up, silastic ring placement in the RYGB resulted in greater weight loss and weight stability and a threefold greater incidence of vomiting. There was no difference in the scores in the quality of life analysis.
Assuntos
Derivação Gástrica/instrumentação , Próteses e Implantes , Vômito/etiologia , Adulto , Dimetilpolisiloxanos , Feminino , Seguimentos , Derivação Gástrica/efeitos adversos , Derivação Gástrica/métodos , Humanos , Masculino , Pessoa de Meia-Idade , Obesidade Mórbida/cirurgia , Estudos Prospectivos , Qualidade de Vida , Redução de Peso , Adulto JovemRESUMO
One thousand healthy recipient gilthead sea bream, Sparus aurata, cohabited with 250 donor fish parasitized by Sparicotyle chrysophrii (Van Beneden and Hesse, 1963) (Monogenea: Polyopisthocotylea), a common parasite of the gills of this fish species. Controls consisted of 1000 healthy fish kept in a separate tank. After 10 weeks, fish were weighed and parasite load, hemoglobin concentration and immunological parameters were assessed. Rather than the absence of parasite, hemoglobin concentration was a better marker of the health status of the fish, because S.chrysophrii had detached from the strongly anemic gills of some animals leaving fish with affected immune system but without parasites. The parasite infection seemed to trigger a cellular response of the fish immune system but to inhibit its humoral components. Thus, parasitized fish may control the parasite infection through the action of reactive oxygen species but they may become more sensitive to potential secondary bacterial or parasitical infections. This phenomenon was demonstrated not only through significant differences between recipient and control fish but also through strong correlations between those parameters and parasite load, fish weight and/or hemoglobin concentration.
Assuntos
Infecções por Cestoides/imunologia , Brânquias/parasitologia , Hemoglobinas/metabolismo , Platelmintos/imunologia , Dourada/imunologia , Animais , Biomarcadores/metabolismo , Peso Corporal/imunologia , Imunidade Celular , Imunidade Humoral , Imunidade Inata , Mediadores da Inflamação/metabolismo , Estresse Oxidativo , Carga Parasitária , Espécies Reativas de Oxigênio/metabolismoRESUMO
Regenerative endodontic procedures are stem cell-based treatments for immature teeth with pulp necrosis. The translation of regenerative endodontic procedures into treating mature teeth depends, among other factors, on the availability and delivery of mesenchymal stem cells (MSCs) into the root canal system. The aim of this clinical study was to evaluate whether evoked bleeding from the periapical tissues elicits the influx of MSCs into the root canal system in mature teeth with apical lesions. Participants included in this study (N = 20) were referred for endodontic treatment of mature teeth with apical lesions. Following chemomechanical debridement, intracanal bleeding from the periapical tissues was achieved, and intracanal blood samples were collected. A positive blood aspirate was also collected in the cartridges during local anesthesia. Total RNA was isolated and used as a template in quantitative reverse transcription polymerase chain reactions using MSC-specific arrays. Data were analyzed with the Wilcoxon signed-rank test, and correlation between gene expression and sex or age was tested with Spearman's rank correlation coefficient test. In addition, MSCs were isolated from an intracanal bleeding sample and subjected to flow cytometry and quantitative osteogenesis assay. Last, the presence and distribution of MSCs within periradicular lesions were evaluated with immunohistochemistry (n = 4). The MSC markers CD73, CD90, CD105, and CD146 were significantly upregulated, with median fold change values of 2.9, 31.7, 4.6, and 6.8, respectively. Conversely, the negative marker for MSCs, CD45, was significantly downregulated (median, -2.7). There was no correlation with age, sex, tooth type, or treatment for any of the evaluated genes. Isolated intracanal cells coexpressed MSC markers and demonstrated robust mineralizing differentiation potential. Finally, immunohistochemical analysis revealed that MSCs were found compartmentalized mainly within vasculature structures located in periapical lesions. Collectively, findings indicate that the evoked-bleeding technique delivers MSCs into the root canal system in mature teeth with apical lesions.
Assuntos
Transplante de Células-Tronco Mesenquimais/métodos , Tecido Periapical/citologia , Tratamento do Canal Radicular/métodos , Adulto , Idoso , Idoso de 80 Anos ou mais , Cavidade Pulpar/cirurgia , Feminino , Citometria de Fluxo , Humanos , Masculino , Microscopia Confocal , Pessoa de Meia-Idade , Osteogênese , Periodontite/cirurgia , Reação em Cadeia da Polimerase em Tempo Real , Adulto JovemRESUMO
The 75 kDa protein nerve growth factor receptor [NGFr(p75)] is a neurotrophin receptor that is able to bind different members of the neurotrophin family of molecules implicated in affecting neuronal survival. Here we describe the light microscopic distribution of NGFr(p75)-immunoreactivity (IR) within the feline trigeminal brainstem sensory nuclear complex and trigeminal ganglion of normal adult subjects and in subjects 10 and 30 days following retrogasserian rhizotomy. Within the trigeminal ganglion of normal subjects, numerous fibers and most of the neuronal cell bodies showed NGFr(p75)-IR that varied in intensity, while cells and fibers with NGFr(p75)-IR were less numerous within the mesencephalic trigeminal nucleus. Within the main sensory and spinal trigeminal nuclei, NGFr(p75)-IR formed a reproducible pattern that varied between the different subnuclei. The NGFr(p75)-IR consisted both of dense pockets and a low level NGFr(p75)-IR that was selective to the trigeminal neuropil. Following rhizotomy, most of the NGFr(p75)-IR was lost from the main sensory and spinal trigeminal nuclei, except in regions where the upper cervical roots and cranial nerves VII, IX, and X project. In contrast, examination of the central root that was still attached to the trigeminal ganglion showed increased NGFr(p75)-IR in fibers and supporting cells, as did the motor root within the peripheral mandibular division. These results indicate that the majority of the NGFr(p75)-IR within the main sensory and spinal trigeminal nuclei originates from primary trigeminal afferents and that retrogasserian rhizotomy leads to an up-regulation of NGFr(p75)-IR in the part of the central root that is contiguous with the ganglion.
Assuntos
Receptores de Fator de Crescimento Neural/metabolismo , Nervo Trigêmeo/metabolismo , Animais , Gatos , Denervação , Imuno-Histoquímica , Bulbo/anatomia & histologia , Bulbo/fisiologia , Mesencéfalo/anatomia & histologia , Mesencéfalo/fisiologia , Neurônios Motores/fisiologia , Neurônios Aferentes/fisiologia , Receptores de Fator de Crescimento Neural/imunologia , Nervo Trigêmeo/anatomia & histologiaRESUMO
We studied the ultrastructure of the synaptic organization in the feline spinal trigeminal nucleus, emphasizing specific neurotransmitter patterns within lamina II of the pars caudalis/medullary dorsal horn. Normal adults were perfused, and Vibratome sections from pars caudalis were processed for electron microscopy. Ultrathin sections were reacted with antibodies for the excitatory neurotransmitter glutamate (Glu) and for the inhibitory neurotransmitter gamma-aminobutyric acid (GABA) by using postembedding immunogold techniques. Both single- and double-labeled preparations were examined. Results with single labeling show that Glu-immunoreactive terminals have round synaptic vesicles and form asymmetric synaptic contacts onto dendrites. GABA-immunoreactive axon terminals and vesicle-containing dendrites have pleomorphic vesicles, and the axon terminals form symmetric contacts onto dendrites and other axons. Double labeling on a single section shows glomeruli with central Glu-immunoreactive terminals that are presynaptic to dendrites, including GABA+ vesicle-containing dendrites. These Glu+ terminals are also postsynaptic to GABA+ axon terminals, and these GABA-immunoreactive terminals may also be presynaptic to the GABA+ vesicle-containing dendrites. Quantitative analyses confirm the specificity of the Glu and GABA immunoreactivities seen in the various glomerular profiles. The results suggest that a subpopulation of Glu-immunoreactive primary afferents (excitatory) may be under the direct synaptic influence of a GABA-immunoreactive intrinsic pathway (inhibitory) by both presynaptic and postsynaptic mechanisms.
Assuntos
Gatos/anatomia & histologia , Ácido Glutâmico/imunologia , Sinapses/química , Núcleos do Trigêmeo/química , Ácido gama-Aminobutírico/imunologia , Animais , Ácido Glutâmico/química , Imuno-Histoquímica , Microscopia Eletrônica , Microscopia Imunoeletrônica , Neurônios/química , Neurônios/ultraestrutura , Sinapses/ultraestrutura , Inclusão do Tecido , Núcleos do Trigêmeo/citologia , Ácido gama-Aminobutírico/químicaRESUMO
Calcitonin gene-related peptide (CGRP) is a neuropeptide that has been implicated in the transmission and modulation of primary afferent nociceptive stimuli. In this study, we describe the light microscopic distribution of CGRP immunoreactivity (IR) within the feline trigeminal ganglion and trigeminal nucleus of normal adult subjects and in subjects 10 and 30 days following complete retrogasserian rhizotomy. Within the trigeminal ganglion of normal subjects, cell bodies and fibers showed CGRP-IR, whereas immunoreactive fibers were rare in the central root region. Within the normal spinal trigeminal and main sensory nuclei, CGRP-IR was seen to form a reproducible pattern that varied between the different nuclei. Following rhizotomy, most, but not all, of the CGRP-IR was lost from the spinal trigeminal and main sensory nuclei, except in regions where the upper cervical roots and cranial nerves VII, IX and X project into the trigeminal nucleus. The pattern seen at 10 days contained more CGRP-IR than that seen at 30 days and suggests that degenerating fibers still show CGRP-IR. In contrast to the decrease seen in the nuclei after rhizotomy, examination of the central root that was still attached to the trigeminal ganglion showed an increase in CGRP-IR within fibers, some of which ended in growth conelike enlargements. Rhizotomy induced a dramatic increase in CGRP-IR within trigeminal motoneurons and their fibers, which was strongest 10 days after rhizotomy and weaker at 30 days, which was still stronger than normal. These results indicate that the majority of CGRP-IR found in the trigeminal nucleus originates from trigeminal primary afferents and that an upregulation of CGRP-IR occurs in trigeminal motoneurons and in regenerating fibers in the part of the central root that was still attached to the ganglion. In addition, the persistence of CGRP-IR fibers in the trigeminal nucleus provides one possible explanation for the preservation of pain in humans following trigeminal rhizotomy.
Assuntos
Peptídeo Relacionado com Gene de Calcitonina/análise , Gatos , Rizotomia , Núcleo Inferior Caudal do Nervo Trigêmeo/citologia , Gânglio Trigeminal/citologia , Nervo Trigêmeo/citologia , Núcleos do Trigêmeo/citologia , Animais , Nervos Cranianos/citologia , Lateralidade Funcional , Imuno-Histoquímica , Fibras Nervosas/ultraestrutura , Valores de Referência , Fatores de Tempo , Núcleo Inferior Caudal do Nervo Trigêmeo/metabolismo , Gânglio Trigeminal/metabolismo , Nervo Trigêmeo/metabolismo , Núcleos do Trigêmeo/metabolismo , Núcleo Espinal do Trigêmeo/citologia , Núcleo Espinal do Trigêmeo/metabolismoRESUMO
Glial cell-lined derived neurotrophic factor (GDNF) has been shown to promote survival of developing mesencephalic dopaminergic neurons in vitro. In order to determine if there is a positive effect of GDNF on injured adult midbrain dopaminergic neurons in situ, we have carried out experiments in which a single dose of GDNF was injected into the substantia nigra following a unilateral lesion of the nigrostriatal system. Rats were unilaterally lesioned by a single stereotaxic injection of 6-hydroxydopamine (6-OHDA; 9 micrograms/4 microliters normal saline with 0.02% ascorbate) into the medial forebrain bundle and tested weekly for apomorphine-induced (0.05 mg/kg s.c.) contralateral rotation behavior. Rats that manifested > 300 turns/hour received a nigral injection of 100 micrograms GDNF, or cytochrome C as a control, 4 weeks following the 6-OHDA lesion. Rotation behavior was quantified weekly for 5 weeks after GDNF. Rats were subsequently anesthetized, transcardially perfused, and processed for tyrosine hydroxylase immunohistochemistry. It was found that 100 micrograms GDNF decreased apomorphine-induced rotational behavior by more than 85%. Immunohistochemical studies revealed that tyrosine hydroxylase immunoreactivity was equally reduced in the striatum ipsilateral to the lesion in both cytochrome C and GDNF-injected animals. In contrast, large increments in tyrosine hydroxylase immunoreactivity were observed in the substantia nigra of animals treated with 100 micrograms of GDNF, with a significant increase in numbers of tyrosine hydroxylase-immunoreactive cell bodies and neurites as well as a small increase in the cell body area of these neurons. The results suggest that GDNF can maintain the dopaminergic neuronal phenotype in a number of nigral neurons following a unilateral nigrostriatal lesion in the rat.
Assuntos
Dopamina/fisiologia , Mesencéfalo/citologia , Fatores de Crescimento Neural/farmacologia , Proteínas do Tecido Nervoso/farmacologia , Neurônios/efeitos dos fármacos , Animais , Apomorfina , Sobrevivência Celular/efeitos dos fármacos , Fator Neurotrófico Derivado de Linhagem de Célula Glial , Processamento de Imagem Assistida por Computador , Imuno-Histoquímica , Masculino , Mesencéfalo/anatomia & histologia , Mesencéfalo/efeitos dos fármacos , Fatores de Crescimento Neural/administração & dosagem , Proteínas do Tecido Nervoso/administração & dosagem , Plasticidade Neuronal/efeitos dos fármacos , Oxidopamina , Ratos , Ratos Endogâmicos F344 , Comportamento Estereotipado/efeitos dos fármacos , Tirosina 3-Mono-Oxigenase/metabolismoRESUMO
Ultrastructural visualization of the horseradish peroxidase-tetramethylbenzidine (HRP-TMB) reaction product within trigeminal ganglion cells and brain stem axons and terminals following HRP injections into the pulpal chambers of cat teeth is enhanced by utilization of a modified osmication procedure that converts the reaction product to a markedly stable and electron-dense form. The results following the use of the modified osmication procedure (pH 5.0 phosphate buffer at 20 degrees C for 12 hours) are compared to results obtained by following Carson's osmication protocol (Carson KA, Mesulam M-M: J Histochem Cytochem 30:425, 1982; Carson KA, Mesulam M-M: In Tracing Neural Connections with Horseradish Peroxidase. Edited by M-M Mesulam. J Wiley, Chichester, England, 1982, p 153-184) (pH 6.0 phosphate buffer at 45 degrees C for 45 min). The results suggest that the conversion of the HRP-TMB reaction product to an electron-dense form during osmication is intimately associated with the pH of the phosphate buffer and the total time of osmication.
Assuntos
Axônios/ultraestrutura , Tronco Encefálico/ultraestrutura , Gânglios/ultraestrutura , Animais , Benzidinas , Gatos , Peroxidase do Rábano Silvestre , Indicadores e Reagentes , Microscopia Eletrônica/métodosRESUMO
The ultrastructure of transganglionic transport of horseradish peroxidase (HRP) from the inferior alveolar (IA) nerve to the brainstem is being studied in the cat. The IA nerve was soaked in an HRP solution and following a two-day survival the animal was perfused transcardially with a paraformaldehyde-glutaraldehyde solution. The tissue was immediately dissected and postfixed for 1-3 h in perfusate. Sections of 75 micron thickness were cut with a Vibratome and reacted utilizing tetramethyl benzidine (TMB) as the chromagen. Optimum results for electron microscopy were obtained by osmication in a pH 6.0, 1% osmium tetroxide solution for 45 min at 45 degrees C, followed by rapid dehydration and embedment in Epon. The resulting HRP-TMB reaction product was characterized and identified ultrastructurally in ganglion cells, peripheral and central axons and in brainstem terminals. The HRP-TMB reaction product varied in density but had consistent crystalline-like laminations of a repeating unit and characterized by a membrane 4-5 nm in diameter. Some of the HRP-TMB reaction product found in terminals and axons was below the limit of resolution of the light microscope.
Assuntos
Peroxidase do Rábano Silvestre , Neuroanatomia/métodos , Peroxidases , Coloração e Rotulagem/métodos , Nervo Trigêmeo/ultraestrutura , Núcleos do Trigêmeo/ultraestrutura , Animais , Benzidinas , Gatos , Microscopia Eletrônica , Terminações Nervosas/ultraestruturaRESUMO
Previous studies have demonstrated the presence of nerve growth factor receptor [NGFr(p75)]-immunoreactivity (IR) in the spinal trigeminal nucleus of both 8-10 week-old kittens and mature cats. Most of the NGFr(p75)-IR is lost following retrogasserian rhizotomy, indicating that the majority of the NGFr(p75)-IR within the spinal trigeminal nucleus is of trigeminal primary afferent origin. Here, we examined the ultrastructural localization of NGFr(p75)-IR within lamina II outer of pars caudalis/medullary dorsal horn in the mature cat. Lamina II outer represents a location where dense NGFr(p75)-IR is seen with the light microscope. The NGFr(p75)-IR identified with the electron microscope was located within small thinly myelinated and unmyelinated axons and within axon terminals. The terminals with NGFr(p75)-IR typically formed asymmetric synaptic specializations onto dendritic profiles and at times were postsynaptic to other axon terminals at symmetric synaptic specializations. The terminals with NGFr(p75)-IR were either simple (associated with a single profile) or more complex, such as those that typically formed the central element in synaptic glomeruli. The NGFr(p75)-IR in terminals was especially prominent on microtubules and the plasmalemma and these findings are consistent with proposed roles for NGFr(p75) in axoplasmic/neuronal transport and as a membrane protein, respectively. The profiles with NGFr(p75)-IR seen with the electron microscope indicate a primary afferent origin and show some similarities when compared to other markers of primary afferent fibers such as calcitonin gene-related peptide. In addition, a possible role for NGFr(p75) in the transmission of nociceptive stimuli is also discussed.
Assuntos
Receptores de Fator de Crescimento Neural/análise , Medula Espinal/química , Animais , Gatos , Microscopia Eletrônica , Peso Molecular , Terminações Pré-Sinápticas/químicaRESUMO
Toxic ricin was used to study the central distribution of dental afferents in the cat. Following intrapulpal ricin injections ganglion cell degeneration is seen in the II and III ganglion divisions. Central argyrophilic degeneration occurs in the dorsal portion of all ipsilateral trigeminal nuclei. Ventral degeneration is seen in the pars interpolaris and pars caudalis. No contralateral degeneration was observed. The results are discussed with regard to previous studies of the central location of dental afferents.
Assuntos
Polpa Dentária/inervação , Nervo Trigêmeo/anatomia & histologia , Núcleos do Trigêmeo/anatomia & histologia , Animais , Gatos , Peroxidase do Rábano Silvestre , Neuroanatomia/métodos , Neurônios Aferentes , Ricina , Núcleo Inferior Caudal do Nervo Trigêmeo/anatomia & histologia , Gânglio Trigeminal/anatomia & histologiaRESUMO
OBJECTIVE: To test the hypotheses that there is a two-factor aspect of cellular damage during cryopreservation that occurs in human sperm (osmotic effects versus intracellular ice formation) and that there is a cooling rate by warming rate interaction related to this damage. DESIGN: Ejaculates from healthy men were cooled at 0.1, 1.0, 10, 175, or 800 degrees C/min to -80 degrees C in a solution of 0.85 M glycerol and plunged into liquid nitrogen. Samples were warmed at 400 degrees C/min (experiment 1) or either 1 degrees C or 400 degrees C/min (experiment 2). After warming, sperm were assessed for survival using motility as the endpoint in experiment 1 and motility, plasma membrane integrity, and mitochondrial function in experiment 2. RESULTS: In experiment 1, over the various cooling rates with a standard 400 degrees C/min warming rate, a plot of motility versus cooling rate produced a classical inverted U-shaped curve (n = 6) with maximum motility at the 10 degrees C/min cooling rate. In experiment 2, over the various cooling rates, both 1 and 400 degrees C/min warming rates produced similar but shifted plots of motility, plasma membrane integrity, and mitochondrial function versus cooling rate, which also produced inverted U-shaped patterns (n = 11). Maximal survival for each of the three endpoints occurred at 10 degrees C/min cooling rate for the rapidly warmed sperm and at 1 degree C/min for the slowly warmed sperm. CONCLUSIONS: These data support the hypotheses that a two-factor hypothesis of cryodamage applies to human spermatozoa and that an interaction exists between cooling rate and warming rate. These data also suggest that motility, plasma membrane integrity, and mitochondrial function are not differently affected by cooling and warming during cryopreservation.
Assuntos
Criopreservação , Mitocôndrias/metabolismo , Motilidade dos Espermatozoides , Espermatozoides , Adulto , Membrana Celular/ultraestrutura , Criopreservação/métodos , Citometria de Fluxo/métodos , Humanos , Masculino , Espermatozoides/fisiologia , Espermatozoides/ultraestrutura , Fatores de TempoRESUMO
Retrogasserian trigeminal rhizotomy was used to study the central projections and patterns of degeneration in the spinal trigeminal nucleus (STN). At survival times of 3-20 days, reduced silver stains show extensive degeneration throughout the ipsilateral STN and in addition, well delineated degeneration was identified in the periobex region of the contralateral STN that varied with survival time. The results suggest that primary afferents may contribute to this contralateral projection.
Assuntos
Degeneração Neural , Nervo Trigêmeo/fisiologia , Núcleos do Trigêmeo/fisiologia , Animais , Gatos , Microscopia Eletrônica , Terminações Nervosas/ultraestrutura , Neurônios Aferentes/ultraestrutura , Coloração e Rotulagem , Nervo Trigêmeo/ultraestrutura , Núcleos do Trigêmeo/ultraestruturaRESUMO
The effect of lesions on nerve growth factor receptor (NGFr) immunoreactivity (IR) in motoneurons within the mature feline trigeminal motor nucleus was investigated. Ten days following complete unilateral retrogasserian trigeminal rhizotomy including transection of the trigeminal motor root, motoneurons within the ipsilateral trigeminal motor nucleus showed NGFr-IR. In contrast, motoneurons within the contralateral trigeminal motor nucleus and within both trigeminal motor nuclei in unoperated control subjects did not show NGFr-IR. It is suggested that the appearance of NGFr-IR in motoneurons within the ipsilateral trigeminal motor nucleus after rhizotomy represents an attempts towards recovery and may be associated with the regrowth of its cut axons.
Assuntos
Neurônios Motores/metabolismo , Receptores de Superfície Celular/metabolismo , Raízes Nervosas Espinhais/fisiologia , Gânglio Trigeminal/fisiologia , Núcleo Espinal do Trigêmeo/metabolismo , Animais , Gatos , Denervação , Imuno-Histoquímica , Fatores de Crescimento Neural/metabolismo , Receptores de Fator de Crescimento NeuralRESUMO
We treated three patients (a 72-year-old man, a 68-year-old woman, and a 60-year-old man) with aphakic glaucoma with flat anterior chamber by administering laser burns to the ciliary processes (11 ciliary processes in one case, 27 in the second, and three in the third). There was some improvement in all three cases; in one case the intraocular pressure decreased from more than 50 mm Hg to less than 20 mm Hg. In each case the anterior chamber deepened immediately without administration of cycloplegics.