The p21-activated kinases in neural cytoskeletal remodeling and related neurological disorders

The serine/threonine p21-activated kinases (PAKs), as main effectors of the Rho GTPases Cdc42 and Rac, represent a group of important molecular switches linking the complex cytoskeletal networks to broad neural activity. PAKs show wide expression in the brain, but they differ in specific cell types, brain regions, and developmental stages. PAKs play an essential and differential role in controlling neural cytoskeletal remodeling and are related to the development and fate of neurons as well as the structural and functional plasticity of dendritic spines. PAK-mediated actin signaling and interacting functional networks represent a common pathway frequently affected in multiple neurodevelopmental and neurodegenerative disorders. Considering specific small-molecule agonists and inhibitors for PAKs have been developed in cancer treatment, comprehensive knowledge about the role of PAKs in neural cytoskeletal remodeling will promote our understanding of the complex mechanisms underlying neurological diseases, which may also represent potential therapeutic targets of these diseases.

Wnt-5a-regulated miR-101b controls COX2 expression in hippocampal neurons

BACKGROUND: Wnt-5a is a member of the WNT family of secreted lipoglycoproteins, whose expression increases during development; moreover, Wnt-5a plays a key role in synaptic structure and function in the adult nervous system. However, the mechanism underlying these effects is still elusive. MicroRNAs (miRNAs) are a family of small non-coding RNAs that control the gene expression of their targets through hybridization with complementary sequences in the 3' UTR, thereby inhibiting the translation of the target proteins. Several evidences indicate that the miRNAs are actively involved in the regulation of neuronal function. RESULTS: In the present study, we examined whether Wnt-5a modulates the levels of miRNAs in hippocampal neurons. Using PCR arrays, we identified a set of miRNAs that respond to Wnt-5a treatment. One of the most affected miRNAs was miR-101b, which targets cyclooxygenase-2 (COX2), an inducible enzyme that converts arachidonic acid to prostanoids, and has been involved in the injury/inflammatory response, and more recently in neuronal plasticity. Consistent with the Wnt-5a regulation of miR-101b, this Wnt ligand regulates COX2 expression in a time-dependent manner in cultured hippocampal neurons. CONCLUSION: The biological processes induced by Wnt-5a in hippocampal neurons, involve the regulation of several miRNAs including miR-101b, which has the capacity to regulate several targets, including COX-2 in the central nervous system

Sobrevida de pacientes con cáncer gástrico en el Perú, 2009-2010 / Survival rate of patients with stomach cancer in Peru, 2009-2010

OBJETIVO: Determinar la sobrevida de pacientes con diagnóstico de cáncer gástrico en 2009-2010 en el Perú. MÉTODOS: Se realizó un estudio de tipo cohorte retrospectivo de pacientes con diagnóstico de cáncer gástrico registrados en el Sistema Nacional de Vigilancia Epidemiológica (SNVE) de la Dirección General de Epidemiología (DGE) y del Registro de Hechos Vitales (RHV) de la Oficina General de Estadística e Informática (OGEI) para los años 2009-2010. RESULTADOS: Se incluyeron 3 568 pacientes del SNVE, 51,5% eran hombres y 48,5% eran mujeres; la media de edad fue 63,9 años, 60,07% tenían 60 años o más. Se halló que 33,6% tenía adenocarcinoma de tipo intestinal, 18,7% tenía carcinoma de tipo difuso y 4,1% tenía linfoma gástrico primario. La sobrevida global fue de 29,7 ± 0,8 meses y fue mejor para los menores de 60 años (P = 0,034), para las mujeres (P = 0,014) y para el adenocarcinoma de tipo intestinal (P < 0,001). No hubo diferencias (P = 0,713) entre la sobrevida de los pacientes con linfoma gástrico y aquellos con adenocarcinoma. Para evaluar la tasa de mortalidad se incluyeron 6 069 registros de pacientes del RHV, la tasa de mortalidad nacional fue de 10,3 por cada 100 000 habitantes y las regiones con mayor mortalidad fueron Huánuco, Huancavelica y Junín. CONCLUSIONES: La sobrevida general fue de 29,7 ± 0,8 meses, las mujeres, los menores de 60 años y los pacientes con adenocarcinoma de tipo intestinal tienen mejor sobrevida. La mayor mortalidad por cáncer gástrico se concentra en las regiones más pobres del Perú, donde es probable que las condiciones de vida faciliten la alta transmisibilidad de Helicobacter pylori.

OBJECTIVE: Determine the survival rate of patients diagnosed with stomach cancer in 2009-2010 in Peru. METHODS: A retrospective cohort study was conducted of patients diagnosed with stomach cancer registered in the National Epidemiological Surveillance System (SNVE) of the Directorate General of Epidemiology (DGE) and the Register of Vital Statistics (RHV) of the General Office of Statistics and Information (OGEI) for the years 2009-2010. RESULTS: 3 568 patients of the SNVE were included; 51.5% were men and 48.5% were women; the average age was 63.9 years; 60.07% were 60 years old or older. It was found that 33.6% had intestinal type adenocarcinoma, 18.7% had diffuse type carcinoma, and 4.1% had primary gastric lymphoma. The overall survival rate was 29.7 ± 0.8 months and was better for those under 60 years (P = 0.034), for women (P = 0.014) and for intestinal type adenocarcinoma (P< 0.001). There was no difference (P = 0.713) between the survival rate of gastric lymphomas and adenocarcinomas. In order to evaluate mortality, 6 069 patient records from the RHV were included; national mortality was 10.3 per 100 000 population; the regions with the highest mortality were Huánuco, Huancavelica, and Junín. CONCLUSIONS: The general survival rate was 29.7 ± 0.8 months; women, those under 60 years, and patients with intestinal type adenocarcinoma had better survival rates. The highest mortality from stomach cancer is concentrated in the poorest regions of Peru, where it is probable that living conditions facilitate the high communicability of Helicobacter pylori.

Doença de depósito lisossomal induzida pelo consumo de Ipomoea verbascoidea (Convolvulaceae) em caprinos no semiárido de Pernambuco / Lysosomal storage disease induced by the consumption of Ipomoea verbascoidea (Convolvulaceae) in goats at northeastern Brazil

The aim of this paper was to reproduce the poisoning of Ipomoea verbascoidea in goats and describe the epidemiological, clinical and pathological aspects of spontaneous poisoning by this plant in Pernambuco. For this, we studied the epidemiology of the disease in seven municipalities in the semiarid region of the State. Three spontaneously poisoned goats were examined and then euthanized and necropsied (Group I). To reproduce the disease, the dried leaves of I. verbascoidea containing 0.02% swainsonine were supplied at doses of 4g/kg (0.8mg swainsonine/kg) to two groups of three animals. The goats in Group II received daily doses of the plant during 40 days and were euthanized on the 41st day of the experiment. Goats from Group III received daily doses of the plant during 55 days and were euthanized on the 120th day of the experiment. Other three goats constituted the control group (Group IV). In experimental groups, the brain lesions were evaluated by histopathology; additionally the cerebellar lesions were evaluated by morphometry, by measuring the molecular layer thickness, the number of Purkinje cells and the area of the cell bodies of these cells. The main clinical signs and microscopic lesions in goats poisoned were similar to those reported by swainsonine containing plants. In goats of GII and GIII, the first nervous signs were observed between 22th and 29th days; clinically, the disease developed by these animals was similar to the spontaneous cases. The goats of GIII did not recover from the neurologic signs. These results show that the consumption of the plant by 26-28 days after observation of the first clinical signs is enough to cause irreversible damage. By morphometric analysis, the molecular layer of the cerebellum of the goats of Group I and III were thinner than those of goats in the control group, and Purkinje neurons were atrophic. It is suggested that these changes are responsible for the neurological picture observed in goats that stop eating the plant and have sequelae of poisoning.

O objetivo deste trabalho foi reproduzir a intoxicação por Ipomoea verbascoidea em caprinos e descrever os aspectos epidemiológicos, clínicos e histopatológicos da intoxicação espontânea por essa planta no Estado de Pernambuco. Para isso, realizou-se o acompanhamento da epidemiologia da doença em sete municípios do semiárido pernambucano. Três caprinos espontaneamente intoxicados foram examinados e, em seguida eutanasiados e necropsiados (Grupo I). Para reproduzir experimentalmente a doença, as folhas secas de I. verbascoidea contendo 0,02% de swainsonina, foram fornecidas na dose de 4g/kg (0,8mg de swainsonina/kg) a dois grupos de três animais. Os caprinos do Grupo II receberam a planta diariamente por 40 dias e foram eutanasiados no 41º dia de experimento. Os caprinos do Grupo III receberam a planta diariamente por 55 dias e foram eutanasiados no 120º dia de experimento. Outros três caprinos constituíram o grupo controle (Grupo IV). Nos grupos experimentais, as lesões encefálicas foram avaliadas por histopatologia e adicionalmente avaliaram-se as lesões cerebelares por morfometria, mediante mensuração da espessura da camada molecular, do número de neurônios de Purkinje e da área dos corpos celulares dessas células. Os principais sinais clínicos e lesões microscópicas foram semelhantes aos previamente reportados em animais intoxicados por plantas que contem swainsonina. Nos caprinos do GII e GIII, os primeiros sinais clínicos foram observados entre o 22º e 29º dia de experimento; clinicamente a doença desenvolvida por esses animais foi semelhante aos casos espontâneos. Nenhum dos caprinos do GIII se recuperou dos sinais neurológicos. Esse resultado evidencia que o consumo da planta por 26-28 dias após a observação dos primeiros sinais clínicos é suficiente para provocar lesões irreversíveis. Pela análise morfométrica, a camada molecular do cerebelo dos caprinos do Grupo I e III eram mais delgadas que às dos caprinos do grupo controle, e os neurônios de Purkinje estavam atróficos. Sugere-se que essas alterações sejam responsáveis pelo quadro clínico neurológico observado nos caprinos que deixam de ingerir a planta e apresentam seqüelas da intoxicação.

Hypoxic-ischemic injury decreases anxiety-like behavior in rats when associated with loss of tyrosine-hydroxylase immunoreactive neurons of the substantia nigra

Neonatal Sprague-Dawley rats were randomly divided into normal control, mild hypoxia-ischemia (HI), and severe HI groups (N = 10 in each group at each time) on postnatal day 7 (P7) to study the effect of mild and severe HI on anxiety-like behavior and the expression of tyrosine hydroxylase (TH) in the substantia nigra (SN). The mild and severe HI groups were exposed to hypoxia (8 percent O2/92 percent N2) for 90 and 150 min, respectively. The elevated plus-maze (EPM) test was performed to assess anxiety-like behavior by measuring time spent in the open arms (OAT) and OAT percent, and immunohistochemistry was used to determine the expression of TH in the SN at P14, P21, and P28. OAT and OAT percent in the EPM were significantly increased in both the mild (1.88-, 1.99-, and 2.04-fold, and 1.94-, 1.51-, and 1.46-fold) and severe HI groups (1.69-, 1.68-, and 1.87-fold, and 1.83-, 1.43-, and 1.39-fold, respectively; P < 0.05). The percent of TH-positive cells occupying the SN area was significantly and similarly decreased in both the mild (17.7, 40.2, and 47.2 percent) and severe HI groups (16.3, 32.2, and 43.8 percent, respectively; P < 0.05). The decrease in the number of TH-positive cells in the SN and the level of protein expression were closely associated (Pearson correlation analysis: r = 0.991, P = 0.000 in the mild HI group and r = 0.974, P = 0.000 in the severe HI group) with the impaired anxiety-like behaviors. We conclude that neonatal HI results in decreased anxiety-like behavior during the juvenile period of Sprague-Dawley rats, which is associated with the decreased activity of TH in the SN. The impairment of anxiety and the expression of TH are not likely to be dependent on the severity of HI.

Quantitative analysis of the neurons from the myenteric plexus in the ileum of rats submitted to severe protein deficiency / Análise quantitativa dos neurônios do plexo mientérico do íleo de ratos submetidos a intensa carência de proteínas

The effects of protein malnutrition on the quantitative aspects of the myenteric plexus in the ileum of adult Rattus norvegicus were assessed. Thirty 90-day-old rats were divided into two groups: Control Group (CG, n=15) and Experimental Group (EG, n=15). The CG received 26 percent protein chow and the EG received 4 percent protein chow for 90 days. At the end of the experiment, the animals from the CG weighed 369.63±26.33, and the ones from the EG 215.34±56.31. The ileum was submitted to Giemsa, NADH- and NADPH-diaphorase technique in order to evidence nervous cells in the whole-mount preparations. Animals from the EG presented a 41.75 percent body weight loss in relation to the CG as well as 17.6 percent length reduction for the ileum-jejunum. Moreover, the organ was 41 percent lighter for the EG. Giemsa-stained neurons were 17.02 percent more concentrated in the EG (p>0.05). NADH-diaphorase-stained neurons were 26.6 percent more concentrated in the EG (p<0.05), while the NADPH-diaphorase were 26.28 percent more concentrated in this group (p<0.05).

Avaliou-se o efeito da desnutrição protéica sobre o número de neurônios mientéricos do íleo de ratos adultos. Foram utilizados 30 animais (90 dias de idade), divididos em dois grupos: controle (GC, n=15) e experimental (GE, n=15), sendo oferecido ao GC ração com teor protéico de 26 por cento e, para o GE, ração com 4 por cento de proteína, durante 90 dias. Os animais do grupo controle pesaram 369,63±26,33g e o experimental 215,34±56,31g. Preparados de membrana do íleo foram submetidos à técnica de Giemsa, NADH- e NADPH-diaforase. Os animais do GE apresentaram perda de peso de 41,75 por cento, em relação ao GC e redução do comprimento do jejuno-íleo de 17,6 por cento, além disso, o órgão apresentou-se 41 por cento mais leve no GE. Os neurônios corados com a técnica de Giemsa apresentaram-se 17,02 por cento mais concentrados no GE (p>0,05). Os neurônios NADH-diaforase apresentaram-se 26,60 por cento mais concentrados no GE (p<0,05). E os neurônios NADPH-diaforase apresentaram-se 26,28 por cento mais concentrados neste grupo (p<0,05).

Effects of the ascorbic acid supplementation on NADH-diaphorase myenteric neurons in the duodenum of diabetic rats

We assessed the ascorbic acid (AA) supplementation on the myenteric neurons in the duodenum of rats. Fifteen rats with 90 days of age were divided into three groups: control (C), diabetics (D) and ascorbic acid treated diabetics (DA). After 120 days of daily treatment with AA, the duodenum was submitted to the NADH-diaphorase (NADH-d) histochemical technique, which allowed us to evaluate theneuronal density in an area of 8.96 mm2 for each duodenum, and also to measure the cellular profile area of 500 neurons per group. The supplementation promoted an increase on AA levels. The neuronal density (p < 0.05) was higher in the group DA when compared to group D. There were no significant differences in the neuronal areas, when we compared groups C (204 +/- 16.5) and D (146.3 +/- 35.84) to groups D and DA (184.5 +/- 5.6 ) (p > 0.05). The AA-supplementation avoided the density reduction of the NADHd myenteric neurons in the duodenum of diabetic rats.

The dynamically changes of COX-2 and the gene expression of COX-2 mRNA after traumatic brain injury in rats / 法医学杂志

OBJECTIVE@#To illuminate the pathology of traumatic brain injury(TBI) and to better understand the relationship between the expression of COX-2 and the time course of TBI.@*METHODS@#Immunocytochemical staining, double-labeled with the anti-COX-2 antibody and either the neuron-specific antibody NeuN or the astroglial-specific marker GFAP, in situ hybridization and computer image analysis were used.@*RESULTS@#Results from immunohistochemistry indicated time-dependent staining changes of neuronal plasma. The immunostained cells were faint at control cortex, mostly were neurons. The immunostained cells appeared to be darkly stained 30 min after TBI for extended periods of time and reached the maximum at 2 d after injury, reached another peak (P < 0.05) at 4 d post-injury. The darker cells persisted in a high level, significant differences (P < 0.05) even presented between control and 15 d post-injury. The COX-2 mRNA expression was faint at control cortex. The expressions of COX-2 mRNA appeared to be darkly stained 15 min after TBI for extended periods of time and reached the maximum (P < 0.05) at 1 d post-injury, reached another peak (P < 0.05) at 3 d post-injury, and significant differences (P < 0.05) even presented between control and 7 d post-injury, but not 15 d post-injury.@*CONCLUSION@#The results of this study indicated that the expression of COX-2 mRNA and protein had a possible relationship with the extended periods of time after TBI. It might have some relationship between the expression of COX-2 and secondary brain injury after TBI.

Study on the changes of ncNOS in chronic heroin dependence and spontaneous withdrawal in rats / 法医学杂志

OBJECTIVE@#To study the changes and actions of neuronal constructive nitric oxide synthase(ncNOS) in heroin drug abuse.@*METHODS@#The expression of ncNOS and ncNOS mRNA in neurons of cerebral cortex, periaqueductal gray matter and the ventral tegmental area was observed by immunohistochemistry, in situ hybridization and image analysis technique after heroin dependence and spontaneous withdrawal in rats.@*RESULTS@#The quantity of ncNOS and ncNOS mRNA rised clearly and the number of ncNOS and ncNOS mRNA positive cells increased greatly in heroin dependence and withdrawal. The changes of ncNOS and ncNOS mRNA in spontaneous withdrawal were more clear than ones of dependence. Heroin dependence and withdrawal led to alterations in ncNOS and ncNOS mRNA expression in important regions implicated in the physical tolerance and dependence.@*CONCLUSION@#The ncNOS plays an important role in heroin dependence and withdrawal. The ncNOS immunohistochemical changes observed in the present study might be useful for the forensic pathological diagnosis of heroin drug abuse.

Immunohistochemical study of constitutive neuronal and inducible nitric oxide synthase in the central nervous system of goat with natural listeriosis

The expression of both constitutive and inducible forms of nitric oxide synthase (NOS) was investigated by immunohistochemical staining of formalin-fixed paraffin-embedded sections in normal and Listeria monocytogenes-infected brains of goats. In normal control goats, a small number of neurons showed immunoreactivity of both iNOS and nNOS, and the number of iNOS-positive neurons was higher than the number of nNOS-positive neurons. In natural listeriosis, listeria antigens were easily immunostained in the inflammatory cells of microabscesses. In this lesion, the immunoreactivity of iNOS in neurons was more intense than the control, but nNOS was not. In microabscesses, nNOS was weakly visualized in macrophages and neutrophils, while iNOS was expressed in macrophages, but not in neutrophils. These findings suggest that normal caprine brain cells, including neurons, constitutively express iNOS and nNOS, and the expressions of these molecules is increased in Listeria monocytogenes infections. Furthermore, inflammatory cells, including macrophages, expressing both nNOS and iNOS may play important roles in the pathogenesis of bacterial meningoencephalitis in goat.

Neuronal and endothelial nitric oxide synthase gene knockout mice

Targeted disruption of the neuronal nitric oxide synthase (nNOS) and endothelial nitric oxide synthase (eNOS) genes has led to knockout mice that lack these isoforms. These animal models have been useful to study the roles of nitric oxide (NO) in physiologic processes. nNOS knockout mice have enlarged stomachs and defects in the inhibitory junction potential involved in gastrointestinal motility. eNOS knockout mice are hypertensive and lack endothelium-derived relaxing factor activity. When these animals are subjected to models of focal ischemia, the nNOS mutant mice develop smaller infarcts, consistent with a role for nNOS in neurotoxicity following cerebral ischemia. In contrast, eNOS mutant mice develop larger infarcts, and show a more pronounced hemodynamic effect of vascular occlusion. The knockout mice also show that nNOS and eNOS isoforms differentially modulate the release of neurotransmitters in various regions of the brain. eNOS knockout mice respond to vessel injury with greater neointimal proliferation, confirming that reduced NO levels seen in endothelial dysfunction change the vessel response to injury. Furthermore, eNOS mutant mice still show a protective effect of female gender, indicating that the mechanism of this protection cannot be limited to upregulation of eNOS expression. The eNOS mutant mice also prove that eNOS modulates the cardiac contractile response to ß-adrenergic agonists and baseline diastolic relaxation. Atrial natriuretic peptide, upregulated in the hearts of eNOS mutant mice, normalizes cGMP levels and restores normal diastolic relaxation.

Effects of a neuronal nitric oxide synthase inhibitor on lipopolysaccharide-induced fever

It has been demonstrated that nitric oxide (NO) has a thermoregulatory action, but very little is known about the mechanisms involved. In the present study we determined the effect of neuronal nitric oxide synthase (nNOS) inhibition on thermoregulation. We used 7-nitroindazole (7-NI, 1, 10 and 30 mg/kg body weight), a selective nNOS inhibitor, injected intraperitoneally into normothermic Wistar rats (200-250 g) and rats with fever induced by lipopolysaccharide (LPS) (100 µg/kg body weight) administration. It has been demonstrated that the effects of 30 mg/kg of 7-NI given intraperitoneally may inhibit 60 per cent of nNOS activity in rats. In all experiments the colonic temperature of awake unrestrained rats was measured over a period of 5 h at 15-min intervals after intraperitoneal injection of 7-NI. We observed that the injection of 30 mg/kg of 7-NI induced a 1.5oC drop in body temperature, which was statistically significant 1 h after injection (P<0.02). The coinjection of LPS and 7-NI was followed by a significant (P<0.02) hypothermia about 0.5oC below baseline. These findings show that an nNOS isoform is required for thermoregulation and participates in the production of fever in rats.

Protéinas y enzimas como posibles mensajeros en la comunicación química interneuronal: parte II / Proteins and enzymes as chemical messengers regulating neuronal transmission: part II

La neurotransmisión química en el cerebro de los mamíferos y en el de las especies no mamíferas, se caracteriza por la liberación neuronal de una amplia gama de moléculas mensajeras, que incluyen neurotransmisores de naturaleza peptídica y no peptídica que poseen la capacidad de facilitar o inhibir la transmisión sináptica mediante la generación de señales eléctricas de tipo excitatorio o inhibitorio. Además de estas sustancias bioactivas, también hay un extenso grupo de macromoléculas de naturaleza protéica que participan en la comunicación química neuronal, generando y regulando diversas respuestas biológicas en el sistema nervioso

Number and size of myenteric neurons of the duodenum of adult rats with acute diabetes

This study had as its purpose to assess the effects of acute diabetes induced by streptozotocin (35 mg/kg body weight) on the number and size of the myenteric neurons of the duodenum of adult rats considering equally the antimesenteric and intermediate regions of the intestinal circunference. Experimental period extended for a week. Neuronal counts were carried out on the same number of fields of both regions of the duodenal circunference and measurements of neuronal and nuclear areas on equal numbers of cells. Number and size of the myenteric neurons stained with Giensa were not significantly different between groups. On the other hand, the proportion of NADH-positive neurons increased from 18.54 per cent on the controls to 39.33 per cent on the diabetics. The authors discuss that this increased reactivity probably results from a greater NADH/NAD* ratio, described in many tissues of diabetic animals, which has consequences on the modulation of the enzymes that use these cofactors and whose activity is detected by the NADH-diaphorase technique.

Proteínas y enzimas como posibles mensajeros en la comunicación química interneuronal. Parte I / Proteins and enzymes as chemical messangers regulating neuronal transmission. First part

En los últimos 30 años un extenso cúmulo de información ha sido reportada sobre la identificación y caracterización fisiológica y molecular de las proteínas y las enzimas; cuya liberación espontánea e inducida a partir del tejido neuronal y de los tejidos extraneuronales, como las células cromafines de la glándula suprarrenal, han permitido no sólo la subsecuente clonación y aislamiento molecular de estas moléculas protéicas, sino que, además, mediante la aplicación de métodos de hibridación in situ, de la generación de anticuerpos selectivos, conjuntamente con el apoyo de técnicas inmunohistoquímicas, se ha podido detectar el origen, localización y expresión celular de enzimas como la acetilcolinesterasa específica (AChE), y la dopamina-betahidroxilasa (DBH), en diversas regiones del cerebro de distintas especies mamíferas, de donde se ha encontrado su capacidad para ejercer funciones neurotróficas. Asimismo, se han abordado similares planteamientos experimentales para identificar y localizar la expresión celular y subcelular de proteínas como las de la familia de los polipéptidos, representados por las cromograninas y las secretoneurinas, que son polipéptidos, cuya síntesis celular modificaciones postraduccionales, almacenamiento y segregación vesicular, como su exocitosis celular en el tejido neuronal y en los tejidos neuroendocrinos, han sido estudiadas in vivo e in vitro a partir de diferentes preparaciones biológicas, cuyos resultados han sido motivo de múltiples publicaciones. Más aún, estas proteínas parecen ser precursoras de varios péptidos bioactivos, en los que se han observado diversas funciones de tipo autocrino, paracrino, o endocrino, en el tejido neuronal y en los tejidos extraneuronales. Asimismo, una extensa familia de factores tróficos, conocidos como neurotrofinas, incluyendo el Factor de Crecimiento Neuronal (NGF), recientemente se aislaron, identificaron, y caracterizaron molecular y funcionalmente, in vivo e in vitro, con lo que se demostró su capacidad para regular y mantener la supervivencia de diferentes poblaciones de células neuronales en el sistema nervioso central y periférico de múltiples especies de vertebrados e invertebrados, lo que parece mediarse por la activación de diversos subgrupos de receptores membranales, recientemente identificados y localizados en discretas poblaciones neuronales

Isolation and characterization of brain-specific transglutaminases from rat

The relevance of transglutaminases with neural function and several disorders has been emphasized recently. Especially, many polypeptides associated with neurodegenerative diseases are suggested to be putative transglutaminase substrates such as beta amyloid protein of Alzheimer's disease, microtubule-associated proteins and neurofilaments, etc. In addition, the CAG repeated gene products with probable polyglutamine tract, putative transglutaminase substrates, were identified in several neurodegenerative disorders. However, the identity of the brain transglutaminase has not been confirmed, because of enzymic stability and low activity. In the present experiment, we have isolated brain-specific transglutaminases, designated as TGase NI and TGase NII, which are different from other types of transglutaminases in respects of molecular weights (mw. 45 kDa, 29 kDa respectively), substrate affinity, elution profile on ion-exchange chromatography, sensitivity to proteases and ethanol, and immunological properties. The enzymes were localized specifically in the brain tissues but not in the liver tissue. And neural cells such as pheochromocytoma cell, glioma cell, primary neuronal and glial cells were shown to be enriched with TGase NI and TGase NII. The possible biological roles of the enzymes were discussed not only on the aspect of crosslinking activity but also of signal transducing capacity of the enzyme in the brain.

Immunohistochemical expression of phospholipase C in global and focal ischemic encephalopathy in gerbil: relationship with morphological changes

Phospholipase C (PLC) and related enzymes in signal transduction system are closely linked to cellular damage in ischemic encephalopathy. This study was undertaken to elucidate the time sequential changes of PLC isoenzymes (beta and gamma) in vulnerable areas of hippocampus in global ischemia and infarcted area in focal infarction. Mongolian gerbils were used because of their susceptibility to ischemic encephalopathy and divided into the following groups: the bilateral ischemia with various reperfusion periods group, unilateral progressive ischemia group, and focal ischemia group induced by infusion of iron particles through the femoral artery. The changes of PLC isoenzymes were observed immunohistochemically and matched with morphological changes. In the global ischemia with reperfusion group, the changes were most significant in hippocampus. Sequential changes of neurons such as red neurons at an early stage progressed to pknotic neurons at a later stage were noted with typical delayed neuronal damage in the corns ammonis (CA) 1 subfield of hippocampus. Red neurons and pyknotic neurons as well as intracytoplasmic inclusion in 3 to 24 hours of reperfusion showed loss of PLC isoenzymes as well as tubulin. The changes of PLC expression were corresponding to the degeneration of neurons with no discernible time sequential changes in remaining neurons. In the unilateral hemispheric progressive ischemia group, ischemic damage was far more marked and extensive with no selective injury pattern according to time and location. At 1 day, there was diffuse vacuolization and necrosis of neuropil with a loss of neuron. Admixed surviving neurons and vacuolated neuropil showed increased reaction to anti-PLC antibodies, which could be either an evidence of protein synthesis responding to ischemic insult or an artifactual change. Focal ischemia group showed time sequential changes of blood vessels and white blood cells with necrosis of surrounding tissue. Degenerating hippocampal neurons in infarction also showed a strong positive reaction to anti-PLC antibody, which was most likely due to condensation of cytoplasm rather than increased synthesis. This study showed different changes of PLC expression in global ischemic encephalopathy with reperfusion, progressive ischemia, and focal infarction, which suggested different pathophysiologic mechanism between these conditions.

Oxido nítrico en la nocicepción / Nitric oxide and nociception

Las investigaciones en el campo del dolor y de la analgesia se están enfocando hacia los eventos celulares y moleculares subyacentes a los mecanismos de dolor crónico. En particular, mucha atención está recibiendo el óxido nítrico (ON), un nuevo tipo de neurotransmisor. El ON es un radical libre gaseoso e inestable que resulta de la oxidación de la L-arginina a L-citrulina en una reacción catalizada por la sintasa del óxido nítrico. El ON cumple un papel de molécula citotóxica de macrófagos activados y de relajante de músculo liso. Además de estas funciones, el ON actúa como neuromodulador y/o neurotransmisor en el sistema nervioso. Reportes recientes han comenzado a definir el papel del ON en los procesos nociceptivos a nivel de la médula espinal. Asociado a receptores sensibles al N-metil-D-aspartato (NMDA), parece estar involucrado en los mecanismos suyacentes de la hiperalgesia térmica, involucrado en la facilitación de reflejos térmicos. Es más, parece que la producción sostenida del On y la subsecuente activación de la guanilato ciclasa soluble en la médula espinal lumbar, son condiciones requeridas para el mantenimiento de la hiperalgesia térmica producida en modelos del dolor persistente. La inhibición de la sintasa del ON con nitro-L-arginina bloquea la tolerancia a la morfina en ratones. La nitro-L-arginina también es capaz de revertir lentamente tolerancia preexistente, además de reducir la dependencia y de revertir la dependencia a la droga previemente establecida. La acción del ON es selectivo para la tolerancia y dependencia a receptores del subtipo µ

Localization of NADPH diaphorase activity in the human visual cortex

The present report describes the activity of NADPH-diaphorase (NADPHd) in area 17 of autopsied normal human visual cortex. Four human brains from autopsy tissue (4-8 h postmortem) were fixed by immersion in 4 per cent paraformaldehyde in 0.1 M sodium phosphate buffer, pH 7.2-7.4, or in 10 per cent formalin for 24 h. NADPHd histochemistry was done using the malic enzyme indirect method. The neurpile pattern of enzyme activity presented a clear six layer appearance. Cell morphology and the laminar distribution of 73 NADPHd-positive neurons are descrived. All neurons found in area 17 of human cortex were sparsely spiny or smooth cells, located in all cortical layers exept layer 4c. Quantitative analysis of the branching pattern of the dendritic tree was carried out. A symmetrical pattern was observed with no particular dendritic bias except for a few white matter and layer 1 cells. Larger dendritic fields were found in white matter cells when compared to the other corical layers. Comparison of cell densities for gray and white matters showed that 85 per cent of the NADPHd-positive neurons were located in the white matter. NADPH was colocalized with nitric oxide synthase which produces nitric oxide, a short-life neuromediator implicated in synaptic plasticity, neuroprotection, and neurotoxicity. thus, the spatial distribution of the NADPHd cells is important for posterior functional studies of the neuromediators in the brain

Distribution of NADPH-diaphorase-positive neurons in the opossum neocortex

The distribution of NADPH-diaphorase reactive cells was evalutated both in horizontal sections of a flattended cortex and in transversal sections of the opossum (Didelphis marsupialis) neocortex. The tangential distribution of labeled cells behind the orbitalis fissure was denser in the rostral vs caudal regions and in the lateral vs medial regions. Transversal sections revealed that most of the positive neurons are in the grey matter, although 1/4 of this population is located in the underlying white matter. This pattern of neuronal distribution is similar to that previously described in rodents, but quite different from that observed in higher mammals such as the cat and primates