Espinogénesis en motoneuronas de la médula espinal tras la lesión farmacológica de la corteza motora de ratas / Spinogenesis in spinal cord motor neurons following pharmacological lesions to the rat motor cortex

INTRODUCCIÓN: Diversas enfermedades neuropatologías asociadas a la degeneración del tracto corticoespinal muestran deterioro de las funciones motoras. Tales alteraciones neurológicas se asocian a diversos fenómenos plásticos subsecuentes, a nivel tanto presináptico como postsináptico. Sin embargo, no existe evidencia que indique la existencia de modificaciones en la transmisión de información del tracto corticoespinal a las motoneuronas espinales. MÉTODOS: Se indujo una lesión por vía estereotáxica en la corteza motora primaria de ratas hembra adultas con ácido kaínico y, 15 días después, se evaluó el desempeño motor mediante la escala BBB y en un dispositivo Rota-Rod. Paralelamente, se cuantificó la densidad numérica y proporcional de las espinas delgadas, en hongo y gordas, en motoneuronas de un segmento torácico-lumbar de la médula espinal. Así mismo, se registró la expresión de las proteínas espinofilina, sinaptofisina β III-tubulina. RESULTADOS: La lesión farmacológica provocó un desempeño motor deficiente. Así mismo, tanto la densidad de espinas como la proporción de espinas delgadas y gordas fue mayor, al igual que la expresión de las 3 proteínas estudiadas. CONCLUSIÓN: La aparición de los síntomas clínicos de daño neurológico provocado por la degeneración walleriana del tracto corticoespinal se acompaña de respuestas plásticas espontáneas de tipo compensador, a nivel sináptico. Lo anterior indica que durante la rehabilitación temprana de este tipo de pacientes, la plasticidad espontánea constituye un factor que se debe considerar para el diseño de estrategias de intervención más eficientes

INTRODUCTION: Motor function is impaired in multiple neurological diseases associated with corticospinal tract degeneration. Motor impairment has been linked to plastic changes at both the presynaptic and postsynaptic levels. However, there is no evidence of changes in information transmission from the cortex to spinal motor neurons. METHODS: We used kainic acid to induce stereotactic lesions to the primary motor cortex of female adult rats. Fifteen days later, we evaluated motor function with the BBB scale and the rotarod and determined the density of thin, stubby, and mushroom spines of motor neurons from a thoracolumbar segment of the spinal cord. Spinophilin, synaptophysin, and β III-tubulin expression was also measured. RESULTS: Pharmacological lesions resulted in poor motor performance. Spine density and the proportion of thin and stubby spines were greater. We also observed increased expression of the 3 proteins analysed. CONCLUSION: The clinical symptoms of neurological damage secondary to Wallerian degeneration of the corticospinal tract are associated with spontaneous, compensatory plastic changes at the synaptic level. Based on these findings, spontaneous plasticity is a factor to consider when designing more efficient strategies in the early phase of rehabilitation

Spinogenesis in spinal cord motor neurons following pharmacological lesions to the rat motor cortex. / Espinogénesis en motoneuronas de la médula espinal tras la lesión farmacológica de la corteza motora de ratas.

INTRODUCTION: Motor function is impaired in multiple neurological diseases associated with corticospinal tract degeneration. Motor impairment has been linked to plastic changes at both the presynaptic and postsynaptic levels. However, there is no evidence of changes in information transmission from the cortex to spinal motor neurons. METHODS: We used kainic acid to induce stereotactic lesions to the primary motor cortex of female adult rats. Fifteen days later, we evaluated motor function with the BBB scale and the rotarod and determined the density of thin, stubby, and mushroom spines of motor neurons from a thoracolumbar segment of the spinal cord. Spinophilin, synaptophysin, and ß iii-tubulin expression was also measured. RESULTS: Pharmacological lesions resulted in poor motor performance. Spine density and the proportion of thin and stubby spines were greater. We also observed increased expression of the 3 proteins analysed. CONCLUSION: The clinical symptoms of neurological damage secondary to Wallerian degeneration of the corticospinal tract are associated with spontaneous, compensatory plastic changes at the synaptic level. Based on these findings, spontaneous plasticity is a factor to consider when designing more efficient strategies in the early phase of rehabilitation.

Reproductive biology in cassava: stigma receptivity and pollen tube growth.

Knowledge on the reproductive biology of cassava, relevant to breeders and molecular geneticists, is still limited. Therefore, different studies were carried out to determine the duration of stigma receptivity and the rate of pollen tube growth. Inflorescences were covered for up to 3 days after the first opening of the bracts (e.g. anthesis day) to prevent open pollination. Results indicate that fruit and seed set are drastically reduced when flowers were covered for 2 or 3 days. However, fruits and seeds were obtained even from flowers that had been covered for 3 days after anthesis, although at low frequency. The rate of pollen tube growth was assessed in many combinations of female and male progenitors crossed through controlled pollinations and collecting the pistils at varying hours after pollination (HAP). Pollen tube growth is fast during the first 6 HAP reaching the tip of the nucellar beak. The growth slows down thereafter, taking 10 additional hours to reach the end of the beak. The growth of pollen tubes slows down even further until they enter the embryo sac. Only 10% of samples showed pollen tubes entering the embryo sac between 48 and 66 HAP. Although several tubes may reach the nucellar beak, only one was observed entering the embryo sac. Results, across the different experiments, were highly variable suggesting that the timeline of fertilization is influenced both by genotypic and environmental factors as well as the manual manipulation of inflorescences and cyathia.

Efecto de Implantes de Polipirrol Sintetizados por Diferentes Métodos sobre Lesiones de Médula Espinal en Ratas / Effect of Pyrrole Implants Synthesized by Different Methods on Spinal Cord Injuries of Rats

Polypyrrole (PPy) and polypyrrole/polyethylene glycol (PPy/PEG) implants synthesized by chemical, electro-chemical, and plasma polymerization methods were implanted into the injured spinal cord of rats to determine their effect on motor function recovery. Before implantation, the materials were characterized by infrared (IR) spectroscopy. An experimental model of traumatic spinal cord injury (TSCI) by complete transection at thoracic level 9, in rats was used. The polymer implants were inserted immediately after transection. Motor function recovery was evaluated once a week during 5 weeks using the Basso, Beattie and Bresnahan (BBB) motor scale. Histological evaluation was done at the end of the recovery evaluation period using hematoxylin/eosin stain. Results showed that animals implanted with polymers synthesized by plasma had a better integration into the nerve tissue, less inflammatory response and a better functional recovery than animals implanted with polymers synthesized by chemical or electrochemical methods.

En el presente trabajo se comparó el efecto de implantes poliméricos derivados del pirrol (polipirrol o PPy) y del copolímero polipirrol/polietilenglicol (PPy/PEG), obtenidos por diferentes métodos de síntesis: químico, electroquímico y polimerización por plasma con el propósito de determinar si el método de síntesis puede influir sobre el efecto que producen al ser implantados después de una lesión traumática de la médula espinal de ratas. Antes de realizar el implante, las características químicas y estructurales de los polímeros fueron analizadas por espectroscopia de infrarrojo (IR). Se utilizó un modelo experimental de lesión traumática de médula espinal (LTME) por sección completa en ratas. La LTME se realizó a nivel torácico 9 y el polímero fue implantado de inmediato en la zona de lesión. La recuperación de la función motora se evaluó mediante la escala Basso, Beattie y Bresnahan (BBB) una vez por semana durante 5 semanas. La evaluación histológica se realizó al término del seguimiento con la tinción de hematoxilina/eosina. Los resultados muestran que los animales implantados con polímeros sintetizados por plasma se integraron mejor al tejido nervioso, redujeron la respuesta inflamatoria y favorecieron una mayor recuperación funcional en comparación con los animales implantados con materiales sintetizados por métodos químicos o electroquímicos.

Prediction of carotenoids, cyanide and dry matter contents in fresh cassava root using NIRS and Hunter color techniques.

Efforts are currently underway to improve carotenoids content in cassava roots through conventional breeding as a strategy to reduce vitamin A deficiency. However, only few samples can be quantified each day for total carotenoids (TCC) and ß-carotene (TBC) contents, limiting the gains from breeding. A database with >3000 samples was used to evaluate the potential of NIRS and chromameter devices to predict root quality traits. Maximum TTC and TBC were up to 25.5 and 16.6 µg/g (fresh weight basis), respectively. NIRS predictions were highly satisfactory for dry matter content (DMC, R(2): 0.96), TCC (R(2): 0.92) and TBC (R(2): 0.93). NIRS could also distinguish roots with high or low cyanogenic potential (R(2): 0.86). Hunter color parameters could also be used for predictions, but with lower accuracy than NIRS. NIRS or chromameter improve selection protocols, allowing faster gains from breeding. Results also demonstrate that TBC and DMC can be improved simultaneously (required for the adoption of biofortified cassava).

Effect of natural exogenous antioxidants on aging and on neurodegenerative diseases.

Aging and neurodegenerative diseases share oxidative stress cell damage and depletion of endogenous antioxidants as mechanisms of injury, phenomena that are occurring at different rates in each process. Nevertheless, as the central nervous system (CNS) consists largely of lipids and has a poor catalase activity, a low amount of superoxide dismutase and is rich in iron, its cellular components are damaged easily by overproduction of free radicals in any of these physiological or pathological conditions. Thus, antioxidants are needed to prevent the formation and to oppose the free radicals damage to DNA, lipids, proteins, and other biomolecules. Due to endogenous antioxidant defenses are inadequate to prevent damage completely, different efforts have been undertaken in order to increase the use of natural antioxidants and to develop antioxidants that might ameliorate neural injury by oxidative stress. In this context, natural antioxidants like flavonoids (quercetin, curcumin, luteolin and catechins), magnolol and honokiol are showing to be the efficient inhibitors of the oxidative process and seem to be a better therapeutic option than the traditional ones (vitamins C and E, and ß-carotene) in various models of aging and injury in vitro and in vivo conditions. Thus, the goal of the present review is to discuss the molecular basis, mechanisms of action, functions, and targets of flavonoids, magnolol, honokiol and traditional antioxidants with the aim of obtaining better results when they are prescribed on aging and neurodegenerative diseases.

Comparative morphology, biology and histology of reproductive development in three lines of Manihot esculenta Crantz (Euphorbiaceae: Crotonoideae).

BACKGROUND AND AIMS: Cassava (Manihot esculenta), a major food staple in the tropics and subtropics, thrives even in environments undergoing threatening climate change. To satisfy the increasing demand for crop improvement and overcome the limitations of conventional breeding, the introduction of inbreeding techniques such as the production of doubled haploid lines via androgenesis or gynogenesis offers advantages. However, comprehensive studies on cassava flower bud biology or structural development are lacking and precise structural and biological information is a prerequisite to enhance the efficiency of these techniques. METHODOLOGY: The floral biology of three selected cassava lines was studied, focusing on morphology, phenology and pollen biology (quantity, viability and dimorphism). Histological studies were also conducted on microsporogenesis/microgametogenesis and megasporogenesis/megagameto-genesis to generate precise developmental data for these lines. PRINCIPAL RESULTS: Male and female cyathia have distinct developmental phases. Pollen viability was high during immature stages of plant development; however, pollen mortality was common at later stages. Pollen trimorphism in male gametophytes towards the larger or smaller pollen size, as compared with normal size, was observed. Ten characteristic events were identified in male gametogenesis and six in female gametogenesis that were correlated with flower bud diameter. Male gametophyte diameter at different developmental stages was also determined. CONCLUSIONS: Results indicate that the three lines did not differ significantly, except regarding a few morphological aspects such as plant height, flower colour and number of male cyathia. Pollen grains were initially viable, but viability decreased drastically at later stages of growth. Abnormal meiosis or mitosis triggered pollen trimorphism. The demonstrated sequential events of reproductive development generated valuable information at the cellular level, which will help close the current information gap for cassava improvement via breeding programmes and doubled haploid plant production.

CAT and MRI Studies of Spinal Cord Injured Rats Implanted with PPy/I / Estudios CAT y MRI en ratas con lesión de médula espinal con implantes de PPy/I

Polymers synthesized by plasma derived from pyrrole have been recently implanted in rats with spinal cord injuries (SCI) using a complete section model; the polymers contribute to the functional recovery after the injury. In this work, the SCI in rats was studied using noninvasive techniques such as magnetic resonance imaging (MRI). Also computerized axial tomography taken chronologically with and without polymeric implants. 3D reconstructions were used to follow the structural arrangement, the location of the implant and the formation of cysts. MRI shows a clear differentiation between white and gray matter, the implanted material and cysts due to secondary damage after the injury.

Polímeros sintetizados por plasma derivados del pirrol, han sido implantados en ratas con lesión de médula espinal (LME) usando un modelo de lesión por sección completa; los polímeros contribuyen a la recuperación funcional después de la lesión. En este trabajo, la LME se estudió usando técnicas no invasivas como imágenes por resonancia magnética (IRM) y tomografía axial computarizada tomada cronológicamente en sujetos con y sin implante polimérico. Se usaron reconstrucciones 3D para seguir el arreglo estructural, la localización del implante y la formación de quistes. Las IRM muestran una clara diferenciación entre materia gris, materia blanca, material implantado y quistes formados debido a los mecanismos de daño secundario después de la lesión.

Deleterious versus neuroprotective effect of metabolic inhibition after traumatic spinal cord injury.

STUDY DESIGN: This work is an experimental and prospective study in adult, female, Long-Evans rats. OBJECTIVES: The aim of this study was to probe the effect of metabolic inhibition after an acute traumatic spinal cord injury (TSCI) using a standardized contusion model (NYU impactor) to know whether the metabolic inhibition is a 'secondary mechanism of injury' or a mechanism of protection. SETTING: All experimental procedures were carried out in the Mexico City. METHODS: Animals were divided into five groups: one sham and four with TSCI, including no treatment, rotenone (inhibitor of mitochondrial complex I), sodium azide (inhibitor of mitochondrial complex IV) and pyrophosphate of thiamine or non-degradable cocarboxylase as a metabolic reactivator. RESULTS: After TSCI, the metabolic inhibition with sodium azide treatment diminished the lipid peroxidation process (malondialdehyde levels by spectrophotometric procedures) and the damage to the spinal cord tissue (morphometric analysis), and increased the activity of creatine kinase and lactate dehydrogenase enzymes (P<0.05) (measured by spectrophotometric procedures 24 h after TSCI as well as after the functional recovery of the hind limb (evaluated weekly for 2 months by the BBB (Basso, Beattie and Bresnahan) scale)) when compared with the TSCI group without treatment. CONCLUSION: The results show that the partial and transitory inhibition of the aerobic metabolism after an acute TSCI could be a self-protection mechanism instead of being a 'secondary mechanism of injury'.

Hyperthermic seizures and hyperthermia in immature rats modify the subsequent pentylenetetrazole-induced seizures.

It has been described that febrile seizures during infancy increase risk of subsequent non-febrile seizures during the adulthood. However, latency period between febrile seizure and the onset of the first spontaneous seizure has not been evaluated. The present study was designed to investigate the susceptibility to subsequent seizures in immature rats that had experienced early-life hyperthermic seizures and before they achieved the adult age. The results were compared with those induced by hyperthermia alone. Pentylenetetrazol (PTZ) was applied 24h or 20 days after hyperthermic seizures or hyperthermia were induced in 10-day-old rats by a regulated stream of moderately heated air. One day after hyperthermic seizures or hyperthermia, animals demonstrated enhanced latency to the PTZ-induced myoclonic (88% and 53%, respectively), clonic (60% and 60%, respectively) and tonic seizures (233% and 659%, respectively). The incidence of myoclonic and clonic seizures was similar to that in control group (100%). However, hyperthermic seizures reduced (50%) the incidence of tonic seizures. Twenty days after hyperthermic seizures there was an augmented latency to tonic seizures (123%) and reduced incidence for all the PTZ-induced seizures (71% myoclonic; 71% clonic seizures; 57% tonic seizures) when compared with control group (100%). In contrast, hyperthermia enhanced only the latency to myoclonic (133%) and clonic seizures (659%). Our data indicate that hyperthermic seizures or hyperthermia induces a protective effect against PTZ-induced seizures during a latency period. A possible involvement of gamma-aminobutyric acid (GABA) system is discussed.

Hyperthermia-induced seizures modify the GABA(A) and benzodiazepine receptor binding in immature rat brain.

Effects of hyperthermia-induced seizures (HS) on GABA(A) and benzodiazepine (BDZ) receptor binding in immature rat brain were evaluated using in vitro autoradiography. HS were induced in 10-days-old rats by a regulated stream of moderately heated air directed 50 cm above the animals. Rats were killed 30 min, 24 h or 20 days after HS and their brains were used for in vitro autoradiography experiments to determine GABA(A) and BDZ receptor binding. GABA(A) binding was significantly enhanced in all brain areas evaluated 30 min after HS, an effect that endures 24 h and 20 days after seizures. Concerning BDZ receptor binding, a significant increase was detected in entorhinal and perirhinal cortices and decreased in basolateral amygdala 30 min following HS. One day after HS, animals demonstrated enhanced BDZ binding in the cingulate, frontal, posterior parietal, entorhinal, temporal and perirhinal cortices; striatum, accumbens, substantia nigra pars compacta and amygdala nuclei. Twenty days after HS enhanced BDZ binding was restricted in the cingulated, frontal, anterior and posterior parietal cortices, as well as in substantia nigra pars reticulata, whereas decreased values were found in accumbens nucleus and substantia nigra pars compacta. Our data indicate differential effects of HS in GABA(A) and BDZ binding in immature brain. HS-induced GABA(A) and BDZ changes are different from those previously described in experimental models of temporal lobe epilepsy in adult animals.

Epistasis in the expression of relevant traits in cassava (Manihot esculenta Crantz) for subhumid conditions.

There is limited knowledge on the inheritance of agronomic traits in cassava and the importance of epistasis for most crops. A nine-parent diallel study was conducted in subhumid environments. Thirty clones were obtained from each F1 cross. Each clone was represented by six plants, which were distributed in three replications at two locations. Therefore the same 30 genotypes of each F1 cross were planted in the three replications at the two locations. Analysis of variance suggested significant genetic effects for all variables analyzed (reaction to thrips, fresh root and foliage yields, harvest index, dry matter content, and root dry matter yield). Significant epistatic effects were observed for all variables, except harvest index. Dominance variance was always significant, except for dry matter content and dry matter yield. Additive variance was significant only for reaction to thrips. Results suggested that dominance plays an important role in complex traits such as root yield. The significance of epistasis can help us understand the difficulties of quantitative genetics models and QTLs in satisfactorily explaining phenotypic variation in traits with complex inheritance. Significant epistasis would justify the production of inbred parental lines to fix favorable allele combinations in the production of hybrid cassava cultivars.

Spontaneous long-term remyelination after traumatic spinal cord injury in rats.

The capability of the central nervous system to remyelinate axons after a lesion has been well documented, even though it had been described as an abortive and incomplete process. At present there are no long-term morphometric studies to assess the spinal cord (S.C.) remyelinative capability. With the purpose to understand this phenomenon better, the S.C. of seven lesionless rats and the S.C. of 21 rats subjected to a severe weight-drop contusion injury were evaluated at 1, 2, 4, 6, and 12 months after injury. The axonal diameter and the myelination index (MI = axolemmal perimeter divided by myelinated fiber perimeter) were registered in the outer rim of the cord at T9 SC level using a transmission electron microscope and a digitizing computer system. The average myelinated fiber loss was 95.1%. One month after the SC, 64% of the surviving fibers were demyelinated while 12 months later, only 30% of the fibers had no myelin sheath. The MI in the control group was 0.72 +/- 0.07 (X +/- S.D.). In the experimental groups, the greatest demyelination was observed two months after the lesion (MI = 0.90 +/- 0.03), while the greatest myelination was observed 12 months after the injury (MI = 0.83 +/- 0.02). There was a statistical difference (p < 0.02) in MI between 2 and 12 months which means that remyelination had taken place. Remyelination was mainly achieved because of Schwann cells. The proportion of small fibers (diameter = 0.5 micron or less) considered as axon collaterals, increased from 18.45% at 1 month to 27.66% a year after the contusion. Results suggest that remyelination is not an abortive phenomenon but in fact a slow process occurring parallel to other tissue plastic phenomena, such as the emission of axon collaterals.

Alteration of cyclosporin-A pharmacokinetics after experimental spinal cord injury.

The pharmacokinetics of the immunosuppressive agent cyclosporin-A (CsA) were studied in rats submitted to spinal cord (SC) injury. A single CsA 10 mg/kg dose was given either intraperitoneally (i.p.) or orally to rats submitted to experimental SC injury at the T8 level. Twenty four hours after lesion (acute stage of SC injury) i.p. CsA bioavailability was increased, while t1/2 was prolonged. However, oral bioavailability was reduced. Seven weeks after lesion (chronic stage of SC injury) CsA bioavailability, by either route, was not significantly different from control values. Results indicate that parenteral CsA bioavailability is increased during the acute stage of SC lesion, probably due to an impaired elimination. Oral bioavailability, however, is decreased, since there is also an important reduction in gastrointestinal CsA absorption that overrides the effect of impaired elimination. Alterations in CsA pharmacokinetics appear to revert during the chronic stage of SC injury. Changes in CsA bioavailability, depending on the route of administration and on time, must be considered to design an adequate immunosuppressive treatment in SC injury.

Neuroprotection of completely lacerated spinal cord of adult rats by homotopic and heterotopic transplantation.

To evaluate the neuroprotective effect of transplants placed in the lesion zone after a complete spinal cord (SC) laceration, two independent series of experiments were carried out. In the first, allogeneic or xenogeneic fetal SC was transplanted into the gaps of the damaged lower thoracic SC of adult rats. In the transplanted rats the incidence of life-threatening complications was reduced, and the survival rate was increased compared with the control group (lesion, without implant). Histological examination showed less damage to the neighboring SC parenchyma in the transplanted rats. The measurement of this neuroprotective effect was made in a second series of experiments. Using the same model of SC injury, allogeneic fetal SC, autologous peripheral nerve and/or adipose tissue were implanted. Rats with implants of Gelfoam and damaged rats without implants were the controls. The implanted rats of all groups, including the Gelfoam group, showed a better survival rate than the nonimplanted rats. Significantly less damage to the neighboring SC parenchyma was measured in implanted rats with any of the live tissues tested compared with non-implanted rats, although no significant differences were observed between the Gelfoam group and the nonimplanted rats. Histological evidence of tissue implant survival was observed in all corresponding groups. It is concluded that the transplanted tissues tested here have a neuroprotective effect, possibly by acting as a buffer to neurotoxic substance(s) released by the stumps, and/or by exerting trophic effect(s) on the host.