Undernutrition - thirty years of the Regional Basic Diet: the legacy of Naíde Teodósio in different fields of knowledge.

Undernutrition is characterized by an imbalance of essential nutrients with an insufficient nutritional intake, a disorder in which the clinical manifestations in most cases are the result of the economic and social context in which the individual lives. In 1990, the study by the medical and humanitarian Naíde Teodósio (1915-2005) and coworkers, which formulated the Regional Basic Diet (RBD) model for inducing undernutrition, was published. This diet model took its origin from the observation of the dietary habits of families that inhabited impoverished areas from the Pernambuco State. RBD mimics an undernutrition framework that extends not only to the Brazilian population, but to populations in different regions worldwide. The studies based on RBD-induced deficiencies provide a better understanding of the impact of undernutrition on the pathophysiological mechanisms underlying the most diverse prevalent diseases. Indexed papers that are analyzed in this review focus on the importance of using RBD in different areas of knowledge. These papers reflect a new paradigm in translational medicine: they show how the study of pathology using the RBD model in animals over the past 30 years has and still can help scientists today, shedding light on the mechanisms of prevalent diseases that affect impoverished populations.

Chlorella vulgaris functional alcoholic beverage: Effect on propagation of cortical spreading depression and functional properties.

Recent advances in microalgae biotechnology have proven that these microorganisms contain a number of bioactive molecules, that can be used as food additives that help prevent disease. The green microalga Chlorella vulgaris presents several biomolecules, such as lutein and astaxanthin, with antioxidant capacity, which can play a protective role in tissues. In this study, we produced and analyzed a C. vulgaris functional alcoholic beverage (produced using a traditional Brazilian alcoholic beverage, cachaça, and C. vulgaris biomass). Assays were conducted in vitro by radical scavenging tests, and in vivo, by modeling cortical spreading depression in rat brains. Scavenging radical assays showed that consumption of the C. vulgaris alcoholic beverage had a DPPH inhibition of 77.2%. This functional alcoholic beverage at a concentration of 12.5 g L-1 significantly improved cortical spreading depression velocity in the rat brains (2.89 mm min-1), when compared with cachaça alone (3.68 mm min-1) and control (distilled water; 3.25 mm min-1). Moreover, animals that consumed the functional beverage gained less weight than those that consumed just alcohol and the control groups. These findings suggest that the C. vulgaris functional alcoholic beverage plays a protective physiologic role in protecting brain cells from the effects of drinking ethanol.

Neonatal pyridoxine administration long lastingly accelerates cortical spreading depression in male rats, without affecting anxiety-like behavior.

Objectives: Pyridoxine plays a key role in the development of the human nervous system. Several reports suggest that administration of high doses of pyridoxine can be helpful in improving disturbances such as anxiety and pyridoxine-dependent epilepsy, although it has also been associated with a proconvulsive action. In this study, we investigated in developing rats the effects of repeated administration of various doses of pyridoxine on anxiety-like behavior and the brain excitability-related phenomenon known as cortical spreading depression (CSD).Methods: From postnatal day (P) 7 to P27, Wistar rat pups received per gavage pyridoxine hydrochloride (1 mg/kg/day, or 5 mg/kg/day, or 10 mg/kg/day). On P60-70, the animals were tested in the elevated plus maze (EPM) to evaluate anxiety-like behavior. On P71-80, we recorded the CSD (4-hour recording session).Results: Compared with naïve (gavage-free) and saline-treated controls, pyridoxine-treated groups displayed a significant (p < 0.001) increase in CSD propagation velocity and amplitude of the CSD negative direct-current (DC)-shift, and a decrease in the CSD DC-shift duration. These effects were long-lasting and dose-dependent. In the EPM, no significant pyridoxine-associated effect was observed.Discussion: Our data demonstrate a novel action of pyridoxine on an electrical activity-related phenomenon (CSD) in the developing brain, confirming the hypothesis that the chronic treatment with pyridoxine early in life modulates CSD. Data on CSD propagation suggest that pyridoxine at a high dose might act as a prooxidant agent in the developing brain, a hypothesis that deserves further testing.

Anxiety behavior is reduced, and physical growth is improved in the progeny of rat dams that consumed lipids from goat milk: an elevated plus maze analysis.

The goat milk contains conjugated linoleic acid (CLA), which can influence physical growth and brain development. This study investigated the impact of a diet containing goat milk fat (GMF) on physical parameters of gestating (G) and/or lactating (L) rat dams, and their progeny's physical growth, and anxiety behavior. In the dams, body weight was evaluated during gestation and lactation. Maternal physical parameters, thoracic and abdominal circumference and liver weight were measured at weaning. In the progeny, indicators of somatic development, and consolidation of reflex responses (palm grasp, righting, free-fall righting, vibrissa placing, auditory startle response, negative geotaxis and cliff avoidance) were determined. Anxiety behavior was tested on the elevated plus maze (EPM). Compared to the controls, GMF-pups presented higher body weight and tail length at days 18 and 21 (groups G+L and L). In the L-group, cliff avoidance and free-fall righting responses were respectively delayed, and accelerated. Fur appearance was anticipated in G+L pups. On postnatal day 35, the EPM responses of the G group indicated less anxiety than in the controls. Data show developmental and behavioral modifications in the progeny of dams fed the GMF-rich diet consumed during gestation and lactation, suggesting the involvement of CLA in such effects.

Differential vulnerability of substantia nigra and corpus striatum to oxidative insult induced by reduced dietary levels of essential fatty acids.

Oxidative stress (OS) has been implicated in the etiology of certain neurodegenerative disorders. Some of these disorders have been associated with unbalanced levels of essential fatty acids (EFA). The response of certain brain regions to OS, however, is not uniform and a selective vulnerability or resilience can occur. In our previous study on rat brains, we observed that a two-generation EFA dietary restriction reduced the number and size of dopaminergic neurons in the substantia nigra (SN) rostro-dorso-medial. To understand whether OS contributes to this effect, we assessed the status of lipid peroxidation (LP) and anti-oxidant markers in both SN and corpus striatum (CS) of rats submitted to this dietary treatment for one (F1) or two (F2) generations. Wistar rats were raised from conception on control or experimental diets containing adequate or reduced levels of linoleic and α-linolenic fatty acids, respectively. LP was measured using the thiobarbituric acid reaction method (TBARS) and the total superoxide dismutase (t-SOD) and catalase (CAT) enzymatic activities were assessed. The experimental diet significantly reduced the docosahexaenoic acid (DHA) levels of SN phospholipids in the F1 (~28%) and F2 (~50%) groups. In F1 adult animals of the experimental group there was no LP in both SN and CS. Consistently, there was a significant increase in the t-SOD activity (p < 0.01) in both regions. In EF2 young animals, degeneration in dopaminergic and non-dopaminergic neurons and a significant increase in LP (p < 0.01) and decrease in the CAT activity (p < 0.001) were detected in the SN, while no inter-group difference was found for these parameters in the CS. Conversely, a significant increase in t-SOD activity (p < 0.05) was detected in the CS of the experimental group compared to the control. The results show that unbalanced EFA dietary levels reduce the redox balance in the SN and reveal mechanisms of resilience in the CS under this stressful condition.

Conjugated linoleic acid in the maternal diet differentially enhances growth and cortical spreading depression in the rat progeny.

BACKGROUND: Conjugated linoleic acids (CLA) are fatty acids that are found in the lipids from goat milk, and appear to protect neurons from excitotoxicity. METHODS: We investigated in developing rats the effects of a maternal CLA-rich diet (containing 7% lipids from goat milk) on body development and cerebral electrical activity of the progeny from dams receiving the CLA diet during gestation (G), lactation (L) or both periods (G+L). RESULTS: Compared to a control group (C) receiving a diet with 7% soybean oil, body weight increased at 14, 21 and 28 days, but not at 35-45 days, in L and G+L groups (P<0.05). No intergroup difference was found on body and brain weights, body length, abdominal and thoracic circumferences, body mass index and abdominal to thoracic circumference ratio at 35-45 days. In contrast, at this later age the CSD velocities of propagation were significantly higher (P<0.05) in L as compared with the C and G group, and in the L+G, as compared with the C, G and L groups, suggesting a long-lasting brain effect. CONCLUSION: These data indicate that a maternal CLA-rich diet can differentially influence body weight increment (short-term effect), and CSD propagation (long-term effect) in the progeny, and the lactation is the most critical period for such diet actions. GENERAL SIGNIFICANCE: The facilitating effect of the lipids from goat milk on an excitability-related phenomenon in the brain (CSD) can be of clinical relevance, since CSD has been associated to neurological disturbances like migraine and epilepsy.

The impairing effect of acute ethanol on spreading depression is antagonized by astaxanthin in rats of 2 young-adult ages.

BACKGROUND: Ethanol (EtOH) abuse and insufficient ingestion of antioxidants are external factors that can alter brain electrophysiology. Our previous studies have demonstrated that the excitability-related brain electrophysiological phenomenon known as cortical spreading depression (CSD) was facilitated by chronic EtOH intake, and chronic treatment with carotenoids attenuated this effect. Here, we investigated the acute effect of a single EtOH administration on CSD in young and adult rats previously (1 hour) treated with 10 µg/kg of astaxanthin. METHODS: Male Wistar rats (5 young- and 5 adult groups, 60 to 80 and 150 to 180 days of age, respectively) were treated by 2 gavage procedures at 1-hour interval as follows: groups 1 and 2 received astaxanthin in gavage I combined with EtOH (group 1) or water (group 2) in gavage II; groups 3 and 4 received olive oil (the vehicle in which astaxanthin was dissolved) in gavage I combined with EtOH (group 3) or water (group 4) in gavage II; group 5 received water in gavage I combined with EtOH in gavage II. CSD was recorded on the cortical surface for 4 hours. RESULTS: Compared to the respective water and oil controls (groups 2 and 4; CSD velocities: 3.73 ± 0.09 and 3.78 ± 0.07 mm/min in the young groups; 2.99 ± 0.10 and 3.05 ± 0.19 mm/min in the adult groups), a single dose of EtOH (groups 3 and 5) decreased CSD propagation velocities (3.29 ± 0.23 and 3.16 ± 0.10 mm/min in the young groups; 2.71 ± 0.27 and 2.75 ± 0.31 mm/min in the adult groups). Astaxanthin antagonized the impairing effect of acute EtOH on CSD (group 1; mean velocity: 3.70 ± 0.19 and 3.13 ± 0.16 mm/min for the young and adult groups, respectively). CONCLUSIONS: The results showed an antagonistic effect of acute EtOH treatment on CSD propagation that was reverted by astaxanthin. The EtOH-astaxanthin interaction was not influenced by the age, as it was found in both young and adult animals.

Chronic treatment with ascorbic acid enhances cortical spreading depression in developing well-nourished and malnourished rats.

Ascorbic acid (AA) is an antioxidant molecule that is highly concentrated in the brain and can exert both anticonvulsant and proconvulsant effects in distinct models of experimental seizures. Herein, we investigated whether chronic AA administration alters cortical excitability as indexed by the cortical spreading depression (CSD). Well-nourished (W) and malnourished (M) rats were treated, by gavage, with 60mg/kg/day of l-AA from postnatal days 7-28, and CSD propagation was analyzed at 30-40 days. Compared to the W groups, M rats presented higher (p<0.05) CSD amplitudes and velocities of propagation. In both nutritional conditions, AA-treatment significantly increased CSD amplitudes and propagation velocities (p<0.05), as compared to non-treated ('naïve'; Nv) and saline-treated (Sal) controls. The mean±standard deviation CSD velocities of propagation (in mm/min) for the Sal, AA and Nv groups were respectively 3.75±0.03, 4.26±0.08 and 3.81±0.04 for the W condition and 4.29±0.08, 4.51±0.04 and 4.30±0.04 for the M groups. The results demonstrate a CSD-facilitation by AA regardless of nutritional status. They also suggest that, at the dose of 60mg/kg/day chronically administered during brain development, AA may act as a prooxidant in brain, in view of the contrasting effect as compared with other antioxidants, which reduce CSD.

Propagation of spreading depression inversely correlates with cortical myelin content.

OBJECTIVE: Cortical myelin can be severely affected in patients with demyelinating disorders of the central nervous system. However, the functional implication of cortical demyelination remains elusive. In this study, we investigated whether cortical myelin influences cortical spreading depression (CSD). METHODS: CSD measurements were performed in rodent models of toxic and autoimmune induced cortical demyelination, in neuregulin-1 type I transgenic mice displaying cortical hypermyelination, and in glial fibrillary acidic protein-transgenic mice exhibiting pronounced astrogliosis. RESULTS: Cortical demyelination, but not astrogliosis or inflammation per se, was associated with accelerated CSD. In contrast, hypermyelinated neuregulin-1 type I transgenic mice displayed a decelerated CSD propagation. INTERPRETATION: Cortical myelin may be crucially involved in the stabilization and buffering of extracellular ion content that is decisive for CSD propagation velocity and cortical excitability, respectively. Our data thus indicate that cortical involvement in human demyelinating diseases may lead to relevant alterations of cortical function.

Effects of transcranial direct current stimulation coupled with repetitive electrical stimulation on cortical spreading depression.

We have recently shown that two techniques of brain stimulation - repetitive electrical stimulation (ES) (that mimics transcranial magnetic stimulation) and transcranial direct current stimulation (tDCS) - modify the velocity of cortical spreading depression (CSD) significantly. Herein we aimed to study the effects of these two techniques combined on CSD. Thirty-two Wistar rats were divided into four groups according to the treatment: sham tDCS/sham ES, sham tDCS/1 Hz ES, anodal tDCS/1 Hz ES, cathodal tDCS/1 Hz ES. Our findings show that 1 Hz ES reduced CSD velocity, and this effect was modified by either anodal or cathodal tDCS. Anodal tDCS induced larger effects than cathodal tDCS. Hereby CSD velocity was actually increased significantly after anodal tDCS/1 Hz ES. Our results show that combining two techniques of brain stimulation can modify significantly the effects of ES alone on cortical excitability as measured by the neurophysiological parameter of cortical spreading depression and therefore provide important insights into the effects of this new approach of brain stimulation on cortical activity.

After-effects of transcranial direct current stimulation (tDCS) on cortical spreading depression.

Abnormal cortical excitability influences susceptibility to cortical spreading depression (CSD) in migraine. Because transcranial direct current stimulation (tDCS) is capable of inducing lasting changes of cortical excitability, we investigated the after-effects of tDCS on the propagation velocity of CSD in the rat. Twenty-five anesthetised rats received either anodal, cathodal or sham tDCS. The stimulation was applied for 20 min at a current strength of 200 microA after the recording of three baseline CSD measurements. Starting 5 min after tDCS, a further three CSDs were elicited and CSD velocity recorded at intervals of 20 min. tDCS and CSD recording was performed under anaesthesia with chloralose and urethane. As compared to the baseline velocity of 3.14 mm/min, anodal tDCS induced a significant increase of propagation velocity during the first post-tDCS recording (3.49 mm/min). In contrast to anodal tDCS, neither cathodal tDCS nor sham tDCS, which consisted of an initial ramped DC stimulation lasting only 20 s, showed a significant effect on CSD propagation velocity. As anodal tDCS is known to induce a lasting increase of cortical excitability in the clinical setting, our results support the notion that CSD propagation velocity reflects cortical excitability. Since cortical excitability and susceptibility to CSD is elevated in migraine patients, anodal tDCS - by increasing cortical excitability - might increase the probability of migraine attack in these patients, even beyond the end of its application.

L-arginine treatment early in life influences NADPH-diaphorase neurons in visual cortex of normal and early-malnourished adult rats.

This study investigated the effects of repeated l-arginine administration during lactation, combined with different suckling conditions, on morphometric parameters of primary visual cortex NADPH-diaphorase-positive neurons. Wistar rat pups reared in "normal-size litters" or "large litters" (N- and L-conditions; litters formed by 6 and 12 pups, respectively) received, from postnatal day 7 to 28, either arginine (300 mg/kg/day, per gavage) or distilled water (control). At 90-120 days of life, they were perfused with saline + formaldehyde, and their brains were processed for histochemical reaction to reveal NADPH-diaphorase-positive neurons (malic enzyme indirect method). Compared to the normal-size litters, L-rats had lower body weights (P < 0.05), confirming the effectiveness of the L-condition in affecting pup development. Concerning NADPH-d histochemistry, arginine treatment was associated with increased (P < 0.05) density of dendrite varicosities and of dendrite branching frequency, suggesting a plastic response of the developing brain to that treatment, even in previously malnourished rats. No difference was seen, however, in dendrite orientation, total number of neurons, soma area and perimeter, as well as dendrite bifurcation points, fractal dimension, and area and volume of dendrite field, suggesting that NADPH-d cells are resistant to arginine and nutritional changes, regarding these features. Data are considered of interest for studies of synaptic plasticity during neural development and its relationships to aggressive agents like malnutrition.

Lasting accelerative effects of 1 Hz and 20 Hz electrical stimulation on cortical spreading depression: relevance for clinical applications of brain stimulation.

Clinical applications of brain stimulation have been increasing during the last decade; however, the mechanisms of action remain unknown. One proposed mechanism of action is that repetitive stimulation modulates cortical excitability. Herein, we explore the question of whether repetitive electric stimulation increases cortical excitability as indexed by the cortical spreading depression. Twenty-four Wistar rats were divided into three groups according to the treatment: sham, 1-Hz and 20-Hz stimulation. Stimulation was applied to the left frontal cortex through a pair of epidurally implanted silver-wire electrodes. The cortical spreading depression-features were analysed at three time points (one day before, one day after and 2 weeks after treatment) in both the stimulated and unstimulated hemisphere. A 3 x 2 x 3 factorial anova with repeated measures showed significant differences in the main effect of time (P < 0.0001), hemisphere (P = 0.0002) and stimulation group (P = 0.008). The interaction between time vs. hemisphere vs. stimulation group was also significant (P < 0.0001). Posthoc analysis demonstrated that 1-Hz and 20-Hz repetitive electrical stimulation significantly increased the velocity of cortical spreading depression in the stimulated hemisphere. Furthermore, 20-Hz stimulation showed a greater effect on cortical spreading depression compared to 1-Hz stimulation. The results show that 1-Hz and 20-Hz repetitive electrical stimulation results in an increase in cortical spreading depression velocity that is associated with the frequency and the hemisphere of stimulation. Furthermore, the effects are found to be long lasting. We believe that these findings have strong relevance to support the clinical application of therapies involving electrical stimulation for diseases of reduced cortical excitability.

Application of potassium chloride to the surface of the rat visual cortex differentially affects the expression of presumptive neuroprotective molecules.

The purpose of this study was to investigate the effects of topical application of 3 M KCl for 20 minutes to the surface of the rat visual cortex upon the expression of presumptive neuroprotective molecules. Conventional immunohistochemical and immunoblotting techniques revealed a marked reduction of the expression of calbindin and parvalbumin after application of KCl, whereas calretinin, nitric oxide synthase, and the alpha7 subunit of the nicotinic acetylcholine receptor clearly increased following that procedure. Such effects were quantified both in terms of the number of immunoreactive perikarya and optical density of immunoblottings, and were observed as soon as 1 h, and to last at least until 15 days, following KCl application. These results suggest that calbindin and parvalbumin may not be able to exert an appreciable neuroprotective effect in the presence of KCl. On the other hand, calretinin, nitric oxide synthase, and the alpha7-containing nicotinic receptors are possibly upregulated to protect the neurons in which they are expressed.

Malnutrition and brain function: experimental studies using the phenomenon of cortical spreading depression

Depending on its intensity and duration, nutritional deficiency can disrupt the structure and function of the nervous system of humans and other mammals, with consequences more or less devastating for the whole organism, particularly in the early postnatal life, when body growth is very rapid and the need for proteins, calories and other nutrients is greatest. In this review, electrophysiological data are presented regarding the use of the phenomenon of cortical spreading depression (CSD) to study effects of malnutrition on the brain. Several conditions of clinical importance and that are known to alter brain function are shown also to influence CSD features in experimental animals. Some of these conditions, (e.g., phamacological manipulation of neurotransmitter systems, dietary treatment with Lithium, acute hyperglycemia, hypothyroidism, aging and environmental stimulation) decrease CSD susceptibility, while other conditions increase it, as, for example, systemic reduction of extracellular chloride levels, deprivation of REM-sleep, acute hypoglycemia, treatment with diazepam, consumption of ethanol and malnutrition. Particular emphasis is laid on the effect of early environmental enrichment on CSD in normal and malnourished animals. Our results suggest that such effect is more evident in the malnourished brain, as compared to the wellnourished one. The data also show that malnutrition alters the brain responsivity to some CSD-facilitatory or inhibitory agents. The underlying mechanisms to explain the observed effects are discussed.