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Spatial memory is responsible for encoding spatial information to form a path, storing this mental representation, and evaluating and recovering spatial configurations to find a target location in the environment. It is mainly supported by the hippocampus and its interaction with other structures, such as the prefrontal cortex, and emerges in rodents around postnatal day (PND) 20. Sex differences in spatial tasks have been found in adults, with a supposedly better performance in males. However, few studies have examined sex differences in orientation throughout postnatal development. This study aimed to analyse the performance of juvenile (PND 23) male (n = 18) and female (n = 21) Wistar rats in a spatial reference memory task in the Morris water maze (MWM) with two different training regimes in the acquisition phase, and their subjacent metabolic brain activity. Based on sex, subjects were assigned to two different groups: one that performed four learning trials per day (n = 9 males and n = 8 females) and the other that was submitted to two trials per day (n = 9 males and n = 13 females). After the behavioural protocols, metabolic activity was evaluated using cytochrome c oxidase histochemistry. Results showed no metabolic brain or behavioural differences in the four-trial protocol performance, in which both sexes reached the learning criterion on the fourth day. By contrast, the two-trial protocol revealed an advantage for females, who reached the learning criterion on day four, whereas males needed more training and succeeded on day six. The female group showed lower metabolic activity than the male group in the cingulate and prelimbic cortex. These results suggest a faster consolidation process in the female group than the male group. Further research is needed to understand sex differences in spatial memory at early stages., Competing Interests: Declaration of Competing Interest The authors declare that they have no conflict of interest to disclose., (Copyright © 2023. Published by Elsevier Inc.)

Background: Although there has developed an increased interest in the vaccines BNT1622b2 (Pfizer/BioNTech), mRNA-1273 (Moderna/NIAID), and ChAdOx1 nCoV-19 (AstraZeneca/University of Oxford), there are still few reports describing the immune response induced by different vaccine platforms in real-world settings of low-income countries. Here, we proposed to analyse the humoral immune response elicited by the primary vaccines used in Argentina from July-December 2021., Methods: Anti-SARS-CoV-2-Spike-RBD IgG and neutralising antibodies were assayed by ELISA in a total of 871 serum samples obtained from 376 volunteers from an educational staff. The individuals were vaccinated with BBIBP-CorV (Sinopharm), ChAdOx1 nCoV-19 (AstraZeneca/University of Oxford, AZ), Gam-COVID-Vac (Sputnik V, SpV) or combined vaccines (mostly SpV and mRNA-1273, Moderna). The antibody response was analysed several days after the initial vaccination (20, 40, 120 and 180 days)., Results: After receiving at least one dose of the COVID-19 vaccine, we detected 93.34% of seroprevalence. Previously SARS-CoV-2 infected showed higher antibody concentrations compared with naïve vaccinees. Six months after the initial vaccination, combined vaccination induced higher anti-SARS-CoV-2 antibody levels than the other vaccines in naïve volunteers. However, we did not find differences in the neutralising responses after any vaccine from naïve vaccines or between the naïve and previously infected volunteers on day 120 after vaccination., Conclusions: Our long-term analysis of volunteers from the educational system provides data in a real-world context, showing the benefits of a boost dose still in previously infected volunteers, and suggesting the advantages of a heterologous prime-boost schedule., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

The retrieval of spatial memories does not always occur in an environment with the same stimuli configuration where the memory was first formed. However, re-exposure to a partial portion of the previously encountered cues can elicit memory successfully. Navigation with contextual changes has received little attention, especially in females. Thus, we aimed to assess memory retrieval using the Morris Water Maze spatial reference protocol in female adult Wistar rats. Rats were trained with five allocentric cues, and retrieval was explored one week later either with the same cues, or with four removed, or with three added cues. We studied the underlying brain oxidative metabolism of the hippocampus, prefrontal, parietal, retrosplenial, entorhinal, and perirhinal cortices through cytochrome c oxidase (CCO) histochemistry. Neither cue removal nor cue addition impaired retrieval performance. Retrieval with a degraded subset of cues led to increased prefrontal, hippocampal, retrosplenial, parietal, and perirhinal CCO activity. Retrieval with extra cues led to an enhancement of CCO activity in the hippocampus and retrosplenial cortex. Different patterns of network intercorrelations were found. The cue-removal group presented a closed reciprocal network, while the group with extra cues had separate parallel networks. Both groups showed a simpler network than the group with no cue modifications. Future research is needed to delve into behavioral and brain-related functions of spatial memory processes under modified environmental conditions., Competing Interests: Declaration of interest All the authors declare that there are no actual or potential conflicts of interest, including any financial, personal, or other relationships with other people or organizations that could inappropriately influence this work. The work described has not been submitted for publication elsewhere, in its entirety or in part, and all the authors listed have approved the enclosed manuscript., (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)

Memory involves a complex network system of interconnected brain areas in which labile trace memories are transformed into enduring ones and reorganized in a time-dependant manner. Although it has been observed that remote memories are less prone to destabilizing, they can become fragile and lead to behavioural decline. We explored the behavioural outcomes of male and female rats in response to the reactivation of a previously acquired allocentric spatial reference memory, under conditions in which animals have shown a retrieval decay. In addition, we assessed their brain metabolic activity through cytochrome c oxidase (CCO) histochemistry. Our results show that a spatial memory amnesia-like behaviour with a time interval of 45 days can be recovered after re-exposure to the environmental configuration with the reinforced contingencies. Moreover, we observed that, following reactivation, male rats reveal a decrease in metabolic activity in septal nuclei and thalamic structures, whereas female rats add a metabolic reduction in the hippocampus, amygdala, mPFC, and retrosplenial, parietal and rhinal cortices, suggesting that they efficiently employ these brain areas when reactivation a memory that has suffered a decay with time. Finally, although male and female rats perform the behavioural task equally, we found sex differences at the brain metabolism level, revealing the differential contribution of brain limbic system energy demands by sex, even when their performance is similar. In conclusion, our work provides behavioural and brain data about remote spatial retrieval and memory reactivation processes., (Copyright © 2021 IBRO. Published by Elsevier Ltd. All rights reserved.)

Photobiomodulation (PBM) or the use of red to near-infrared irradiation spectrum, is a non-invasive intervention that produces neurostimulatory effects and reaches benefits in several pathologies as well as in healthy subjects. The main objective of this study was to evaluate and compare the effects of PBM in a rat brain network on basal state and functional activity during the execution of a reversal task. Twenty-eight rats were divided into four groups: control group (n = 7), control photobiomodulation group (n = 8), behavioural group (n = 6) and behavioural photobiomodulation group (n = 7). Reversal memory was assessed using a Morris water maze and cytochrome c oxidase (CCO) was used as a brain metabolic activity marker. After five days of PBM, the control photobiomodulation group showed a decrease of CCO levels in the striatum, medial septum, entorhinal, hippocampus, amygdala, thalamus, mammillary nuclei and VTA. Both behavioural groups performed the task correctly, however, the behavioural photobiomodulation group displayed CCO reduction in some regions involved in the execution of the reversal task: septum, entorhinal, CA1, CA3, central amygdala and supramammilar, along with higher levels in accumbens. These results could show the effect of PBM on active brain networks. Further studies will be necessary to elucidate its effects in different brain networks that are involved in the execution of other memory tasks., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Repetitive transcranial magnetic stimulation (rTMS) is a non-invasive neuromodulation technique capable of producing changes in the electrical potential of neurons. Currently, the application of rTMS in clinical practice and as a neurophysiological tool is increasing. However, the exact cellular mechanisms underlying rTMS-based therapies are not completely clear. Additionally, glial cells have been studied less. Our aim was to investigate the effect of three days of high-frequency rTMS on neuronal metabolism and neuronal activation, in addition to its effect on glial cells. For this purpose, we performed histochemistry and immunohistochemistry procedures: the histochemistry of cytochrome oxidase (COx) to assess neuronal metabolic activity, and the immunohistochemistry of c-Fos (marker of neuronal activity), GFAP (marker of astrocytic reactivity), and Iba1 (selective marker of reactive microglia). Our results showed enhanced metabolic activity after rTMS in the retrosplenial and parietal cortex and CA1 and CA3 subfields of the hippocampus. Moreover, higher c-Fos activity was found in the agranular retrosplenial cortex. Finally, we did not find changes between groups in the induction of astrocyte and microglia reactivity in any of the immunostained regions. In conclusion, we can assume that three days of high-frequency rTMS applied in healthy rats does not alter astroglia reactivity or inflammatory responses, such as microglia proliferation. Because we have shown an upregulation of neuronal metabolic activity in many limbic brain structures, in addition to higher c-Fos levels in the nearest cortical area to the rTMS, our work provides novel insight into the effectiveness and safety of rTMS as a brain modulation therapy., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Early life stress comprises a wide range of adverse events that can occur in the subject's early developmental stages: from child abuse to rodent repeated maternal separation models. This study used young adult male Wistar rats that were maternally raised (AFR), maternally separated from post-natal day (PND) 1 to PND10 (MS10), or maternally separated from PND1 to PND21 (MS21), to assess the effects of maternal separation on spatial learning and memory using the Morris Water Maze (MWM). We also performed quantitative cytochrome oxidase (COx) histochemistry on some selected brain areas in order to find out whether maternal separation affects brain energy metabolism. We obtained a similar spatial learning pattern in maternally raised and maternally separated subjects, with MS10 showing faster acquisition; however, different brain networks with different energy consumptions were used by each group. MS10 spent more energy to get the same behavioral outputs, whereas MS21 resembled AFR more in term of energy demands., (Copyright © 2017 Elsevier Inc. All rights reserved.)

Background & Aims: Minimal hepatic encephalopathy (mHE) has been shown to affect daily functioning, quality of life, driving and overall mortality. However, little is known about treating or diagnosing early impairments in mHE. We studied one of its precipitating factors, portal hypertension which is driving the inflammatory process behind mHE. The purpose was to describe an indirect diagnostic method able to detect the pathology at early stages based on the study of the vascularization and mast cells conjunctival hyperplasia as secondary inflammatory response associated to portal hypertension. Finally, we correlated the presence of histological changes in the eye in mHE with deficits in behavioral task acquisition., Methods: Rats were trained on a stimulus-response task and a spatial working memory task using the Morris water maze. Two groups of animals were used: a SHAM (sham-operated) group (n=10) and a portal hypertension (HT) group (n=10). The triple portal vein ligation method was used to create an animal model of mHE. Latencies to reach the platform, number of glial fibrillary acidic protein-immunoreactive astrocytes (GFAP-IR), mast cell expression and presence/absence of blood and lymphatic vessels were examined., Results: There were differences in stimulus-response behavioral performance, with a deficit in the acquisition in the HT group. However, no differences between groups were found on the spatial working memory task. At the same time, differences between groups were found in the GFAP-IR density, which was lower in the HT group, and in the number of mast cells and the presence of vessels, which were higher in the HT group., Conclusions: In this study, we provide the first preliminary insight into the validity of exploring the eye as a possible tool to assess the diagnosis of mHE conditions., (Copyright © 2017 Elsevier Inc. All rights reserved.)

Hepatic encephalopathy (HE) is a neurological complication observed in patients with liver disease. Patients who suffer from HE present neuropsychiatric, neuromuscular and behavioral symptoms. Animal models proposed to study HE resulting from cirrhosis mimic the clinical characteristics of cirrhosis and portal hypertension, and require the administration of hepatotoxins such as thioacetamide (TAA). The aim of this study was to assess the effects of a high protein diet on motor function, anxiety and memory processes in a model of cirrhosis induced by TAA administration. In addition, we used cytochrome c-oxidase (COx) histochemistry to assess the metabolic activity of the limbic system regions. Male rats were distributed into groups: control, animals with cirrhosis, Control rats receiving a high protein diet, and animals with cirrhosis receiving a high protein diet. Results showed preserved motor function and normal anxiety levels in all the groups. The animals with cirrhosis showed an impairment in active avoidance behavior and spatial memory, regardless of the diet they received. However, the animals with cirrhosis and a high protein diet showed longer escape latencies on the spatial memory task. The model of cirrhosis presented an under-activation of the dentate gyrus and CA3 hippocampal subfields and the medial part of the medial mammillary nucleus. The results suggest that a high protein intake worsens spatial memory deficits shown by the TAA-induced model of cirrhosis. However, high protein ingestion has no influence on the COx hypoactivity associated with the model., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Although often not considered clinically relevant and, therefore, not diagnosed or treated, minimal hepatic encephalopathy (MHE) has been shown to affect daily functioning, quality of life, driving and overall mortality. To discover early impairments involved in MHE, we studied one of its precipitating factors, portal hypertension. Rats were trained on a stimulus-response task using the Morris water maze. Two groups of animals were used: a SHAM (sham-operated) group (n= 13) and a portal hypertension (PH) group (n= 13). The triple portal vein ligation method was used to create an animal model of an early developmental phase of HE. Brain metabolic activity was studied with cytochrome c-oxidase histochemistry (C.O.). Neuronal nuclear volume was assessed by nucleator probe; the number of glial fibrillary acidic protein-immunoreactive astrocytes (GFAP-IR) and proinflammatory mediators was measured. The results revealed that the PH group was not able to reach the behavioural criterion, in contrast to the SHAM group. The metabolic brain consumption revealed decreased C.O. activity in the ventral striatum. The PH group showed lower density of GFAP-IR and an increase in the tumour necrotic factor-α (TNF-α). The PH group showed decreased neuronal nuclear volume in the dorsal striatum. On the contrary, increased neuronal nuclear volume was found in the ventral striatum. For the first time, a relationship has been established between inflammation, astrocytic and neural damage, and brain metabolic impairment in a model of MHE. Disruption of the striatum and related structures was highlighted as the main target in early stages of HE. Finally, a simple task was presented to assess the subtle impairments found in the clinic, which could provide fresh insights into the development of new tools for the assessment of MHE.

In rodents, many studies have been carried out using novelty-preference paradigms. The results show that the perirhinal cortex and the hippocampus are involved in the recognition of a novel object, "what", and its new position, "where", respectively. We employed these two variants of a novelty-preference paradigm to assess whether the expression of the immediate-early gene c-fos in the dorsal hippocampus and perirhinal cortex correlates with the performance discrimination ratio (d2), on the respective versions of the novelty preference tests. A control group (CO) was added to explore c-fos activation not specific to recognition. The results showed different patterns of c-Fos protein expression in the hippocampus and perirhinal cortex. The Where Group presented more c-Fos positive nuclei than the What and CO groups in the CA1 and CA3 regions, whereas in the perirhinal cortex, the What Group showed more c-Fos positive nuclei than the Where and CO groups. The correlation results indicate that levels of c-Fos in the CA1 area and perirhinal cortex correlate with effective exploration, d2, on the respective versions of the novelty preference tests, novel place and novel object recognition. These data suggest that the hippocampal CA1 and perirhinal cortex are specifically related to the level of recognition of place and objects, respectively., (Copyright © 2015 Elsevier Inc. All rights reserved.)

The hippocampus has been established as a site of plasticity during the acquisition of spatial memory. The memory for spatial locations is impaired in patients who develop hepatic encephalopathy (HE). We wondered how the hippocampus can manage different hippocampal-dependent tasks in a type B model of the early evolutive phases of HE induced by triple portal vein ligation. We used a one-trial object-place recognition task that involves making judgements about whether a stimulus was encountered before in that location and a reversal learning task performed in the Morris water maze that involves reward-guided behavior and decision making. Our behavioral results showed impairments in the acquisition of both tasks by the portal hypertension group compared with the sham-operated group. To label brain areas related to these tasks, we marked the expression of the c-Fos protein and revealed high c-Fos immunoreactivity in cornu ammonis 1 (CA1), cornu ammonis 3 (CA3) and entorhinal (Ent) cortex of the PH group compared with the SHAM group in the object-place recognition task and a decrease in c-Fos-positive cells in the reversal task in the CA1, CA3, dentate gyrus (DG), cingulate (CG), prelimbic (PL), and infralimbic (IL) cortices in the PH group compared with the SHAM group. In conclusion, the study corroborated the pivotal role of the hippocampus in spatial memory deficits found in the early stages of type B HE and noted its differential contribution in each of the tasks., (Copyright © 2014 IBRO. Published by Elsevier Ltd. All rights reserved.)

We have studied the performance of a spatial reference memory task, the navigation strategy and the changes in the cytochrome c oxidase activity (COx) in different brain regions in exercised (forced exercise, 10 consecutive days, 15min/day) and non-exercised adult Wistar rats. The spatial learning task was carried out in the radial-arm water maze (RAWM) for four days with six daily trials, and on the fifth day, a probe session was run, in which we rotated the position of the distal cues 90° in a clockwise direction. During the four days of training, the exercised group showed shorter latency and distance traveled to find the platform, as well as fewer memory errors and reduced use of non-appropriate navigation strategies according to the protocol of the task (egocentric). Interestingly, the rotation of the cues did not affect the performance of the exercised group, in contrast to the non-exercised group, which spent more time in the center of the maze and traveled longer distance to find the platform. Finally, higher COx activity in the cingulate and the retrosplenial cortices, as well as in the dorsal CA1 and CA3 was found in the exercised group. All in all, it seems that the exercise favored the configuration of an efficient and accurate cognitive map of the environment, which was supported by our finding that the rotation of the cues, without altering their overall configuration, did not affect performance. The brain regions with higher COx activity in the exercised group seem to be involved in this function., (Copyright © 2014 IBRO. Published by Elsevier Ltd. All rights reserved.)

Our aim was to assess the changes in the Synapsin I and glucocorticoid receptor (GR) expression induced by behavioral testing in the dorsal and ventral hippocampi of standard and enriched aged Wistar rats. The environmental enrichment (EE) was carried out 3h/day over a period of two months and then, the rats were tested in the elevated zero-maze (EZM) and radial-arm water maze (RAWM). Behavioral results showed that, even at an advanced age, EE was able to reduce anxiety-related behaviors and improve the performance in the RAWM. Regarding the neurobiological data, Synapsin I expression in the dorsal CA3, but not in the ventral, was enhanced both in enriched and standard rats when they performed the behavioral testing. Interestingly, the EE exposure was enough to increase Synapsin I in the ventral CA3. The analysis of GR in the dorsal hippocampus showed an increase of this receptor in the dDG both in enriched and standard rats when they performed the behavioral testing, whereas in the dCA1 and dCA3, the effect of the testing depended on the previous housing condition. In the ventral region, we found that the effects of EE were higher because on the one hand, the GR expression induced by the behavioral testing was enhanced in the dSUB, vCA1 and vCA3 when the rats were previously enriched and on the other hand, EE, regardless of the behavioral testing, increased the GR expression in the vDG and vSUB. Therefore, our results suggest that the effect of the behavioral testing on the neurobiological mechanisms studied is different depending on the previous housing condition of aged rats., (Copyright © 2014 Elsevier Inc. All rights reserved.)

Patients with minimal hepatic encephalopathy exhibit early impairments in their ability to shift attentional set. We employed a task-switching protocol to evaluate brain network changes. Strategy switching requires the modification of both the relevant stimulus dimension and the required memory system. Rats were trained in an allocentric (A) and a cue-guided (C) task using a four-arm maze. To examine priming, we changed the order in which the tasks were presented. Five groups of animals were used: a SHAM (sham-operated) A-C group (n=10), a SHAM C-A group (n=8), a PH (portal hypertension) A-C group (n=8), PH C-A group (n=8), and a naïve group (n=10). The triple portal vein ligation method was used to create an animal model of the early evolutive phase of PH. The animals were tested in the four-arm radial water maze in a single 10-trial session each day for six days (three days for the allocentric task and three days for the cue-guided task). The metabolic activities of the brains were studied with cytochrome oxidase histochemistry, and brain network changes were assessed with principal component analysis. The behavioural results revealed significant increases in the numbers of correct choices across training days in all groups studied, and facilitation of the acquisition of the second task was present in the C-A groups. Moreover, different brain network activities were found; in the experimental groups, the performance of A-C switch involved the prefrontal cortex, and the key structures involved in the C-A switch in the other groups were the dentate gyrus of the dorsal hippocampus and the basolateral and central amygdala. These networks have a common nucleus of structures (i.e., the parietal cortex and the dorsal and ventral striatum), whereas other structures were specifically involved in each type of strategy, suggesting that these regions are part of both circuits and may interact with one another during learning., (Copyright © 2014 Elsevier Inc. All rights reserved.)

Environmental enrichment (EE) produces a remarkable degree of structural and functional plasticity in the hippocampus and possible mediators of these changes, such as glucocorticoid receptors (GRs), are of considerable interest. GRs are richly expressed in the hippocampus and they are involved in the adaptation to stressors and facilitate active coping in anxious situations. In this study, we assessed the effect of an EE protocol (24h/day during 69days) in adult Wistar rats on the activity in the elevated-zero maze (EZM), performance in the holeboard task (HB) and we also examined the changes in the glucocorticoid receptors (GRs) expression in the dorsal hippocampus (CA1, CA3 and DG). Our EE protocol reduced anxious behaviors in the EZM, so the animals spent more time and made more entries into the open sections. In the HB task, the enriched group showed more explorative behavior, a reduction of anxiety-related behaviors and a better cognitive performance compared to non-enriched animals. With regard to the GR expression, the EE condition produced an increase in the number of immunopositive cells for GRs in CA1, CA3 and DG. These results suggest that the better performance of enriched animals could be mediated in part by the increase of GRs in the dorsal hippocampus, which may alter the hippocampal neuronal function and accordingly, the anxiety levels, the spatial memory performance and the exploration levels in these animals., (Copyright © 2014 Elsevier Inc. All rights reserved.)

The present investigation aimed to develop magnetoliposome nanoparticles loaded with 5-fluorouracil by following a reproducible thin film hydration technique. The physicochemical characterization (including electron microscopy analysis, dynamic light scattering, infrared spectrometry, X-ray diffractometry, electrophoresis, and surface thermodynamics) suggested that superparamagnetic magnetite nuclei were successfully embedded into a multilamellar lipid vesicle. Magnetic responsiveness of these nanocomposites was quantitatively analyzed by determining the hysteresis cycle and qualitatively confirmed by microscopic visualizations. A high frequency alternating electromagnetic field was further used to define their heating properties. The absence of cytotoxicity in human colon fibroblast CCD-18 and in human colon carcinoma T-84 cell lines and excellent hemocompatibility of these core/shell particles were demonstrated. Additionally, 5-fluorouracil incorporation was investigated by two procedures: (i) entrapment into the nanoparticulate matrix and (ii) surface deposition onto already formed magnetoliposome particles. The former method reported greater drug loading values and a sustained release profile. Interestingly, 5-fluorouracil release was also triggered by the heating properties of the nanoparticles (hyperthermia-triggered drug release). Hence, we put forward that magnetoliposome particles hold important properties, that is, magnetically targeted delivery, hyperthermia inducing capability, high 5-fluorouracil loading capability, and hyperthermia-triggered burst drug release, suggestive of their potential for a combined antitumor therapy against colon cancer., (Copyright © 2013 Elsevier B.V. All rights reserved.)

We assessed the effect of 3h of environmental enrichment (EE) exposure per day started at different ages (3 and 18months old) on the performance in a spatial memory task and on brain regions involved in the spatial learning (SPL) process using the principal component analysis (PCA). The animals were tested in the four-arm radial water maze (4-RAWM) for 4days, with six daily trials. We used cytochrome c oxidase (COx) histochemistry to determine the brain oxidative metabolic changes related to age, SPL and EE. Behavioural results showed that the enriched groups, regardless of their age, achieved better performance in the spatial task. Interestingly, in the case of the distance travelled in the 4-RAWM, the effect of the EE was dependent on the age, so the young enriched group travelled a shorter distance compared to the aged enriched group. Respect to COx histochemistry results, we found that different brain mechanisms are triggered in aged rats to solve the spatial task, compared to young rats. PCA revealed the same brain functional network in both age groups, but the contribution of the brain regions involved in this network was slightly different depending on the age of the rats. Thus, in the aged group, brain regions involved in anxiety-like behaviour, such as the amygdala or the bed nucleus of the stria terminalis had more relevance; whereas in the young enriched group the frontal and the hippocampal subregions had more contribution., (Copyright © 2013 IBRO. Published by Elsevier Ltd. All rights reserved.)

We have studied the effect of an environmental enrichment (EE) protocol in adult Wistar rats on the activity in the elevated zero-maze (EZM), performance in the radial-arm water maze (RAWM) and we have also examined the changes in the neuronal metabolic activity of several brain regions related to anxiety response and spatial memory through cytochrome c oxidase histochemistry (COx). Our EE protocol had anxiolytic effect in the EZM; the animals spent more time and made more entries into the open quadrants, they had lower latency to enter into the open quadrant and lower levels of defecation. Also, the EE group showed fewer working memory and reference memory errors, as well as lesser distance travelled in the first day of the spatial training. In relation to the neuronal metabolic activity, EE reduced the COx activity in brain regions related to anxiety response, such as the infralimbic cortex, the paraventricular thalamic and hypothalamic nucleus, the basolateral amygdala, and the ventral hippocampus. Interestingly, there were no significant differences between groups in the dorsal hippocampus, more related to spatial cognition. These results suggest a beneficial effect of EE on spatial memory as a result of reducing anxiety levels and the COx activity in brain regions involved in anxiety response. We also found a differential pattern of activation inside the hippocampus, suggesting that the dorsal hippocampus has a preferential involvement in spatial learning and memory, whereas the ventral hippocampus has a role in anxiety response., (Copyright © 2013 Elsevier Inc. All rights reserved.)

The involvement of different brain regions in place- and response-learning was examined using a water cross-maze. Rats were trained to find the goal from the initial arm by turning left at the choice point (egocentric strategy) or by using environmental cues (allocentric strategy). Although different strategies were required, the same maze and learning conditions were used. Using cytochrome oxidase histochemistry as a marker of cellular activity, the function of the 13 diverse cortical and subcortical regions was assessed in rats performing these two tasks. Our results show that allocentric learning depends on the recruitment of a large functional network, which includes the hippocampal CA3, dentate gyrus, medial mammillary nucleus and supramammillary nucleus. Along with the striatum, these last three structures are also related to egocentric spatial learning. The present study provides evidence for the contribution of these regions to spatial navigation and supports a possible functional interaction between the two memory systems, as their structural convergence may facilitate functional cooperation in the behaviours guided by more than one strategy. In summary, it can be argued that spatial learning is based on dynamic functional systems in which the interaction of brain regions is modulated by task requirements., (Copyright © 2012 IBRO. Published by Elsevier Ltd. All rights reserved.)

The specific brain regions and circuits involved in the acquisition and expression of contextual fear conditioning are still a matter of debate. To address this issue, regional changes in brain metabolic capacity were mapped during the acquisition and expression of contextual fear conditioning using cytochrome oxidase (CO) quantitative histochemistry. In comparison with a group briefly exposed to a conditioning chamber, rats that received a series of randomly presented footshocks in the same conditioning chamber (fear acquisition group) showed increased CO activity in anxiety-related brain regions like the ventral periaqueductal gray, the ventral hippocampus, the lateral habenula, the mammillary bodies, and the laterodorsal thalamic nucleus. Another group received randomly presented footshocks, and it was re-exposed to the same conditioning chamber one week later (fear expression group). The conditioned group had significantly higher CO activity as compared with the matched control group in the following brain regions: the ventral periaqueductal gray, the central and lateral nuclei of the amygdala, and the bed nucleus of the stria terminalis. In addition, analysis of functional brain networks using interregional CO activity correlations revealed different patterns of functional connectivity between fear acquisition and fear expression groups. In particular, a network comprising the ventral hippocampus and amygdala nuclei was found in the fear acquisition group, whereas a closed reciprocal dorsal hippocampal network was detected in the fear expression group. These results suggest that contextual fear acquisition and expression differ as regards to the brain networks involved, although they share common brain regions involved in fear, anxiety, and defensive behavior., (Copyright © 2011 IBRO. Published by Elsevier Ltd. All rights reserved.)

Portal hypertension is a major complication of cirrhosis that frequently leads to a neuropsychiatric disorder that affects cognition. We compared the performance of 18-month-old prehepatic portal hypertensive rats (PH) and 18-month-old normal rats (CO) in spatial short-term and reference memory tasks in the Morris water maze and in active avoidance task. The PH group showed worse spatial short-term memory than the CO group. Also, the PH group tended to perform worse than the CO group in the reference memory task, but it presented a correct acquisition of the active avoidance task. We assessed the brain metabolic activity of the animals by means of cytochrome c-oxidase (COx) histochemistry. We found that the PH group developed prefrontal dysfunction characterized by increased COx activity in this region compared to the CO group. Similar results were found in the medial mammillary nucleus and dentate gyrus, whereas the CA1 area, bed nucleus of the stria terminalis, and supramammillary nucleus showed lower COx activity in the PH group as compared to the CO group. We conclude that the 18-month-old portal hypertensive rats present spatial memory impairment without alteration of implicit learning. This deficit could be related to the alteration of the metabolic activity of the brain regions involved in the processing of spatial memories., (Copyright 2010 Elsevier Inc. All rights reserved.)

The sex differences in the functional contribution of brain substrates were explored following acquisition of a spatial working memory task using quantification of c-Fos protein. Rats of both sexes were trained during adolescence and adulthood in Morris water maze using a hidden escape platform with different daily location. Two control groups for each sex and age were added to explore the c-Fos activation not specific to the memory process. These were a free-swimming group (yoked control) and a handled control (CO) group. Behaviorally, no age differences were found in number of days required by males to acquire the task, but females showed a delay in acquisition during adolescence (P30) that improved in adulthood (P90). Both sexes showed a learning-related increase in Fos immunoreactivity in the anterodorsal and anteroventral thalamus and medial and lateral mammillary nuclei during adolescence. Higher levels of learning-related Fos immunoreactivity were found in the infralimbic cortex, CA3 and CA1 only in females. During adulthood the common activated region was the prelimbic cortex with the addition of the infralimbic cortex in the male group and the lateral mammillary nucleus in the female group. These results indicated sex and age differences in brain functioning following working memory task. However, they could not be necessarily linked with differences in performance since similar results were found between males and females during adulthood. The activation of common and interrelated structures suggests that these structures are involved in spatial processing but it also highlights the relevance of developmental changes for understanding the memory process.

Several works have shown that males and females differ in the ability to learn spatial locations in mazes. In this study, we used the Morris water maze to assess the acquisition of a spatial working memory (WM) task in adult male and female Wistar rats. The task consisted of a paired sample procedure made up of two daily identical trials, sample and retention. To study the oxidative metabolic activity of some brain limbic system regions after the WM task, we applied the cytochrome oxidase (COx) histochemistry. In addition to the experimental groups, free swimming control groups and untrained naïve groups were added to explore the COx changes not specific to the learning process. Similar spatial performances were found between sexes as only one more sample and retention trials were needed in males to reduce the escape latencies significantly. Males showed decreased COx activity as compared to control groups in the medial prefrontal cortex (prelimbic and infralimbic regions) as well as in the lateral septum and dentate gyrus. Regarding females, an increase in COx activity was found in nucleus accumbens, ventral tegmental area and supramammillary region in relation to control groups. Overall, these findings suggest that the acquisition of the spatial WM task is mediated by different subsystems in a sex-dependent manner that points to the hippocampus as the central structure in males whereas other structures would be central in females.

Hepatic encephalopathy is a neurological complication observed in patients with liver disease. Subjects with hepatic encephalopathy can develop memory alterations. In order to investigate brain oxidative metabolism in an animal model of chronic cirrhosis and its modification after spatial working memory task, we determined the neural metabolic activity of several brain limbic system regions by cytochrome oxidase (COx) histochemistry and assessed the spatial working memory in the Morris water maze of rats with cirrhosis by administration of thioacetamide. This COx histochemistry was done in cirrhotic and control rats under basal conditions and after the spatial working memory task. The histochemical results showed differences in basal COx activity between control and cirrhotic rats in hippocampal and thalamic regions. In cirrhotic rats basal COx activity was increased in the CA1 and CA3 areas of the hippocampus and reduced in the anterodorsal and anteroventral thalamic nuclei. We found impaired spatial working memory in animals with cirrhosis. These animals showed absence of metabolic activation of the CA3 hippocampal subfield and the lateral mammillary nucleus and disturbance of COx activity in the medial mammillary nucleus and the anteroventral thalamus. These findings suggest that cirrhotic rats show spatial working memory deficits that could be related to the alteration of metabolic activity of neural regions thought to be involved in the processing of spatial memories.

Hepatic encephalopathy (HE) is a frequent neurological complication observed in patients with liver malfunction. Previous studies have shown memory impairment in these patients. In order to investigate brain substrates of spatial working memory impairment in chronic HE, neuronal expression of c-Fos protein was studied in an experimental model of cirrhosis. Control and cirrhotic rats were trained on a spatial working memory task in the Morris water maze (MWM). Differences between groups were found in the working memory task. Cirrhotic rats were unable to locate the platform in the retention trial. Neuronal activation, measured by c-Fos protein, was compared between groups. No differences were found in c-Fos expression of control and cirrhotic rats that were not tested in the MWM. Working memory task produced increase in c-Fos positive cells in dorsal hippocampus, CA1 and CA3, and prefrontal cortex in control group compared to thioacetamide group or naïve, which only swam in the maze during a similar time. These findings suggest that cirrhotic rats show spatial working memory impairment that could be linked to dysfunction in neuronal activity in prefrontal cortex and hippocampus.

The aim of our study was to examine the supramammillary (SuM) area involvement in spatial memory. Sprague-Dawley rats with chronically implanted cannula in the supramammillary area were trained in two spatial memory tasks with different memory demands: reference and working memory. In the spatial reference memory task, the rats received microinjections in the SuM area of tetrodotoxin (TTX) (0.5 ng diluted in 0.5 microL of saline) or saline (0.5 microL). The microinjections were administered 30 min before the spatial training (day 4) (to assess the effect on acquisition) and on the following two days (days 5 and 6) the training was conducted without microinjections (to study the effect on consolidation). On the last training day (day 7), in order to assess the retrieval of spatial information, the rats received the microinjections 30 min before the spatial training. The spatial working memory used was a delayed-matching-to-position (DMTP) task. Spatial training was performed for seven days. During the first three days of the spatial training, the rats achieved a good spatial knowledge and learnt the working memory rule necessary to solve the spatial task. On days 4 and 6, the rats received microinjections to study involvement of the SuM area in working memory. The results showed that temporary inactivation of SuM area impairs both the rat's ability to solve a spatial working memory task with DMTP demands and the recovery of spatial information in a spatial reference memory task. We suggest that SuM area is involved in the rearrangement of spatial information during spatial working memory tasks with DMTP memory demands.

In this article, a reproducible emulsion polymerization process is described to prepare core/shell colloidal nanospheres, loaded with 5-Fluorouracil, and consisting of a magnetic core (magnetite) and a biodegradable polymeric shell [poly(ethyl-2-cyanoacrylate), poly(butylcyanoacrylate), poly(hexylcyanoacrylate), or poly(octylcyanoacrylate)]. The heterogeneous structure of these carriers can confer them both the possibility of being used as drug delivery systems and the responsiveness to external magnetic fields, allowing an active drug targeting without a concurrent systemic distribution. Zeta potential determinations as a function of ionic strength showed that the surface behaviour of the core/shell particles is similar to that of pure cyanoacrylate particles. The first magnetization curve of both magnetite and magnetite/polymer particles demonstrated that the polymer shell reduces the magnetic responsiveness of the particles, but keeps unchanged their ferrimagnetic character. Two drug loading mechanisms were studied: absorption or entrapment in the polymeric network, and surface adsorption. We found that the acidity of the medium had significant effects on the drug absorption per unit mass of polymer, and needs to be controlled to avoid formation of macroaggregates and to reach significant 5-Fluorouracil absorption. The type of polymer and the drug concentration are also main factors determining the drug incorporation to the core/shell particles. 5-Fluorouracil release evaluations showed a biphasic profile affected by the type of polymeric shell, the type of drug incorporation and the amount of drug loaded.

The aim of the present study was to provide further evidence on the role of particular subdivisions of the mammillary bodies, anterior thalamus and dorsal hippocampus to contextual and auditory fear conditioning. We used c-Fos expression as a marker of neuronal activation to compare rats that received tone-footshock pairings in a distinctive context (conditioned group) to rats being exposed to both the context and the auditory CS without receiving footshocks (unconditioned group), and naïve rats that were only handled. Fos immunoreactivity was significantly increased only in the anterodorsal thalamic nucleus and the lateral mammillary nucleus of the conditioned group. However, the dorsal hippocampus showed the highest density of c-Fos positive nuclei in the naïve group as compared to the other groups. Together, our data support previous studies indicating a particular involvement of the mammillary bodies and anterior thalamus in fear conditioning.

Although the hippocampus has been shown to be essential for spatial memory, the contribution of associated brain regions is not well established. Wistar rats were trained to find a hidden escape platform in the water maze during eight days. Following training, the oxidative metabolism in different brain regions was evaluated using cytochrome oxidase histochemistry. Metabolic activations were found in the prelimbic cortex, cornu ammonis (CA) 1 subfield of the dorsal hippocampus and the anterior thalamic nuclei, relative to yoked swim controls and naïve rats. In addition, many cross-correlations in brain metabolism were observed among the latter regions. These results support the implication of a hippocampal-prefrontal-thalamic system to spatial memory in rats.

Background: Regulations concerning the use of x-rays in medical diagnoses were published in Mexico in 1997. In this work, we evaluate technical aspects of mammography services in the Mexico City area and radiation dose and coincidence between the radiological interpretation by the institution radiologist and by a panel of experts., Methods: Following methodology proposed by the American College of Radiology and the European Community among others, we have evaluated the performance of six mammography systems in Mexico City public and private services. The studied services carry out approximately one half of the mammography studies in the capital's metropolitan area., Results: The systems comply with 53-82% of a total of 31 applied quality control tests and measurements, which include the mammography unit, x-ray generation, collimation, automatic exposure control, compression devices, grid and image receptor, film processing, darkroom, viewboxes, dose, film rejection, and image quality. The elements that most frequently fail are film processing, darkroom, and light boxes; average ACR phantom score is 11.2 (9.5, 12.0); mean average glandular dose measured with the phantom is 1.00 (0.71-1.15) mGy, and measured in patients is 1.75 (0.3, 4.9) mGy; coincidence between radiologic reports (BI-RADS) by the institution radiologist and a panel of experts is obtained in 35% of studied cases., Conclusions: Statistical analysis of results indicated that the level of equipment performance is correlated with image quality, image quality estimated by the panel of radiologists is correlated with phantom score, and coincidence in clinical mammography reports is not correlated with equipment performance and appears to depend on the radiologist's experience.

Micro- and nano-testing methods have been explored to study the thin calcium phosphate coatings with high adhesive strength. The pulsed laser deposition (PLD) technique was utilised to produce calcium phosphate coatings on metal substrates, because this type of coatings exhibit much higher adhesive strength with substrates than conventional plasma-sprayed coatings. Due to the limitations of the conventional techniques to evaluate the mechanical properties of these thin coatings (1 microm thick), micro-scratch testing has been applied to evaluate the coating-to-substrate adhesion, and nano-indentation to determine the coating hardness and elastic modulus. The test results showed that the PLD produced amorphous and crystalline HA coatings are more ductile than titanium substrates, and the PLD coatings are not delaminated from the substrates by scratch. Also, the results showed that the crystalline HA coating is superior in internal cohesion to the amorphous one, even though the lower elastic modulus of amorphous coating could be more mechanically compatible with natural bone.

To investigate brain substrates of spatial memory, neuronal expression of c-Fos protein was studied. Two groups of rats were trained in two spatial memory tasks in the Morris water maze, where the rats have to apply a reference memory rule or a working memory rule. In addition to the experimental groups, two control groups were used to study c-fos activation not specific to the memory processes studied. After immunohistochemical procedures, the number of c-Fos positive neuronal nuclei was quantified in the mammillary body (MB) region (medial mammillary nucleus [MMn] and supramammillary nucleus [SuM]). The results have shown that some MMn neurons expressed c-Fos nuclear immunoreactivity related to spatial working memory but not to spatial reference memory. The increased number of c-Fos immunoreactive neuronal nuclei in the SuM was related to spatial training but not to either working or reference memory demands of the tasks.

Chronic alcohol consumption induces morphological changes in the central nervous system and withdrawal does not reverse these changes. It is well known that the hippocampal formation is one of the brain regions most sensitive to prolonged alcohol ingestion. The aim of our study was to evaluate the transcriptional neuronal activity by measuring the argyrophilic nucleolar organizer regions (AgNORs) in the dentate gyrus, CA3, and CA1 hippocampal areas from adult male rats receiving chronic administration of ethanol (ALC) and after withdrawal (WDL). The parameters evaluated were the number and area of AgNORs, together with the area of nucleus and the proportion between AgNOR and nuclear areas (ratio). The animals from ALC and WDL groups showed a reduction in the number of AgNOR per cell as compared to the control group. CA3 was the hippocampal area most affected by chronic alcohol intake. No improvement was observed in animals after withdrawal. Our data support the idea that the chronic intake of alcohol decreases protein synthesis in hippocampal neurons at an early age. This decrease may explain the memory impairment showed by rats receiving chronic treatment with alcohol because, both in humans and rats, it is associated with a reduction in the number of cholinergic neurons in the basal forebrain that would in turn affect the hippocampal function.

The aim of this work was to determine the spatial memory impairments induced by chronic alcohol consumption in rats. The alcoholization process began on the 21st postnatal day and alcohol concentrations were gradually increased to reach a concentration of 20% that was maintained for 4 mon. Behavioral tests were performed in the Morris Water Maze (MWM). The first study assessed the effects of chronic alcohol intake on two reference memory tasks (a place learning with multiple trials and a new place learning carried out in the same experimental context). Alcohol-treated animals presented no overall impairment in their ability to process spatial information. Deficits were restricted to reduced behavioral flexibility in spatial strategies. The second study assessed working memory in two tasks in which information about platform location was only valid for one trial. In the first working memory task, the animals had to perform one trial per day and in the second task they were submitted to four trials per day. At the end of the second experiment, all animals were trained in a visual-cued task. In the second experiment, the most important deficits in alcohol-treated animals occur in spatial working memory tasks, and this impairment was independent of the intertrial interval used. In the second spatial working memory task, performance of the alcohol-treated animals in the earlier trials affected their performance in subsequent trials, suggesting that a process of proactive interference had taken place. The visual-cued task demonstrated that these behavioral impairments were produced without visuoperceptive impairments.

In this study, we demonstrate that aging does not provoke any changes in neuronal number or in the glial cells of the medial mammillary nucleus (MMN) in humans. Three age groups were used: young (age 17-35), adult (age 50-57), and aged (age 70-88). Furthermore, no age-dependent volumetric changes were observed in the MMN. All the estimations were performed with stereological methods: an optical fractionator and Cavalier's principle. The total number of neurons cells was estimated using an optical fractionator and amounted to 32x10(3) in the young group, 24x103 in the adult group, and 29x103 in the aged group. The number of glial cells was 164x10(3), 187x103, 185x103, respectively. Thus, all three age groups had a neuron/glial ratio of about 1:5, 1:8, and 1:6, respectively. The MMN volume was estimated using the Cavalier's principle. The total volume was 6.98 mm3 in the young group, 6.66 mm3 in the adult group, and 6.80 mm3 in the aged group. We have demonstrated that neither the total number of neurons and glial cells nor the volume of MMN are affected by age.

Calcium phosphate coatings were produced by pulsed laser deposition from targets of non-stoichiometric hydroxyapatite (Ca/P = 1.70) at a substrate temperature of 485 degrees C and different processing pressures of water vapour: 0.15, 0.30, 0.45, 0.60 and 0.80 mbar. The physicochemical properties of these coatings were studied using Fourier-transform IR spectroscopy (FT-IR) and energy dispersive X-ray analysis (EDX). A minimum pressure of water vapour was necessary in order to obtain a crystalline coating, as deduced from the FT-IR spectroscopy of these coatings. This analysis also revealed that when the deposition pressure of water vapour was further increased, the coatings were less crystalline and the content of hydroxyl groups, the carbonate substitution for phosphate, and the Ca/P ratio, as measured by EDX, were lower. These effects can be explained by a combined substitution of carbonate and HPO4(2-) for phosphate, being predominant the carbonate substitution at low pressures and the HPO4(2-) substitution at high pressures.

During recent decades, the existence of cerebral and behavioural asymmetries has been manifested in many different animal species. In this work, the asymmetric spatial preference recorded in the water T-maze has been analysed. Fifty-five male rats were tested in this maze for 8 consecutive days. Spatial preference, the number of times each arm was chosen, and the percentages of animals that showed right, left, or no spatial preference were calculated. The results show the evolution of this asymmetric behaviour. Most of the animals tested preferred the right arm (populational laterality). Recently, the influence of factors such as stress and practice in the process of acquiring this behaviour has been investigated. We used 40 male rats divided into two groups with two different stress levels: water temperature at 21 degrees C and 14 degrees C. Our results, based on these two criteria, hypothesize that both variables, stress and practice, are involved in the acquisition of spatial preference behaviour.

A new method for quantitative determination of cytochrome oxidase (C.O.) activity was applied to diencephalic structures of the limbic system that are closely connected anatomically, that is, the mammillary bodies (MB) and the anterior nucleus of the thalamus (AT). This method makes it possible to easily evaluate the oxidative metabolic capacity of brain regions, an index of their functionality. By using this technique, we studied the postnatal development of both structures in Wistar rats of 14, 21, 30, and 120 days of age. Furthermore, animals of 730 days were included in order to evaluate the effects of aging on C.O. activity of these structures. The results showed a significant increase in the C.O. activity of the subdivisions of the AT, its levels remaining constant until the adult age, with a significant decrease in its activity in aged animals. In the MB, only the increase in C.O. activity of the medial mammillary nucleus (pars medialis) was significant until the adult age. A decrease of C.O. values with aging was significant only in the lateral mammillary nucleus. These data suggest that there is a wide heterogeneity in the maturation and aging of brain oxidative metabolism in diencephalic structures.