These properties are very promising devices for gene therapy of n

These properties are very promising devices for gene therapy of new age (Cytoplasmic Gene Therapy) because of its genotoxicity-free nature. Further, it is non-pathogenic for humans. Since Sendai virus is a murine parainfluenza virus (PIV) with certain homologies to human PIV, it was tested

as xenotropic vaccine in African Green monkeys and humans without any significant adverse reactions [34] and [35]. Recombinant SeV vector carrying human PIV was also tested in rats [36] and [37]. Further, recombinant SeV vaccine for human immunodeficiency virus (HIV) infection is going to be tested in humans (http://www.dnavec.co.jp/en/index.html). Thus, safety of Sendai virus vector is gradually established. We inserted mouse IL-10 cDNA to construct rSeV-Aβ with the aim of helping antibodies productions and suppressing Th1 type T cell activations. Nasal administration of rSeV-Aβ without IL-10 had less effect to remove Epacadostat Aβ depositions (data not shown). Recently, soluble Aβ oligomers, but not fibrils nor Lonafarnib purchase monomers, have been considered responsible for cognitive dysfunction prior to the formation of Aβ plaques [22], and Aβ*56, a 56-kDa soluble Aβ dodecamer was found responsible in Tg2576 mice [29]. Our nasal vaccine efficiently reduced not only senile plaque amyloid but also the contents of Aβ*56 oligomer without changing sAPPα and improved cognitive dysfunction in water

maze, Y-maze and contextual fear test which could evaluate hippocampus-related cognition. Thus, our vaccine, if applicable, can be given at the stage of mild cognitive impairment or earlier. Aβ is released from presynaptic sites and deposited in PAK6 extracellular plaques [38], and APP and synaptophysin are co-localized at the growth cones of developing neurons in culture [39]. These reports have indicated that Aβ deposition plays an important role in degeneration of presynaptic structures. In addition, it is reported that Aβ oligomers

directly disrupt synaptic structures [40]. In our study, synaptophysin staining showed amelioration of presynaptic degeneration following our nasal vaccine at 24 months old, suggesting prevention of synaptic degeneration or repair of synaptic structures after removal of Aβ deposits including Aβ oligomers. Our next plan is to see whether Tg2576 mice show improvement of cognitive functions by eliminating senile plaque amyloid even at 24 months old. In conclusion, a new vaccine using Sendai virus vector with Aβ and IL-10 cDNA was developed. A nasal administration of this vaccine reduced amyloid burden including Aβ oligomers significantly in AD mice and improved cognitive functions without causing side effects such as brain inflammation. This vaccine can be used to treat and prevent Alzheimer disease. Authors are grateful to Dr. Y. Noda at Meijo University, Dr. T. Nagai at Nagoya University, and Dr. M.

On the other hand, with WHO prequalification, a user-friendly del

On the other hand, with WHO prequalification, a user-friendly delivery system and an affordable vaccine, we expect to be able to offer LAIV to United Nations agencies for inclusion by developing countries in their national immunization programme (the WHO technology transfer grant stipulates that at least 10% of our pandemic influenza selleck chemicals production must be made available to this channel). In this way, we hope to be able to sell sufficient vaccine to sustain our manufacturing activity. Given that LAIV will be new to most countries, we also expect the need

for awareness-building over at least a year before the vaccine will be taken up. To this end, SII proposes to undertake further studies on LAIV, for example to elucidate immunological correlates of protection. To understand better the mechanisms of LAIV protection with homologous as well as drifted strains, SII would like to explore a human challenge trial using LAIV and carry out well controlled experiments to collect more data on cell-mediated immunity and other immunological parameters. However, this research would require additional financial and scientific support.

The opportunity to work on influenza vaccine has opened up a new era of South–South cooperation. For example, SII and the Government Pharmaceutical Organization (GPO) in Thailand have been collaborating on the development of LAIV ever since seed strains became available from IEM. Among other initiatives, the GPO team visited SII to acquire the techniques and skills for their own development of LAIV. In a health crisis such as an influenza pandemic, Quisinostat order science should override commerce and SII is committed to such collaborations. The WHO project to build capacity in developing countries to manufacture pandemic influenza

vaccine has provided India with the critical skills needed to help protect its 1.2 billion population from a deadly influenza pandemic. The technical inputs and excellent coordination by the WHO team were of immense help in resolving several technical issues and enabling swift and pivotal decision-making. Our ability to develop and market a pandemic LAIV in such record ALOX15 time was partly due to our long-standing experience in vaccine manufacture, our qualified staff, and this WHO collaboration. However, with hindsight, this would not have happened without the exceptional ingenuity and commitment of the SII team, who subdivided into independent virological, formulation, analytical methods and clinical development groups, and achieved their defined goals in the face of stringent time constraints. In the future, LAIV and tissue culture may be the way forward, and SII will continue its research and development efforts to remain at the forefront of providers of solutions to major public health priorities.

0003) was elicited by the MC 6M OMV vaccine ( Fig 2B) However,

0003) was elicited by the MC.6M OMV vaccine ( Fig. 2B). However, the 2.0 μg dose of this vaccine induced significantly higher titres (p = 0.032) than the same dose of the FM OMV vaccine. A significant dose response in SBA (p = 0.004) was also found for the two doses of the FM OMV vaccine in a separate experiment (data not shown). The titres obtained with the 2.0 μg dose Akt targets of both vaccines were significantly higher (p < 0.001) than those of the saline control group, whereas no significant differences

were observed between the saline controls and 0.5 μg of the MC.6M OMV vaccine ( Fig. 2B) or 0.5 μg of the FM OMV vaccine (data not shown). Specific antibody levels in immunized mice to the major OMPs were measured on immunoblots using the MC.6M OMV as antigen (data not shown). The 2.0 μg dose

of both vaccines induced similar Ig levels to Omp85, PorA, PorB, RmpM, OpcA, and OpaJ129 which were the main immunogenic bands on the blots. Significantly lower levels (p = 0.001–0.046) to these antigens were induced by 0.5 μg of the MC.6M vaccine. The FM OMV vaccine also gave significant dose responses (p ≤ 0.001) to the OMPs determined with FM OMVs as blotting antigen (data not shown). Antibodies to PorA contributed markedly to the bactericidal activity of the murine sera as there was a significant correlation between the Ig binding intensity to PorA on the blots and the bactericidal titres with both doses of each OMV vaccine (range of Pearson product moment correlation or Spearman rank order PD98059 clinical trial correlation coefficients 0.580–0.856; p = 0.0004–0.048). The DIGE method was used to investigate differences in protein content between the OMV preparations prepared using different culture media. A total of 2005 spots were common amongst six gels from the three batches of OMVs from each medium.

The level of expression of about 97% of the protein spots did not change between the two OMV preparations (Table 1B). Only 3.2% (64 spots) exhibited a greater than 1.1-fold difference in the amount of protein (p = 0.00023–0.049). Forty-one proteins were more abundant in OMVs produced in MC.6M, whereas 23 were more abundant in OMVs produced in FM. Most of the spots that differed between the OMVs from the two media were in the basic region and included a range of molecular masses ( Fig. 3). High abundance spots, identified as the major Phosphatidylinositol diacylglycerol-lyase OMPs, i.e. PorA, PorB and OpcA, were excluded from accurate quantitative comparison due to their saturation in fluorescence intensity which exceeded the linear range of scanning conditions. Table 2 shows the details of 10 protein spots that were differentially expressed by meningococci grown in each of the media and in sufficient abundance to be identified by MS analysis. Lipoprotein NMB1126/NMB1164, hypothetical protein NMB2134 (2 spots), NspA, TonB-dependent receptor TdfH (2 spots), OMP NMB0088, MafA and OpcA (2 spots) were amongst the proteins that were more abundant in MC.6M OMVs.

002) ( Table 3) In the control group, among the 20 pneumococcal

002) ( Table 3). In the control group, among the 20 pneumococcal isolates recovered from multiple carriers during

May 2001, four serotypes were identified, of which VT serotypes (6B, 19F, and 23F) represented 95% of the isolates ( Table 3). In June 2001, two serotypes were identified among the 10 pneumococcal isolates, with VT serotypes increasing from 95 to 100%, while NVT isolates decreased from 5 to 0% (P = 1) ( Table 3). Among the vaccinated group, in May 2001, co-colonization with VT isolates was detected in five out of seven multiple carriers, of which four presented the VT as the dominant serotype. In June 2001, co-colonization with VT isolates was detected selleck chemicals llc in four out of six multiple carriers, with the VT being identified always as a minor serotype (Fig. 1, children A to K). Regarding the control, in May 2001, co-colonization with VT isolates was detected in two children who presented

Smad inhibitor VTs as the dominant serotypes. In June 2001, co-colonization was detected only once and two VT serotypes were found in association (23F—dominant serotype; 19F—minor serotype) (Fig. 1, children L and M). Serotype 6A was the most common serotype found among multiple carriers—it was found co-colonizing with 19F (three occasions), 6B (two occasions), and 14, 19A and non-typeable isolates (one occasion). Overall we compared 174 PFGE profiles of representative isolates of each of the serotypes found among the vaccinated (124 isolates) and control (50 isolates) groups and no capsular switch phenomenon was detected. In the group where the vaccine pressure was present, no vaccine escapee recombinant isolate was observed and the NVT PFGE profiles were found to differ from the preceding VT serotypes. A few examples of the PFGE profiles analyzed are shown in Fig. 2. By observing the colonization pattern change from May to June 2001 among children of the vaccinated and control groups, we were able to assess the number of isolates that were cleared, de novo acquired, unmasked or maintained

( Fig. 3). Bearing in mind that PCV7 targets directly VT and indirectly NVT isolates, the effect of the vaccine on pneumococcal carriage was Idoxuridine explored based on three potential mechanisms: prevention of VT de novo acquisition, enhancement of VT clearance, and enhancement of NVT unmasking. We compared these three mechanisms capable of affecting pneumococcal colonization between vaccinated and control groups to identify those that could explain the vaccine’s effect. Serotype clearance was similar between VT and NVT isolates among the vaccinated and control groups (P = 0.635). VT and NVT isolates were equally probable to be cleared in both groups (OR, 1.12; 95% confidence interval (CI), 0.68–1.84) ( Table 4).

Another limitation of the study is that those not educated within

Another limitation of the study is that those not educated within the state system were not involved with the NCMP and so it was not possible to consider those who were home or privately educated. There were

some differences in the characteristics of the sample analysed for this study compared with that analysed by Procter et al. (2008); notably Devon is much less ethnically selleck chemical diverse than Leeds. However, the similarity between our findings within any year, and those of Procter et al. (2008) would suggest that the methods employed were not sensitive to differing sample characteristics and hence the approach has some external validity. The problems associated with the reliability of league tables are well documented (Goldstein and Spiegelhalter, 1996 and Marshall and Spiegelhalter, 1998) and yet they remain in regular use in health, education and other areas of political interest (Marshall et al., 2004). Marshall and Spiegelhalter (1998) in examining in vitro fertilisation clinics found that ‘[e]ven when there

are substantial differences between institutions, ranks are extremely unreliable statistical summaries of performance and change in performance’ (p. 1701). Phenomena such as regression towards the mean are responsible for the instability of league tables and control chart methods have been proposed as a more robust alternative ( Marshall et al., 2004). Further work is needed to establish whether control charts could reliably identify schools which are ‘hot’ and ‘cold’ spots for obesity. However, the failure to find patterns among the rankings of individual schools over the five years studied indicates that individual

GSK1349572 mouse schools were not differentially affecting pupil weight status, suggesting that school-based ‘hot’ and ‘cold’ spots for obesity may not exist and therefore are not appropriate targets for resources. In conclusion, this study found that estimates of individual school impacts on pupil weight status were small and labile across SB-3CT the five-year study period, refuting the hypothesis of a systematic differential impact of primary schools on pupil weight status. Furthermore, this suggests that ranking schools into ‘obesogenic league tables’ using current value-added methods is not a reliable approach to the identification of schools requiring targeted resources. As with previous studies (e.g. Harrison et al., 2011 and Townsend et al., 2012), only a small proportion of the variation in pupil weight status was found to be attributed to schools (Table 1). The marked changes in the impact of individual schools on pupil weight status from year-to-year bring into question whether the argument that small population level changes can reflect significant changes for individuals, proposed by Rose and Day (1990) is still a valid justification for school-based obesity prevention. It would appear that interventions intended to affect pupil weight status need to influence the wider environment and not just the school in isolation.

HPV16/18 prevalence pre- and post-immunisation among 16–18 year o

HPV16/18 prevalence pre- and post-immunisation among 16–18 year olds was

(i) 19.1% vs. 6.2% (68% reduction) (ii) 19.1% vs. 7.4% (61% reduction), (iii) 38.6% vs. 13.8% in chlamydia positives (64% reduction) and 16.7% vs. 5.9% in chlamydia negatives (65% reduction), and (iv) 19.7% vs. 4.8% in the GP clinics (76% reduction), 18.4% vs. 6.7% in community sexual health services (64% reduction) and 19.6% vs. 8.9% in Youth clinics (55% reduction), respectively. The detected prevalence of non-vaccine HR HPV types was slightly higher in the post-immunisation period than pre-immunisation Selleck SNS032 for each age group (Fig. 3). There was no clear change in the pattern of age-specific prevalence, nor trend in the adjusted odds ratio by age group (Table 2). These increases combined with the decreases in HPV 16/18 resulted in similar prevalence of all HR HPV (i.e. vaccine and non-vaccine types) among 16–18 year olds in both periods (post-immunisation 34.1% (95% Sirolimus solubility dmso CI 31.4–36.9): pre-immunisation 34.1% (95% CI 31.1–37.3) p-value = 0.998). The detected prevalence of three HR HPV types against which cross-protection has been reported from clinical trials, HPV 31, 33 and 45 [11] and [12] was slightly lower overall post-immunisation, but with no clear change in the pattern of age-specific

prevalence (data not shown), nor trend in the adjusted odds ratio by age group (Table 2). Multiple infections remained common in this age group, albeit somewhat reduced in the immunised ages in line with reduced prevalence of HPV 16/18 (36.8% of HR HPV positive 16–18 year olds with more than one HR HPV vs. 52 7% in 2008). As in 2008, non-vaccine HR HPV types were found in over half of the HPV 16/18 positives. These findings are an early indication that the national HPV immunisation programme is successfully

L-NAME HCl preventing HPV 16/18 infection in sexually active young women in England. There was a clear change in the pattern of age-specific HPV 16/18 prevalence and the prevalence amongst females eligible for immunisation was considerably lower than previously measured in 2008 prior to immunisation. Lower HPV16/18 prevalence was associated with higher immunisation coverage. These surveillance data show the impact of a high coverage immunisation programme within the targeted, and slightly older, population. Without vaccination status, we could not report the effectiveness amongst those immunised, however that would likely be heavily influenced by biases in vaccine uptake in these catch-up cohorts. The finding of no fall in HPV 16/18 prevalence between time periods among females above the age of HPV immunisation, and no change in the age-specific pattern of non-vaccine HR prevalence argues against the HPV 16/18 changes being solely due to selection biases or time trends and supports their attribution to the impact of the immunisation programme. In fact, the known changes in selection of subjects (e.g.

Out of the 50 eyes

Out of the 50 eyes selleck screening library with retinal hemorrhages, only 1 (2%) lacked either a subdural or intrascleral hemorrhage. Within these, 33 (66%) had both subdural and intrascleral hemorrhages, while 15 (30%) had a subdural without intrascleral hemorrhage, and 1 (2%) had an intrascleral without subdural hemorrhage. Subdural hemorrhage was present in 58 eyes (97%), of which 33 (57%) also had retinal and intrascleral hemorrhages. Only 6 of these eyes

(10%) positive for subdural hemorrhage had neither retinal nor intrascleral hemorrhages, while 15 (26%) had retinal hemorrhage of any kind without intrascleral hemorrhage, and 4 (6.9%) had intrascleral hemorrhage without retinal hemorrhage. Therefore, 10 eyes (17%) had subdural hemorrhage without retinal hemorrhage, of which 6 had unilateral retinal hemorrhages and 4 lacked retinal hemorrhages bilaterally. Intrascleral hemorrhage was present in 38 eyes (63%): selleck compound 33 of those eyes (87%) also had subdural and retinal hemorrhages, 4 (11%) had subdural without retinal hemorrhages, and 1 (2.6%) had retinal without subdural hemorrhage. Intrascleral hemorrhage always accompanied a retinal or subdural hemorrhage. Vitreoretinal interface abnormalities were seen in 51 abusive head trauma eyes (85%) (Figure 1, Right panel). ILM tear in isolation was the most common observation in 22 eyes (37%). The incidence of ILM tear with a perimacular ridge and cherry hemorrhage

was 20 (33%), while incidence of only ILM tear and a perimacular ridge was 5 (8%) and of only cherry hemorrhage with ILM tear was 4 (6.7%). Every eye with a perimacular ridge or cherry hemorrhage had a torn ILM. In eyes with ILM tear, 20 (39%) also had a cherry hemorrhage and a perimacular ridge, 5 (10%) had a perimacular ridge without a cherry hemorrhage, 4 (7.8%) had a cherry hemorrhage without a perimacular ridge, and 22 (43%) did not have an accompanying perimacular ridge

or a cherry hemorrhage. In total, 24 (40%) eyes had a cherry hemorrhage: 20 (83%) also had ILM tears and a perimacular ridge, while not 4 (17%) had an ILM tear without a perimacular ridge. There were 25 (42%) eyes out of 60 with perimacular ridges: 20 (80%) also had both cherry hemorrhages and ILM tears, while 5 (20%) had a torn ILM without a cherry hemorrhage. Subdural hemorrhage at the optic nerve has a bluish hue externally. In cross-section, the blood is visible inside the dura (Figure 2, Left). Microscopically, intrascleral hemorrhage is found surrounding ruptured intrascleral vessels at the junction of the optic nerve and sclera (Figure 2, Right). Intrascleral bleeding is often continuous with the subdural space. Typical perimacular ridges are elevated, circular retinal folds with a canopy of ILM above, torn away from retina, with fibrin-hemorrhage debris below. Often a portion of the perimacular ridge can be seen clinically, surrounding hemorrhage at the macula (Figure 3, Top left).

The dynamics of the lesions’ healing process following laser trea

The dynamics of the lesions’ healing process following laser treatment after week 1 suggest a strong dependency on the loss of initial fluence at each specific laser spot, presumably attributable to small media opacities and overlying retinal edema. The overall persistence of polarization-scrambling columns over the course

of 3 months indicates a much more intense healing reaction and proliferation of RPE cells than previously shown in rodent studies. These findings might support the hypothesis that the beneficial effect of grid and focal photocoagulation is driven by an increase in metabolically active RPE Fasudil mw tissue. This study was limited by its small sample size, its short follow-up period, and the use of only 1 laser system. Nevertheless, the setting is adequate to demonstrate the ability of polarization-sensitive SD-OCT to identify and automatically segment the retinal pigment epithelium in different stages of healing following photocoagulation, in contrast to current SD-OCT devices. Considering further development of minimal-damage photocoagulation, such as subthreshold or selective retinal treatment,28, 30 and 31 polarization-sensitive OCT is a new modality to investigate the therapeutically induced changes of defined retinal Bax protein layers in the human eye over time. All authors have completed and submitted the ICMJE Form for Disclosure Bumetanide of Potential

Conflicts of Interest. M. Pircher, E. Götzinger, and C.K. Hitzenberger have received research

support from Canon, Tokyo, Japan. C.K. Hitzenberger has received lecture fees from National Institute of Health, Bethesda, Maryland. Publication of this article was financially supported by the Austrian Science Fund (FWF grant no. P19624-B02, Vienna, Austria) and the European Union (project FUN OCT, FP7 HEALTH, Contract No. 201880). The high-definition OCT system was provided by Heidelberg Engineering. Polarization-sensitive OCT was constructed and provided by the Center for Biomedical Engineering and Physics, Medical University of Vienna, Vienna, Austria. Contributions of authors: design and conduct of the study (J.L., M.B.); data collection (J.L., M.G.); management (J.L., M.B.); analysis and interpretation of the data (J.L., M.G.); design and construction of polarization-sensitive OCT device (B.B., M.P., E.G., C.H.); and review and approval of the manuscript (M.B., C.H., U.E.). The authors thank Ferdinand Schlanitz and Christopher Schütze for helping with recording the polarization-sensitive OCT images and Robert Blum for English proofreading. All three are members of the Department of Ophthalmology at the Medical University of Vienna, Vienna, Austria. For further information on members and mission statement of the Diabetic Retinopathy Research Group (DRRG), Vienna, please visit: http://www.meduniwien.ac.

Logs (one per week) were handed to the participants

Logs (one per week) were handed to the participants XL184 to record unguided

mental practice behaviour. In principle, a maximum of six logs could be completed. The main goal of the mental practice intervention was to improve locomotor tasks like walking, standing up from a chair or the floor. Therapists were trained to teach and monitor mental practice according to the framework in which four steps are distinguished: explaining the concept, developing imagery techniques, applying mental practice, and consolidating (Braun et al 2008). Figure 1 presents the time frame over which these four stages were utilised. Unlike a fixed treatment regimen, the mental imagery framework allowed the physiotherapist to tailor the content to each participant’s abilities and preferences. Examples of tailoring are the chosen view and the ratio of actual to imagined attempts at movements. Participants were told

that imagery inherently involved a point of view. They were advised to try first person (as if looking through their own eyes) and third person (as if looking at oneself from a distance), and were then allowed to choose whichever view they preferred (Milton et al 2008). Sotrastaurin molecular weight During therapy, imagery attempts and overt movements were combined, ie, movements were performed to generate sensory information. This information was then embedded in the imagery attempts to make them as vivid as possible. The proportions of actual movements and imagery attempts were based on individual preferences (Malouin Calpain et al 2004). The ratio of actual to imagined attempts could change over time or differ depending on the task or its difficulty. The success of a participant in imagining the actions correctly and vividly was judged by the therapist in several ways: self-report by the participant, comparing the time taken to perform a task mentally against the time in reality, and by checking that the participant

could recite the order of actions correctly. The control therapy was used to control for attention and consisted of treatment according to the national Dutch guidelines (Keus et al 2004) with relaxation therapy being incorporated into each session. The amount of relaxation incorporated matched the amount of mental practice in the experimental group. Relaxation was chosen to enable comparison with the trial by Tamir and colleagues and followed the principles of progressive muscle relaxation according to Jacobson (Gessel 1989). Participants were encouraged to do relaxation homework outside of therapy as well, using unguided progressive muscle relaxation or by listening to a relaxation CD. Improvement in walking was assessed with a visual analogue scale (Donnelly and Carswell 2002, Stratford et al 1995, Wewers and Lowe 1990). Participants and therapists were asked to score on a scale from 0 to 10 how well they thought the participant walked with 0 being ‘poor’ and 10 being ‘excellent’.

The films were scanned and bands intensities were analyzed using

The films were scanned and bands intensities were analyzed using Image J software (developed at the US National Institutes of Health and available on the web site (http://rsb.info.nih.gov/nih-image/).

In order to determine the adequate amount of protein to be assayed, different protein concentrations were carried out in the same gel for each antibody tested. Perfusion and fixation of the brain from 4 animals/group were performed 24 h after the end of seizures period through transcardiac perfusion with 4% paraformaldehyde and 0.25% glutaraldehyde, followed by cryoprotection selleck products in 30% sucrose solution overnight. Brain was sectioned (50 μm coronal sections) using a Leica VT1000S microtome (Leica Microsystems, São Paulo, Brazil). Coronal sections were separated in 4 series throughout the dorsal hippocampus with 300 μm interval between

each section and collected in PBS. Free-floating sections of rat brain were processed for immunohistochemistry against the neuronal specific protein neuronal nuclei (NeuN) and glial fibrillary acidic protein (GFAP), using a primary mouse anti-NeuN (1: 500, Chemicon International, São Paulo/SP, Brazil) as well as rabbit anti-GFAP antibodies (1:500, Dako, Denmark A/S). Antibodies were diluted in Tris buffer saline (TBS, 0.5 M NaCl and 30 mM Tris, Bosutinib pH 7.4) containing 0.2% Triton X-100 and 10% normal goat serum and incubated for 48 h at 4 °C. After incubation, sections were rinsed 4 times for 10 min in TBS and subsequently incubated with secondary fluorescent antibodies overnight: Alexa fluor anti-rabbit 488 and anti-mouse 594 (1:500, Invitrogen, Porto Alegre/RS), in 0.1 M TBS containing 0.2% Triton

X-100 and 10% normal goat serum for 24 h at 4 °C. After rinsing 4 times for 10 min in TBS, the sections were mounted on slides coated with 2% gelatin with chromium and potassium sulfate. The slices were mounted in a Vectashield mounting medium containing the nuclear marker DAPI (4′-6-diamidino-2-phenylindole dilactate) (Vector Laboratories, São Edoxaban Paulo/SP, Brazil). The CA1, CA3 and dentate gyrus (DG) subfields of each hippocampus were examined in the Olympus FluorView 1000 system and the fluorescence was quantified using ImageJ software. The images were captured and a square region of interest (ROI) was created considering the pyramidal layer size. The ROI square of 8019 μm2 was overlaid on the analyzed subfields with blood vessels and other artifacts being avoided, using a magnification of 20x. Six ROI were analyzed per subfield. Rats (60-day-old) were exposed to the elevated plus-maze apparatus that consisted of a central platform (10 cm × 10 cm) with 2 open and 2 closed arms (45 cm × 10 cm), arranged in such a way that the 2 arms of each type were opposite to each other.