Innovative statistical approaches: the use of neural networks reduces the sample size in the splenectomy-MCAO mouse model.

AIM: To compare the effectiveness of artificial neural network (ANN) and traditional statistical analysis on identical data sets within the splenectomy-middle carotid artery occlusion (MCAO) mouse model. METHODS: Mice were divided into the splenectomized (SPLX) and sham-operated (SPLX-sham) group. A splenectomy was conducted 14 days before middle carotid artery occlusion (MCAO). Magnetic resonance imaging (MRI), bioluminescent imaging, neurological scoring (NS), and histological analysis, were conducted at two, four, seven, and 28 days after MCAO. Frequentist statistical analyses and ANN analysis employing a multi-layer perceptron architecture were performed to assess the probability of discriminating between SPLX and SPLX-sham mice. RESULTS: Repeated measures ANOVA showed no significant differences in body weight (F (5, 45)=0.696, P=0.629), NS (F (2.024, 18.218)=1.032, P=0.377) and brain infarct size on MRI between the SPLX and SPLX-sham groups post-MCAO (F (2, 24)=0.267, P=0.768). ANN analysis was employed to predict SPLX and SPL-sham classes. The highest accuracy in predicting SPLX class was observed when the model was trained on a data set containing all variables (0.7736±0.0234). For SPL-sham class, the highest accuracy was achieved when it was trained on a data set excluding the variable combination MR contralateral/animal mass/NS (0.9284±0.0366). CONCLUSION: This study validated the neuroprotective impact of splenectomy in an MCAO model using ANN for data analysis with a reduced animal sample size, demonstrating the potential for leveraging advanced statistical methods to minimize sample sizes in experimental biomedical research.

suMRak: a multi-tool solution for preclinical brain MRI data analysis.

Introduction: Magnetic resonance imaging (MRI) is invaluable for understanding brain disorders, but data complexity poses a challenge in experimental research. In this study, we introduce suMRak, a MATLAB application designed for efficient preclinical brain MRI analysis. SuMRak integrates brain segmentation, volumetry, image registration, and parameter map generation into a unified interface, thereby reducing the number of separate tools that researchers may require for straightforward data handling. Methods and implementation: All functionalities of suMRak are implemented using the MATLAB App Designer and the MATLAB-integrated Python engine. A total of six helper applications were developed alongside the main suMRak interface to allow for a cohesive and streamlined workflow. The brain segmentation strategy was validated by comparing suMRak against manual segmentation and ITK-SNAP, a popular open-source application for biomedical image segmentation. Results: When compared with the manual segmentation of coronal mouse brain slices, suMRak achieved a high Sørensen-Dice similarity coefficient (0.98 ± 0.01), approaching manual accuracy. Additionally, suMRak exhibited significant improvement (p = 0.03) when compared to ITK-SNAP, particularly for caudally located brain slices. Furthermore, suMRak was capable of effectively analyzing preclinical MRI data obtained in our own studies. Most notably, the results of brain perfusion map registration to T2-weighted images were shown, improving the topographic connection to anatomical areas and enabling further data analysis to better account for the inherent spatial distortions of echoplanar imaging. Discussion: SuMRak offers efficient MRI data processing of preclinical brain images, enabling researchers' consistency and precision. Notably, the accelerated brain segmentation, achieved through K-means clustering and morphological operations, significantly reduces processing time and allows for easier handling of larger datasets.

Perspectives on the COVID-19 Vaccination Rollout in 17 Countries: Reflexive Thematic and Frequency Analysis Based on the Strengths, Weaknesses, Opportunities, and Threats (SWOT) Framework.

BACKGROUND: As the SARS-CoV-2 virus created a global pandemic and rapidly became an imminent threat to the health and lives of people worldwide, the need for a vaccine and its quick distribution among the population was evident. Due to the urgency, and on the back of international collaboration, vaccines were developed rapidly. However, vaccination rollouts showed different success rates in different countries and some also led to increased vaccine hesitancy. OBJECTIVE: The aim of this study was to identify the role of information sharing and context sensitivity in various vaccination programs throughout the initial COVID-19 vaccination rollout in different countries. Moreover, we aimed to identify factors in national vaccination programs related to COVID-19 vaccine hesitancy, safety, and effectiveness. Toward this end, multidisciplinary and multinational opinions from members of the Navigating Knowledge Landscape (NKL) network were analyzed. METHODS: From May to July 2021, 25 completed questionnaires from 27 NKL network members were collected. These contributors were from 17 different countries. The responses reflected the contributors' subjective viewpoints on the status and details of the COVID-19 vaccination rollout in their countries. Contributors were asked to identify strengths, weaknesses, opportunities, and threats (ie, SWOT) of the respective vaccination programs. The responses were analyzed using reflexive thematic analysis, followed by frequency analysis of identified themes according to the represented countries. RESULTS: The perspectives of NKL network members showed a link between organizational elements of the vaccination rollout and the accompanying societal response, both of which were related to strengths and weaknesses of the process. External sociocultural variables, improved public communication around vaccination-related issues, ethical controversies, and the spread of disinformation were the dominant themes related to opportunities and challenges. In the SWOT 2×2 matrix, Availability and Barriers emerged as internal categories, whereas Transparent communication and promotion and Societal divide emerged as key external categories. CONCLUSIONS: Inventory of themes and categories inspired by elements of the SWOT framework provides an informative multidisciplinary perspective for effective implementation of public health strategies in the battle against COVID-19 or any future pandemics of a similar nature.

Elderly rats fed with a high-fat high-sucrose diet developed sex-dependent metabolic syndrome regardless of long-term metformin and liraglutide treatment.

Aim/Introduction: The study aimed to determine the effectiveness of early antidiabetic therapy in reversing metabolic changes caused by high-fat and high-sucrose diet (HFHSD) in both sexes. Methods: Elderly Sprague-Dawley rats, 45 weeks old, were randomized into four groups: a control group fed on the standard diet (STD), one group fed the HFHSD, and two groups fed the HFHSD along with long-term treatment of either metformin (HFHSD+M) or liraglutide (HFHSD+L). Antidiabetic treatment started 5 weeks after the introduction of the diet and lasted 13 weeks until the animals were 64 weeks old. Results: Unexpectedly, HFHSD-fed animals did not gain weight but underwent significant metabolic changes. Both antidiabetic treatments produced sex-specific effects, but neither prevented the onset of prediabetes nor diabetes. Conclusion: Liraglutide vested benefits to liver and skeletal muscle tissue in males but induced signs of insulin resistance in females.

Reactivation of corticogenesis-related transcriptional factors BCL11B and SATB2 after ischemic lesion of the adult mouse brain.

The aim of this study was to characterize expression of corticogenesis-related transcription factors BCL11B and SATB2 after brain ischemic lesion in the adult mice, and to analyze their correlation to the subsequent brain recovery. Ischemic brain lesion was induced by transient middle cerebral artery occlusion followed by reperfusion, and the animals with ischemic lesion were compared to the sham controls. Progression of the brain damage and subsequent recovery was longitudinally monitored structurally, by magnetic resonance imaging, and functionally, by neurological deficit assessment. Seven days after the ischemic injury the brains were isolated and analyzed by immunohistochemistry. The results showed higher expression in the brain of both, BCL11B and SATB2 in the animals with ischemic lesion compared to the sham controls. The co-expression of both markers, BCL11B and SATB2, increased in the ischemic brains, as well as the co-expression of BCL11B with the beneficial transcriptional factor ATF3 but not its co-expression with detrimental HDAC2. BCL11B was mainly implicated in the ipsilateral and SATB2 in the contralateral brain hemisphere, and their level in these regions correlated with the functional recovery rate. The results indicate that the reactivation of corticogenesis-related transcription factors BCL11B and SATB2 is beneficial after brain ischemic lesion.

Facts and doubts on the beginning of human life - scientific, legal, philosophical and religious controversies.

It is very complicated to give correct answer to the question "How to define human life?" Nowadays dilemmas consider the respect of human life from the birth to death involve not just biology but also other sciences like philosophy, theology, sociology, psychology, law and politics. These sciences evaluate the topic from different points of view. Integration of all of these perspectives could result with a proper definition. The principal purpose of this paper is to try to determine when a human individual begins. If this proves to be too difficult, we might have to settle for a specific stage in the reproductive process before which it would be impossible to say with any plausibility that a human individual exists. It is necessary to return the moral dimension of observation to the science of life. The point is to reconcile the universal ethical principles concerning the absolute value of life with the everyday challenges and dilemmas. It is our deepest conviction that life has an absolute value and that there always remains something indestructible and substantial in life, which may neither be evaluated by anything final, nor completely reduced to the material biological equivalent and the genetic substratum.

Proposed practical protocol for flow cytometry analysis of microglia from the healthy adult mouse brain: Systematic review and isolation methods' evaluation.

The aim of our study was to systematically analyze the literature for published flow cytometry protocols for microglia isolation and compare their effectiveness in terms of microglial yield, including our own protocol using sucrose for myelin removal and accutase for enzymatic digestion. For systematic review, the PubMed was searched for the terms "flow cytometry," "microglia," "brain," and "mice." Three different myelin removal methods (Percoll, sucrose, and no removal) and five protocols for enzymatic digestion (accutase, dispase II, papain, trypsin, and no enzymatic digestion) were tested for the effectiveness of microglia (CD11b+CD45int cell population) isolation from the adult mouse brain using flow cytometry. Qualitative analysis of the 32 selected studies identified three most commonly used myelin removal protocols: Percoll, the use of myelin removal kit, and no removal. Nine enzymatic digestion protocols were identified, from which we selected dispase II, papain, trypsin, and no enzymatic digestion. A comparison of these myelin removal methods and digestion protocols showed the Percoll method to be preferable in removal of non-immune cells, and superior to the use of sucrose which was less effective in removal of non-immune cells, but resulted in a comparable microglial yield to Percoll myelin removal. Digestion with accutase resulted in one of the highest microglial yields, all while having the lowest variance among tested protocols. The proposed protocol for microglia isolation uses Percoll for myelin removal and accutase for enzymatic digestion. All tested protocols had different features, and the choice between them can depend on the individual focus of the research.

The effects of splenectomy in murine models of ischemic stroke: a systematic review and meta-analysis.

BACKGROUND: The spleen, a substantial reservoir of non-differentiated monocytes, may play a crucial role in the pathophysiology of post-ischemic inflammation and influence outcomes after ischemic stroke. AIM OF THE STUDY: To analyze splenectomy as a preclinical intervention in murine models of ischemic stroke. METHODS: Following systematic searches of PubMed, Scopus and Web of Science, a qualitative synthesis of study characteristics was performed, and the effect of splenectomy estimated by a three-level random-effects meta-analysis of infarct volumes and a conventional two-level random-effects meta-analysis of neurological deficit scores. RESULTS: Database searches identified a total of 14 studies, 13 of which were used for meta-analysis. The ischemic lesion volumes were reduced in splenectomized animals compared to the control groups (difference in standardized mean differences: - 1.42; 95% CI [- 1.98, - 0.85]; 95% PI [- 2.03, - 0.80]; I2(2) = 19.04%; 95% CI [0.00%, 65.49%]; I2(3) = 47.24%; 95% CI [0.00%, 85.23%]) and neurological deficit scores were improved (- 1.20; 95% CI [- 2.20, - 0.20]; 95% PI [- 4.58, 2.18]; I2 = 77.5%; 95% CI [50.0%, 89.9%]). A subgroup analysis for infarct volumes showed that splenectomy performed prior to ischemia achieved a higher reduction of the ischemic lesion than when splenectomy was performed immediately prior or after stroke. Although the overall effect size of splenectomy could be classified as large, there was a significant presence of risks of bias, study heterogeneity, and a potential presence of publication bias. CONCLUSION: Despite limitations related to heterogeneity, risks of bias, and potential publication bias, this meta-analysis points to the spleen and its functional cell populations as promising targets for the therapeutic modulation of post-stroke inflammation.

Novel micro-MRI approach for subchondral trabecular bone analysis in patients with hip osteoarthritis is comparable to micro-CT approach.

AIM: To test the agreement between a newly developed micro-magnetic resonance imaging (MRI) analysis of the subchondral bone and the micro-computed tomography (CT) approach. METHODS: Samples obtained from 10 patients with osteoarthritis undergoing total hip arthroplasty were scanned with a 7.0 T micro-MRI. Proton density-weighted images and proton density-weighted images with fat suppression were obtained. The results were validated with a micro-CT device. Micro-MRI and micro-CT scans of the same sample were aligned, and regions of interest were delineated on equal areas of the sample. Bone volume fraction was calculated by using in-house plugins. The agreement between the methods was tested with Bland-Altman analysis. RESULTS: The agreement between the methods was good, with average difference of 2.167%. The differences between the methods were not significant (P=0.272, t test). CONCLUSION: The novel micro-MRI approach could be used for subchondral bone analysis. With further optimization for clinical MRI machines, the approach can be also used in the diagnostics of hip osteoarthritis.

Lentivirus- or AAV-mediated gene therapy interventions in ischemic stroke: A systematic review of preclinical in vivo studies.

Due to the limited therapeutic options after ischemic stroke, gene therapy has emerged as a promising choice, especially with recent advances in viral vector delivery systems. Therefore, we aimed to provide the current state of the art of lentivirus (LV) and adeno-associated virus (AAV) mediated gene interventions in preclinical ischemic stroke models. A systematic analysis including qualitative and quantitative syntheses of studies published until December 2020 was performed. Most of the 87 selected publications used adult male rodents and the preferred stroke model was transient middle cerebral artery occlusion. LV and AAV vectors were equally used for transgene delivery, however loads of AAVs were higher than LVs. Serotypes having broad cell tropism, the use of constitutive promoters, and virus delivery before the stroke induction via stereotaxic injection in the cortex and striatum were preferred in the analyzed studies. The meta-analysis based on infarct volume as the primary outcome confirmed the efficacy of the preclinical interventions. The quality assessment exposed publication bias and setbacks in regard to risks of bias and study relevance. The translational potential could increase by using specific cell targeting, post-stroke interventions, non-invasive systematic delivery, and use of large animals.

Prognostic Significance of BRAF V600E Mutation and CPSF2 Protein Expression in Papillary Thyroid Cancer.

The early-stage diagnosis of papillary thyroid cancer (PTC) has significantly increased in incidence worldwide without any beneficial impact on survival. In order to improve the risk assessment in PTC, we have conducted a retrospective study in which we analyzed the BRAF V600E mutation and CPSF2 protein expression as prognostic markers on archival tissue samples of 49 patients without (control group) and 97 patients with (study group) PTC metastases in the cervical lymph nodes at the time of initial diagnosis. Our aim was to correlate the BRAF V600E mutation and the expression of CPSF2 protein with the clinical and pathological features of PTC. The expression of CPSF2 protein was evaluated via immunohistochemistry and graded semi-quantitatively. The presence of the BRAF V600E mutation was determined via real-time polymerase chain reac-tion (PCR). CPSF2 protein < 3+ intensity expression was correlated with more frequent recurrences (Fisher-Freeman-Halton exact test; p = 0.010; 95% CI: 1.26−22.03), and patients who presented with the BRAF V600E mutation and CPSF2 protein expression < 3+ intensity had shorter disease-free survival (log-rank test; 105.0 months vs. 146.6 months; p < 0.001; HR 8.32, 95% CI: 2.91−23.83), whereas patients with PTC who had CPSF2 expression 3+ had longer disease-free survival in correlation with other lower intensity expressions of CPSF2 protein (log-rank test; 139.7 months vs. 129.6 months; p = 0.008). The multivariate analysis showed that younger patients with CPSF2 protein expression <3+ and the BRAF V600E mutation are at an increased risk for recurrence and require more intensive monitoring (Cox proportional hazards regression model; X2 = 17.5, df = 10, p = 0.025). Our results correlate the BRAF V600E mutation and CPSF2 protein expression with recurrence and disease-free survival as relevant prognostic factors for PTC.

Menstrual Cycle Control - A Controversial Example of Human Enhancement in Relation to Women's Body and Psyche.

By discussing the position of bio-conservatism and transhumanism, we question if the women menstrual cycle control would represent a new way toward a more responsive relation with one's own physical and mental health, a choice of freedom from undesired physiological conditions, a medicalization of a natural physiological event or an innovative carrier of social stigma against the women. We argue that the advancement of medical science may allow women a choice if to regulate own menstrual cycle, offering them as well a right to intervene responsibly on their own body and psyche. Accordingly, a post-human society could provide a suitable coexistence between women who claim menstruation as the biological essence and those who claim it as an option.

Self-Perceived Mental Health Status, Digital Activity, and Physical Distancing in the Context of Lockdown Versus Not-in-Lockdown Measures in Italy and Croatia: Cross-Sectional Study in the Early Ascending Phase of the COVID-19 Pandemic in March 2020.

The novelty of the coronavirus disease 2019 (COVID-19) pandemic is that it is occurring in a globalized society enhanced by digital capabilities. Our aim was to analyze the psychological and emotional states of participants in different pandemic-related contexts, with a focus on their digital and physical distancing behaviors. The online survey was applied during the ascending phase of the pandemic in March 2020 in two neighboring EU countries: Italy and Croatia. The study subjects involved four groups, two directly affected by epidemiological measures and two serving as controls-(1) participants from Italy who were in lockdown (Italy group), (2) participants from Croatia who were not in lockdown but who were in direct contact with an infected person and underwent epidemiological measures (CRO-contact group), (3) participants from Croatia who were in an analogous situation but not near the same infected person (CRO-no contact group), and (4) participants from Croatia who were not aware of any infected person (CRO-unrelated group). The survey consisted of validated scales of psychological and emotional states, and custom-made questionnaires on the digital (online) and physical (off-line) behavior of the participants. The Italy group in lockdown had higher self-perceived scores for depression, stress, post-traumatic intrusion, and avoidance, as well as the highest digital activity and physical distancing than the not-in-lockdown Croatian groups. The insight into the extent of online activities and off-line isolation allowed for the introduction of Digital Activity and Physical Distancing Scores. Self-perceived post-traumatic avoidance was higher in both the Italy and CRO-contact groups than the control CRO-no contact and CRO-unrelated groups, and higher avoidance correlated with higher Digital Activity and Physical Distancing Scores. Being in direct contact with the infected person, the CRO-contact group had no other alterations than unexpectedly lower post-traumatic hyperarousal when compared with the Italy group. The Italy group in lockdown demonstrated higher self-perceived psychological toll together with higher digital activity and physical distancing than Croatian groups not in lockdown, even when compared with the affected CRO-contact group. The study outcomes suggest that the general emergency measures influenced citizens in lockdown more than exposure to the virus through direct contact with an infected person.

RANK/RANKL/OPG Signaling in the Brain: A Systematic Review of the Literature.

Together with its dominant immunological and bone remodeling involvement, RRO axis, comprising of receptor activator of nuclear factor-κB (RANK), RANK ligand (RANKL), and osteoprotegerin (OPG) signaling, is as well-implicated in CNS functioning and corresponding pathologies. The CNS aspects of RANKL/RANK/OPG (RRO) axis were systematically reviewed. With search 10 databases, and 7 additional resources from first article publication to July 2019, resulted in total 2,222 hits, from which 33 relevant articles were selected. The elements of RRO axis in CNS include cells involved in neuroinflammation, predominantly in microglia, but as well in resident macrophages and inflammatory cells migrating across the blood-brain barrier. The expression in neurons and oligodendrocytes is mainly confined to processes of differentiation and cell death. RRO axis tunes the neuroinflammatory response, depending on the molecular, cellular and pathological context. RANK/RANKL signaling is neuroprotective in TLR-mediated inflammation, while OPG seems detrimental in stroke, but beneficial in multiple sclerosis. The levels of RRO axis elements can serve as biomarkers in the blood and cerebrospinal fluid. They act as neuroprotectant after brain damage even being implicated in body weight- and thermo-regulation. As derivatives of RRO axis already exist as therapeutic agents in bone remodeling, it would be intriquing to see if these or new RRO-based pharmaceuticals would appear effective in CNS therapies.

The innate immune toll-like-receptor-2 modulates the depressogenic and anorexiolytic neuroinflammatory response in obstructive sleep apnoea.

The increased awareness of obstructive sleep apnoea's (OSA) links to Alzheimer's disease and major psychiatric disorders has recently directed an intensified search for their potential shared mechanisms. We hypothesised that neuroinflammation and the microglial TLR2-system may act as a core process at the intersection of their pathophysiology. Moreover, we postulated that inflammatory-response might underlie development of key behavioural and neurostructural changes in OSA. Henceforth, we set out to investigate effects of 3 weeks' exposure to chronic intermittent hypoxia in mice with or without functional TRL2 (TLR2+/+, C57BL/6-Tyrc-Brd-Tg(Tlr2-luc/gfp)Kri/Gaj;TLR2-/-,C57BL/6-Tlr2tm1Kir). By utilising multimodal imaging in this established model of OSA, a discernible neuroinflammatory response was demonstrated for the first time. The septal nuclei and forebrain were shown as the initial key seed-sites of the inflammatory cascade that led to wider structural changes in the associated neurocircuitry. Finally, the modulatory role for the functional TLR2-system was suggested in aetiology of depressive, anxious and anorexiolytic symptoms in OSA.

Wildfire-Like Effect of a WhatsApp Campaign to Mobilize a Group of Predominantly Health Professionals With a University Degree on a Health Issue: Infodemiology Study.

BACKGROUND: Online interactions within a closed WhatsApp group can influence the attitudes and behaviors of the users in relation to health issues. OBJECTIVE: This study aimed to analyze the activity of the members of a WhatsApp group initiated to raise awareness of the possible health effects of 5G mobile networks and mobilize members to sign the related petition. METHODS: We retrospectively analyzed data from the WhatsApp group of 205 members that was active during 4 consecutive days in August 2019. The messages exchanged were collected, anonymized, and analyzed according to their timing and content. RESULTS: The WhatsApp group members were invited to the group from the administrator's contacts; 91% (187/205) had a university degree, 68% (140/205) were medical professionals, and 24% (50/205) held academic positions. Approximately a quarter of the members (47/205, 23%) declared in their messages they signed the corresponding petition. The intense message exchange had wildfire-like features, and the majority of messages (126/133, 95%) were exchanged during the first 26 hours. Despite the viral activity and high rate of members openly declaring that they signed the petition, only 8 (8/133, 6%) messages from the group members, excluding the administrator, referred to the health issue, which was the topic of the group. No member expressed an opposite opinion to those presented by the administrator, and there was no debate in the form of exchanging opposite opinions. CONCLUSIONS: The wildfire-like activity of the WhatsApp group and open declaration of signing the petition as a result of the mobilization campaign were not accompanied by any form of a debate related to the corresponding health issue, although the group members were predominantly health professionals, with a quarter of holding academic positions.

Face Masks During the COVID-19 Pandemic: A Simple Protection Tool With Many Meanings.

Wearing face masks is recommended as part of personal protective equipment and as a public health measure to prevent the spread of coronavirus disease 2019 (COVID-19) pandemic. Their use, however, is deeply connected to social and cultural practices and has acquired a variety of personal and social meanings. This article aims to identify the diversity of sociocultural, ethical, and political meanings attributed to face masks, how they might impact public health policies, and how they should be considered in health communication. In May 2020, we involved 29 experts of an interdisciplinary research network on health and society to provide their testimonies on the use of face masks in 20 European and 2 Asian countries (China and South Korea). They reflected on regulations in the corresponding jurisdictions as well as the personal and social aspects of face mask wearing. We analyzed those testimonies thematically, employing the method of qualitative descriptive analysis. The analysis framed the four dimensions of the societal and personal practices of wearing (or not wearing) face masks: individual perceptions of infection risk, personal interpretations of responsibility and solidarity, cultural traditions and religious imprinting, and the need of expressing self-identity. Our study points to the importance for an in-depth understanding of the cultural and sociopolitical considerations around the personal and social meaning of mask wearing in different contexts as a necessary prerequisite for the assessment of the effectiveness of face masks as a public health measure. Improving the personal and collective understanding of citizens' behaviors and attitudes appears essential for designing more effective health communications about COVID-19 pandemic or other global crises in the future.    To wear a face mask or not to wear a face mask?    Nowadays, this question has been analogous    to the famous line from Shakespeare's Hamlet:    "To be or not to be, that is the question."    This is a bit allegorical,    but certainly not far from the current circumstances    where a deadly virus is spreading amongst us... Vanja Kopilas, Croatia.

Impact of High Salt Diet on Cerebral Vascular Function and Stroke in Tff3-/-/C57BL/6N Knockout and WT (C57BL/6N) Control Mice.

High salt (HS) dietary intake leads to impaired vascular endothelium-dependent responses to various physiological stimuli, some of which are mediated by arachidonic acid (AA) metabolites. Transgenic Tff3-/- gene knockout mice (Tff3-/-/C57BL/6N) have changes in lipid metabolism which may affect vascular function and outcomes of stroke. We aimed to study the effects of one week of HS diet (4% NaCl) on vascular function and stroke induced by transient occlusion of middle cerebral artery in Tff3-/- and wild type (WT/C57BL/6N) mice. Flow-induced dilation (FID) of carotid artery was reduced in WT-HS mice, but not affected in Tff3-/--HS mice. Nitric oxide (NO) mediated FID. NO production was decreased with HS diet. On the contrary, acetylcholine-induced dilation was significantly decreased in Tff3-/- mice on both diets and WT-HS mice. HS intake and Tff3 gene depletion affected the structural components of the vessels. Proteomic analysis revealed a significant effect of Tff3 gene deficiency on HS diet-induced changes in neuronal structural proteins and acute innate immune response proteins' expression and Tff3 depletion, but HS diet did not increase the stroke volume, which is related to proteome modification and upregulation of genes involved mainly in cellular antioxidative defense. In conclusion, Tff3 depletion seems to partially impair vascular function and worsen the outcomes of stroke, which is moderately affected by HS diet.

D-mannose-Coating of Maghemite Nanoparticles Improved Labeling of Neural Stem Cells and Allowed Their Visualization by ex vivo MRI after Transplantation in the Mouse Brain.

Magnetic resonance imaging (MRI) of superparamagnetic iron oxide-labeled cells can be used as a non-invasive technique to track stem cells after transplantation. The aim of this study was to (1) evaluate labeling efficiency of D-mannose-coated maghemite nanoparticles (D-mannose(γ-Fe2O3)) in neural stem cells (NSCs) in comparison to the uncoated nanoparticles, (2) assess nanoparticle utilization as MRI contrast agent to visualize NSCs transplanted into the mouse brain, and (3) test nanoparticle biocompatibility. D-mannose(γ-Fe2O3) labeled the NSCs better than the uncoated nanoparticles. The labeled cells were visualized by ex vivo MRI and their localization subsequently confirmed on histological sections. Although the progenitor properties and differentiation of the NSCs were not affected by labeling, subtle effects on stem cells could be detected depending on dose increase, including changes in cell proliferation, viability, and neurosphere diameter. D-mannose coating of maghemite nanoparticles improved NSC labeling and allowed for NSC tracking by ex vivo MRI in the mouse brain, but further analysis of the eventual side effects might be necessary before translation to the clinic.