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The purpose of this research is to learn which plant species can best survive in polluted urban environments. Increasing the number of parks and other green areas has been found to greatly enhance air quality. Depending on a number of criteria, including the climatic conditions of the region and the amount and density of vegetation, green spaces have the potential to minimize industrially produced pollutants, and the use of vegetation cover can increase environmental quality. Growing more parks and other green areas in cities can help cut down on pollution caused by cars and factories. Important implications for selecting plant species for vegetative cover may be found in this study. The creation of green spaces and the use of afforestation in public locations within cities are two examples of the nature-based measures that many countries are embracing to reduce air pollution. The purpose of this essay was to investigate how tolerant various plant species are to air pollution in suburban and urban environments. Different plant species play a substantial role in lowering environmental pollutants such as NO2 and O3. Plants have been shown to minimize air pollution, although this benefit may come at the expense of the plants themselves. Planners are paying more attention to greenery planting as a technique of mitigating air pollution due to the proliferation of pollution sources. Trees and shrubs in these landscapes are chosen for their ability to thrive in the region's unique climatic, soil, and water conditions, as well as their other desirable qualities, such as their height, crown shape, root type, leaf structure, heat and cold resistance, and adaptability to a range of conditions. The hazards associated with pollution emissions are reduced, and air quality is enhanced, thanks to this.

Fungal keratitis is a rare yet severe infection of the cornea. Fungal species distribution depends on the climate and socioeconomic status and can show regional variation. This retrospective single-center study was conducted at a tertiary eye care center and the collaborating Institute of Medical Microbiology in Switzerland. On investigating all fungal-positive corneal scrapings and contact lens assessments of patients with keratitis from January 2012 to December 2023, 206 patients were identified, of which 113 (54.9%) were female. The median age was 38 (IQR 29.8, [18–93]), and 154 (74.8%) applied contact lenses. The most commonly found pathogen was Candida spp., followed by Fusarium spp. Molds were 1.8 times more common than yeasts. Linear regression showed no significant increase or decrease in the infection rate over time (p = 0.5). In addition, 10 patients (4.9%) were found to have coinfections with Acanthamoeba, 11 (5.3%) with HSV-1, none with HSV-2, and 4 (1.9%) with VZV. This study provides a long-term overview of fungal-positive corneal scrapings and contact lens specimens of patients with fungal keratitis. Based on our results, coinfections with Acanthamoeba, HSV, and VZV are frequent, especially in patients wearing contact lenses. Thus, wearing contact lenses may facilitate coinfection in fungal keratitis.

Ophthalmologists’ diagnostic and treatment competence in Acanthamoeba keratitis varies widely. This investigator-initiated, retrospective, single-center chart review examined the electronic patient files regarding PCR-positive Acanthamoeba keratitis. We included corneal and contact lens assessments. We further reviewed the patient’s medical history, corneal scraping results regarding viral or fungal co-infections, and the duration from symptom onset to final diagnosis. We identified 59 eyes of 52 patients from February 2010 to February 2023, with 31 of 52 (59.6%) being female patients. The median (IQR, range) patient age was 33 (25.3 to 45.5 [13 to 90]) years, and the mean (SD, range) time to diagnosis after symptom onset was 18 (10.5 to 35 [3 to 70]) days. Overall, 7 of 52 (7.7%) patients displayed a bilateral Acanthamoeba infection, and 48 (92.3%) used contact lenses at symptom onset. Regarding other microbiological co-infections, we found virologic PCR testing in 45 of 52 (86.5%) patients, with 3 (6.7%) positive corneal scrapings. Fungal cultures were performed in 49 of 52 (94.2%) patients, with 5 (10.2%) positive corneal scrapings. The medical treatment success rate was 45/46 (97.8%). This study raises awareness of patient education in contact lens handling and screens for further microbial co-infections in suspected Acanthamoeba cases.

The environmental behavior of rock and oil pollutants in soil has long been a focus of environmental protection research. In this research, a laboratory model was established to study the oil pollutant transfer process in two stages. Two samples from porous media of sandy soil and agricultural soil (organic) were used with oil as a pollutant. The oil is pumped through a system consisting of two pipes with a length of 4 m and a diameter of 0.07 m. Each pipe is assigned to a specific type of soil. The results showed that the organic soil needs a long time for the pollutants to travel over greater distances, while the sandy soil showed its ability to absorb and drain the pollutants and get rid of them faster. However, it was also found that both soils contain a small percentage of the pollutant after the water washing process, which lasts for more than (5 hours). The remaining oil percentage is estimated at about (0.1 ml), which is a very small percentage, and it is possible to get rid of it by increasing the washing time. It was found that the physical properties of the soil such as permeability and porosity have a significant effect on slowing down the speed of the pollutant and its transmission through the porous medium.

Computer networks form much of the infrastructure supporting day-to-day life in this digital age. Computer networks, however, are prone to attack and therefore require intrusion detection systems. Intrusion detection systems provide a mechanism to detect network attacks at an early stage and generate alerts. These systems, however, are far from a panacea. Rather, they tend to overwhelm their operators with alerts, which in more than 90% of cases can be false positives. As such, the problem of false positives in intrusion detection systems is a costly issue. This paper presents research to design a hierarchical network intrusion detector, using deep learning, which protects against raising vast numbers of false positives through the design and implementation of a hierarchical NIDS. This paper presents a valuable advancement in performance by reducing the occurrence of false alarms by 87.52%. The research contained in this paper presents three contributions to knowledge. The first of these is the comparison between hierarchical systems and non-hierarchical systems to understand which would yield fewer false alarms. The second contribution is the formulation of a hierarchical approach, which was able to reduce false alarms by 87.52%. Lastly, the proposed hierarchical model was deployed in a live IoT environment, exposed to genuine threats, and the performance in this environment was analysed.

Objectives: The COVID-19 pandemic is a major health crisis that emerged at the end of the first quarter of the 21st century and changed the lives of millions of people globally. Nurses have close contact with infectious patients; therefore, nurses need to obtain sufficient knowledge in this regard. They need to assess their level of knowledge about COVID-19 and explore their needs to enhance and improve their knowledge in order to be more qualified to fight this virus. This study intends to assess nurses’ knowledge regarding COVID-19 and to find out the differences in nurses’ knowledge about COVID-19 with regard to their demographic variables. Methods: A descriptive (cross-sectional design) study was conducted to assess nurses’ knowledge regarding COVID-19 in Al-Hilla teaching hospitals in Al-Hilla City, Iraq, from September 25, 2020, to February 20, 2022. Using a convenient sampling method, a sample of 200 nurses working at epidemic hospitals was selected. The data collection process began with a questionnaire, which nurses answered as a self-report (questionnaire). Then the questionnaires were collected from the respondents. The average time to fill out the questionnaire was between 10 and 20 minutes. Results: The findings indicate that most nurses (58%) had poor knowledge about COVID-19. There were significant differences in nurses’ knowledge about COVID-19 with regard to their educational levels (P0.05), but there was a significant difference between nurses’ knowledge about COVID-19 with regard to their source of information about COVID-19 (P

Abstract Background Cognitive ergonomics design of patient monitoring may reduce human factor errors in high-stress environments. Eye-tracking is a suitable tool to gain insight into the distribution of visual attention of healthcare professionals with patient monitors, which may facilitate their further development. Methods This prospective, exploratory, high-fidelity simulation study compared anesthesia personnel’s visual attention (fixation count and dwell-time) to 15 areas of interest on the patient monitor during non-critical and critical anesthesia situations. Furthermore, we examined the extent to which participants’ experience influenced visual attention and which vital signs displayed on the patient monitor received the most visual attention. We used mixed zero-inflated Poisson regression and mixed linear models to analyze the data. Results Analyzing 23 ten-minute scenarios, we found significantly more fixations to the areas of interest on the patient monitor during critical than non-critical situations (rate ratio of 1.45; 95% CI 1.33 to 1.59; p

Using multimodal imaging, the literature proposed the following risk factors for choroidal nevus growth into melanoma: increased tumor thickness, subretinal fluid, decreased visual acuity, presence of orange pigment, ultrasound acoustic hollowness, and increased tumor diameter. This study investigated the presence of the mentioned risk factors in choroidal nevi, choroidal melanomas, and indeterminate choroidal melanocytic lesions. This retrospective, single-center chart review assessed choroidal melanocytic tumors with multimodal imaging. We defined our primary outcome as the cumulative presence of mentioned risk factors. Further, we evaluated various optical coherence tomography (OCT), ultrasound, and autofluorescence findings. We analyzed 51 tumors from 49 patients during the period from April 2008 to June 2021. The median (IQR) age was 64.0 (56.0 to 70.5) years, with 23 of 49 (46.9%) patients being female. The follow-up time for all tumors was median (IQR) 25.0 (12.0 to 39.0) months. The choroidal nevi had a median (range) risk score of 0.0 (0.0 to 3.0), and the choroidal melanoma of 5.0 (3.0 to 6.0), with statistically significant different ratings (p < 0.001). Multimodal imaging creates a score that may help to distinguish choroidal nevi from choroidal melanomas objectively.

Abstract Background Acute bleeding is an omnipresent challenge for all physicians. Uncontrolled hemorrhage is the most common preventable cause of death after trauma worldwide. In different surgical disciplines, hemorrhage represents an independent risk factor for increased postoperative morbimortality, directly affecting patients’ outcomes. This study asked anesthesiologists about their personal perceived challenges when treating bleeding patients. Methods This investigator-initiated, prospective, international, dual-center, mixed qualitative and quantitative study interrogated anesthesiologists about what they found easy and what difficult in treating acutely bleeding patients. Following the template approach for qualitative research, we identified major and minor topics through free inductive coding and word count. In a second step, we derived ten statements from the participants’ answers. Using a field survey, we then asked the participants to rate their level of agreement with the derived statements. We analyzed the answers using one sample Wilcoxon test and the Mann-Whitney test. Results We included a total of 84 physicians in the qualitative interrogations and a different group of 42 anesthesiologists in the quantitative part. We identified 11 major topics and 19 associated subtopics. The main topics and the degree of agreement (here as agree or strongly agree) were as follows: “Complexity of the topic” (52.4% agreed to find the topic complex), “Cognitive aids” (92.9% agreed to find them helpful), “Time management” (64.3% agreed to feeling time pressure), “Human factors” (95.2% agreed that human factors are essential), “Resources” (95.2% agreed that resources are essential), “Experience” and “Low frequency of cases” (57.1% agreed to lack practice), “Diagnostic methods” (31.0% agreed that the interpretation of test results is difficult), “Anticoagulation” (85.7% agreed to it being difficult), “Treatment” (81.0% agreed to knowing the first therapeutic steps), and “Nothing”. Conclusions Anesthesiologists in two large tertiary care facilities in different parts of the world found coagulation management, especially in anticoagulated patients, complex. We identified the delayed diagnostic test results and their interpretation as challenges. Resources, treatment protocols and human factors such as team communication were perceived to facilitate management. Future studies should explore the challenges in smaller hospitals and other parts of the world and test new technologies addressing the identified difficulties.

Purpose: To compare retinal changes in young adults with previous SARS-CoV-2 infection with healthy young controls using optical coherence tomography (OCT) and optical coherence tomography angiography (OCTA). Methods: This prospective single-center study was conducted at the University Hospital of Zurich, Zurich, Switzerland. Participants were imaged from May to November 2021 using the SOLIX device (Visionix International SAS, Pont-de-l’Arche, France). We performed 12 mm × 12 mm, 6.4 mm × 6.4 mm, 6 mm × 6 mm and 3 mm × 3 mm OCT and OCTA scans, as well as fundus photography of each participant’s eyes. Results: In total, 466 participants were imaged. Of these, 233 were healthy controls with negative RT-PCR tests for SARS-CoV-2, 168 were young adults who had a SARS-CoV-2 infection at least 180 days previously, 19 were participants who had a SARS-CoV-2 infection < 180 days previously, and 46 were participants with asymptomatic SARS-CoV-2 infection (i.e., serologically positive but with no symptoms). Compared with healthy controls, statistically significant differences were found for OCTA recordings of the optic disc for the whole image (WI) and WI capillary vessel density, with both being higher in the SARS-CoV-2 group. Conclusion: Statistically significant results were only observed for selected variables, and in parts, only unilaterally, with relatively large p values (p = 0.02–0.03). Thus, we did not interpret these as clinically significant, leading to the conclusion that young and otherwise healthy individuals (mainly men) seem to recover from mild COVID-19 infections with no ophthalmological residues.

BackgroundPatient safety during anesthesia is crucially dependent on the monitoring of vital signs. However, the values obtained must also be perceived and correctly classified by the attending care providers. To facilitate these processes, we developed Visual-Patient-avatar, an animated virtual model of the monitored patient, which innovatively presents numerical and waveform data following user-centered design principles. After a high-fidelity simulation study, we analyzed the participants’ perceptions of 3 different monitor modalities, including this newly introduced technique. ObjectiveThe aim of this study was to collect and evaluate participants’ opinions and experiences regarding 3 different monitor modalities, which are Visual-Patient-avatar, Split Screen (avatar and Conventional monitor alongside each other), and Conventional monitor after using them during simulated critical anesthetic events. MethodsThis study was a researcher-initiated, single-center, semiquantitative study. We asked 92 care providers right after finishing 3 simulated emergency scenarios about their positive and negative opinions concerning the different monitor modalities. We processed the field notes obtained and derived the main categories and corresponding subthemes following qualitative research methods. ResultsWe gained a total of 307 statements. Through a context-based analysis, we identified the 3 main categories of “Visual-Patient-avatar,” “Split Screen,” and “Conventional monitor” and divided them into 11 positive and negative subthemes. We achieved substantial interrater reliability in assigning the statements to 1 of the topics. Most of the statements concerned the design and usability features of the avatar or the Split Screen mode. ConclusionsThis study semiquantitatively reviewed the clinical applicability of the Visual-Patient-avatar technique in a high-fidelity simulation study and revealed the strengths and limitations of the avatar only and Split Screen modality. In addition to valuable suggestions for improving the design, the requirement for training prior to clinical implementation was emphasized. The responses to the Split Screen suggest that this symbiotic modality generates better situation awareness in combination with numerical data and accurate curves. As a subsequent development step, a real-life introduction study is planned, where we will test the avatar in Split Screen mode under actual clinical conditions.

BackgroundInadequate situational awareness accounts for two-thirds of preventable complications in anesthesia. An essential tool for situational awareness in the perioperative setting is the patient monitor. However, the conventional monitor has several weaknesses. Avatar-based patient monitoring may address these shortcomings and promote situation awareness, a prerequisite for good decision making. ObjectiveThe spatial distribution of visual attention is a fundamental process for achieving adequate situation awareness and thus a potential quantifiable surrogate for situation awareness. Moreover, measuring visual attention with a head-mounted eye-tracker may provide insights into usage and acceptance of the new avatar-based patient monitoring modality. MethodsThis prospective eye-tracking study compared anesthesia providers' visual attention on conventional and avatar-based patient monitors during simulated critical anesthesia events. We defined visual attention, measured as fixation count and dwell time, as our primary outcome. We correlated visual attention with the potential confounders: performance in managing simulated critical anesthesia events (task performance), work experience, and profession. We used mixed linear models to analyze the results. ResultsFifty-two teams performed 156 simulations. After a manual quality check of the eye-tracking footage, we excluded 57 simulations due to technical problems and quality issues. Participants had a median of 198 (IQR 92.5-317.5) fixations on the patient monitor with a median dwell time of 30.2 (IQR 14.9-51.3) seconds. We found no significant difference in participants' visual attention when using avatar-based patient monitoring or conventional patient monitoring. However, we found that with each percentage point of better task performance, the number of fixations decreased by about 1.39 (coefficient –1.39; 95% CI –2.44 to –0.34; P=.02), and the dwell time diminished by 0.23 seconds (coefficient –0.23; 95% CI: –0.4 to –0.06; P=.01). ConclusionsUsing eye tracking, we found no significant difference in visual attention when anesthesia providers used avatar-based monitoring or conventional patient monitoring in simulated critical anesthesia events. However, we identified visual attention in conjunction with task performance as a surrogate for situational awareness.

Oil palm agro-industry is a major revenue earner for Malaysia with the country being one of the major producers of crude palm oil (CPO) and oil palm products. Its growth has, however, led to massive water consumption and high generation of highly polluting palm oil mill effluent (POME). The inadequacies of ponding system adopted by most palm oil mills (POMs) for the treatment of POME to alleviate environmental and public health concerns are quite alarming. Fenton advanced oxidation technologies are a current research area providing viable alternatives for POME treatment, recovery and managing high demand for water. Its major setback is the generation of a large amount of unwanted sludge of iron (III) complexes thus increasing the costs of sludge management, treatment, and disposal. The salient and promising features of this technique for industrial applications motivate researchers to find ways to overcome its inherent drawbacks. This brief review aimed at discussing and evaluating the performances of the various Fenton oxidation processes, including homogeneous, heterogeneous, photo-Fenton, electro-Fenton, sono-Fenton etc., for POME treatment. Discussions on the future direction of these Fenton processes points towards the utilization of abundant magnetically separable heterogeneous composites as catalysts with high stability, activity, recyclability, and cost-effectiveness for decontamination of POME and other agro-industrial effluents from recalcitrant organic pollutants. The low deployment of such composite catalyst coupled with scarce literature on POME treatment in this regard offers a vast opportunity for research exploration.

Liquid-liquid two-phase flow has been studied widely by many authors. Two phase flow exist in petroleum industries, nuclear power generation and chemical plants. In the present study a review on two phase flow such as oil/water was discussed, also the patterns of the flow and types of measurement are presented. Also a review on the new trends using the computational fluid dynamic (CFD) to predict the liquid-liquid two phase are viewed.

Brain Tumour is a severe condition caused due to abnormal growth of cells in the brain. Brain Tumour is broadly classified into two categories namely Malignant (Cancerous) and Benign (Non-Cancerous). As tumour grows, the pressure within the skull can increase which can damage the brain and be life-threatening. Early detection and classification of the brain tumours is important as it helps to select the most appropriate treatment for saving the patient’s life. Usually, Brain Tumour Detection can be done manually by the doctors or use machine learning models in case of MRI images of the brain. In literature, it is identified that deep learning techniques such as CNN, DCNN and RNN show good results in image processing applications. This paper aims to detect and classify the Brain Tumours effectively using CNN deep learning techniques. The dataset is collected from Kaggle. The proposed method achieved an accuracy of 93.5% and 98.4% with CNN and Resnet50 respectively.

As the interpretation of viscoelastic coagulation test results remains challenging, we created Visual Clot, an animated blood clot aiming to facilitate raw rotational thromboelastometry (ROTEM) parameters. This study investigated anesthesia personnel’s cognitive processing in managing simulated bleeding scenarios using eye-tracking technology. This multicenter, international, computer-based study across five large, central European hospitals included 35 participants with minimal to no prior experience interpreting viscoelastic test results. Using eye-tracking technology and an iPad tagged with quick response codes, we defined the time to treatment decision and the time on screen surface in seconds of correctly solved scenarios as our outcomes. The median time to treatment decision was 52 s for Visual Clot and 205 s for ROTEM (p < 0.0001). The probability of solving the scenario correctly was more than 8 times higher when using Visual Clot than when using ROTEM (Hazard ratio [HR] 8.54, 95% CI from 6.5 to 11.21; p < 0.0001). Out of 194 correctly answered scenarios of participants with the eye-tracker, 154 (79.4%) were solved with Visual Clot and 40 (20.6%) with ROTEM. Participants spent on average 30 s less looking at the screen surface with Visual Clot compared to ROTEM (Coefficient −30.74 s, 95% CI from −39.27 to −22.27; p < 0.0001). For a comparison of the two modalities in terms of information transfer, we calculated the percentage of time on the screen surface of the overall time to treatment decision, which with Visual Clot was 14 percentage points shorter than with ROTEM (Coefficient −14.55, 95% CI from −20.05 to −9.12; p < 0.0001). Visual Clot seems to improve perception and detection of coagulopathies and leads to earlier initiation of the appropriate treatment. In a high-pressure working environment such as the operating and the resuscitation room, correct and timely decisions regarding bleeding management may have a relevant impact on patients’ outcomes.

Visual-Patient-avatar, an avatar-based visualisation of patient monitoring, is a newly developed technology aiming to promote situation awareness through user-centred design. Before the technology’s introduction into clinical practice, the initial design used to validate the concept had to undergo thorough examination and adjustments where necessary. This mixed qualitative and quantitative study, consisting of three different study parts, aimed to create a design with high user acceptance regarding perceived professionalism and potential for identification while maintaining its original functionality. The first qualitative part was based on structured interviews and explored anaesthesia personnel’s first impressions regarding the original design. Recurrent topics were identified using inductive coding, participants’ interpretations of the vital sign visualisations analysed and design modifications derived. The second study part consisted of a redesign process, in which the visualisations were adapted according to the results of the first part. In a third, quantitative study part, participants rated Likert scales about Visual-Patient-avatar’s appearance and interpreted displayed vital signs in a computer-based survey. The first, qualitative study part included 51 structured interviews. Twenty-eight of 51 (55%) participants mentioned the appearance of Visual-Patient-avatar. In 23 of 51 (45%) interviews, 26 statements about the general impression were identified with a balanced count of positive (14 of 26) and negative (12 of 26) comments. The analysis of vital sign visualisations showed deficits in several vital sign visualisations, especially central venous pressure. These findings were incorporated into part two, the redesign of Visual-Patient-avatar. In the subsequent quantitative analysis of study for part three, 20 of 30 (67%) new participants agreed that the avatar looks professional enough for medical use. Finally, the participants identified 73% (435 of 600 cases) of all vital sign visualisations intuitively correctly without prior instruction. This study succeeded in improving the original design with good user acceptance and a reasonable degree of intuitiveness of the new, revised design. Furthermore, the study identified aspects relevant for the release of Visual-Patient-avatar, such as the requirement for providing at least some training, despite the design’s intuitiveness. The results of this study will guide further research and improvement of the technology. The study provides a link between Visual-Patient-avatar as a scientific concept and as an actual product from a cognitive engineering point of view, and may serve as an example of methods to study the designs of technologies in similar contexts.

The influence of variable reaction time (tr) on surface/textural properties (surface area, total pore volume, and pore diameter) of carbon-encapsulated magnetite (Fe3O4@C) nanocomposites fabricated by a hydrothermal process at 190 °C for 3, 4, and 5 h was studied. The properties were calculated using the Brunauer–Emmett–Teller (BET) isotherms data. The nanocomposites were characterised using Fourier transform infrared spectroscopy, X-ray diffraction analysis, thermogravimetry, and scanning and transmission electron microscopies. Analysis of variance shows tr has the largest effect on pore volume (F value = 1117.6, p value < 0.0001), followed by the surface area (F value = 54.8, p value < 0.0001) and pore diameter (F value = 10.4, p value < 0.001) with R2-adjusted values of 99.5%, 88.5% and 63.1%, respectively. Tukey and Fisher tests confirmed tr rise to have caused increased variations in mean particle sizes (11–91 nm), crystallite sizes (5–21 nm), pore diameters (9–16 nm), pore volume (0.017–0.089 cm3 g−1) and surface area (7.6–22.4 m2 g−1) of the nanocomposites with individual and simultaneous confidence limits of 97.9 and 84.4 (p-adj < 0.05). The nanocomposites’ retained Fe–O vibrations at octahedral (436 cm−1) and tetrahedral (570 cm−1) cubic ferrite sites, modest thermal stability (37–60 % weight loss), and large volume-specific surface area with potential for catalytic application in advanced oxidation processes.

Visual Patient technology is a situation awareness-oriented visualization technology that translates numerical and waveform patient monitoring data into a new user-centered visual language. Vital sign values are converted into colors, shapes, and rhythmic movements—a language humans can easily perceive and interpret—on a patient avatar model in real time. In this review, we summarize the current state of the research on the Visual Patient, including the technology, its history, and its scientific context. We also provide a summary of our primary research and a brief overview of research work on similar user-centered visualizations in medicine. In several computer-based studies under various experimental conditions, Visual Patient transferred more information per unit time, increased perceived diagnostic certainty, and lowered perceived workload. Eye tracking showed the technology worked because of the way it synthesizes and transforms vital sign information into new and logical forms corresponding to the real phenomena. The technology could be particularly useful for improving situation awareness in settings with high cognitive demand or when users must make quick decisions. This comprehensive review of Visual Patient research is the foundation for an evaluation of the technology in clinical applications, starting with a high-fidelity simulation study in early 2020.

Agaricus bisporus and Agaricus bitorquis are among most favorite mushrooms of the world. Mineral contents, proximate composition and fatty acid profile of these mushrooms were determined for evaluation of their dietary value. Na, K and Li contents were high, while Cr, Cu, Pb, Co, Zn, Mn and Ni concentrations were low in both Agaricus bisporus and Agaricus bitorquis. Proximate analysis showed that both mushrooms have sufficient quantity of protein, lipids, carbohydrates and cellulose. Gas chromatographic analysis of fatty acid methyl esters revealed that linoleic acid (44.19 %) and oleic acid (40.13 %) were the dominant fatty acids in Agaricus bisporus and Agaricus bitorquis respectively

Abstract Background The secretory leukocyte protease inhibitor (SLPI) exerts wide ranging effects on inflammatory pathways and is upregulated in EAE but the biological role of SLPI in EAE, an animal model of multiple sclerosis is unknown Methods To investigate the pathophysiological effects of SLPI within EAE, we induced SLPI-neutralizing antibodies in mice and rats to determine the clinical severity of the disease. In addition we studied the effects of SLPI on the anti-inflammatory cytokine TGF-β. Results The induction of SLPI neutralizing antibodies resulted in a milder disease course in mouse and rat EAE. SLPI neutralization was associated with increased serum levels of TGF-β and increased numbers of FoxP3+ CD4+ T cells in lymph nodes. In vitro, the addition of SLPI significantly decreased the number of functional FoxP3+ CD25hi CD4+ regulatory T cells in cultures of naive human CD4+ T cells. Adding recombinant TGF-β to SLPI-treated human T cell cultures neutralized SLPI's inhibitory effect on regulatory T cell differentiation. Conclusion In EAE, SLPI exerts potent pro-inflammatory actions by modulation of T-cell activity and its neutralization may be beneficial for the disease.

Aim: The aim of this study was to follow the instruments' pathways and cost each segment to calculate whether reusable or disposable ophthalmic instruments offer better value for money for intravitreal injections., Methods: The cycles and costs of reusable and single-use disposable instruments used for intravitreal injections were mapped out, including purchase costs, transport to and from the place of use, opening and disposal, sterilisation, replacement, salary costs of staff involved, etc. results: The cost of using reusable instruments for intravitreal injections (NZ$29.00) was lower than the cost of using disposable instruments ($30.51) by $1.51 per patient., Conclusions: Intravitreal injections performed with reusable instruments offer better value for money than when performed with disposable instruments. This equates to a beneficial financial saving just for this one low-complexity case. Such savings can multiply significantly when considering the instruments used in a wider variety of ophthalmic procedures. There are of course trade-offs between safety, quality, cost and sustainability., Competing Interests: The authors declare no competing/conflicts of interest. Originally presented as a video poster at the 2022 Royal Australian and New Zealand College of Ophthalmologists New Zealand Branch Annual Scientific Meeting., (© PMA.)

Background: Mesenchymal stem cell-neural progenitors (MSC-NPs) are a bone marrow mesenchymal stem cell (MSC)-derived ex vivo manipulated cell product with therapeutic potential in multiple sclerosis (MS). The objective of this study was to determine efficacy of intrathecal (IT) MSC-NP treatment in patients with progressive MS., Methods: The study is a phase II randomized, double-blind, placebo-controlled clinical trial with a compassionate crossover design conducted at a single site. Subjects were stratified according to baseline Expanded Disability Status Scale (EDSS) (3.0-6.5) and disease subtype (secondary or primary progressive MS) and randomized into either treatment or placebo group to receive six IT injections of autologous MSC-NPs or saline every two months. The primary outcome was EDSS Plus, defined by improvement in EDSS, timed 25-foot walk (T25FW) or nine-hole peg test. Secondary outcomes included the individual components of EDSS Plus, the six-minute walk test (6MWT), urodynamics testing, and brain atrophy measurement., Results: Subjects were randomized into MSC-NP (n = 27) or saline (n = 27) groups. There was no difference in EDSS Plus improvement between the MSC-NP (33%) and saline (37%) groups. Exploratory subgroup analysis demonstrated that in subjects who require assistance for ambulation (EDSS 6.0-6.5) there was a significantly higher percentage of improvement in T25FW and 6MWT in the MSC-NP group (3.7% ± 23.1% and - 9.2% ± 18.2%) compared to the saline group (-54.4% ± 70.5% and - 32.1% ± 30.0%), (p = 0.030 and p = 0.036, respectively). IT-MSC-NP treatment was also associated with improved bladder function and reduced rate of grey matter atrophy on brain MRI. Biomarker analysis demonstrated increased MMP9 and decreased CCL2 levels in the cerebrospinal fluid following treatment., Conclusion: Results from exploratory outcomes suggest that IT-MSC-NP treatment may be associated with a therapeutic response in a subgroup of MS patients., Trial Registration: ClinicalTrials.gov NCT03355365, registered November 14, 2017, https://clinicaltrials.gov/study/NCT03355365?term=NCT03355365&rank=1 ., (© 2024. The Author(s).)

Mesenchymal stem cell-neural progenitors (MSC-NP) are a neural derivative of MSCs that are being investigated in clinical trials as an autologous intrathecal cell therapy to treat patients with secondary progressive (SP) or primary progressive (PP) multiple sclerosis (MS). MSC-NPs promote tissue repair through paracrine mechanisms, however which secreted factors mediate the therapeutic potential of MSC-NPs and how this cell population differs from MSCs remain poorly understood. The objective of this study was to define the transcriptional profile of MSCs and MSC-NPs from MS and non-MS donors to better characterize each cell population. MSCs derived from SPMS, PPMS, or non-MS bone marrow donors demonstrated minimal differential gene expression, despite differences in disease status. MSC-NPs from both MS and non-MS-donors exhibited significant differential gene expression compared to MSCs, with 2,156 and 1,467 genes upregulated and downregulated, respectively. Gene ontology analysis demonstrated pronounced downregulation of cell cycle genes in MSC-NPs compared to MSC consistent with reduced proliferation of MSC-NPs in vitro. In addition, MSC-NPs demonstrated significant enrichment of genes involved in cell signaling, cell communication, neuronal differentiation, chemotaxis, migration, and complement activation. These findings suggest that increased cell signaling and chemotactic capability of MSC-NPs may support their therapeutic potential in MS., Competing Interests: I have read the journal’s policy and the authors of this manuscript have the following competing interests: Authors Violaine Harris and Saud Sadiq are listed as inventors on US patent #US 8,642,331, which is related to the study presented in the manuscript. The patent is issued to the Tisch MS Research Center of New York and is considered a non-financial competing interest. This does not alter our adherence to PLOS ONE policies on sharing data or materials. Furthermore, Violaine Harris and Saud Sadiq have no other relevant declarations related to employment, consultancy, products in development, etc. The remaining 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: © 2023 Harris et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Background/aims: Retinal microvascular ischaemia may produce localised middle retinal disruption with corresponding scotoma, a phenomenon termed paracentral acute middle maculopathy (PAMM). Small chronic middle retinal atrophic lesions termed retinal ischaemic perivascular lesions (RIPLs) appear qualitatively similar to PAMM lesions and have recently been hypothesised to result specifically from PAMM. However, no studies have quantitatively demonstrated an ischaemic origin of RIPLs. We quantitatively investigated the pathophysiology of RIPLs and their relationship with PAMM with swept-source optical coherence tomography angiography (SS-OCTA)., Methods: A total of 14 controls and 25 patients being evaluated for carotid artery stenosis (CAS) were enrolled. SS-OCTA imaging of each eye was taken. Projection-resolved en face 6 mm × 6 mm superficial capillary plexus (SCP) and deep capillary plexus (DCP) images were quantitatively analysed with two algorithms for changes in vessel linear density (VLD) and vessel tortuosity (VT) at RIPLs relative to both the immediately surrounding macula and the entire macula, as well as between eyes with RIPLs and eyes without RIPLs., Results: All controls and 22 of 25 CAS patients were included in the analysis. RIPLs demonstrated a localised decrease in DCP VLD in CAS patients and controls. RIPLs tended to show a localised decrease in SCP VLD in CAS patients but a localised increase in controls. No changes in VT were found. Eyes with RIPLs had VLD and VT similar to their RIPL-free fellow eyes., Conclusion: RIPLs are associated with quantifiable local, but not global, ischaemia, supporting the idea of shared pathophysiology with classic PAMM lesions along a continuum of ischaemia severity., Competing Interests: Competing interests: None declared., (© Author(s) (or their employer(s)) 2023. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.)

Multiple sclerosis patients treated with anti-CD20 therapy (aCD20-MS) are considered especially vulnerable to complications from SARS-CoV-2 infection due to severe B-cell depletion with limited viral antigen-specific immunoglobulin production. Therefore, multiple vaccine doses as part of the primary vaccination series and booster updates have been recommended for this group of immunocompromised individuals. Even though much less studied than antibody-mediated humoral responses, T-cell responses play an important role against CoV-2 infection and are induced efficiently in vaccinated aCD20-MS patients. For individuals with such decoupled adaptive immunity, an understanding of the contribution of T-cell mediated immunity is essential to better assess protection against CoV-2 infection. Here, we present results from a prospective, single-center study for the assessment of humoral and cellular immune responses induced in aCD20-MS patients (203 donors/350 samples) compared to a healthy control group (43/146) after initial exposure to CoV-2 spike antigen and subsequent re-challenges. Low rates of seroconversion and RBD-hACE2 blocking activity were observed in aCD20-MS patients, even after multiple exposures (responders after 1st exposure = 17.5%; 2nd exposure = 29.3%). Regarding cellular immunity, an increase in the number of spike-specific monofunctional IFNγ + -, IL-2 + -, and polyfunctional IFNγ + /IL-2 + -secreting T-cells after 2nd exposure was found most noticeably in healthy controls. Nevertheless, a persistently higher T-cell response was detected in aCD20-MS patients compared to control individuals before and after re-exposure (mean fold increase in spike-specific IFNγ + -, IL-2 + -, and IFNγ + /IL-2 + -T cells before re-exposure = 3.9X, 3.6X, 3.5X/P< 0.001; after = 3.2X, 1.4X, 2.2X/P = 0.002, P = 0.05, P = 0.004). Moreover, cellular responses against sublineage BA.2 of the currently circulating omicron variant were maintained, to a similar degree, in both groups (15-30% T-cell response drop compared to ancestral). Overall, these results highlight the potential for a severely impaired humoral response in aCD20-MS patients even after multiple exposures, while still generating a strong T-cell response. Evaluating both humoral and cellular responses in vaccinated or infected MS patients on B-cell depletion therapy is essential to better assess individual correlations of immune protection and has implications for the design of future vaccines and healthcare strategies., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Alfonso-Dunn, Lin, Lei, Liu, Roche, De Oliveira, Raisingani, Kumar, Kirschner, Feuer, Malin and Sadiq.)

Multiple sclerosis is clinically characterized by relapses and remissions (relapsing-remitting multiple sclerosis) that over time may evolve to a progressive course (secondary progressive multiple sclerosis) or as having a progressive course from disease onset (primary progressive multiple sclerosis). At present, it is not definitively known whether these clinical entities constitute a single pathological disease or whether these manifestations represent two distinct disease entities sharing inflammatory demyelination as a pathological feature. Here we show using a novel mouse model that CSF of primary progressive multiple sclerosis patients is unique in its capacity to induce motor disability and spinal cord pathology including demyelination, impaired remyelination, reactive astrogliosis and axonal damage. Notably, removal of immunoglobulin G from primary progressive multiple sclerosis CSF via filtration or immunodepletion attenuates its pathogenic capacity. Furthermore, injection of recombinant antibodies derived from primary progressive multiple sclerosis CSF recapitulates the pathology. Our findings suggest that the clinical and pathological features of primary progressive multiple sclerosis are antibody-mediated and pathogenically distinct from relapsing-remitting and secondary progressive multiple sclerosis. Our study has potentially important implications for the development of specific therapies for patients with primary progressive multiple sclerosis., (© The Author(s) 2023. Published by Oxford University Press on behalf of the Guarantors of Brain.)

Multiple sclerosis (MS) is an auto-immune inflammatory disorder affecting the central nervous system. The cause of the disease is unknown but both genetic and environmental factors are implicated in the pathogenesis. We derived cerebral organoids from induced pluripotent stem cells (iPSC) of healthy control subjects as well as from primary progressive MS (PPMS), secondary progressive MS (SPMS) and relapsing remitting MS (RRMS) patients to better understand the pathologic basis of the varied clinical phenotypic expressions of MS. In MS organoids, most notably in PPMS, we observed a decrease of proliferation marker Ki67 and a reduction of the SOX2+ stem cell pool associated with an increased expression of neuronal markers CTIP2 and TBR1 as well as a strong decrease of oligodendrocyte differentiation. This dysregulation of the stem cell pool is associated with a decreased expression of the cell cycle inhibitor p21. Our findings show that the genetic background of a patient can directly alter stem cell function, provides new insights on the innate cellular dysregulation in MS and identifies p21 pathway as a new potential target for therapeutic strategies in MS., Competing Interests: Competing interests The authors declare no competing or financial interests., (© 2023. Published by The Company of Biologists Ltd.)

Background: Mesenchymal stem cell-derived neural progenitor cell (MSC-NP) therapy is an experimental approach to treat multiple sclerosis. The influence of MSC-NPs on microglial activation was investigated. Methods: Microglia were stimulated in the presence of MSC-NP-conditioned media, and proinflammatory or proregenerative marker expression was assessed by quantitative PCR and ELISA. Results: Microglia stimulated in the presence of MSC-NP-conditioned media displayed reduced expression of proinflammatory markers including CCL2, increased expression of proregenerative markers and reduced phagocytic activity. The paracrine effects of MSC-NPs from multiple donors correlated with TGF-β3 gene expression and was reversed by TGF-β signaling inhibition. Conclusion: MSC-NPs promote beneficial microglial polarization through secreted factors. This study suggests that microglia are a potential therapeutic target of MSC-NP cell therapy.

Amyotrophic lateral sclerosis is a fatal neurodegenerative disease characterized by motor neuron degeneration. Approximately 90% of cases occur sporadically with no known cause while 10% are familial cases arising from known inherited genetic mutations. In vivo studies have predominantly utilized transgenic models harbouring amyotrophic lateral sclerosis-associated gene mutations, which have not hitherto elucidated mechanisms underlying motor neuron death or identified therapeutic targets specific to sporadic amyotrophic lateral sclerosis. Here we provide evidence demonstrating pathogenic differences in CSF from patients with sporadic amyotrophic lateral sclerosis and familial amyotrophic lateral sclerosis patients with mutations in SOD1, C9orf72 and TARDBP . Using a novel CSF-mediated animal model, we show that intrathecal delivery of sporadic amyotrophic lateral sclerosis patient-derived CSF into the cervical subarachnoid space in adult wild-type mice induces permanent motor disability which is associated with hallmark pathological features of amyotrophic lateral sclerosis including motor neuron loss, cytoplasmic TDP-43 translocation, reactive astrogliosis and microglial activation. Motor impairments are not induced by SOD1, C9orf72 or TARDBP CSF, although a moderate degree of histopathological change occurs in C9orf72 and TARDBP CSF-injected mice. By conducting a series of CSF filtration studies and global proteomic analysis of CSF, we identified apolipoprotein B-100 in sporadic amyotrophic lateral sclerosis CSF as the putative agent responsible for inducing motor disability, motor neuron degeneration and pathological translocation of TDP-43. Apolipoprotein B-100 alone is sufficient to recapitulate clinical and pathological outcomes in vivo and induce death of human induced pluripotent stem cell-derived motor neurons in vitro . Targeted removal of apolipoprotein B-100 from sporadic amyotrophic lateral sclerosis CSF via filtration or immunodepletion successfully attenuated the neurotoxic capacity of sporadic amyotrophic lateral sclerosis CSF to induce motor disability, motor neuron death, and TDP-43 translocation. This study presents apolipoprotein B-100 as a novel therapeutic target specific for the predominant sporadic form of amyotrophic lateral sclerosis and establishes proof-of-concept to support CSF pheresis as a therapeutic strategy for mitigating neurotoxicity in sporadic amyotrophic lateral sclerosis., (© The Author(s) 2022. Published by Oxford University Press on behalf of the Guarantors of Brain.)

Immunocompromised individuals, including multiple sclerosis (MS) patients on certain immunotherapy treatments, are considered susceptible to complications from severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection and specific vaccination regimens have been recommended for suitable protection. MS patients receiving anti-CD20 therapy (aCD20-MS) are considered especially vulnerable due to acquired B-cell depletion and impaired antibody production in response to virus infection and COVID-19 vaccination. Here, the humoral and cellular responses are analyzed in a group of aCD20-MS patients (n=43) compared to a healthy control cohort (n=34) during the first 6 months after a 2-dose cycle mRNA-based COVID-19 vaccination. Both IgG antibodies recognizing receptor binding domain (RBD) from CoV-2 spike protein and their blocking activity against RBD-hACE2 binding were significantly reduced in aCD20-MS patients, with a seroconversion rate of only 23.8%. Interestingly, even under conditions of severe B-cell depletion and failed seroconversion, a significantly higher polyfunctional IFNγ + and IL-2 + T-cell response and strong T-cell proliferation capacity were detected compared to controls. Moreover, no difference in T-cell response was observed between forms of disease (relapsing remitting- vs progressive-MS), anti-CD20 therapy (Rituximab vs Ocrelizumab) and type of mRNA-based vaccine received (mRNA-1273 vs BNT162b2). These results suggest the generation of a partial adaptive immune response to COVID-19 vaccination in B-cell depleted MS individuals driven by a functionally competent T-cell arm. Investigation into the role of the cellular immune response is important to identifying the level of protection against SARS-CoV-2 in aCD20-MS patients and could have potential implications for future vaccine design and application., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Alfonso-Dunn, Lin, Kirschner, Lei, Feuer, Malin, Liu, Roche and Sadiq.)

Previous studies have shown retinal vein occlusion (RVO) is associated with changes in vessel density visible on swept-source optical coherence tomography angiography (ss-OCTA). This study aimed to characterize retinal changes on ss-OCTA among RVO patients stratified by the need for continuous anti-VEGF therapy. This cross-sectional study of 24 RVO patients ≥ 18 years were imaged with SS-OCT-A. Patients were categorized into continuous vs. limited therapy (≥1 vs. no injections in previous 12 months) based on recurrence of intraretinal fluid (IRF) on OCT. Images were analyzed using ImageJ. T-tests were used to compare vessel density of the macula and peripheral retina. Overall, RVO patients undergoing continuous therapy ( n = 14) had higher diabetes prevalence, worse baseline visual acuity, and higher baseline macular thickness compared to the limited ( n = 10) therapy group. Continuous therapy was associated with lower macular VD in the combined retina layer and the superficial capillary plexus (SCP), but not in the deep capillary plexus (DCP). Further, the continuous therapy group exhibited lower peripheral VD in the combined retina layer, and no difference in the SCP and DCP layers when analyzed separately. In conclusion, RVO patients requiring continuous anti-VEGF injections demonstrate reduced VD of the macula and in the periphery on SS-OCTA imaging. SS-OCTA may be valuable for monitoring and prognosticating treatment for RVO patients.

Objective: To determine the long-term safety and efficacy of repeated intrathecal (IT) administration of autologous mesenchymal stem cell-derived neural progenitors (MSC-NPs) in patients with progressive MS by evaluating subjects 2 years after treatment., Methods: Twenty subjects were enrolled as part of a phase I, open-label single-arm study of 3 IT injections of MSC-NPs spaced 3 months apart. Subjects were evaluated for adverse events and disability outcomes including the Expanded Disability Status Scale (EDSS) and the timed 25-foot walk (T25FW). Long-term evaluation was conducted 2 years after the third treatment. CSF was collected before and 3 months after treatment., Results: Eighteen of the 20 study participants completed the full 2-year follow-up protocol. There were no long-term adverse events associated with repeated IT-MSC-NP treatment. Seven subjects showed sustained improvement in EDSS after 2 years, although the degree of improvement was not maintained in 5 of the subjects. Three of the 10 ambulatory subjects showed sustained improvement in the T25FW after 2 years. CSF biomarker analysis revealed a decrease in C-C motif chemokine ligand 2 (CCL2) and an increase in interleukin 8, hepatocyte growth factor, and C-X-C motif chemokine ligand 12 (CXCL12) after treatment., Conclusions: Safety and efficacy of repeated IT-MSC-NP treatment was sustained for 2 years; however, the degree of disability reversal was not sustained in a subset of patients. CSF biomarkers altered in response to IT-MSC-NP treatment may reflect specific immunoregulatory and trophic mechanisms of therapeutic response in MS., Classification of Evidence: This study provides Class IV evidence that for patients with progressive MS, IT administration of MSC-NPs is safe and effective. The study is rated Class IV because of the absence of a non-IT-MSC-NP-treated control group., Clinicaltrialsgov Identifier: NCT01933802., (Copyright © 2020 The Author(s). Published by Wolters Kluwer Health, Inc. on behalf of the American Academy of Neurology.)

Background: There are few treatment options for multiple sclerosis (MS) patients with advanced disability [expanded disability status scale (EDSS) ⩾ 6.0]. In 2010, we reported initial results of using intrathecal methotrexate (ITMTX) in patients with progressive MS. We now report on long-term use of ITMTX. We performed a retrospective chart analysis of patients who have had 18 or more treatments to establish the ongoing safety and tolerability of ITMTX. Thus, the objective of this study was to establish the safety and tolerability of long-term therapy with (ITMTX) in patients with treatment-resistant, progressive forms of MS., Methods: We studied 83 patients (67 secondary and 16 primary progressive) who received ITMTX 12.5 mg every 8-11 weeks for 3-10 years (range: 18-57 treatments). All patients were evaluated neurologically, and their EDSS was assessed at every treatment. In addition, all adverse events, frequency of infections, and any hospitalizations, were noted., Results: There were no deaths, hospitalizations, or other serious adverse effects related to ITMTX. Headaches occurred at least once in 12% of patients, and transient fatigue occurred in 53% of patients. As determined by EDSS, there was no significant change from baseline status to post-treatment scores in both primary progressive MS (PPMS) and secondary progressive (SPMS) patients., Conclusions: Pulsed ITMTX was well tolerated for up to 10 years in PPMS patients with no serious adverse effects. Although this was an open-label, retrospective analysis, and efficacy could not be studied, there was evidence of disease stabilization in many patients receiving ITMTX. It appears that long-term ITMTX is a safe therapeutic option in advanced progressive MS., Competing Interests: Conflict of interest statement: The authors declare that there is no conflict of interest., (© The Author(s), 2019.)

Background: Multiple sclerosis (MS) is an immune-mediated demyelinating disease of the central nervous system and is one of the leading causes of disability in young adults. Cell therapy is emerging as a therapeutic strategy to promote repair and regeneration in patients with disability associated with progressive MS., Methods: We conducted a phase I open-label clinical trial investigating the safety and tolerability of autologous bone marrow mesenchymal stem cell-derived neural progenitor (MSC-NP) treatment in 20 patients with progressive MS. MSC-NPs were administered intrathecally (IT) in three separate doses of up to 1 × 10 7 cells per dose, spaced three months apart. The primary endpoint was to assess safety and tolerability of the treatment. Expanded disability status scale (EDSS), timed 25-ft walk (T25FW), muscle strength, and urodynamic testing were used to evaluate treatment response. This trial is registered with ClinicalTrials.gov, number NCT01933802., Findings: IT MSC-NP treatment was safe and well tolerated. The 20 enrolled subjects completed all 60 planned treatments without serious adverse effects. Minor adverse events included transient fever and mild headaches usually resolving in <24 h. Post-treatment disability score analysis demonstrated improved median EDSS suggesting possible efficacy. Positive trends were more frequently observed in the subset of SPMS patients and in ambulatory subjects (EDSS ≤ 6.5). In addition, 70% and 50% of the subjects demonstrated improved muscle strength and bladder function, respectively, following IT MSC-NP treatment., Interpretation: The possible reversal of disability that was observed in a subset of patients warrants a larger phase II placebo-controlled study to establish efficacy of IT MSC-NP treatment in patients with MS., Funding Source: The Damial Foundation., (Copyright © 2018 The Authors. Published by Elsevier B.V. All rights reserved.)

Introduction: Multiple sclerosis (MS) disease activity is associated with blood-brain barrier (BBB) disruption, which is mediated by inflammatory cytokines released by CD4+ lymphocytes. To assess the effects of adenosine A2A receptors on BBB permeability in vitro and in vivo., Methods: A2A receptor expression was detected by immunostaining in experimental autoimmune encephalomyelitis (EAE) C57BL/6 mice immunized with myelin oligodendrocyte glycoprotein (MOG) 35-55 , and human MS brain. F-actin and the tight junction protein Claudin-5 were assessed in endothelial cells treated with an A2A receptor specific agonist (CGS-21680) after Th1 cytokine stimulation. EAE mice were divided into control and CGS-21680 (50 µg/kg, i.p., daily) groups. Disease scores were recorded daily to evaluate neurological impairment. The effects of A2A receptor on inflammation and demyelination were assessed after euthanasia by immunostaining or histology; BBB permeability was measured by sodium fluoride (Na-F) and FITC-dextran amounts., Results: Endothelial A2A receptor was detected in demyelination areas of MS brain samples. In EAE lesions, A2A receptor was expressed in the endothelium in association with immune cell infiltration. Treatment with CGS-21680 counteracted the effects of Th1 cytokines on endothelial cells in vitro, preventing the reduction of tight junction protein expression and stress fiber formation. The effects of A2A receptor activation were correlated with MLCK phosphorylation signaling repression. In EAE, A2A receptor agonist decreased BBB permeability and inhibited EAE neurologic deficiency in mice., Conclusions: A2A receptor activation at EAE onset helps reduce the effects of Th1 stimulation on BBB permeability, indicating that A2A receptor mediates BBB function in CNS demyelinated disease., (© 2017 The Authors. Immunity, Inflammation and Disease Published by John Wiley & Sons Ltd.)

Fetuin-A is a biomarker of disease activity in multiple sclerosis. Our aim was to investigate whether Fetuin-A plays a direct role in the neuroinflammatory response in the mouse EAE model. Peak Fetuin-A expression in the CNS and in peripheral lymphoid tissue correlated with peak EAE disease activity. Fetuin-A-deficient mice showed reduced EAE severity associated with an accumulation of splenic monocyte and dendritic cell populations, increased IL-12p40, ASC1, and IL-1β expression, and an increase in T regulatory cells. The upregulation of Fetuin-A in LPS-stimulated dendritic cells and microglia further supports an intrinsic role of Fetuin-A in regulating innate immune activation during EAE.

Multiple sclerosis (MS) is an autoimmune disease affecting the brain and spinal cord that is associated with chronic inflammation leading to demyelination and neurodegeneration. With the recent increase in the number of available therapies for MS, optimal treatment will be based on a personalized approach determined by an individual patient's prognosis and treatment risks. An integral part of such therapeutic decisions will be the use of molecular biomarkers to predict disability progression, monitor ongoing disease activity, and assess treatment response. This review describes current published findings within the past 3 years in biomarker research in MS, specifically highlighting recent advances in the validation of cerebrospinal fluid biomarkers such as neurofilaments (light and heavy chains), chitinases and chitinase 3-like proteins, soluble surface markers of innate immunity, and oligoclonal immunoglobulin M antibodies. Current research in circulating miRNAs as biomarkers of MS is also discussed. Continued validation and testing will be required before MS biomarkers are routinely applied in a clinical setting., Competing Interests: Disclosure The authors report no conflicts of interest in this work.

Background Aims: There is a critical unmet need to develop regenerative therapies for the demyelinating disease multiple sclerosis (MS). We previously characterized the immunoregulatory and trophic properties of neural progenitors derived from bone marrow mesenchymal stromal cells (MSC-NPs) and established that cells derived from MS and non-MS patients alike were therapeutically viable. In an experimental model of MS, intrathecal MSC-NP injection resulted in disease amelioration with decreased T-cell infiltration, and less severe lesion pathology associated with recruitment of resident progenitors to inflammatory sites. In this pilot feasibility study, we investigated safety and dosing of intrathecal MSC-NP therapy in six patients with MS., Methods: Patients with progressive MS and advanced disability who were refractory to all other conventional MS treatments were enrolled in the study. For each dose, MSC-NP cells were cultured from autologous MSCs and tested for quality control before intrathecal administration. Patients were evaluated for adverse events and neurological status to assess safety of the treatment., Results: Six patients with progressive MS were treated with between 2 and 5 intrathecal injections of escalating doses of autologous MSC-NPs and were followed an average of 7.4 years after initial injection. There were no safety concerns noted, no serious adverse events, and the multiple dosing regimen was well tolerated. Four of the six patients showed a measurable clinical improvement following MSC-NP treatment., Discussion: This pilot study supports preliminary first-in-human safety and tolerability of autologous MSC-NP treatment for MS., (Copyright © 2016 International Society for Cellular Therapy. Published by Elsevier Inc. All rights reserved.)

Hyaluronan (HA) may have proinflammatory roles in the context of CNS autoimmunity. It accumulates in demyelinated multiple sclerosis (MS) lesions, promotes antigen presentation, and enhances T-cell activation and proliferation. HA facilitates lymphocyte binding to vessels and CNS infiltration at the CNS vascular endothelium. Furthermore, HA signals through Toll-like receptors 2 and 4 to stimulate inflammatory gene expression. We assessed the role of HA in experimental autoimmune encephalomyelitis (EAE), an animal model of MS by administration of 4-methylumbelliferone (4MU), a well established inhibitor of HA synthesis. 4MU decreased hyaluronan synthesis in vitro and in vivo. It was protective in active EAE of C57Bl/6 mice, decreased spinal inflammatory infiltrates and spinal infiltration of Th1 cells, and increased differentiation of regulatory T-cells. In adoptive transfer EAE, feeding of 4MU to donor mice significantly decreased the encephalitogenicity of lymph node cells. The transfer of proteolipid protein (PLP)-stimulated lymph node cells to 4MU-fed mice resulted in a delayed EAE onset and delayed spinal T-cell infiltration. Expression of CXCL12, an anti-inflammatory chemokine, is reduced in MS patients in CSF cells and in spinal cord tissue during EAE. Hyaluronan suppressed production of CXCL12, whereas 4MU increased spinal CXCL12 in naive animals and during neuroinflammation. Neutralization of CXCR4, the most prominent receptor of CXCL12, by administration of AMD3100 diminished the protective impact of 4MU in adoptive transfer EAE. In conclusion, hyaluronan exacerbates CNS autoimmunity, enhances encephalitogenic T-cell responses, and suppresses the protective chemokine CXCL12 in CNS tissue. Inhibition of hyaluronan synthesis with 4MU protects against an animal model of MS and may represent an important therapeutic option in MS and other neuroinflammatory diseases., (© 2014 by The American Society for Biochemistry and Molecular Biology, Inc.)

Objective: To investigate the potential of 8-iso-prostaglandin F2α (8-iso-PGF2α) as a biomarker for disease activity and oxidative stress in the CSF of patients with multiple sclerosis (MS)., Methods: The isoprostane 8-iso-PGF2α is an established biomarker for in vivo oxidative stress and lipid peroxidation. We measured CSF 8-isoPGF2α levels in 231 patients with MS (74 with relapsing-remitting MS, 67 with primary progressive MS, and 90 with secondary progressive MS [SPMS]) and 40 controls using a competition ELISA., Results: We found increased CSF levels of 8-iso-PGF2α in patients with MS compared to controls, with the most striking values in a subgroup of patients with SPMS. Furthermore, the increase in 8-iso-PGF2α correlated with other parameters of lipid peroxidation as well as with a decrease in the total antioxidant status in the MS CSF samples., Conclusions: Our study demonstrates that CSF levels of 8-iso-PGF2α may serve as a biomarker of oxidative stress in MS. Further investigation will help establish the pathologic and clinical significance of our preliminary findings.

We previously showed that intrathecal administration of methotrexate slowed disability progression in multiple sclerosis (MS) patients with progressive disease. In general MS patients with progressive disease respond poorly to anti-inflammatory therapies. In order to better understand the mechanism by which methotrexate is protective in progressive MS, we analyzed its impact on the non-inflammatory cuprizone-induced demyelination model. When low-dose methotrexate was administered intracerebroventricularly it reduced demyelination and accumulation of GFAP+ reactive astrocytes in the corpus callosum. Administration of methotrexate after the withdrawal of cuprizone neither delayed remyelination nor influenced the number of astrocytes in the corpus callosum suggesting that methotrexate does not interfere with repair processes in the CNS. Moreover, methotrexate increased the expression of IGF1 in vitro and in vivo, a factor known to protect oligodendrocytes and limit the activation of astrocytes. Our studies show that methotrexate has an impact on pathogenic process in a demyelination model whose pathophysiological basis is not primarily related to inflammatory mechanisms, similar to neurodegenerative mechanisms associated with progressive MS. The pronounced inhibitory influence of methotrexate on the accumulation of astrocytes in the corpus callosum suggests that intrathecal methotrexate modulates astroglial activation in progressive MS possibly by promoting CNS production of IGF1.

Introduction: In long-standing facial palsy, muscles on the normal side overcontract causing difficulty in articulation, eating, drinking, cosmetic embarrassment, and psychological effects as patients lack confidence in public., Methods: We injected botulinum toxin A (BTXA) into the normal contralateral smile muscles to weaken them and restore symmetry to both active and passive movements by neutralising these overacting muscles., Results: A total of 14 patients received BTXA (79% women, median age 47 years, average length of palsy 8 years). They were all difficult cases graded between 2 and 6 (average grade 3 House-Brackmann). All 14 patients reported improved facial symmetry with BTXA (dose altered in some to achieve maximum benefit). Average dose was 30 units, but varied from 10 to 80 units. Average time to peak effect was 6 days; average duration of effect was 11 weeks. Three patients had increased drooling (resolved within a few days)., Conclusion: The improvement in symmetry was observed by both patient and examining doctor. Patients commented on increased confidence, being more likely to allow photographs taken of themselves, and families reported improved legibility of speech. Younger patients have more muscle tone than older patients; the effect is more noticeable and the benefit greater for them. BTXA improves symmetry in patients with facial palsy, is simple and acceptable, and provides approximately 4 months of benefit. The site of injection depends on the dynamics of the muscles in each individual patient.

Bone marrow mesenchymal stem cell-derived neural progenitors (MSC-NPs) are a potential therapeutic source of cells that have been shown to be efficacious in a preclinical model of multiple sclerosis (MS). To examine the feasibility of using MSC-NPs as an autologous source of cells to promote central nervous system (CNS) repair in MS, this study characterized human MSC-NPs from a panel of both MS and non-MS donors. Expanded MSCs showed similar characteristics in terms of growth and cell surface phenotype, regardless of the donor disease status. MSC-NPs derived from all MSCs showed a consistent pattern of gene expression changes that correlated with neural commitment and increased homogeneity. Furthermore, the reduced expression of mesodermal markers and reduced capacity for adipogenic or osteogenic differentiation in MSC-NPs compared with MSCs suggested that MSC-NPs have reduced potential of unwanted mesodermal differentiation upon CNS transplantation. The immunoregulatory function of MSC-NPs was similar to that of MSCs in their ability to suppress T-cell proliferation and to promote expansion of FoxP3-positive T regulatory cells in vitro. In addition, MSC-NPs promoted oligodendroglial differentiation from brain-derived neural stem cells that correlated with the secretion of bioactive factors. Our results provide a set of identity characteristics for autologous MSC-NPs and suggest that the in vitro immunoregulatory and trophic properties of these cells may have therapeutic value in the treatment of MS.

Objectives: To analyse the usefulness of a multidisciplinary facial function clinic (FFC)., Design: Retrospective case-note review., Setting: The FFC was established in July 2006 at the Manchester Royal Eye Hospital with attending consultants from Ophthalmology, Skull-Base Otolaryngology, Plastic Surgery, and Physiotherapy., Participants: We retrospectively reviewed the case notes for 59 consecutive patients seen at the FFC from July 2006 to February 2009., Main Outcome Measures: We documented demographic data, including distance travelled and average journey time., Results: The 59 patients (mean age 46 years) made a total of 106 clinical visits (mean 1.8). The mean distance travelled by a patient was 31.9 miles with an estimated journey time of 47 min, each way. At presentation the average House-Brackmann grade of facial nerve weakness was IV. Ophthalmologist's advice was needed for 58 (98.3%), otolaryngologist's for 57 (96.6%), plastic surgeon for 49 (83.0%), physiotherapist for 58 (98.3%), and 4 (6.8%) were referred for psychological counselling. In all, 47 (79.7%) of our patients needed input from all four consultants during their visit at the FFC. By combining the presence of several consultants in one clinic, we saved an average of 5.1 visits (325.4 miles; 8 h travel time) for each patient., Conclusion: We and our patients feel our multidisciplinary facial function clinic has been an effective service and has continued to work.

Aims: Psychological distress is well documented in people with facial disfigurement. However, the prevalence of psychological distress in patients with facial palsy has not been studied. This study aims to establish the prevalence of psychological distress and the extent of anxiety and depression in a sample of facial palsy patients from the Northwest of England., Method: A total of 103 participants with facial palsy completed a questionnaire pack comprising the Illness Perception Questionnaire-Revised (IPQ-R), a demographic questionnaire, and the Hospital Anxiety and Depression Scale (HADS). The severity of participants' facial palsy was measured by the House-Brackmann scale., Results: In all, 32.7 and 31.3% of the sample had significant levels of anxiety and depression, respectively. The mean age of participants was 59, and 35.9% had grade 6 facial palsy. Significant associations were found between participants' perception of consequences, duration, timeline, and the level of distress. No significant associations were found between clinical severity of facial palsy and levels of distress. Females had significantly higher levels of anxiety compared with males., Conclusions: There was a significant level of distress in this study group. The levels of psychological distress were higher than the levels found in other outpatient attenders. There were significant associations between participants' illness perceptions and their level of distress.

We reviewed 40 cases with non-union of the humeral shaft. There were 31 men and nine women patients with an average age of 38.5 (35-65) years. Thirty-four non-unions were of the atrophic type. Non-union was most often found at the transition of the middle third to the lower third of the humeral shaft. Six fractures were classified as open at the time of the initial injury. All patients were treated by open reduction and internal fixation with a dynamic compression plate (DCP). Cancellous bone graft was used in all atrophic non-unions. In one patient an additional fibular graft was used. The average follow-up was 13 (6-18) months. Final results were available for 34 patients. Thirty-one fractures (91%) healed in an average of 4.5 (3-9) months. Main complications were temporary radial nerve palsy in two patients and deep infection in one.

A case of acute transient flaccid quadriparesis after a low voltage electric shock is reported. The patient recovered completely with in three days.