Genetic testing, another important tool in presurgical evaluation of focal epilepsies in childhood.

Epilepsy surgery may be a curative therapy for patients with drug-resistant epilepsies when focal lesions or foci are identified. Genetic testing is not yet routinely included in many presurgical evaluation programs although recent evidence support that finding a germline genetic mutation could help to better delineate the patient candidacy to surgery and provide valuable information on the expected surgery outcome. In this study, we report nine patients presenting drug-resistant focal epilepsy enrolled in presurgical evaluation. We show how the identification of genetic pathogenic variant in epilepsy known genes led to the interruption of the presurgical work-up and ruled out surgery in 7 of them. We observed that the co-existence of some recurrent clinical characteristics as early seizures' onset, frequent precipitating factors including fever, and developmental delay or intellectual disability may be useful markers for germline genetic pathogenic variants. In this group, genetic assessment should be mandatory during presurgical work up, mainly in patients with negative magnetic resonance imaging (MRI) or doubtful structural lesions. The integration of next generation targeted sequencing into the presurgical evaluation can improve the selection of candidates for resective surgery and fosters a personalized medicine approach with a better outcome. PLAINE LANGUAGE ABSTRACT: Genetic testing is not yet systematically included in the pre-surgical assessment of patients with drug-resistant focal epilepsies. In this study, through the description of nine patients, we underline how the integration of genomics into the presurgical work up can help in evaluating the patient candidacy to surgery and provide valuable information on expected surgery outcome.

Repurposed Drugs That Activate Autophagy in Filarial Worms Act as Effective Macrofilaricides.

Onchocerciasis and lymphatic filariasis are two neglected tropical diseases caused by filarial nematodes that utilize insect vectors for transmission to their human hosts. Current control strategies are based on annual or biannual mass drug administration (MDA) of the drugs Ivermectin or Ivermectin plus Albendazole, respectively. These drug regimens kill the first-stage larvae of filarial worms (i.e., microfilariae) and interrupt the transmission of infections. MDA programs for these microfilaricidal drugs must be given over the lifetime of the filarial adult worms, which can reach 15 years in the case of Onchocerca volvulus. This is problematic because of suboptimal responses to ivermectin in various endemic regions and inefficient reduction of transmission even after decades of MDA. There is an urgent need for the development of novel alternative treatments to support the 2030 elimination goals of onchocerciasis and lymphatic filariasis. One successful approach has been to target Wolbachia, obligatory endosymbiotic bacteria on which filarial worms are dependent for their survival and reproduction within the human host. A 4-6-week antibiotic therapy with doxycycline, for example, resulted in the loss of Wolbachia that subsequently led to extensive apoptosis of somatic cells, germline, embryos, and microfilariae, as well as inhibition of fourth-stage larval development. However, this long-course regimen has limited use in MDA programs. As an alternative approach to the use of bacteriostatic antibiotics, in this study, we focused on autophagy-inducing compounds, which we hypothesized could disturb various pathways involved in the interdependency between Wolbachia and filarial worms. We demonstrated that several such compounds, including Niclosamide, an FDA-approved drug, Niclosamide ethanolamine (NEN), and Rottlerin, a natural product derived from Kamala trees, significantly reduced the levels of Wolbachia in vitro. Moreover, when these compounds were used in vivo to treat Brugia pahangi-infected gerbils, Niclosamide and NEN significantly decreased adult worm survival, reduced the release of microfilariae, and decreased embryonic development depending on the regimen and dose used. All three drugs given orally significantly reduced Wolbachia loads and induced an increase in levels of lysosome-associated membrane protein in worms from treated animals, suggesting that Niclosamide, NEN, and Rottlerin were effective in causing drug-induced autophagy in these filarial worms. These repurposed drugs provide a new avenue for the clearance of adult worms in filarial infections.

Negative Association of Lignan and Phytosterol Intake with Stress Perception during the COVID-19 Pandemic-A Polish Study on Young Adults.

BACKGROUND: There has been an increasing global prevalence of depression and other psychiatric diseases in recent years. Perceived stress has been proven to be associated with psychiatric and somatic symptoms. Some animal and human studies have suggested that consuming foods abundant in lignans and phytosterols may be associated with lower levels of stress, depression, and anxiety. Still, the evidence is not yet strong enough to draw firm conclusions. Thus, we investigated the association between dietary intake of these phytochemicals and the level of stress experienced by adult individuals. METHODS: Diet was assessed using self-reported 7-day dietary records. The intakes of lignans and phytosterols were estimated using databases with their content in various food products. The Perceived Stress Scale (PSS) was implemented to measure the level of perceived stress. A logistic regression analysis was used to test for associations. RESULTS: The odds of elevated PSS were negatively associated with dietary intake of total phytosterols, stigmasterol, and ß-sitosterol, with evidence of a decreasing trend across tertiles of phytochemicals. The analysis for doubling the intake reinforced the aforementioned relationships and found protective effects against PSS for total lignans, pinoresinol, and campesterol. CONCLUSIONS: Habitual inclusion of lignans and phytosterols in the diet may play a role in psychological health. To address the global outbreak of depression and other mental health issues triggered by stress, it is important to take a holistic approach. There is a need to develop effective strategies for prevention and treatment, among which certain dietary interventions such as consumption of products abundant in lignans and phytosterols may play a substantial role.

Molecular Dynamics of Steroidal Rotors Probed by Theoretical, Spectroscopic and Dielectric Methods.

Our study focuses on molecular rotors with fast-moving rotators and their potential applications in the development of new amphidynamic crystals. Steroidal molecular rotors with a dipolar fluorine-substituted phenyl group as the rotator were synthesized and characterized. Three different rotors were investigated with varying numbers of fluorine atoms. A comprehensive analysis was performed using vibrational spectroscopy (Raman, FT-IR), electronic circular dichroism (ECD), and dielectric response to understand the behavior of the investigated model rotors. The results were supported by theoretical calculations using Density Functional Theory (DFT) methods. The angle-dependent polarized Raman spectra confirmed the crystallinity of the samples. Nearly frequency and temperature-independent permittivity suggest low-frequency librational motion of stators. An in-depth analysis of ECD spectra revealed high conformational flexibility in solution, resulting in low ECD effects, while in the solid-state with restricted rotation, significant ECD effects were observed. These findings shed light on the conformational behavior and potential applications of the studied steroidal molecular rotors.

Interhemispheric Vertical Hemispherotomy: Technique, Outcome, and Pitfalls-A Bicentric Retrospective Case Series of 39 Cases.

BACKGROUND AND OBJECTIVES: When seizure onset affects a whole hemisphere, hemispheric disconnections are efficient and safe procedures. However, both lateral peri-insular hemispherotomy and vertical paramedian hemispherotomy approaches report a failure rate around 20%, which can be explained by residual connections giving rise to persistent seizures. In this study, we present the interhemispheric vertical hemispherotomy (IVH), a technical variation of the vertical paramedian hemispherotomy approach, that aims to increase seizure control avoiding residual connections while exposing the corpus callosum. METHODS: This is a retrospective study of IVH in two centers, with analysis of clinical and MRI data and outcomes. A detailed description of the technique is provided with a video. RESULTS: IVH was performed in 39 children. The mean age at surgery was 7.2 years, and etiologies were as follows: malformations of cortical development (n = 14), Rasmussen's encephalitis (n = 10), stroke (n = 10), post-traumatic (3), and Sturge-Weber Syndrome (2). Hemispheric disconnection was complete on postoperative MRI in 34 cases. There was no mortality, hydrocephalus occurred in one case, and subdural collection occurred in four cases. A second surgery was performed in four cases because of seizure relapse (n = 3) and/or incomplete disconnection on MRI (n = 4). With a mean follow-up of 3.2 years, International League Against Epilepsy class I epilepsy outcome was obtained for 37/39 patients. CONCLUSION: IVH is a safe and effective variation of the vertical approaches for hemispheric disconnection. It allows a good exposure and anatomic control of the corpus callosum, which is a frequent site of incomplete disconnection. IVH may be limited by the thalamic volume and the ventricular size, notably in hemimegalencephaly cases.

Association between acute complications in PMM2-CDG patients and haemostasis anomalies: Data from a multicentric study and suggestions for acute management.

OBJECTIVES: Patients with PMM2-CDG develop acute events (stroke-like episodes (SLEs), thromboses, haemorrhages, seizures, migraines) associated with both clotting factors (factor XI) and coagulation inhibitors (antithrombin, protein C and protein S) deficiencies. The aim of the study was to correlate acute events to haemostasis and propose practical guidelines. METHODS: In this multicentric retrospective study, we evaluated clinical, radiological, haemostasis and electroencephalography data for PMM2-CDG patients hospitalized for acute events. Cerebral events were classified as thrombosis, haemorrhage, SLE, or "stroke mimic" (SM: normal brain imaging or evoking a migraine). RESULTS: Thirteen patients had a total of 31 acute episodes: 27 cerebral events with 7 SLEs, 4 venous thromboses, 4 haemorrhages (3 associated with thrombosis), 15 SMs at a mean age of 7.7 years; 4 non-cerebral thromboses, one of which included bleeding. A trigger was frequently involved (infection, head trauma). Although sometimes normal at baseline state, factor XI, antithrombin and protein C levels decreased during these episodes. No correlation between haemostasis anomalies and type of acute event was found. DISCUSSION: Acute events in PMM2-CDG are not negligible and are associated with haemostasis anomalies. An emergency protocol is proposed for their prevention and treatment (https://www.filiere-g2m.fr/urgences). For cerebral events, brain Magnetic Resonance Imaging with perfusion weight imaging and diffusion sequences, electroencephalogram and haemostasis protein levels guide the treatment: anticoagulation, antithrombin or fresh frozen plasma supplementation, antiepileptic therapy. Preventing bleeding and thrombosis is required in cases of surgery, prolonged immobilization, hormone replacement therapy. CONCLUSION: Acute events in PMM2-CDG are associated with abnormal haemostasis, requiring practical guidance.

Influence of sustainable packaging material and packaging conditions on physicochemical, microbiological, and sensorial properties of cheeses.

The aim of this study was to evaluate the influence of different packaging materials [standard foil: BOPP (biaxially oriented polypropylene)/PET (polyester)/PE (polyethylene) for upper layer, and APET (polyethylene terephthalate)/PE for bottom layer; foil 1: PP (polypropylene)/PET/PE/EVOH (ethylene-vinyl alcohol copolymer)/PE upper layer, and PP/PE/EVOH/PE bottom layer; foil 2: PP/PET/PE/EVOH/PE upper layer, and PA (polyamide)/EVOH/PE bottom layer; foil 3: PP/PET/PE upper layer, and PA/EVOH/PE bottom layer; foil 4: PP/PET/PE upper layer, and PA/PE bottom layer; foil 5: PP upper layer, and PP/PP bottom layer] on the quality of 3 different ripening rennet cheeses packed under different modified atmosphere (MAP) conditions as reflected in particular physicochemical, microbiological, and sensorial changes. The changes were monitored during a period of 90 d of storage at 2°C or 8°C. For Gouda cheese, CO2 content of the headspace of the packages was in the range 35% to 45%, whereas for Maasdamer and Sielski Klasyczny cheeses it was 55% to 65%. Three-way ANOVA showed that the foil type influenced the moisture content of Gouda cheese stored for 90 d at 2°C and for Sielski Klasyczny cheese at 8°C, whereas the moisture content was not dependent on MAP conditions during storage. Moreover, the foil type had a significant effect on free fatty acid changes for Gouda and Sielski Klasyczny cheeses stored at 2°C for 90 d. Sensory attributes changed significantly over storage time at 2°C for all studied cheeses as affected by foil type, whereas there was no effect of MAP conditions. In general, the cheeses packed in standard foil and foil 4 were characterized by the highest values of mean sensory attributes. Time was the most significant factor influencing most changes in physicochemical and sensory attributes of cheeses stored at 2°C and 8°C. The storage temperature did not affect the moisture of the samples during storage. In general, we found an increase in the pH value during storage regardless of storage temperature. It was possible to decrease the thickness of the packaging material from initial 103 and 250 µm (standard foil; lid and bottom, respectively) to 98 and 100 µm (foil 4) without affecting sensory attributes of the product.

Interpersonal memory failure in the workplace: The effect of memory and hierarchy on employee's affective commitment.

The current work examined whether being forgotten or remembered by a boss or a coworker affects employee's interpersonal closeness to that person and, in turn, affective organizational commitment (AOC). A first correlational study examined these possibilities in an employed student (1a) and general employed (1b) samples. Perceived memory by both bosses and coworkers was a significant predictor of closeness to the boss or coworker and, in turn, of AOC. The indirect effect of perceived memory on AOC was stronger for boss memory than coworker memory, but only when memory ratings were supported by specific examples of memory. Study 2 provided additional support for the direction of effects posited in Study 1 using vignettes depicting memory and forgetting in the workplace. Overall, these findings suggest that perceptions of boss and coworker memory have an effect on employee's AOC through interpersonal closeness, and that this indirect effect is stronger for boss memory.

Clinical and radiological description of 120 pediatric stroke-like episodes.

BACKGROUND AND PURPOSE: Stroke-like episodes (SLEs) are defined as acute onset of neurological symptoms mimicking a stroke and radiological lesions non-congruent to vascular territory. We aimed to analyze the acute clinical and radiological features of SLEs to determine their pathophysiology. METHODS: We performed a monocenter retrospective analysis of 120 SLEs in 60 children over a 20-year period. Inclusion criteria were compatible clinical symptoms and stroke-like lesions on brain magnetic resonance imaging (MRI; performed for all 120 events) with focal hyperintensity on diffusion-weighted imaging in a non-vascular territory. RESULTS: Three groups were identified: children with mitochondrial diseases (n = 22) involving mitochondrial DNA mutations (55%) or nuclear DNA mutations (45%); those with other metabolic diseases or epilepsy disorders (n = 22); and those in whom no etiology was found despite extensive investigations (n = 16). Age at first SLE was younger in the group with metabolic or epilepsy disorders (18 months vs. 128 months; p < 0.0001) and an infectious trigger was more frequent (69% vs. 20%; p = 0.0001). Seizures occurred in 75% of episodes, revealing 50% episodes of SLEs and mainly leading to status epilepticus (90%). Of the 120 MRI scans confirming the diagnosis, 28 were performed within a short and strict 48-h period and were further analyzed to better understand the underlying mechanisms. The scans showed primary cortical hyperintensity (n = 28/28) with decreased apparent diffusion coefficient in 52% of cases. Systematic hyperperfusion was found on spin labeling sequences when available (n = 18/18). CONCLUSION: Clinical and radiological results support the existence of a vicious circle based on two main mechanisms: energy deficit and neuronal hyperexcitability at the origin of SLE.

Preoperative Detection of Subtle Focal Cortical Dysplasia in Children by Combined Arterial Spin Labeling, Voxel-Based Morphometry, Electroencephalography-Synchronized Functional MRI, Resting-State Regional Homogeneity, and 18F-fluorodeoxyglucose Positron Emission Tomography.

BACKGROUND: Focal cortical dysplasia (FCD) causes drug-resistant epilepsy in children that can be cured surgically, but the lesions are often unseen by imaging. OBJECTIVE: To assess the efficiency of arterial spin labeling (ASL), voxel-based-morphometry (VBM), fMRI electroencephalography (EEG), resting-state regional homogeneity (ReHo), 18F-fluorodeoxyglucose (FDG) positron emission tomography (PET), and their combination in detecting pediatric FCD. METHODS: We prospectively included 10 children for whom FCD was localized by surgical resection. They underwent 3T MR acquisition with concurrent EEG, including ASL perfusion, resting-state BOLD fMRI (allowing the processing of EEG-fMRI and ReHo), 3D T1-weighted images processed using VBM, and FDG PET-CT coregistered with MRI. Detection was assessed visually and by comparison with healthy controls (for ASL and VBM). RESULTS: Eight children had normal MRI, and 2 had asymmetric sulci. Using MR techniques, FCD was accurately detected by ASL for 6/10, VBM for 5/10, EEG-fMRI for 5/8 (excluding 2 with uninterpretable results), and ReHo for 4/10 patients. The combination of ASL, VBM, and ReHo allowed correct FCD detection for 9/10 patients. FDG PET alone showed higher accuracy than the other techniques (7/9), and its combination with VBM allowed correct FCD detection for 8/9 patients. The detection efficiency was better for patients with asymmetric sulci (2/2 for all techniques), but advanced MR techniques and PET were useful for MR-negative patients (7/8). CONCLUSION: A combination of multiple imaging techniques, including PET, ASL, and VBM analysis of T1-weighted images, is effective in detecting subtle FCD in children.

Periodic electroencephalographic discharges and epileptic spasms involve cortico-striatal-thalamic loops on Arterial Spin Labeling Magnetic Resonance Imaging.

Periodic discharges are a rare peculiar electroencephalogram pattern, occasionally associated with motor or other clinical manifestations, usually observed in critically ill patients. Their underlying pathophysiology remains poorly understood. Epileptic spasms in clusters and periodic discharges with motor manifestations share similar electroencephalogram pattern and some aetiologies of unfavourable prognosis such as subacute sclerosing panencephalitis or herpes encephalitis. Arterial spin labelling magnetic resonance imaging identifies localizing ictal and inter-ictal changes in neurovascular coupling, therefore assumed able to reveal concerned cerebral structures. Here, we retrospectively analysed ictal and inter-ictal arterial spin labelling magnetic resonance imaging in patients aged 6 months to 15 years (median 3 years 4 months) with periodic discharges including epileptic spasms, and compared these findings with those of patients with drug-resistant focal epilepsy who never presented periodic discharges nor epileptic spasms as well as to those of age-matched healthy controls. Ictal electroencephalogram was recorded either simultaneously with arterial spin labelling magnetic resonance imaging or during the close time lapse of patients' periodic discharges, whereas inter-ictal examinations were performed during the patients' active epilepsy but without seizures during the arterial spin labelling magnetic resonance imaging. Ictal arterial spin labelling magnetic resonance imaging was acquired in five patients with periodic discharges [subacute sclerosing panencephalitis (1), stroke-like events (3), West syndrome with cortical malformation (1), two of them also had inter-ictal arterial spin labelling magnetic resonance imaging]. Inter-ictal group included patients with drug-resistant epileptic spasms of various aetiologies (14) and structural drug-resistant focal epilepsy (8). Cortex, striatum and thalamus were segmented and divided in six functional subregions: prefrontal, motor (rostral, caudal), parietal, occipital and temporal. Rest cerebral blood flow values, absolute and relative to whole brain, were compared with those of age-matched controls for each subregion. Main findings were diffuse striatal as well as cortical motor cerebral blood flow increase during ictal examinations in generalized periodic discharges with motor manifestations (subacute sclerosing panencephalitis) and focal cerebral blood flow increase in corresponding cortical-striatal-thalamic subdivisions in lateralized periodic discharges with or without motor manifestations (stroke-like events and asymmetrical epileptic spasms) with straight topographical correlation with the electroencephalogram focus. For inter-ictal examinations, patients with epileptic spasms disclosed cerebral blood flow changes in corresponding cortical-striatal-thalamic subdivisions (absolute-cerebral blood flow decrease and relative-cerebral blood flow increase), more frequently when compared with the group of drug-resistant focal epilepsies, and not related to Vigabatrin treatment. Our results suggest that corresponding cortical-striatal-thalamic circuits are involved in periodic discharges with and without motor manifestations, including epileptic spasms, opening new insights in their pathophysiology and new therapeutical perspectives. Based on these findings, we propose a model for the generation of periodic discharges and of epileptic spasms combining existing pathophysiological models of cortical-striatal-thalamic network dynamics.

Effect of COVID-19 Lockdowns on Eye Emergency Department, Increasing Prevalence of Uveitis and Optic Neuritis in the COVID-19 Era.

Background: The COVID-19 pandemic led to the reorganization of the health care system. A decline in health- and life-saving procedures has been reported in various medical specialties. However, data on ophthalmic emergencies during lockdowns is limited. Methods: We conducted a retrospective, observational, case-control study of 2351 patients registered at the ophthalmic emergency department of a tertiary hospital in Poland during three national COVID-19 lockdowns (March/April 2020, November 2020, and March/April 2021) and corresponding months in 2019. Results: The total number of visits declined from a mean of 720/month in the non-COVID era to 304/month during COVID-19 lockdowns (p < 0.001). Ocular trauma incidence dropped significantly from 2019 (non-COVID months) to 2020/2021 (COVID group mean 201 vs. 97 patients monthly, respectively, p = 0.03). Of note, the percentage of foreign bodies removal was significantly higher during lockdowns than corresponding time in the non-COVID era. A downward trend for vitreous detachment and macular disorders cases was observed between COVID and non-COVID time. Uveitis and optic neuritis patients were seen more often during lockdowns (p < 0.001 and p = 0.0013, respectively). In contrast, the frequency of conjunctivitis and keratitis, potentially COVID-related problems, decreased significantly in COVID-19 time (mean 138 vs. 23 per month in non-COVID vs. COVID lockdowns, respectively, p < 0.001). Conclusions: The overall number of eye emergency visits declined during COVID-19 lockdowns. Conjunctivitis and keratitis rates dropped during the lockdowns. Interestingly, the frequency of immune-mediated ocular conditions (uveitis, optic neuritis) increased significantly which might be triggered by SARS-CoV2 infection.

Value of short exercise and short exercise with cooling tests in diagnosis of recessive form of myotonia congenita (Becker disease) - are sex differences important?

INTRODUCTION: In myotonia congenita (MC), activation with exercise or cooling can induce transient changes in compound motor action potential (CMAP) parameters, thus providing a guide to genetic analysis. MATERIAL AND METHODS: We performed the short exercise test (SET) and the short exercise test with cooling (SETC) in 30 patients with genetically confirmed Becker disease (BMC) to estimate their utility in the diagnosis of BMC. RESULTS: Although we observed a significant decrease in CMAP amplitude immediately after maximal voluntary effort in both tests in the whole BMC group, in men this decline was significantly smaller than in women, especially in SET. Clinical implications/future directions: In men with a clinical suspicion of BMC, a small decrease in CMAP amplitude in SET together with a typical decline in SETC does not exclude the diagnosis of BMC. Our results show a sex-specific difference in chloride channel function in BMC, which needs further investigation.

Can the biological stage of a mechanical-biological treatment plant that is designed for mixed municipal solid waste be successfully utilized for effective composting of selectively collected biowaste?

Although the requirements for overall recycling rates can only be met when organic recycling is not overlooked, information is scarce regarding adaption to biowaste composting of existing mechanical-biological treatment (MBT) plants originally designed for stabilization of organic municipal solid waste (OFMSW). Thus, this study aimed to assess the suitability of the operational conditions in the biological part of a full-scale MBT plant now used for stabilization of OFMSW (working line: closed-module-covered-pile-open-pile) with a view to producing compost from biowaste. Temperatures above 75 °C were maintained in the closed module and reached again in the covered pile, indicating that intensive organic-matter mineralization occurred in both stages. In the covered pile, the temperature sharply decreased, indicating depletion of easily biodegradable organic matter. An aerobic 4-day respiration test (AT4) value below 10 mg O2/g dry matter, the cut-off for assessing compost stability, was obtained after 8 weeks. However, a high content of humic substances (HS), reflecting compost maturity, was obtained only after 120 days. The increase in HS content proceeded in two phases. In the first phase (45-84 day), the rate constant and the rate of HS formation were lower than in the second phase (84-120 day) (0.072 vs. 0.087 day-1, 1.97 vs. 3.06 mg C/(g organic matter·d)). All the above-mentioned indicators and the nutrient content (N, P, K, Mg, Ca) in the compost indicates that the biological stage of an MBT plant can successfully treat biowaste. This is in accordance with a circular economy and will contribute to increasing recycling rates.

Recurrent de novo mutations in CLDN5 induce an anion-selective blood-brain barrier and alternating hemiplegia.

Claudin-5 is the most enriched tight junction protein at the blood-brain barrier. Perturbations in its levels of expression have been observed across numerous neurological and neuropsychiatric conditions; however, pathogenic variants in the coding sequence of the gene have never been reported previously. Here, we report the identification of a novel de novo mutation (c.178G>A) in the CLDN5 gene in two unrelated cases of alternating hemiplegia with microcephaly. This mutation (G60R) lies within the first extracellular loop of claudin-5 and based on protein modelling and sequence alignment, we predicted it would modify claudin-5 to become an anion-selective junctional component as opposed to a purely barrier-forming protein. Generation of stably transfected cell lines expressing wild-type or G60R claudin-5 showed that the tight junctions could still form in the presence of the G60R mutation but that the barrier against small molecules was clearly attenuated and displayed higher Cl- ion permeability and lower Na+ permeability. While this study strongly suggests that CLDN5 associated alternating hemiplegia is a channelopathy, it is also the first study to identify the conversion of the blood-brain barrier to an anion-selective channel mediated by a dominant acting variant in CLDN5.

Novel approach to antiangiogenic factors in age-related macular degeneration therapy.

Age-related macular degeneration (AMD) is a leading cause of irreversible vision loss among the population above 85 worldwide. There are two main types of AMD: neovascular and dry AMD. Neovascular AMD leads to macular changes resulting from abnormal choroidal neovascularization. Untreated neovascular AMD leads to scar formation and irreversible sight deterioration. Dry AMD in consequence leads to atrophic changes of the macula. The last decades brought a breakthrough in the therapy of neovascular age-related macular degeneration by introduction of, firstly, photodynamic therapy and, later, anti-VEGF agents administered intravitreally in order to stop neoangiogenesis. However, the treatment of dry AMD is still challenging. Among the directions in dry AMD treatment, the most promising are complement cascade inhibitors and complement cascade targeted gene therapy. In the article we outline the main directions in up-to-date experimental and practical approaches to wet and dry AMD therapy with the emphasis on antiangiogenic factors and gene therapy focused on the inhibition of pathological angiogenesis.

Consequences of diabetes and pre-diabetes and the role of biochemical parameters of carbohydrate metabolism for the functioning of the prefrontal cortex in obese patients.

BACKGROUND: The role of executive functions (EF) is to maintain particular behaviours in order to achieve intended goals. EF are crucial in management of pre-diabetes, diabetes and obesity which are grievous diseases and can lead to severe complications. The aims of our study were to: assess EF in group of obese subject with carbohydrate disorders, evaluate whether biochemical factors and comorbidities related to metabolic disorders have adverse effect on EF in this group of patients. METHODS: The study included 185 obese patients (146 women; 39 men) who were divided on three groups: pre-diabetic, diabetic and control subgroup. Patient underwent Wisconsin Card Sorting Test (WCST) to evaluate EF. Assessed biochemical factors included C-peptide, fasting plasma glucose (FPG) and glycosylated hemoglobin A1c (HbA1c). RESULTS: Diabetic patients showed the worst WCST scores among the rest of groups. Pre-diabetic individuals did not differ in EF performance from control subgroup. We observed significant correlations between FPG and HbA1c and worse WCST scores in pre-diabetic subgroup. In diabetic patients C-peptide correlated with poorer EF. Depressive symptoms and hypertension significantly correlated with non-perseverative errors in WCST. CONCLUSIONS: The subgroup of diabetic patients were the most obese and had the worst glycemia parameters. They also showed the worst EF in WCST. According to obtained results, hyperglycemia positively correlated with poor EF in pre-diabetes. However, in diabetic subjects cognitive deterioration may results from insulin resistance rather than hyperglycemia. In obese individuals with carbohydrate disorders both hypertension and depressive symptoms significantly contributed to EF dysfunction.

Canaloplasty - Efficacy and Safety in an 18-Month Follow Up Period, and Analysis of Outcomes in Primary Open Angle Glaucoma Pigmentary Glaucoma and Pseudoexfoliative Glaucoma.

PURPOSE: Open-angle glaucoma (OAG), accounting for 90% of all glaucoma cases, is a progressive optic nerve neuropathy. It may lead to irreversible loss of visual field and complete blindness. When conservative treatment becomes insufficient to stop OAG progression, a surgical intervention is considered. Currently, canaloplasty procedure is being introduced instead of conventional trabeculectomy for invasive OAG treatment. The aim of the study is to asses safety and efficacy of canaloplasty. METHODS: This prospective study included 67 eyes that received 360° canaloplasty with placement of a tensioning suture. Primary OAG (n = 35), secondary OAG in pseudoexfoliative syndrome (n = 13), and pigmentary glaucoma (n = 19) patients were included. Control check-ups were conducted pre-operatively and in a 18-month follow-up time. Study endpoints involved reduction in IOP values and in the number of glaucoma medications after the intervention. RESULTS: The intervention led to a significant 38% reduction in IOP value from the preoperative baseline to 18 months after the intervention. The number of medications decreased significantly by 89%. At 18 months postoperative, 79% eyes did not require any glaucoma medications. The incidence of complications after canaloplasty was low, and none of the adverse effects were vision threatening. A surgically-induced astigmatism was the most frequent complication. Pigmentary glaucoma patients were the most beneficial subgroup, with 50% reduction in IOP, the highest success rate, and 98% reduction in the number of medications used. CONCLUSION: This study proved that canaloplasty is an efficient and safe procedure in OAG eyes.

High-throughput cell-free screening of eukaryotic membrane protein expression in lipidic mimetics.

Membrane proteins play essential roles in cellular function and metabolism. Nonetheless, biophysical and structural studies of membrane proteins are impeded by the difficulty of their expression in and purification from heterologous cell-based systems. As an alternative to these cell-based systems, cell-free protein synthesis has proven to be an exquisite method for screening membrane protein targets in a variety of lipidic mimetics. Here we report a high-throughput screening workflow and apply it to screen 61 eukaryotic membrane protein targets. For each target, we tested its expression in lipidic mimetics: two detergents, two liposomes, and two nanodiscs. We show that 35 membrane proteins (57%) can be expressed in a soluble fraction in at least one of the mimetics with the two detergents performing significantly better than nanodiscs and liposomes, in that order. Using the established cell-free workflow, we studied the production and biophysical assays for mitochondrial pyruvate carrier (MPC) complexes. Our studies show that the complexes produced in cell-free are functionally competent in complex formation and substrate binding. Our results highlight the utility of using cell-free systems for screening and production of eukaryotic membrane proteins.