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Abstract Cladribine tablets (CladT) has been available for therapeutic use in France since March 2021 for the management of highly active relapsing multiple sclerosis (RMS). This high-efficacy disease-modifying therapy (DMT) acts as an immune reconstitution therapy. In contrast to most high-efficacy DMTs, which act via continuous immunosuppression, two short courses of oral treatment with CladT at the beginning of years 1 and 2 of treatment provide long-term control of MS disease activity in responders to treatment, without the need for any further pharmacological treatment for several years. Although the labelling for CladT does not provide guidance beyond the initial treatment courses, real-world data on the therapeutic use of CladT from registries of previous clinical trial participants and patients treated in routine practice indicate that MS disease activity is controlled for a period of years beyond this time for a substantial proportion of patients. Moreover, this clinical experience has provided useful information on how to initiate and manage treatment with CladT. In this article we, a group of expert neurologists from France, provide recommendations on the initiation of CladT in DMT-naïve patients, how to switch from existing DMTs to CladT for patients with continuing MS disease activity, how to manage patients during the first 2 years of treatment and finally, how to manage patients with or without MS disease activity in years 3, 4 and beyond after initiating treatment with CladT. We believe that optimisation of the use of CladT beyond its initial courses of treatment will maximise the benefits of this treatment, especially early in the course of MS when suppression of focal inflammation in the CNS is a clinical priority to limit MS disease progression.

A series of skulls (n = 27) of the American mink (Neogale vison) from the south-east of Belarus (Gomel region, floodplain of the Sozh River) was studied. The animals were caught by different hunters in 2000–2004. The sex of individuals was not determined. When examining the skull, only the most pronounced morpho-anatomical changes that can be diagnosed confidently as deviations from the norm were taken into account. In all cases, lamellar deposition of calcium salts in the area of tentorium cerebelli osseum inside the cranial vault was detected. The growing plate length reached half of the arch height in some individuals. These traits (considerable area of bone plates; presence of a sharp spine growing in different plains) allow suggesting that the analysed growths are of pathological origin. This pathology can considerably affect the viability and physiological status of individuals as it disrupts the functioning of the central nervous system. It is difficult to identify the cause of intracranial calcifications due to the possible effect of factors of various nature. Some degree of calcification of the opisthion region of foramen magnum was found. The changes occurred in the foramen shape cannot be considered phenetic variability. In most individuals, the thinning of maxillary bone in the teeth roots area is observed. However, we believe that the identified degree of bone tissue thinning is not critical and therefore does not affect the life expectancy of individuals. Two adult individuals have swelling of the maxillary bone. In one case, an extensive bone tissue excavation was identified on the left lower jaw, which led to the loss of the canine tooth. The analysed pathomorphological change is not of traumatic nature because in case of post-traumatic osteomyelitis sequesters (separating fragments) are formed. It is necessary to further analyse the American mink skulls available at scientific collections of Belarus and to identify the degree of calcium salt deposits and their impact on the foramen magnum phenotypes. We consider it necessary to create an annotated catalogue of pathologies and anomalies of the skull of the American mink in the south-east of Belarus and adjacent territories of Ukraine, since the morphological method is essential in the diagnosis of bone tissue diseases.

The studies were carried out in 2021 near the village of Yushevichi, Nesvizh Raion, Minsk Oblast (Belarus). The content of pellets of three bird species were analysed: Buteo buteo, Buteo lagopus, and Corvus corax. In total, 129 intact and about 30 destructed pellets were processed. Skeletal elements of five small mammal species belonging to five genera of two orders (Rodentia and Eulipotyphla) were found, including remains of nine speckled ground squirrels. The remains of seven of the nine ground squirrel individuals found in the pellets were found in raven pellets, however, the authors believe that this fact does not yet prove a greater influence of the raven on the local ground squirrel population. The pellets analysis and observations of feeding behaviour of the three bird species prove that the speckled ground squirrel is constantly included in the diet of both diurnal birds of prey (genus Buteo) and of atypical predators with a mixed nutrition (raven). Finding of intact skulls of Spermophilus suslicus in pellets of the mentioned bird species is unlikely, which is determined by its considerable size. This circumstance does not allow obtaining most of the craniometric characters, but does not prevent the detection of pathomorphological changes. To compare the craniological characters, we used the skulls of dead ground squirrels (n = 5) found in the field near the village of Yushevichi. These individuals had visible injuries (lacerated wounds, etc.) presumably left both by predators and fights between ground squirrels. The most striking pathomorphological changes were revealed: osteoporosis and osteolysis of dental alveoli of the upper and lower jaws and initial osteomyelitis of the cranial vault (in the parietal and frontal bones). We believe that the speckled ground squirrel’s conservation status should be raised to at least Category II (according to the system of categories adopted in Belarus) for the following reasons: over the past six years, out of 12 known colonies, only four colonies have survived, two of which are practically unviable; stable for 10 years reduction in the number; and range fragmentation (single localities). According to the IUCN classification, it corresponds to category CR (critically endangered).

Abstract Cladribine tablets (CladT) is a highly active oral disease-modifying therapy (DMT) for the management of relapsing multiple sclerosis (RMS). CladT acts as an immune reconstitution therapy, in that two short courses of treatment 1 year apart have been shown to suppress disease activity for a prolonged period in most patients, without need for continued DMT. Each course of CladT induces a profound reduction in B lymphocytes that recovers over months, and serious lymphopenia (Grade 3–4) is uncommon. Smaller reductions in levels of T lymphocytes occur slightly later: on average, these remain within the normal range and repopulate progressively. A larger effect occurs on CD8 vs. CD4 cells. Reactivation of latent or opportunistic infections (e.g. varicella zoster, tuberculosis) is mostly associated with very low lymphocyte counts ( 800/mm3 (if necessary) are important for avoiding infections and higher-grade lymphopenia. There was no demonstrable or apparent effect of CladT on the efficacy of vaccinations, including against Covid-19. Adverse events consistent with drug-induced liver injury (DILI) represent a rare but potentially serious complication of CladT therapy in spontaneous adverse event reporting; patients should be screened for liver dysfunction before starting treatment. Ongoing hepatic monitoring is not required, but CladT must be withdrawn if signs and symptoms of DILI develop. There was a numerical imbalance for malignancies when comparing cladribine to placebo in the clinical programme, particularly in short-term data, but recent evidence shows that the risk of malignancy with CladT is similar to the background rate in the general population and to that with other DMTs. Overall, CladT is well tolerated with a favorable safety profile appropriate for the management of RMS.

Diabetes mellitus is a widely prevalent chronic disease, and effective treatments for it remain limited. Allicin has anti-diabetic effect, however, the detailed mechanism has not been well investigated. The diabetic rats were treated with allicin for 6 weeks, and then the fecal samples, serum and tissues were collected for measurement of gut microbiota (GM), bile acids (BAs) and other indexes. Allicin decreased the blood glucose levels, serum lipid levels, and alleviated hepatic lipid deposition of the diabetic rats. Simultaneously, allicin changed the composition of GM and increased secondary BAs in serum. Interestingly, allicin significantly increased the glucagon-like peptide-1 (GLP-1) content in colon homogenate, and reduced the protein expression of fibroblast growth factor 15 (FGF15) in ileum. Moreover, allicin increased 7α-hydroxylase 1 (CYP7A1) expression in liver. The study demonstrated that allicin ameliorates metabolisms in the diabetic rats though modulating gut microbiota, bile acids, and inhibiting enterohepatic FGF15-CYP7A1 signaling.

The history of theriological research in the territory of Polissia from the 19th century to the present is considered (authors, works, and contributions to the study of the mammal fauna of the region). A review of the most significant publications on the theriofauna of the Polissia region is presented. The material is considered in chronological order, in four sections: ‘The period before systematic collectings’ (from the 19th century to 1920), ‘The period of field expeditionary research’ (1920–1940), ‘The resumption of theriological research at a new level’ (1945–1970), ‘Period of detailed stationary studies’ (1971–2000), ‘Modern stage of research’ (from 2001 to the present). A list of the most famous researchers of Polissia is given in accordance with the proposed periodization. The authors conclude that the number of studies and publications for the analysed period is connected, first, with state projects for the economic development of this territory (the construction of the Brest–Moscow railway at the end of the 19th century, large-scale land reclamation of Polissia throughout almost the entire 20th century). The key role of A. V. Fedyushin and I. N. Serzhanin for the development of theriological research is noted. The role of nature reserves created in the Chernobyl territory in the study of the mammal fauna of this region is discussed. The complete list of Polissia mammals including 80 species is given. It is concluded that the Polissia theriofauna has not been fully studied (insectivores, bats, and rodents). The theriological studies in the regions of Polissia were carried out unevenly: from single visits and local expeditions to stationary and long-term monitoring studies. The insufficient level of theriological monitoring and, as a result, the lack of proper analysis of changes in mammal assemblages are indicated. The existing scientific collections, where specimens of mammals from Polissia are stored, are not catalogued into a single database and are difficult to study. In Belarus, theriologists are divided and do not have a common platform for the exchange of information and experience. The positive experience of Ukrainian colleagues in the creation and maintenance of the long-term activity of the Theriological School is presented.

Abstract Background Triggering receptor expressed on myeloid cells 2 (Trem2) plays a protective role in neurodegenerative diseases. By contrast, Trem2 functions can exacerbate tissue damage during respiratory viral or liver infections. We, therefore, investigated the role of Trem2 in a viral encephalomyelitis model associated with prominent Th1 mediated antiviral immunity leading to demyelination. Methods Wild-type (WT) and Trem2 deficient (Trem2 −/−) mice were infected with a sublethal glia tropic murine coronavirus (MHV–JHM) intracranially. Disease progression and survival were monitored daily. Leukocyte accumulation and pathological features including demyelination and axonal damage in spinal cords (SC) were determined by flow cytometry and tissue section immunofluorescence analysis. Expression of select inflammatory cytokines and chemokines was measured by RT-PCR and global myeloid cell gene expression in SC-derived microglia and infiltrated bone-marrow-derived macrophages (BMDM) were determined using the Nanostring nCounter platform. Results BMDM recruited to SCs in response to infection highly upregulated Trem2 mRNA compared to microglia coincident with viral control. Trem2 deficiency did not alter disease onset or severity, but impaired clinical recovery after onset of demyelination. Disease progression in Trem2 −/− mice could not be attributed to altered virus control or an elevated proinflammatory response. A prominent difference was increased degenerated myelin not associated with the myeloid cell markers IBA1 and/or CD68. Gene expression profiles of SC-derived microglia and BMDM further revealed that Trem2 deficiency resulted in impaired upregulation of phagocytosis associated genes Lpl and Cd36 in microglia, but a more complex pattern in BMDM. Conclusions Trem2 deficiency during viral-induced demyelination dysregulates expression of other select genes regulating phagocytic pathways and lipid metabolism, with distinct effects on microglia and BMDM. The ultimate failure to remove damaged myelin is reminiscent of toxin or autoimmune cell-induced demyelination models and supports that Trem2 function is regulated by sensing tissue damage including a dysregulated lipid environment in very distinct inflammatory environments.

The purpose of the study was to analyse the impact of the COVID-19 pandemic on the healthcare of the orthopaedic patient, i.e. numbers of hip and knee replacement surgeries, 90-day mortality, waiting times and outpatient clinic visits.

SPX receptors regulate plant phosphate response via PHR transcription factors. Here, based on crystal structure analysis of rice PHR2 complexes, the authors propose that SPX2 regulates PHR2 by preventing DNA binding and oligomerisation of the PHR2 CC domain.

Abstract This study investigates effects of atmosphere‐ocean coupling on MJO precipitation and eastward propagation, and upper ocean conditions during and after MJO passage. To explore pathways for improving MJO prediction, three model experiments are conducted using the Unified Wave Interface‐Coupled Model at convection‐permitting (4 km) resolution: (a) uncoupled atmosphere‐only, (b) coupled atmosphere‐ocean, and (c) coupled atmosphere‐ocean with improved air‐sea flux algorithm simulations. The model simulations are compared with observations from the DYNAMO field campaign in 2011. Both coupled atmosphere‐ocean simulations produced eastward propagation of the MJO where the uncoupled, atmosphere‐only simulation did not. The uncoupled model overestimates both precipitation and surface winds associated with the MJO, while coupled model simulations substantially reduce model bias. Improved air‐sea fluxes lead to systematic improvements in precipitation, winds, sea surface temperature, and the ocean mixed layer when compared to the original coupled simulation. This leads to further improvement of the MJO's eastward propagation speed compared with observations. Despite these improvements, the regional coupled simulations still have difficulty representing the extent of convectively suppressed conditions in the Indian Ocean after MJO passage, which indicates the importance of the large‐scale environment from lateral boundary conditions. Coupled model simulations also reveal some issues in the representation of upper ocean stratification in the ocean model, especially errors in salinity, which result in overestimation of the mixed layer depth after MJO passage.

Abstract Despite recent advancements in numerical models, prediction of the Madden‐Julian Oscillation (MJO) remains a major challenge in numerical weather prediction and climate modeling. This study explores pathways for improving MJO prediction through systematic investigation of the effects of model resolution and moist physics on simulations of the MJO in Part 1, followed by effects of atmosphere‐ocean coupling in Part 2. The Unified Wave Interface—Coupled Model (UWIN‐CM) experiments with different cumulus parameterizations (CP) and explicit microphysics with convection‐permitting resolution are used to study the MJO convective initiation and eastward propagation. Observations of the atmosphere and ocean from the Dynamics of the MJO (DYNAMO) field campaign in 2011 are used to evaluate coupled model simulations. At lower resolution (12 km), the simulation with the Tiedtke CP produced an eastward propagating MJO event, while the simulation with the Kain‐Fritsch CP did not. The main difference between the two low‐resolution simulations is in the vertical structure of the atmospheric boundary layer which impacts the large‐scale MJO circulation and precipitation. The Kain‐Fritsch CP produced a persistent cloudy boundary layer that decoupled the tropospheric circulation from the surface and failed to produce the MJO. At higher resolution (4 km), convection‐permitting simulations are robust in capturing the observed eastward propagation of MJO precipitation and surface westerly winds. Increasing resolution improves the boundary layer structure and convective systems, which results in more realistic vertical structures of wind and moisture throughout the troposphere, as well as an improved eastward propagation of the MJO.

(1) Background: antimicrobial resistance is becoming a dramatic problem for public health, and the design of new antimicrobial agents is an active research area. (2) Methods: based on our previous work, we designed an improved version of the crabrolin peptide and characterized its functional and structural properties with a wide range of techniques. (3) Results: the newly designed peptide, crabrolin21, is much more active than the previous ones and shows specific selectivity towards bacterial cells. (4) Conclusions: crabrolin21 shows interesting properties and deserves further studies.

Genska terapija postaja čedalje bolj zanimiva tudi v onkologiji. Med aplikacijami je morda najzanimivejša imunostimulacija. Pripravimo lahko plazmidno DNA, ki nosi zapis za različne imunostimulatorne molekule, ki jih vnesemo v celice tumorjev ali normalnih tkiv. Ta tkiva postanejo proizvajalci teh molekul, ki lahko delujejo lokalno ali pa se izločajo tudi sistemsko v krvni obtok. Ker plazmidna DNA ne prehaja celične membrane, so potrebni dostavni sistemi, virusni ali nevirusni. V naših študijah uporabljamo predvsem nevirusni dostavni sistem – elektroporacijo. Interlevkin 12 (IL-12) je eden od zanimivih citokinov, za katerega je znano protitumorsko delovanje s spodbujanjem imunskega odziva in antiangiogenim delovanjem. Namen projekta SmartGene.si je bil pripraviti plazmid z zapisom za interlevkin 12 (plazmid phIL12) in pripraviti vse potrebno za njegovo klinično testiranje za zdravljenje kožnih tumorjev. V konzorciju smo združili moči s partnerji z akademskega in industrijskega področja. Treba je bilo pripraviti plazmid za uporabo v humani onkologiji po zahtevah Evropske agencije za zdravila (EMA). Za prijavo klinične študije na Javno agencijo za zdravila in medicinske pripomočke (JAZMP) smo morali izvesti tudi vse neklinične raziskave o varnosti in učinkovitosti zdravila. Nato je bilo treba razviti postopek priprave zdravila, zagotoviti primerne prostore za pripravo in izvedbo postopka priprave zdravila. V treh letih smo dosegli vse te zastavljene cilje in dobili dovoljenje za izvajanje klinične študije na kožnih tumorjih, ki ga je izdala JAZMP na osnovi pozitivnega mnenja Komisije Republike Slovenije za medicinsko etiko. Zdaj poteka klinična študija faze I preizkušanja plazmida phIL12 na kožnih tumorjih glave in vratu z namenom preveriti varnost in sprejemljivost genskega elektroprenosa plazmida v tumorje. Cilj študije je prav tako določiti primeren odmerek zdravila, ki bi ga v nadaljnji klinični študiji uporabili kot adjuvantno zdravljenje k ablativnim terapijam, kot sta radioterapija ali elektrokemoterapija.

The article discusses the species and taxonomic composition of the long-eared owl (Asio otus) preys based on the analysis of pellets (n = 209) collected in the winter-spring period in 2016 in the Malarytsky district (Lozitsa village) and the Brest region district center. The distance between Malaryta town and Lozitsa village is about 10 km. Parts of the skull of 512 small mammals (2.45 individuals per pellet) and one bird were found. Feeding on birds for the long-eared owl is episodic. Representatives of 2 orders, 10 genera and 12 species of small mammals (5 species of shrews and 7 rodents) became preys of the owl. The proportion of rodents is 98.24 % of all preys. The absolute dominant among prey species is Microtus arvalis (85.16 % of all victims), which is consistent with numerous work carried out in other regions. Significant portions are of Apodemus agrarius (4.10 %), Muscardinus avellanarius (2.54 %), Sylvaemus tauricus (1.76 %), and Alexandromys oeconomus (1.56 %). The list of preys is presented by meadow-field, synanthropic and different species actively moving from adjacent forests in the Malaryta river floodplain and canal systems (Sylvaemus tauricus, Sorex araneus, S. minutus, Neomys fodiens). The occurence of two shrew species Crocidura leucodon and C. suaveolens in the city of Malaryta has been proved, which corresponds to similar information for the city of Brest. This suggests that C. suaveolens inhabits the entire territory of the Belarusian Polesie at present. The occurence of the non-abundant species Sicista betulina in vicinities of the town of Malaryta was confirmed. The results obtained confirm the significant trophic effect of the long-eared owl on the local population of the hazel dormouse and also indicate the relatively high abundance of this rodent in the study area. Seven species were identified in pellets of the long-eared owl living near the village of Lozitsa, and 12 species of small mammals were identified in the town of Malaryta. The diversity of the landscape of the town of Malaryta determines the large number of prey species.

Management of locoregionally recurrent head and neck squamous cell carcinomas (HNSCC) is challenging due to potential radioresistance. Pulsed low-dose rate (PLDR) irradiation exploits phenomena of increased radiosensitivity, low-dose hyperradiosensitivity (LDHRS), and inverse dose-rate effect. The purpose of this study was to evaluate LDHRS and the effect of PLDR irradiation in isogenic HNSCC cells with different radiosensitivity.

Targeting the tumor vasculature through specific endothelial cell markers involved in different signaling pathways represents a promising tool for tumor radiosensitization. Two prominent targets are endoglin (CD105), a transforming growth factor β co-receptor, and the melanoma cell adhesion molecule (CD1046), present also on many tumors. In our recent in vitro study, we constructed and evaluated a plasmid for simultaneous silencing of these two targets. In the current study, our aim was to explore the therapeutic potential of gene electrotransfer-mediated delivery of this new plasmid in vivo, and to elucidate the effects of combined therapy with tumor irradiation. The antitumor effect was evaluated by determination of tumor growth delay and proportion of tumor free mice in the syngeneic murine mammary adenocarcinoma tumor model TS/A. Histological analysis of tumors (vascularization, proliferation, hypoxia, necrosis, apoptosis and infiltration of immune cells) was performed to evaluate the therapeutic mechanisms. Additionally, potential activation of the immune response was evaluated by determining the induction of DNA sensor STING and selected pro-inflammatory cytokines using qRT-PCR. The results point to a significant radiosensitization and a good therapeutic potential of this gene therapy approach in an otherwise radioresistant and immunologically cold TS/A tumor model, making it a promising novel treatment modality for a wide range of tumors.

The article analyses the new record of Neomys anomalus in the lakes Dolzhin (geographic coordinates of the place of capture 55°06'33,8"N, 28°36'03.1"E) and Vechelye (55°07'55.3"N, 28°36'38.6"E and 55°07'55.4"N, 28°36'37.2"E) of Ushachsky district, Vitebsk region, Belarus. The material was collected in July 2019. Captured individuals (n = 3) differed by individual external characteristics (a grey-white spot around the eye but not behind it; non-contrasting transition of colouration between the back and belly) from individuals captured in 2018 (n = 4) on Lake Borkovshchina and its ducts. The revealed morphological differences between individuals trapped in different years confirm the known data on phenetic variability of N. anomalus. The body weight (7.14–8.03 g) and the main parameters (for example, the ratio of tail length to body length was 0.65–0.68) did not differ significantly. Also individuals of N. anomalus did not differ significantly by craniometrical characters (height of coronoid process was 4.04–4.17 mm). In 2018–2019, individuals of the Mediterranean water shrew were trapped in three interconnected lakes, the total length of which with the channels is about 8 km. The shallow and densely overgrown with trees and shrubs channels between the lakes contribute to the dispersal of individuals. According to the results of the 2018 survey, the relative abundance of N. anomalus on Lake Borkovshchina and its channels was 4 individuals / 100 trap-days, and according to 2019 data on Lake Dolzhina and Lake Vechelye — 5.0 and 4.4, respectively. The findings give a reason to suggest the stability of the local population. One of the factors contributing to this phenomenon is the reduction in flood water. To maintain the abundance of the species, it is necessary, first of all, to preserve the shoreline, riparian and aquatic vegetation in lakes. It is advisable to include lakes into the system of protected areas with the status of reserves.

Metabolic dysfunction-associated fatty liver disease (MAFLD) is a complex disorder that is implicated in dysregulations in multiple biological pathways, orchestrated by interactions between genetic predisposition, metabolic syndromes and environmental factors. The limited knowledge of its pathogenesis is one of the bottlenecks in the development of prognostic and therapeutic options for MAFLD. Moreover, the extent to which metabolic pathways are altered due to ongoing hepatic steatosis, inflammation and fibrosis and subsequent liver damage remains unclear. To uncover potential MAFLD pathogenesis in humans, we employed an untargeted nuclear magnetic resonance (NMR) spectroscopy- and high-resolution mass spectrometry (HRMS)-based multiplatform approach combined with a computational multiblock omics framework to characterize the plasma metabolomes and lipidomes of obese patients without (n = 19) or with liver biopsy confirmed MAFLD (n = 63). Metabolite features associated with MAFLD were identified using a metabolome-wide association study pipeline that tested for the relationships between feature responses and MAFLD. A metabolic pathway enrichment analysis revealed 16 pathways associated with MAFLD and highlighted pathway changes, including amino acid metabolism, bile acid metabolism, carnitine shuttle, fatty acid metabolism, glycerophospholipid metabolism, arachidonic acid metabolism and steroid metabolism. These results suggested that there were alterations in energy metabolism, specifically amino acid and lipid metabolism, and pointed to the pathways being implicated in alerted liver function, mitochondrial dysfunctions and immune system disorders, which have previously been linked to MAFLD in human and animal studies. Together, this study revealed specific metabolic alterations associated with MAFLD and supported the idea that MAFLD is fundamentally a metabolism-related disorder, thereby providing new perspectives for diagnostic and therapeutic strategies.

Abstract Background Treatment options for recurrent head and neck tumours in the previously irradiated area are limited, including re-irradiation due to radioresistance of the recurrent tumour and previous dose received by surrounding normal tissues. As an in vitro model to study radioresistance mechanisms, isogenic cells with different radiosensitivity can be used. However, they are not readily available. Therefore, our objective was to establish and characterize radioresistant isogenic human pharyngeal squamous carcinoma cells and to evaluate early radiation response in isogenic parental, radioresistant and radiosensitive cells. Methods Radioresistant cells were derived from parental FaDu cells by repeated exposure to ionizing radiation. Radiosensitivity of the established isogenic radioresistant FaDu-RR cells was evaluated by clonogenic assay and compared to isogenic parental FaDu and radiosensitive 2A3 cells. Additional phenotypic characterization of these isogenic cells with different radiosensitivity included evaluation of chemosensitivity, cell proliferation, cell cycle, radiation-induced apoptosis, resolution of DNA double-strand breaks, and DNA damage and repair signalling gene expression before and after irradiation. Results In the newly established radioresistant cells in response to 5 Gy irradiation, we observed no alteration in cell cycle regulation, but delayed induction and enhanced resolution of DNA double-strand breaks, lower induction of apoptosis, and pronounced over-expression of DNA damage signalling genes in comparison to parental cells. On the other hand, radiosensitive 2A3 cells were arrested in G2/M-phase in response to 5 Gy irradiation, had a prominent accumulation of and slower resolution of DNA double-strand breaks, and no change in DNA damage signalling genes expression. Conclusions We concluded that the emergence of the radioresistance in the established radioresistant isogenic cells can be at least partially attributed to the enhanced DNA double-strand break repair, altered expression of DNA damage signalling and repair genes. On the other hand, in radiosensitive isogenic cells the reduced ability to repair a high number of induced DNA double-strand breaks and no transcriptional response in DNA damage signalling genes indicate on a lack of adaptive response to irradiation. Altogether, our results confirmed that these isogenic cells with different radiosensitivity are an appropriate model to study the mechanisms of radioresistance.

Abstract Background Septic shock is the most severe phase of sepsis and is associated with high rates of mortality. However, early stage prediction of septic shock outcomes remains difficult. Metabolomic techniques have emerged as a promising tool for improving prognosis. Methods Orthogonal projections to latent structures-discriminant analysis (OPLS-DA) models separating the serum metabolomes of survivors from those of non-survivors were established with samples obtained at the intensive care unit (ICU) admission (H0) and 24 h later (H24). For 51 patients with available H0 and H24 samples, multi-level modeling was performed to provide insight into different metabolic evolutions that occurred between H0 and H24 in the surviving and non-surviving patients. Relative quantification and receiver operational characteristic curves (ROC) were applied to estimate the predictability of key discriminatory metabolites for septic shock mortality. Results Metabolites that were involved in energy supply and protein breakdown were primarily responsible for differentiating survivors from non-survivors. This was not only seen in the H0 and H24 discriminatory models, but also in the H0-H24 paired models. Reanalysis of extra H0-H24 paired samples in the established multi-level model demonstrated good performance of the model for the classification of samplings. According to the ROC results, nine discriminatory metabolites defined consistently from the unpaired model and the H0-H24 time-trend change (ΔH24-H0) show good prediction of mortality. These results suggest that NMR-based metabolomic analysis is useful for a better overall assessment of septic shock patients. Conclusions Dysregulation of the metabolites identified by this study is associated with poor outcomes for septic shock. Evaluation of these compounds during the first 24 h after ICU admission in the septic shock patient may be helpful for estimating the severity of cases and for predicting outcomes. Trial registration All human serum samples were collected and stored, provided by the “center of biologic resources for liver disease”, in Jean Verdier Hospital, Bondy, France (BB-0033-00027).

Malignant gliomas are rapidly growing tumours that extensively invade the brain and have bad prognosis. Our study was performed to assess the metabolic effects of bevacizumab on the glioma cells carrying the IDH1 mutation, a mutation, associated with better prognosis and treatment outcome. Bevacizumab is known to inhibit tumour growth by neutralizing the biological activity of vascular endothelial growth factor (VEGF). However, the direct effects of bevacizumab on tumour cells metabolism remain poorly known.

Abstract Background Tumor necrosis factor (TNF) is associated with several neurodegenerative disorders including multiple sclerosis (MS). Although TNF-targeted therapies have been largely unsuccessful in MS, recent preclinical data suggests selective soluble TNF inhibition can promote remyelination. This has renewed interest in regulation of TNF signaling in demyelinating disease, especially given the limited treatment options for progressive MS. Using a mouse model of progressive MS, this study evaluates the effects of sustained TNF on oligodendrocyte (OLG) apoptosis and OLG precursor cell (OPC) differentiation. Methods Induction of experimental autoimmune encephalomyelitis (EAE) in transgenic mice expressing a dominant-negative interferon-γ receptor under the human glial fibrillary acidic protein promoter (GFAPγR1Δ) causes severe non-remitting disease associated with sustained TNF. Therapeutic effects in GFAPγR1Δ mice treated with anti-TNF compared to control antibody during acute EAE were evaluated by assessing demyelinating lesion size, remyelination, OLG apoptosis, and OPC differentiation. Results More severe and enlarged demyelinating lesions in GFAPγR1Δ compared to wild-type (WT) mice were associated with increased OLG apoptosis and reduced differentiated CC1+Olig2+ OLG within lesions, as well as impaired upregulation of TNF receptor-2, suggesting impaired OPC differentiation. TNF blockade during acute EAE in GFAPγR1Δ both limited OLG apoptosis and enhanced OPC differentiation consistent with reduced lesion size and clinical recovery. TNF neutralization further limited increasing endothelin-1 (ET-1) expression in astrocytes and myeloid cells noted in lesions during disease progression in GFAPγR1Δ mice, supporting inhibitory effects of ET-1 on OPC maturation. Conclusion Our data implicate that IFNγ signaling to astrocytes is essential to limit a detrimental positive feedback loop of TNF and ET-1 production, which increases OLG apoptosis and impairs OPC differentiation. Interference of this cycle by TNF blockade promotes repair independent of TNFR2 and supports selective TNF targeting to mitigate progressive forms of MS.

Tumor irradiation combined with adjuvant treatments, either vascular targeted or immunomodulatory, is under intense investigation. Gene electrotransfer of therapeutic genes is one of these approaches. The aim of this study was to determine, whether gene electrotransfer of plasmid encoding shRNA for silencing endoglin, with vascular targeted effectiveness, can radiosensitize melanoma B16F10 tumors.

Translationally Controlled Tumor Protein (TCTP) controls growth by regulating the G1/S transition during cell cycle progression. Our genetic interaction studies show that TCTP fulfills this role by interacting with CSN4, a subunit of the COP9 Signalosome complex, known to influence CULLIN-RING ubiquitin ligases activity by controlling CULLIN (CUL) neddylation status. In agreement with these data, downregulation of CSN4 in Arabidopsis and in tobacco cells leads to delayed G1/S transition comparable to that observed when TCTP is downregulated. Loss-of-function of AtTCTP leads to increased fraction of deneddylated CUL1, suggesting that AtTCTP interferes negatively with COP9 function. Similar defects in cell proliferation and CUL1 neddylation status were observed in Drosophila knockdown for dCSN4 or dTCTP, respectively, demonstrating a conserved mechanism between plants and animals. Together, our data show that CSN4 is the missing factor linking TCTP to the control of cell cycle progression and cell proliferation during organ development and open perspectives towards understanding TCTP's role in organ development and disorders associated with TCTP miss-expression.

The central nervous system (CNS) is vulnerable to several viral infections including herpes viruses, arboviruses and HIV to name a few. While a rapid and effective immune response is essential to limit viral spread and mortality, this anti-viral response needs to be tightly regulated in order to limit immune mediated tissue damage. This balance between effective virus control with limited pathology is especially important due to the highly specialized functions and limited regenerative capacity of neurons, which can be targets of direct virus cytolysis or bystander damage. CNS infection with the neurotropic strain of mouse hepatitis virus (MHV) induces an acute encephalomyelitis associated with focal areas of demyelination, which is sustained during viral persistence. Both innate and adaptive immune cells work in coordination to control virus replication. While type I interferons are essential to limit virus spread associated with early mortality, perforin, and interferon-γ promote further virus clearance in astrocytes/microglia and oligodendrocytes, respectively. Effective control of virus replication is nonetheless associated with tissue damage, characterized by demyelinating lesions. Interestingly, the anti-inflammatory cytokine IL-10 limits expansion of tissue lesions during chronic infection without affecting viral persistence. Thus, effective coordination of pro- and anti-inflammatory cytokines is essential during MHV induced encephalomyelitis in order to protect the host against viral infection at a limited cost.

Targeting tumor vasculature through specific endothelial cell markers represents a promising approach for cancer treatment. Here our aim was to construct an antibiotic resistance gene-free plasmid encoding shRNAs to simultaneously target two endothelial cell markers, CD105 and CD146, and to test its functionality and therapeutic potential in vitro when delivered by gene electrotransfer (GET) and combined with irradiation (IR). Functionality of the plasmid was evaluated by determining the silencing of the targeted genes using qRT-PCR. Antiproliferative and antiangiogenic effects were determined by the cytotoxicity assay tube formation assay and wound healing assay in murine endothelial cells 2H-11. The functionality of the plasmid construct was also evaluated in malignant melanoma tumor cell line B16F10. Additionally, potential activation of immune response was measured by induction of DNA sensor STING and proinflammatory cytokines by qRT-PCR in endothelial cells 2H-11. We demonstrated that the plasmid construction was successful and can efficiently silence the expression of the two targeted genes. As a consequence of silencing, reduced migration rate and angiogenic potential was confirmed in 2H-11 endothelial cells. Furthermore, induction of DNA sensor STING and proinflammatory cytokines were determined, which could add to the therapeutic effectiveness when used in vivo. To conclude, we successfully constructed a novel plasmid DNA with two shRNAs, which holds a great promise for further in vivo testing.

In this study, radiotherapy was combined with the gene electrotransfer (GET) of plasmid encoding shRNA against melanoma cell adhesion molecule (pMCAM) with dual action, which was a vascular-targeted effect mediated by the silencing of MCAM and an immunological effect mediated by the presence of plasmid DNA in the cytosol-activating DNA sensors. The effects and underlying mechanisms of therapy were evaluated in more immunogenic B16F10 melanoma and less immunogenic TS/A carcinoma. The silencing of MCAM potentiated the effect of irradiation (IR) in both tumor models. Combined therapy resulted in 81% complete responses (CR) in melanoma and 27% CR in carcinoma. Moreover, after the secondary challenge of cured mice, 59% of mice were resistant to challenge with melanoma cells, and none were resistant to carcinoma. Combined therapy reduced the number of blood vessels; induced hypoxia, apoptosis, and necrosis; and reduced cell proliferation in both tumor models. In addition, the significant increase of infiltrating immune cells was observed in both tumor models but more so in melanoma, where the expression of IL-12 and TNF-α was determined as well. Our results indicate that the combined therapy exerts both antiangiogenic and immune responses that contribute to the antitumor effect. However, tumor immunological status is crucial for a sufficient immune system contribution to the overall antitumor effect.

Multiple Sclerosis (MS) is a chronic inflammatory disease of the central nervous system (CNS) characterized by demyelination and axonal loss. Demyelinating lesions are associated with infiltrating T lymphocytes, bone marrow-derived macrophages (BMDM), and activated resident microglia. Tissue damage is thought to be mediated by T cell produced cytokines and chemokines, which activate microglia and/or BMDM to both strip myelin and produce toxic factors, ultimately damaging axons and promoting disability. However, the relative contributions of BMDM and microglia to demyelinating pathology are unclear, as their identification in MS tissue is difficult due to similar morphology and indistinguishable surface markers when activated. The CD4 T cell-induced autoimmune murine model of MS, experimental autoimmune encephalitis (EAE), in which BMDM are essential for demyelination, has revealed pathogenic and repair-promoting phenotypes associated with BMDM and microglia, respectively. Using a murine model of demyelination induced by a gliatropic coronavirus, in which BMDM are redundant for demyelination, we herein characterize gene expression profiles of BMDM versus microglia associated with demyelination. While gene expression in CNS infiltrating BMDM was upregulated early following infection and subsequently sustained, microglia expressed a more dynamic gene profile with extensive mRNA upregulation coinciding with peak demyelination after viral control. This delayed microglia response comprised a highly pro-inflammatory and phagocytic profile. Furthermore, while BMDM exhibited a mixed phenotype of M1 and M2 markers, microglia repressed the vast majority of M2-markers. Overall, these data support a pro-inflammatory and pathogenic role of microglia temporally remote from viral control, whereas BMDM retained their gene expression profile independent of the changing environment. As demyelination is caused by multifactorial insults, our results highlight the plasticity of microglia in responding to distinct inflammatory settings, which may be relevant for MS pathogenesis.

Methods of the diagnosis of pathophysiological processes in the skulls of hedgehogs are provided. They are based on the identification of functional relationships between the development of the individual parts of skull and evaluation of the potential danger for the changes. Swelling of frontal bones is not a diagnostic feature for diagnostics of the Eastern hedgehog, but a sign (indicator) of pathology. Swelling of frontal bones is not a diagnostic feature, but a sign (indicator) of pathology. Age changes of the hedgehogs skull arch’s inner surface are analyzed.

Periodontitis is characterized by the loss of the supporting tissues of the teeth in an inflammatory-infectious context. The diagnosis relies on clinical and X-ray examination. Unfortunately, clinical signs of tissue destruction occur late in the disease progression. Therefore, it is mandatory to identify reliable biomarkers to facilitate a better and earlier management of this disease. To this end, saliva represents a promising fluid for identification of biomarkers as metabolomic fingerprints. The present study used high-resolution 1H-nuclear magnetic resonance (NMR) spectroscopy coupled with multivariate statistical analysis to identify the metabolic signature of active periodontitis. The metabolome of stimulated saliva of 26 patients with generalized periodontitis (18 chronic and 8 aggressive) was compared to that of 25 healthy controls. Principal Components Analysis (PCA), performed with clinical variables, indicated that the patient population was homogeneous, demonstrating a strong correlation between the clinical and the radiological variables used to assess the loss of periodontal tissues and criteria of active disease. Orthogonal Projection to Latent Structure (OPLS) analysis showed that patients with periodontitis can be discriminated from controls on the basis of metabolite concentrations in saliva with satisfactory explained variance (R2X = 0.81 and R2Y = 0.61) and predictability (Q2Y = 0.49, CV-AUROC = 0.94). Interestingly, this discrimination was irrespective of the type of generalized periodontitis, i.e. chronic or aggressive. Among the main discriminating metabolites were short chain fatty acids as butyrate, observed in higher concentrations, and lactate, γ-amino-butyrate, methanol, and threonine observed in lower concentrations in periodontitis. The association of lactate, GABA, and butyrate to generate an aggregated variable reached the best positive predictive value for diagnosis of periodontitis. In conclusion, this pilot study showed that 1H-NMR spectroscopy analysis of saliva could differentiate patients with periodontitis from controls. Therefore, this simple, robust, non-invasive method, may offer a significant help for early diagnosis and follow-up of periodontitis.

Naso-maxillary suture's variability of Erinaceus concolor (n = 106) from Gomel Polesye was studied. The frequency distribution of the «concolor» morphotype (by Krystufek, 2002) is 8,5 %. It's much higher that in Europe (< 1 %). Length of suture in males is bigger that in females.

Viral infections have long been implicated as triggers of autoimmune diseases, including Multiple Sclerosis (MS), a central nervous system (CNS) inflammatory demyelinating disorder. Epitope spreading, molecular mimicry, cryptic antigen and bystander activation have been implicated as mechanisms responsible for activating self-reactive (SR) immune cells, ultimately leading to organ-specific autoimmune disease. Taking advantage of coronavirus JHM strain of mouse hepatitis virus (JHMV) induced demyelination, this study demonstrates that the host also mounts counteractive measures to specifically limit expansion of endogenous SR T cells. In this model, immune mediated demyelination is associated with induction of SR T cells after viral control. However, their decline during persisting infection, despite ongoing demyelination, suggests an active control mechanism.Antigen-specific IL-10 secreting CD4+ T cells (Tr1) and Foxp3+ regulatory T cells (Tregs), both known to control autoimmunity and induced following JHMV infection, were assessed for their relative in vivo suppressive function of SR T cells. Ablation of Foxp3+ Tregs in chronically infected DEREG mice significantly increased SR CD4+ T cells within cervical lymph nodes (CLN), albeit without affecting their numbers or activation within the CNS compared to controls. In contrast, infected IL-27 receptor deficient (IL-27R-/-) mice, characterized by a drastic reduction of Tr1 cells, revealed that SR CD4+ T cells in CLN remained unchanged, but were specifically increased within the CNS. These results suggest that distinct regulatory T cell subsets limit SR T cells in the draining lymph nodes and CNS to maximize suppression of SR T cell mediated autoimmune pathology. The JHMV model is thus valuable to decipher tissue specific mechanisms preventing autoimmunity.

The alkylpyridinium polymer APS8, a potent antagonist of α7 nicotinic acetylcholine receptors (nAChRs), selectively induces apoptosis in non-small cell lung cancer cells but not in normal lung fibroblasts. To explore the potential therapeutic value of APS8 for at least certain types of lung cancer, we determined its systemic and organ-specific toxicity in mice, evaluated its antitumor activity against adenocarcinoma xenograft models, and examined the in-vitro mechanisms of APS8 in terms of apoptosis, cytotoxicity, and viability. We also measured Ca2+ influx into cells, and evaluated the effects of APS8 on Ca2+ uptake while siRNA silencing of the gene for α7 nAChRs, CHRNA7. APS8 was not toxic to mice up to 5 mg/kg i.v., and no significant histological changes were observed in mice that survived APS8 treatment. Repetitive intratumoral injections of APS8 (4 mg/kg) significantly delayed growth of A549 cell tumors, and generally prevented regrowth of tumors, but were less effective in reducing growth of HT29 cell tumors. APS8 impaired the viability of A549 cells in a dose-dependent manner and induced apoptosis at micro molar concentrations. Nano molar APS8 caused minor cytotoxic effects, while cell lysis occurred at APS8 >3 µM. Furthermore, Ca2+ uptake was significantly reduced in APS8-treated A549 cells. Observed differences in response to APS8 can be attributed to the number of α7 nAChRs expressed in these cells, with those with more AChRs (i.e., A549 cells) being more sensitive to nAChR antagonists like APS8. We conclude that α7 nAChR antagonists like APS8 have potential to be used as therapeutics for tumors expressing large numbers of α7 nAChRs.

A variety of thermodynamic models have been developed to predict inorganic gas-aerosol equilibrium. To achieve computational efficiency a number of the models rely on a priori specification of the phases present in certain relative humidity regimes. Presented here is a new computational model, named UHAERO, that is both efficient and rigorously computes phase behavior without any a priori specification. The computational implementation is based on minimization of the Gibbs free energy using a primal-dual method, coupled to a Newton iteration. The mathematical details of the solution are given elsewhere. The model computes deliquescence behavior without any a priori specification of the relative humidities of deliquescence. Also included in the model is a formulation based on classical theory of nucleation kinetics that predicts crystallization behavior. Detailed phase diagrams of the sulfate/nitrate/ammonium/water system are presented as a function of relative humidity at 298.15 K over the complete space of composition.

INTRODUCTION: Acute-on-chronic liver failure is characterized by acute deterioration of liver function in patients with compensated or decompensated, but stable, cirrhosis. However, there is no accurate definition of acute-on-chronic liver failure and physicians often use this term to describe different clinical entities. Metabolomics investigates metabolic changes in biological systems and identifies the biomarkers or metabolic profiles. Our study assessed the metabolomic profile of serum using proton nuclear magnetic resonance ((1)H-NMR) spectroscopy to identify metabolic changes related to acute-on-chronic liver failure. PATIENTS: Ninety-three patients with compensated or decompensated cirrhosis (CLF group) but stable liver function and 30 patients with cirrhosis and hospitalized for the management of an acute event who may be responsible of acute-on-chronic liver failure (ACLF group), were fully analyzed. Blood samples were drawn at admission, and sera were separated and stored at -80°C until (1)H-NMR spectral analysis. Using orthogonal projection to latent-structure discriminant analyses, various metabolites contribute to the complete separation between these both groups. RESULTS: The predictability of the model was 0.73 (Q(2) Y) and the explained variance was 0.63 (R(2) Y). The main metabolites that had increased signals related to acute-on-chronic liver failure were lactate, pyruvate, ketone bodies, glutamine, phenylalanine, tyrosine, and creatinine. High-density lipids were lower in the ALCF group than in CLF group. CONCLUSION: A serum metabolite fingerprint for acute-on-chronic liver failure, obtained with (1)H-NMR, was identified. Metabolomic profiling may aid clinical evaluation of patients with cirrhosis admitted into intensive care units with acute-on-chronic liver failure, and provide new insights into the metabolic processes involved in acute impairment of hepatic function.

BACKGROUND: Glioma cells not only secrete high levels of vascular endothelial growth factor (VEGF) but also express VEGF receptors (VEGFR), supporting the existence of an autocrine loop. The direct impact on glioma cells metabolism of drugs targeting the VEGF pathway, such as Bevacizumab (Bev) or VEGFR Tyrosine Kinase Inhibitor (TKI), is poorly known. MATERIAL AND METHODS: U87 cells were treated with Bev or SU1498, a selective VEGFR2 TKI. VEGFR expression was checked with FACS flow cytometry and Quantitative Real-Time PCR. VEGF secretion into the medium was assessed with an ELISA kit. Metabolomic studies on cells were performed using High Resolution Magic Angle Spinning Spectroscopy (HR-MAS). RESULTS: U87 cells secreted VEGF and expressed low level of VEGFR2, but no detectable VEGFR1. Exposure to SU1498, but not Bev, significantly impacted cell proliferation and apoptosis. Metabolomic studies with HR MAS showed that Bev had no significant effect on cell metabolism, while SU1498 induced a marked increase in lipids and a decrease in glycerophosphocholine. Accordingly, accumulation of lipid droplets was seen in the cytoplasm of SU1498-treated U87 cells. CONCLUSION: Although both drugs target the VEGF pathway, only SU1498 showed a clear impact on cell proliferation, cell morphology and metabolism. Bevacizumab is thus less likely to modify glioma cells phenotype due to a direct therapeutic pressure on the VEGF autocrine loop. In patients treated with VEGFR TKI, monitoring lipids with magnetic resonance spectroscopic (MRS) might be a valuable marker to assess drug cytotoxicity.