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Nowadays, life is highly influenced by intense growth of various industries, high levels of pollution, and other environmental factors with harmful effects on human health. Therefore, cytogenetic monitoring is essential for detection of changes in the structure of chromosomes, which occur because of the effects of various genotoxic agents. In this review, we shall apprize the theoretical and experimental aspects of several tests for assessment of genotoxicity in humans such as Micronucleus assay, Comet assay, Chromosomal aberrations assessment and Sister chromatid exchanges analysis. These methods are accepted by the World Health Organization as standard tests for genotoxicological screening in humans. The methods are sensitive and confirm the cellular genotoxic effects of various genotoxicants.

We surveyed chestnut stands at 18 sites in 11 locations in Bulgaria in 2005 and 2007 for the presence of chestnut blight. We found chestnut blight in seven locations ( Belogradchik, Berkovitsa, Brezhani, Barziya, Govezhda, Petrich and Petrovo) but not in four others ( Tsaparevo, Kresna, Dupnica and Botevgrad). We successfully isolated Cryphonectria parasitica from cankers on 606 trees with symptoms of chestnut blight and assayed them for vegetative compatibility (vc) types and mating type. Three vc types were identified among the 606 isolates; all three were among the European vc types with known vegetative incompatibility ( vic) genotypes. Vc type EU-12 was the most common, representing 80% of the isolates, and was found at all locations with blight, with the exception of Belogradchik in north-west Bulgaria, where all isolates were vc type EU-2. Only one population ( Barziya) had more than one vc type, with a combination of EU-12 and EU-10 in almost equal frequencies. Similarly, the diversity of mating types was very low. All but three of 536 isolates assayed were in mating-type MAT-1; MAT-2 was only found in one population in the north-west ( Berkovitsa). We inspected 671 bark samples from chestnut blight cankers with stromata of C. parasitica and found perithecia in only 33, of which 28 were from Berkovitsa where MAT-2 was present. We did not detect hypoviruses in any of the 270 isolates screened using the standard double-stranded RNA extraction protocol. Similar to results from previous studies in south-eastern Europe, the diversity of vc types and mating type of C. parasitica in Bulgaria is low, and reproduction of the fungus is mainly asexual. Unfortunately, naturally occurring hypovirulence was not detected. Nevertheless, we observed a small number of superficial cankers typical of those caused by C. parasitica isolates infected with a hypovirus. [ABSTRACT FROM AUTHOR]

Abstract Background Human immunodeficiency virus type 1 (HIV-1) uses cellular proteins and machinery to ensure transmission to uninfected cells. Although the host proteins involved in the transport of viral components toward the plasma membrane have been investigated, the dynamics of this process remain incompletely described. Previously we showed that the double-stranded (ds)RNA-binding protein, Staufen1 is found in the HIV-1 ribonucleoprotein (RNP) that contains the HIV-1 genomic RNA (vRNA), Gag and other host RNA-binding proteins in HIV-1-producing cells. Staufen1 interacts with the nucleocapsid domain (NC) domain of Gag and regulates Gag multimerization on membranes thereby modulating HIV-1 assembly. The formation of the HIV-1 RNP is dynamic and likely central to the fate of the vRNA during the late phase of the HIV-1 replication cycle. Results Detailed molecular imaging of both the intracellular trafficking of virus components and of virus-host protein complexes is critical to enhance our understanding of factors that contribute to HIV-1 pathogenesis. In this work, we visualized the interactions between Gag and host proteins using bimolecular and trimolecular fluorescence complementation (BiFC and TriFC) analyses. These methods allow for the direct visualization of the localization of protein-protein and protein-protein-RNA interactions in live cells. We identified where the virus-host interactions between Gag and Staufen1 and Gag and IMP1 (also known as VICKZ1, IGF2BP1 and ZBP1) occur in cells. These virus-host interactions were not only detected in the cytoplasm, but were also found at cholesterol-enriched GM1-containing lipid raft plasma membrane domains. Importantly, Gag specifically recruited Staufen1 to the detergent insoluble membranes supporting a key function for this host factor during virus assembly. Notably, the TriFC experiments showed that Gag and Staufen1 actively recruited protein partners when tethered to mRNA. Conclusions The present work characterizes the interaction sites of key components of the HIV-1 RNP (Gag, Staufen1 and IMP1), thereby bringing to light where HIV-1 recruits and co-opts RNA-binding proteins during virus assembly.

The effectiveness of modern medical education largely depends on the integration and utilization of digital technologies in teaching various disciplines. In this pilot usability study, we introduced 3D printed anthropomorphic dental models, specifically designed for the elective discipline "Digital and Metal-Free Techniques in Dental Technology" from the curriculum of the Dental Technician specialty in the Medical University of Varna. The evaluation focused on dental technician students' perception of this novel learning environment, its influence on their performance, and the potential for future application of these models and related 3D technologies in their professional practice. A validated satisfaction questionnaire was distributed among 80 students, comprising the total cohort. The results indicated a high acceptance rate, with nearly 95% of participants finding the use of digitally created 3D-printed dental models beneficial. More than 90% believed that exploring digital technologies would enhance their skills. The well-trained instructor's competence in technology use convinced students of its value, with more than 98% expressing a willingness to incorporate these technologies into their future work for improved precision in dental models. However, due to the current high cost of needed equipment, only 10% of participants may practicably introduce this novel technology into their practical work. The use of anatomically accurate 3D printed models is a valuable addition to the current dental technician curriculum in medical colleges.

Background Neuroanatomy is one of the most complex areas of anatomy to teach to medical students. Traditional study methods such as atlases and textbooks are mandatory but require significant effort to conceptualize the three-dimensional (3D) aspects of the neuroanatomical regions of interest. Objectives To test the feasibility of human anatomy teaching medical students in a virtual reality (VR) immersive environment using photorealistic three-dimensional models (PR3DM) of human anatomy, in a limited anatomical body donation program. Methods We used surface scanning technology (photogrammetry) to create PR3DM of brain dissections. The 3D models were uploaded to VR headsets and used in immersive environment classes to teach second-year medical students. Twenty-eight medical students (mean age 20.11, SD 1.42), among which 19 females (n=28/67.9%) and nine males (n=28/32.1%), participated in the study. The students had either none or minimal experience with the use of VR devices. The duration of the study was three months. After completing the curriculum, a survey was done to examine the results. Results The average rating of the students for their overall experience with the method is 4.57/5 (SD=0.63). The "Possibility to study models from many points of view" and "Good Visualization of the models" were the most agreed upon advantages, with 24 students (n=28, 85.7%), and 95% confidence intervals (CI) [0.6643, 0.9532]. The limited availability of the VR headsets was the major disadvantage as perceived by the students, with 11 students (n=28, 39.3%), 95% CI [0.2213, 0.5927] having voted for the option. The majority of the students (25) (n=28, 89.2%, SD=0.31) agreed with the statement that the use of VR facilitated their neuroanatomy education. Conclusion This study shows the future potential of this model of training in limited cadaver dissection options to provide students with modern technological methods of training. Our first results indicate a prominent level of student satisfaction from VR training with minimum negative reactions to the nature of headsets. The proof of concept for the application of photorealistic models in VR neuroanatomy training combined with the initial results of appreciation among the students predisposes the application of the method on a larger scale, adding a nuance to the traditional anatomy training methods. The low number of headsets used in the study limits the generalization of the results but offers possibilities for future perspectives of research., Competing Interests: The authors have declared that no competing interests exist., (Copyright © 2024, Trandzhiev et al.)

Background and Objectives: Intraoperative orientation during microsurgery has a prolonged learning curve among neurosurgical residents. Three-dimensional (3D) understanding of anatomy can be facilitated with realistic 3D anatomic models created from photogrammetry, where a series of 2-dimensional images is converted into a 3D model. This study implements an algorithm that can create photorealistic intraoperative 3D models to exemplify important steps of the operation, operative corridors, and surgical perspectives., Methods: We implemented photograph-based and video-based scanning algorithms for uptakes using the operating room (OR) microscope, targeted for superficial structures, after surgical exposure, and deep operative corridors, in cranial microsurgery. The algorithm required between 30-45 photographs (superficial scanning), 45-65 photographs (deep scanning), or approximately 1 minute of video recording of the entire operative field to create a 3D model. A multicenter approach in 3 neurosurgical departments was applied to test reproducibility and refine the method., Results: Twenty-five 3D models were created of some of the most common neurosurgical approaches-frontolateral, pterional, retrosigmoid, frontal, and temporal craniotomy. The 3D models present important steps of the surgical approaches and allow rotation, zooming, and panning of the model, enabling visualization from different surgical perspectives. The superficial and medium depth structures were consistently presented through the 3D models, whereas scanning of the deepest structures presented some technical challenges, which were gradually overcome with refinement of the image capturing process., Conclusion: Intraoperative photogrammetry is an accessible method to create 3D educational material to show complex anatomy and demonstrate concepts of intraoperative orientation. Detailed interactive 3D models, displaying stepwise surgical case-based anatomy, can be used to help understand details of the operative corridor. Further development includes refining or automatization of image acquisition intraoperatively and evaluation of other applications of the resulting 3D models in training and surgical planning., (Copyright © Congress of Neurological Surgeons 2024. All rights reserved.)

Background: Spinal cord hemangioblastomas are benign, highly vascular neoplasms that affect the brain and, rarely, the spinal cord. They can be solitary or as part of von Hippel-Lindau syndrome. Radiosurgery is not a suitable treatment option. Endovascular embolization can only be adjunct to surgery., Method: We present a detailed approach to resection of a spinal cord hemangioblastoma. A video demonstrates the microsurgical technique and discusses complication avoidance., Conclusion: The pitfalls to consider are preservation of normal spinal cord vessels, protection of the pia-arachnoid cleavage plane, and avoidance of tumor piecemeal removal. Careful microsurgical resection and detailed preoperative planning are key., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

In the 2021 WHO classification of Tumors of the Central Nervous System, additional molecular characteristics have been included, defining the following adult-type diffuse glioma entities: Astrocytoma IDH-mutant, Oligodendroglioma IDH-mutant and 1p/19q-codeleted, and Glioblastoma IDH-wildtype. Despite advances in genetic analysis, precision oncology, and targeted therapy, malignant adult-type diffuse gliomas remain "hard-to-treat tumors", indicating an urgent need for better diagnostic and therapeutic strategies. In the last decades, miRNA analysis has been a hotspot for researching and developing diagnostic, prognostic, and predictive biomarkers for various disorders, including brain cancer. Scientific interest has recently been directed towards therapeutic applications of miRNAs, with encouraging results. Databases such as NCBI, PubMed, and Medline were searched for a selection of articles reporting the relationship between deregulated miRNAs and genetic aberrations used in the latest WHO CNS classification. The current review discussed the recommended molecular biomarkers and genetic aberrations based on the 2021 WHO classification in adult-type diffuse gliomas, along with associated deregulated miRNAs. Additionally, the study highlights miRNA-based treatment advancements in adults with gliomas., Competing Interests: The authors declare that there are no conflicts of interest., (© 2023 The Authors.)

Introduction Three-dimensional (3D) printing is an affordable aid that is useful in neurosurgery. It allows for better visualization and tactile appreciation of the individual anatomy and regions of interest and therefore potentially lowers the risk of complications. There are various applications of this technology in the field of neurosurgery. Materials and methods In this paper, we present a basic methodology for the creation of a 3D printed model using only open-source software for medical image editing, model generation, pre-printing preparation, and analysis of the literature concerning the practical use of this methodology. Results The literature review on the current applications of 3D printed models in neurosurgery shows that they are mostly used for preoperative planning, surgical training, and simulation, closely followed by their use in patient-specific implants and instrumentation and medical education. Materialise TM Mimics is the most frequently used commercial software for a 3D modeling for preoperative planning and surgical simulation, while the most popular open-source software for the same applications is 3D Slicer. In this paper, we present the algorithm that we employ for 3D printing using Horos TM , Blender, and Cura software packages which are all free and open-source. Conclusion Three-dimensional printing is becoming widely available and of significance to neurosurgical practice. Currently, there are various applications of this technology that are less demanding in terms of technical knowledge and required fluency in medical imaging software. These predispositions open the field for further research on the possible use of 3D printing in neurosurgery., Competing Interests: The authors have declared that no competing interests exist., (Copyright © 2022, Vezirska et al.)

At present, brain tumours remain one of the "hard-to-treat" malignancies with minimal improvement in patients' survival. Recently, miRNAs have been shown to correlate with oncogenesis and metastasis and have been investigated as potential biomarkers for diagnosis, prognosis and therapy prediction in different brain malignancies. The aim of the current study was to select an accurate and affordable brain tumour detection and grading approach. In the present study, we analysed the applicability of a restricted miRNA signature that could differentiate among patients with primary as well as metastatic brain tumours. Fresh tumour tissues were collected from Bulgarian patients (n = 38), including high-grade gliomas (n = 23), low-grade gliomas (n = 10) and brain metastases (n = 5) from lung cancer. Total RNAs enriched with microRNAs were isolated and differentially expressed miRNAs were analyzed by RT-qPCR using TaqMan Advanced miRNA assay. We selected a signature of miR-21, miR-10b, miR-7, miR-491 that showed good diagnostic potential in high-grade gliomas, low-grade gliomas and brain metastases compared with normal brain tissues. Our results showed that miR-10b could reliably differentiate brain metastases from high-grade gliomas, while miR-491 could distinguish low-grade from high-grade gliomas and brain metastases from low-grade gliomas. We observed that miR-21 and miR-7 correlated with disease recurrence, survival status and the Karnofsky Performance Status. The selected signature of miR-7, miR-21, miR-10b and miR-491 could be used as a highly accurate diagnostic, grading and prognostic biomarker in differentiating various types of brain tumours. Our data suggest that the 4-miRNAs signature could be further analysed for predicting treatment response and for future miRs-based targeted therapy. The ongoing studies on miRs-based targeted therapy related to our selected miRNA signature are also reviewed., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Background: Next-generation sequencing has facilitated the diagnosis of neurodevelopmental disorders with variable and non-specific clinical findings. Recently, a homozygous missense p.(Asp37Tyr) variant in TRAPPC2L, a core subunit of TRAPP complexes which function as tethering factors during membrane trafficking, was reported in two unrelated individuals with neurodevelopmental delay, post-infectious encephalopathy-associated developmental arrest, tetraplegia and accompanying rhabdomyolysis., Methods: We performed whole genome sequencing on members of an Ashkenazi Jewish pedigree to identify the underlying genetic aetiology of global developmental delay/intellectual disability in three affected siblings. To assess the effect of the identified TRAPPC2L variant, we performed biochemical and cell biological functional studies on the TRAPPC2L protein., Results: A rare homozygous predicted deleterious missense variant, p.(Ala2Gly), in TRAPPC2L was identified in the affected siblings and it segregated with the neurodevelopmental phenotype within the family. Using a yeast two-hybrid assay and in vitro binding, we demonstrate that the p.(Ala2Gly) variant, but not the p.(Asp37Tyr) variant, disrupted the interaction between TRAPPC2L and another core TRAPP protein, TRAPPC6a. Size exclusion chromatography suggested that this variant affects the assembly of TRAPP complexes. Employing two different membrane trafficking assays using fibroblasts from one of the affected siblings, we found a delay in traffic into and out of the Golgi. Similar to the p.(Asp37Tyr) variant, the p.(Ala2Gly) variant resulted in an increase in the levels of active RAB11., Conclusion: Our data fill in a gap in the knowledge of TRAPP architecture with TRAPPC2L interacting with TRAPPC6a, positioning it as a putative adaptor for other TRAPP subunits. Collectively, our findings support the pathogenicity of the TRAPPC2L p.(Ala2Gly) variant., Competing Interests: Competing interests: None declared., (© Author(s) (or their employer(s)) 2021. No commercial re-use. See rights and permissions. Published by BMJ.)

Background: Vitamins and/or mineral food supplements (VMS) are mono- or multi-component products prepared in different pharmaceutical forms and categorized as food supplements. Numerous researchers have investigated the epidemiological predictors of use of VMS in population. Data of this kind in our setting are limited., Aims: This survey aimed to gain information on the prevalence of use, certain sociodemographic predictors and reasons for VMS use in a group of 256 outpatients in Skopje., Materials and Methods: This study was designed as an open-ended, cross-sectional survey. Data on VMS use were collected by survey method with a specially designed questionnaire as an instrument for this research. Appropriate statistical tests were used to analyse the data., Results: Data from 256 outpatients from Skopje were analysed. Of them, one hundred and five (41.02%) reported using VMS. Female outpatients (p<0.01) and participants of Macedonian ethnicity (p<0.01), with a higher educational level (p<0.01) and those having their own income (p<0.05) reported significantly higher consumption of VMS. We found a statistically significant difference between the group of users and non-users of VMS regarding smoking status (p<0.05), alcohol consumption (p<0.05) and presence of a chronic disease (p<0.01). Physicians, magazines and friends were the most common source of information on VMS while maintaining good health and prevention of disease was the most common reasons for the use of these products. Only 63% of the users had disclosed the information on taking VMS to their physicians., Conclusion: This research provided an insight into VMS in our setting. Predictors on the use of VMS are of interest since these products contribute to total intake of vitamins and minerals in the population and they represent a source of valuable information when planning public health activities.

Background: Borrelial infection is characterized by various skin manifestations that are usually classified into three main types: chronic migratory erythema, borrelial lymphocytoma and acrodermatitis chronica atrophicans. We report an unusual case of borrelial cutaneous infection presenting as a mediofacial erythema that cannot be included in any of these three categories., Case Report: A 51-year-old woman presented with infiltrated erythema of the middle of the face extending to the neck and chin. Medical history and physical examination revealed no signs of rosaceae. Infection with Borrelia was suspected on skin biopsy examination, which showed an inflammatory dermal infiltrate containing numerous plasma cells. The diagnosis of B.afzelii infection was confirmed by serology and polymerase chain reaction on the skin biopsy, both of which were positive for B.afzelii., Discussion: Borrelial erythema of the face may represent a special form of cutaneous borrelial infection, which must be considered in the differential diagnosis of facial erythema, especially in areas of endemic borreliosis.

Expanding populations are often less genetically diverse at their margins than at the centre of a species' range. Established, older populations of the chestnut blight fungus, Cryphonectria parasitica, are more variable for vegetative compatibility (vc) types than in expanding populations in southeastern Europe where C. parasitica has colonized relatively recently. To test whether vc types represent clones, we genotyped 373 isolates of C. parasitica from southern Italy, Romania, Bulgaria, Macedonia, Greece and Turkey using 11 sequence-characterized amplified region (SCAR) markers. Ten SCAR loci and six vegetative incompatibility (vic) loci were polymorphic in these samples. These populations are clonal by all criteria tested: (i) among 373 isolates, we found only eight multilocus haplotypes, and the same haplotypes were found in multiple countries, sometimes separated in time by as much as 12 years; (ii) the number of haplotypes observed was significantly less than expected under random mating; (iii) populations are in linkage disequilibrium; (iv) the two sets of independent markers, SCARs and vc types, are highly correlated; and (v) sexual structures of C. parasitica were found only in Bulgaria and Romania. One mating type (MAT-1) was found in 98% of the isolates sampled. In contrast, a population in northern Italy, in the central part of the range in Europe, had 12 multilocus haplotypes among 19 isolates. The spread of a few clones could be the result either of founder effect and restricted migration, or these clones have greater fitness than others and spread because they are better adapted to conditions in southeastern Europe.

The HIV-1 ribonucleoprotein (RNP) contains the major structural protein, pr55(Gag), viral genomic RNA, as well as the host protein, Staufen1. In this report, we show that the nonsense-mediated decay (NMD) factor UPF1 is also a component of the HIV-1 RNP. We investigated the role of UPF1 in HIV-1-expressing cells. Depletion of UPF1 by siRNA resulted in a dramatic reduction in steady-state HIV-1 RNA and pr55(Gag). Pr55(Gag) synthesis, but not the cognate genomic RNA, was efficiently rescued by expression of an siRNA-insensitive UPF1, demonstrating that UPF1 positively influences HIV-1 RNA translatability. Conversely, overexpression of UPF1 led to a dramatic up-regulation of HIV-1 expression at the RNA and protein synthesis levels. The effects of UPF1 on HIV-1 RNA stability were observed in the nucleus and cytoplasm and required ongoing translation. We also demonstrate that the effects exerted by UPF1 on HIV-1 expression were dependent on its ATPase activity, but were separable from its role in NMD and did not require interaction with UPF2.

Investigations were carried out in production conditions on the sources of surface contamination of pork with regard to sanitary-indicative and pathogenic (Salmonellae) organisms. A total of 74 series of experiments were carried out, every series making use of the following: 10 fecal samples taken on the preslaughter premise; the results of studying the skin cover of 3 animals prior to washing, after washing, and after stunning, employing the method of Dryabina in studying skin areas of 25 cm2; as many as three washings taken from within the path from the place where the animals were washed to the place where they were stunned; the results of studying the surface of the meat produced (from 3 animals in each case) and at the site of starting skin removal, with sampling the abdomen muscles and the group along with sampling the meat itself after cleaning and washing it - taking impression preparations, resp., employing the contact method with a 44 cm2 surface area of the agar. No Salmonella bacteria were found in the 1628 analyses and fecal samples of the skin of pigs intended for slaughter. The washing of the animals lowered more than 70 times the total bacterial contamination of the skin cover. It was found that the site of starting skin removal and the muscles of the abdomen and those of the forelimbs were most strongly contaminated.