The Effect of Liquid-Phase Exfoliated Graphene Film on Neurodifferentiation of Stem Cells from Apical Papilla

Background: Dental stem cells, which originate from the neural crest, due to their easy accessibility might be good candidates in neuro-regenerative procedures, along with graphene-based nanomaterials shown to promote neurogenesis in vitro. We aimed to explore the potential of liquid-phase exfoliated graphene (LPEG) film to stimulate the neuro-differentiation of stem cells from apical papilla (SCAP). Methods: The experimental procedure was structured as follows: (1) fabrication of graphene film; (2) isolation, cultivation and SCAP stemness characterization by flowcytometry, multilineage differentiation (osteo, chondro and adipo) and quantitative PCR (qPCR); (3) SCAP neuro-induction by cultivation on polyethylene terephthalate (PET) coated with graphene film; (4) evaluation of neural differentiation by means of several microscopy techniques (light, confocal, atomic force and scanning electron microscopy), followed by neural marker gene expression analysis using qPCR. Results: SCAP demonstrated exceptional stemness, as judged by mesenchymal markers’ expression (CD73, CD90 and CD105), and by multilineage differentiation capacity (osteo, chondro and adipo-differentiation). Neuro-induction of SCAP grown on PET coated with graphene film resulted in neuron-like cellular phenotype observed under different microscopes. This was corroborated by the high gene expression of all examined key neuronal markers (Ngn2, NF-M, Nestin, MAP2, MASH1). Conclusions: The ability of SCAPs to differentiate toward neural lineages was markedly enhanced by graphene film

The evolution of a key segment in the Europe – Adria collision: the Fruška Gora of northern Serbia

The large number of roll-back systems in Mediterranean orogens poses interesting questions concerning interacting extensional back-arc deformation driven by different slabs. One such area characterized by a critical lack of kinematic studies is the connection between the Carpathians and Dinarides, where the Fruška Gora is an isolated inselberg of basement and Mesozoic cover surrounded by Miocene sediments. This area recorded a complex evolution related to the Cretaceous-Paleogene collision between Europe- and Adria-derived tectonic units, the Miocene extension of the Pannonian Basin and its subsequent inversion. This evolution has been analysed in a kinematic study combined with biostratigraphic and Rb-Sr thermochronology of sediments and theirmetamorphism. Results demonstrate a poly-phase tectonic evolution and allowed the discrimination of deformation events and basement affinities. The protolith of the Fruška Gora metamorphic core contains a typical Triassic-Jurassic sequence of the distal Adriatic margin that is overlain by Upper Cretaceous-Paleogene sediments deposited in the Neotethys subduction zone. A part of this basement still records a Late Jurassic (~148 Ma) burial metamorphic event that is associatedwith the coeval structural emplacement of overlying oceanic crust. Three successive deformation eventswere associatedwith the Latest Cretaceous-Early Oligocene contraction. The subsequent exhumation of the Fruška Gora metamorphic core started at ~28 Ma in the footwall of a large extensional detachment and continued by normal faulting during Early-Middle Miocene times. The large-scale extension took place during the extension of the Pannonian Basin and was associated with coeval translations and clockwise rotations of the Fruška Gora. Its present-day antiformal geometry truncated by high-angle reverse faults with S-ward vergence was established during the inversion of the Pannonian Basin, an effect of the late stage Pliocene-Quaternary Adriatic indentation. © 2012 Elsevier B.V. All rights reserved

The balance between orogenic building and subsequent extension during the Tertiary evolution of the NE Dinarides: Constraints from low-temperature thermochronology

One classical example of a back-arc basin is the Pannonian basin of Central Europe that formed duringMiocene times in response to the retreat of a slab situated at the exterior of the Carpathians. Within this overall framework, much less is known on the kinematics and exhumation of the NE Dinarides at their contact with the Pannonian basin. This area is key to understand the mechanics of the formation of the southeastern part of the basin in terms of coupling between basin subsidence and coeval orogenic exhumation. Three areas were selected as key sites for this exhumation study, i.e. Cer, Bukulja, and Fruška Gora Mountains, which are located in the Serbian part of the Dinarides. The low-temperature thermochronological study demonstrates that the formation of the Pannonian basin in this sector of the orogenic chain was associated with coeval large-scale exhumation in the footwall of large scale extensional detachments. This rapid exhumation took place during late Early to early Late Miocene times in response to asymmetric crustal stretching forming the SE part of the Pannonian basin and its southern prolongation along the Morava river valley. In footwalls of detachments, this process has exhumed rocks that were previously buried to intermediate crustal depths by the subduction of the Sava Ocean and the subsequent collision that took place between European- and Adriatic-derived units. This exhumation was associated with decompressionalmelting and local emplacement of genetically related plutons. The top-E extension cannot be associated with an E-ward movement of the intervening Carpathians units in this sector of the chain. The only other possible genetic option is the roll-back of a Dinaridic slab, a speculative hypothesis that would be compatible with the large-scale extension that was recently quantified in this segment of the orogenic chain. © 2012 Elsevier B.V

DNA analysis from human skeletal remains in forensic casework

To assess our laboratory’s success with skeletal remains and provide a benchmark for the forensic community involved in identification of these remains, we retrospectively examined our ability to develop DNA profiles from the remains analyzed in our laboratory in the last 7 years. Between January 2009 and December 2016, 70 DNA extractions were completed on skeletal remains from routine casework. 92% of skeletal remains analyzed were samples submitted for body identifications by law enforcement and only 8% were samples submitted to answer family identity or historical questions. Overall, the ability to obtain a full or partial profile primarily reflects the difference in the average age and the condition of the samples in these two categories and thus, difference in the quantity and quality of the DNA. We describe here the approximate age and type of remains we have received, whether a full, partial, or no profile was obtained, as well as the condition of the samples

Mutation rate at 13 rapidly mutating Y-STR loci in the population of Serbia

Recently, the interest of the forensic community has been focused on new Y-chromosomal short tandem repeats (Y-STRs), termed Rapidly Mutating Y-STRs (RM-YSTRs), which is able to differentiate between close males belonging to the same paternal lineage due to their high mutation rates. In this study, we have estimated a mutation rate for 13 RM-YSTR in 85 pairs of male relatives in the population of Serbia. We analysed 74 father-son pairs, and 11 twin pairs, to evaluate the capacity of distinguishing between male subjects within a single lineage. Each father-son couple was previously confirmed by autosomal STRs testing (AmpFℓSTR® Identifiler Plus™ kit, Applied Biosystems) with paternity probability ≥99.99% and also confirmed monozygotic or dizygotic twins. Results showed that, in the 74 father-son pairs 23 mutations were detected of which 22 were one-step mutations and 1 was two-step mutation, while in the 11 twin pairs 1 mutation was observed in one dizygotic twin pair. Five father-son pairs were found to have mutations at two loci, while one pair at four loci. Overall, the most mutable markers were DYF399S1, DYF387S1, DYF403S1a and DYS612. Our findings are encouraging and concur with previous studies showing that by RM-YSTR typing the discrimination power of male relatives could be considerably increased in comparison to every YSTR markers commonly used in forensic genetics

Rapidly mutating Y-STRs population data in the population of Serbia and haplotype probability assessment for forensic purposes

Recent research with Rapidly Mutating Y-STRs (RM Y-STRs) have shown that these loci provide substantially higher haplotype diversity and haplotype discrimination capacity in worldwide populations when compared with the Y-STRs commonly used in genetic forensics. The aim of this study was to develop an allelic frequency database for the population of Serbia in order to evaluate the resolution power of 13 RM Y-STRs. A total of 279 unrelated males from the population of Serbia were typed with 13 RM Y-STRs loci: DYF387S1, DYF399S1, DYF403S1a/b, DYF404S1, DYS449, DYS518, DYS526a/b, DYS547, DYS570, DYS576, DYS612, DYS626 and DYS627. A high Y-STR haplotype diversity was found (0.999768) in our samples. As expected, the RM Y-STR loci showed high genetics diversity (GD) values (>0.73) in the Serbian population. The highest GD was observed for the locus DYF399S1 (0.991), followed by loci DYF403S1a (0.976), DYF387S1 (0.907) and DYF404S1 (0.899). Based on the results of this study, the RM Y-STR loci showed remarkable haplotype resolution power in the population of Serbia, high genetics diversity and, therefore, demonstrating their usefulness in forensic identification cases

Ageing with HIV - a periodontal perspective

The importance of oral microflora composition in HIV-infected patients is well recognized. However, no studies so far have dealt with age-related changes in periodontal pathogens occurrence in HIV+ individuals. The aim of the present study was to assess and compare temporal changes of bacteria frequency in younger ( lt = 35 years) and older (>= 50 years) HIV-infected and non-infected individuals. Bacterial DNA was isolated from buccal swabs of 30 younger and 30 older subjects in both HIV+ and HIV-groups. By means of PCR the following microorganisms were detected: Aggregatibacter actinomycetemcomitans, Eikenella corrodens, Peptostreptococcus micros, Porphyromonas gingivalis, Prevotella intermedia, Tannerella forsythia and Treponema denticola. Oral and periodontal examinations were performed in all subjects. The prevalence of microorganisms was significantly higher in HIV+ patients compared to controls, and their distribution showed a notable shift. The decreasing incidence in HIV-subjects was: Pi>Pm>Pg>Aa>Ec>Tf>Td whilst in HIV+ it was: Pi>Pm>Ec>Pg>Tf>Aa>Td. Oral manifestations of HIV infection were more frequent in older compared to younger patients. All measured values of clinical periodontal parameters were significantly higher in older compared to younger HIV+ patients. Ageing in HIV+ subjects is accompanied with a substantial increase and rearrangements of periodontal microflora, potentially aggravating oral and systemic health