Ochratoxin A affects oocyte maturation and subsequent embryo developmental dynamics in the juvenile sheep model

The genotoxic and nephrotoxic mycotoxin Ochratoxin A (OTA) has also been reported to have adverse effects on oocyte maturation and embryo development. Previous studies on the effects of OTA on female fertility have used micromolar concentrations, but no information is available to date on effects in a more relevant nanomolar range. This study used a juvenile sheep model to evaluate the effects of oocyte exposure to low levels of OTA on maturation, fertilization, and embryo development. Further, it was investigated whether different mechanisms of action of OTA could be responsible for varying toxic effects at different levels of exposure. Cumulus-oocyte-complexes (COCs) were exposed to 10 μmol/L–0.1 nmol/L OTA during in vitro maturation and evaluated for cumulus viability, oocyte maturation, and bioenergetic/oxidative status. COCs were subjected to in vitro fertilization, embryo culture, and embryo quality assessment via morphology, viability, bioenergetic/oxidative status, and time-lapse monitoring. At micromolar concentrations, OTA induced cytotoxic effects, by reducing cumulus expansion and oocyte maturation. OTA altered temporospatial dynamics of zygote pronuclear formation and embryo morphokinetics. Blastocysts, even morphologically normal, were found to undergo collapse events, which were probably related to boosted blastocyst mitochondrial activity. At nanomolar concentrations, OTA did not affect COC morpho-functional parameters, but impaired oocyte ability to prevent polyspermy and increased blastocyst apoptosis. In conclusion, in the female germ cell, cytotoxic nonspecific effects characterize OTA-induced toxicity at high exposure levels, whereas fine tuning-mode effects, not associated with altered cell viability and integrity, characterize OTA toxic action at low levels

Elemental Analysys of metallic samples by nanopartiles-enhanced laser-ablation ICP-MS

This conventional method of Solid sample analysis is very dirty and produces a lot of chemical waste. Conversely, laser ablation is a green technology because it does not require acid dissolution and does not generate chemical wastes. Moreover, LA-ICPMS technique presents other interesting features such as high spatial resolution, extended linear dynamic range, low detection limits, extremely small sample quantities required for analysis. We propose an analytical strategy to enhance the sensitivity of LA- ICPMS by the deposition of gold nanoparticles (AuNPs) on the surface of investigated samples. The undoubted strengths of this approach are represented by simplicity, low-cost budget and fast performance. The commercial LA-ICPMS set-up has not been changed in any way and laser parameters, type and flow of gas carrier is not modified to improve the ablation of the sample [1]. The sample surfaces have been altered by depositing metallic nanoparticle (NPs) colloidal dispersions, thus increasing their response to laser, but preserving their chemical properties. A consistent lowering of the breakdown threshold and a remarkable increase in the measured signal intensity have been observed. This has allowed to determine traces and ultratraces in many areas (environmental, forensic, clinical, cultural heritage, food analysis, ecc.). In the method developed, the drops of NPs can be put in a controlled manner on the sample surfaces and completely removed before the analysis by laser ablation. This represents a further advantage compared to the classical methods of sample analysis. The first results obtained with this technique in the analysis of metal matrices show enhancement of the signal of one order of magnitude for most of the analyzed metals. [1] Z. Wang, B. Hattendorf and D. Günther, J. Am. Soc. Mass Spectrom. 17 (2006) 641-651

Mapping Apulian Red Figure Pottery by a multitechnique approach

Apulian red-figured pottery, one of the most important examples of ceramic handcraft production in Magna Graecia, dating back to the 5th and 4th centuries BCE and coming from the most relevant sites in Apulia (Southern Italy), has been extensively characterized by our group for several years [1-5]. Our main goals are various and quite ambitious: highlighting technological differences between Apulian red-figured pottery and the most famous Attic one, obtaining valuable knowledge about pottery workshops and painters and defining the nature of coatings and decorations. We have investigated ceramic body, black gloss and overpainting areas of items by different techniques according to issues to be solved and samples availability. The ceramic bodies’ elemental composition has been investigated by inductively coupled plasma mass spectrometry (ICPMS), the mineralogical composition of pastes by polarized-light optical and electron microscopies (OM and SEM-EDS), and X-ray powder diffraction (PXRD). The fruitful combination of results driven from multivariate statistical treatment of compositional data and mineralogical arrangement of pastes allows us to formulate hypotheses about the provenance of items and manufacturing tradition of workshops, starting to make it possible to comprehend the connections among ceramic technology, artistic expression, and workshop practice in the samples analyzed. Also, with regard to the material brought to light during the 19th century, it is known that “antiquarian type” of restoration was the most used (i.e. reconstruction and repainting, following the mimetic taste of the time). From this point of view, our archaeometric investigations have also provide detailed guidelines on the 19th century restoration techniques [6]. All 5th century objects analyzed up to know, nevertheless sites of provenance, show the same features: fine texture of the ceramic body, red figures saved from the ceramic paste and black gloss painted directly on the ceramic body. Regarding the 4th century objects, some highlight features similar to the 5th century ones, whereas others are characterized by a ceramic body with a coarser texture and a layer of ingobbio rosso. This intermediate layer entirely covered the external part of the vase and was reddish than the ceramic body -visually better to obtain red decorations- and the black gloss -when present- was painted on it. The chemical and minero-petrographic results make it possible to discriminate different production technologies of red figured Apulian vases used in Apulia during the 4th century BCE. This technology seems to take shape of a distinctive characteristic of Late Apulian production regardless of sites of provenance. Finally, we selected a consistent number of items to be analyzed by LA-ICPMS (Laser Ablation ICPMS). We compared Apulian samples and Attic ones, obtaining info on major, minor and trace elements. From an archaeometric point of view, the results showed differences both in the black gloss and in the ceramic body raw materials used in Apulia with respect to Attic ones, so providing an objective parameter of regional production discrimination. The comparison carried on leads us to exclude imports of black gloss from Greece, as hypothesized by some scholars [7]. Nevertheless, in order to accomplish conclusive observations, sampling has to be extended, both in terms of numbers and provenance of samples

Morphometric analysis fo the root developement of the third molar by an experimental method using digital ortopantomographs

The aim of the study was to conduct a morphometric analysis of the root development of the third molar, with the purpose of overcoming the limits of an exclusively morphological analysis. The study was divided in two phases. The first one was the verification of the existence of a constant crown-root (C/R) ratio on a sample of 100 third molars, characterized by a complete root development. The value obtained was used in the second phase to predict the final root's length, knowing the crown height. So we have calculated, on a sample of 322 third molars with developing roots, the final ratios between incomplete roots and complete roots. Statistical analysis was then made with 90%, 95%, and 99% confidence intervals. The results showed a significant difference according to the age of the subject and the limit values, lower and upper, for subjects aged 16 and 17 years versus those aged 18 and 19. For each analysis, the width of the class of tolerance and other statistical indicators were calculated. The results we obtained support the advantage of a morphometric study as opposed to an exclusively morphological study, but all the techniques used to determine the age of a living subject can provide only indications of the biological age, but no certainties as regards chronological age

Morphometric Analysis of Third Molar Development: A Comparison of Albanian and Italian Sample Populations

The goal of this study was to investigate the differences between third molar root development in Italian and Albanian populations when determining the age of adults. This presentation will impact the forensic science community by underlining the importance of using morphometric analysis in age estimation while also taking into account the differences that exist between various ethnic groups Introduction: Determination of adult age by tooth analysis is an important issue in forensics and has significant implications in determining criminal liability. It also plays a critical role in issues regarding young illegal immigrants and refugee children. Moreover, the results of such analyses play a substantial role in areas related to school attendance, social benefits, adoption procedures, employment, and marriage as related to international protections guaranteed by the United Nations High Commissioner for Refugees (UNHCR). The study was conducted on digital orthopantomographs (OPG) and was based on identification criteria using morphometric analysis with the goal of overcoming the limits associated with using morphological analysis alone. This investigation also served to verify the existence of differences in third molar development among Italian and Albanian populations. The goal of the study was to test the possibility of applying the results of a previous morphometric analysis conducted on an Italian sample population (Forensic Sci. 2008 Jul; 53(4): 904-9) to an Albanian sample population: Albanians are the second most populous ethnic group in Italy after Italians. Materials and Methods: OPGs were obtained by systematic digital analysis using specialized dental software. In the first phase of the study, the confidence intervals obtained from the Italian sample were applied to a sample of 140 Albanians whose OPG ages were between 16 and 19 years. This was done in order to ascertain the feasibility of using these confidence intervals. Preliminary results revealed the risk of age overestimation in 15% of the sample when these confidence intervals were applied to the Albanian sample. As a result, another analysis was carried out on a sample of 100 third molars with fully developed roots, all belonging to Albanians, in order to investigate the existence of a constant crown-to-root ratio (C/R). After recording the measurements, a statistical analysis aimed at identifying the arithmetic mean of the ratios, and standard deviation was carried out. This resulted in an average C⁄R ratio value equal to 0.560, slightly higher than the average values obtained from the Italian population, and a standard deviation of 0.07. In the second phase of our study, 494 third molars were analyzed with developing roots belonging to Albanian individuals (from Tirana and Valona) aged between 16 and 19 years, and subdivided by sex. The values obtained from the first phase of the study were used in the subsequent phase, thus establishing that when the crown height (h) of the third molar, with developing roots, is known, the root length (lt) of the molar, when it is fully developed, may also be known. After establishing the length of the developing root, and based on the constant ratio established in the first phase, the lengths of the incomplete roots (ir) were calculated and the complete roots according to growth estimates (cr). Final Ratio (fr): ir ⁄ cr incomplete root ⁄ complete root beginning with root size values, a preliminary macro identification was made for both sexes that distinguished minors from those whose ages were 18 years or older. Analysis was made with 90%, 95%, and 99% confidence intervals, obtaining two limit values (lower and upper) of the ir ⁄ cr ratios in subjects under 18 (i.e. 16 and 17 years old), and over 18 in our sample, according to sex. A comparison of the results obtained from the two populations was carried out. Results: The Albanian sample showed higher developmental asymmetry of the left and right inferior third molars than the Italian sample. The Albanian sample showed a higher crown-to-root ratio as well as a higher growth index than the Italian sample. There was not an over estimation of the predicted root length in any of the cases. This is an important consideration when seeking to avoid the risk of over-estimating age

Chemical-phisical and morpho-mineralogical methodologies as tools for a fresh viewpoint on medieval graffita pottery

Shards of medieval glazed pottery with negative decoration, usually called graffita, coming from three different archaeological sites in Apulia werw examined by OM, SEM, EDS, ICP-MS. The results obtained, based on similarities/differences of raw materials and technological features, allowed to enclose objects in two groups, reflecting the shared tendencies in taste in the medieval era across the Mediterranean area (11 th - 13 th centuries: Islamic style and Bizantine-styl