This study uses a combination of digital microscopic analysis and experimental archaeology to assess stone tool cut marks on animal bones. We used two un-retouched flint flakes and two burins to inflict cut marks on fresh, boiled, and dry ungulate bones. The experiment produced three series of three engravings on each bone with each of the experimental tools. The first series involved one single stroke; the second, two strokes in the same direction; and the third, multiple strokes using a to-and-fro movement. We analyzed the striations using a Hirox 3D digital microscope (KH-7700) and collected metric and profile data on the morphology of the cut marks. In order to describe the shape of each cross section, we calculated the ratio between the breadth at the top and the breadth at the floor of cut marks. Preliminary results show that both the tool type and the method of creating the cut mark influence the shape of the resulting groove. In our experiment, morphological parameters can be used to differentiate between marks produced using un-retouched flint flakes and those produced using burins. However, neither morphological nor morphometric analysis allows us to identify the mechanical motion used to produce the cuts, nor the state of the bone (fresh, boiled, or dry) at the moment of marking.
Abstract. Quercetin (Q) and rutin (R) are two natural flavonoids with antioxidant properties. We evaluated the toxicity of Q and R at 20μM, 30μM, 50μM, 100μM, 200μM, 400μM in swim up selected human sperm. The antioxidant activity (Q and R 20μM and 30μM) was tested on lipid peroxidation (LPO) induced by tert-butylhydroperoxide in human sperm. LPO was evaluated using the C11-BODIPY581/591 probe and sperm structural damages were assessed by transmission electron microscopy in samples incubated with and without Q and R. A significant dose dependent toxic effect was observed for both compounds on sperm viability (Q and R: r= -0.98 P<0.001), on sperm progressive motility (Q: r=-0.98, R: r=-0.97; P<0.001), and on non progressive motility (Q; r=-0.58, P<0.01; R: r=-0.50, P<0.05). Both flavonoids, used at 20μM and 30μM, showed antioxidant properties on LPO induced in human sperm and a general protective effect against ultrastructural damages of LPO. In conclusion, we observed that Q exhibited a little higher toxicity than R; on the other hand R is little low protective on induced LPO. Our preliminary results demonstrated the scavenger properties of these flavonoids in vitro on human sperm.
The analysis of bone-surface modifications (BSM), such as butchering marks, is necessary to better understand how the exploitation of animal resources by past hominins influenced their biological and cultural evolution. In this paper, we try to quantify to what extent the depth of the cut marks influences the shape of their cross sections. This is of crucial importance for a valid interpretation of the shape data collected on archaeological BSMs. Two groups of slicing cut-mark cross sections were experimentally produced with two flint burins on a defleshed cattle innominate, and a set of butchering marks were produced with an unretouched flint flake. These were analysed by means of 3D microscopy and geometric morphometrics. The resulting sets of striae show different depths and different cross-sectional shapes. Shallower cross sections display less steep walls and, consequently, a wider opening angle. When the characteristics of the burin cutting edges were investigated, it was clear that the difference in shape between the two groups of striations was probably a function of the way in which the tool penetrated the bone. These results are taphonomically relevant since similar differences in cross-sectional shapes have been found in marks produced with different tools.
Objective: To assess the antioxidant activity of Resveratrol (3,5,4'-trihydroxystilbene, RES) after induction of lipid peroxidation (LPO) in human spermatozoa and in immature rat germinal cells. Materials and Methods: Ejaculated human spermatozoa, selected by swim up, have been incubated with tert-Butylhydroperoxide and tert-Butylhydroperoxide-RES. The localization of LPO has been performed using the probe C11-BODIPY 581/591 . The same assays were carried out on pachytene spermatocytes and round spermatids obtained from three Wistar rats 35 days of age. The two cellular fractions were achieved after enzymatic digestion with collagenase and subsequent fractionation on bovine serum albumin 0.5-3% gradient (STAPUT). The ultrastructure of all samples was assessed by transmission electron microscopy (TEM). Results: The midpiece of sperm tail and the whole plasma membrane of germ cells were the target of LPO. TEM analysis of sperm, quantitatively elaborated by a mathematical formula, showed a significantly lower percentage of necrosis in the samples treated with RES (P<0.01); as regards rat germinal cells, necrosis features (cytoplasmic vacuoles, disrupted chromatin and broken plasma membrane) were mainly evident in the meiotic fraction without RES. Conclusions: RES, found in the skins of grape, reduces the damage induced by oxidative stress in human sperm and rat testicular germ cells; in particular spermatids appeared to be less sensitive to oxidative damage compared with spermatocytes.
Humanin (HN) is a peptide showing neuroprotective properties against damage associated with Alzheimer’s disease. In male reproductive system HN is expressed in human and rat testis. This preliminary study aimed to localize HN by immunocytochemistry (ICC) directly in human, mouse and rabbit sperm. Semen samples were processed for the ICC analyses using two different rabbit polyclonal anti-HN antibodies. Western blotting analyses were performed using fresh human semen samples. Normal human, rabbit and mouse sperm, showed HN labeling in sub-acrosomal and mid-piece regions, whereas human abnormally shaped sperm were stained also at acrosomal level and particularly in the flagellum. Western blotting analysis revealed the presence of HN in analyzed human semen samples. HN was localized directly on human, rabbit and mouse spermatozoa. In human samples, we demonstrated a different localization pattern in normally shaped sperm compared to abnormal sperm probably due to a protective effect of HN in pathological sperm
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