Applications of Electron Backscatter Diffraction (EBSD) in Archaeology

Electron backscattered diffraction (EBSD) has been used to study samples of archaeological gold, silver, copper and bronze prepared using simple metallographic etches. EBSD maps of orientation and local misorientation revealed the metals’ microstructures and deformation substructures, and from this, combined in some cases with information on the crystallographic texture (also determined by EBSD), the methods of manufacture and working of the artefacts were determined. EBSD was also used to examine discontinuous precipitation at grain boundaries in a silver alloy at a high resolution

Inhibition of NO-synthase and degranulation of rat omental mast cells in vitro

Mast cell amines, platelet-activating factor (PAF), thromboxanes and leukotrienes have been shown to be released during nitric oxide-synthase inhibition in the rat intestine. Mast cells in rat isolated omentum (OMCs) or isolated from the rat peritoneal cavity (PMCs) have been used here to investigate the relationship(s) between these agents. N-nitro-L-arginine methyl ester (L-NAME, 100 μM) caused some degranulation of OMCs, but no enhancement of histamine release from PMCs. PAF (5 μM) and U46619 (1 μM) degranulated OMCs and enhanced histamine release from PMCs. Pre-treatment of the omentum with BN52021 (10 μM) inhibited degranulation of OMCs in response to L-NAME, PAF or U46619. Pretreatment with 1-benzylimidazole (5 or 50 μM) inhibited the effect of L-NAME but not that of PAF. Indomethacin (1 μM) or sodium nitroprusside (10 μM) also inhibited the effects of L-NAME, but nordihydroguaiaretic acid (30 μM) did not. In PMCs BN52021 inhibited PAF-induced, but not U46619-induced, release of histamine. These results suggest that inhibition of nitric oxidesynthase in the omentum by L-NAME allows thromboxanes to release PAF, which in turn degranulates and releases histamine from OMCs

Possible bi-directional link between ETA receptors and protein kinase C in rat blood vessels

Possible links have been investigated between activation of protein kinase C (PKC) and endothelin (ET) production by small blood vessels. Perfusion pressures were recorded from rat isolated mesenteric artery, with or without the small intestine attached, before and after addition to the perfusate of either ET-1, ET-3 or the PKC activator 12-deoxyphorbol 13-phenylacetate (DOPPA). Rises in perfusion pressure in response to ET-1 (10−8 M)or DOPPA (10−6 M) were reduced significantly by pre-treatment with either the ETA receptor antagonist PD151242 (10−6 M) or the PKC inhibitor Ro 31-8220 (10−6 M). ET-3 (10−8 M) had a significant, albeit small, effect only when the gut was still attached to the mesentery. Inthis latter preparation ET-1 and DOPPA increased the permeability of villi microvessels to colloidal carbon in the perfusate. This effect of DOPPA was reduced by pre-treatment with either PD151242 or Ro 31-8220, but the effects of ET-1 were reduced significantly only by Ro 31-8220. ET-3 (10−8 M) was without effect. The results suggest a possible bi-directional link between ETA receptors and PKC in the intestinal vasculature

Rate of perfusion modulates colloidal carbon leakage from rat intestinal microvessels in vitro

In order to investigate the effects of varying the rate of flow on endothelial integrity the rat isolated small intestinal vasculature was perfused at 1, 5, 10 or 20 ml/min with a gelatin-containing physiological salt solution (GPSS), followed by an injection of colloidal carbon suspension (CC). Significantly greater microvascular CC leakage occurred at 1 or 5 ml/min than at 10 or 20 ml/ mitt. CC leakage at the two slower rates of flow was reduced by adding red blood cells to the GPSS, suggesting that the microvascular endothelium became hypoxic when perfused with GPSS at 1 or 5 ml/min. After perfusion at 20 ml/min with GPSS containing resiniferatoxin (1 μM) or 5-hydroxytryptamine (100 μM), CC leakage was significantly lower than after similar perfusion at 10 ml/min. Two nitric oxide (NO) synthesis blockers, N-nitro-L-arginine methyl ester (L-NAME, 100 μM) and methylene blue (20 μM), and an NO scavenger CPTIO (100 μM) each increased CC leakage. This suggests that NO was being produced at perfusion rates of 10 or 20 ml/min. Sodium nitroprusside (10 μM), 8-bromo-cGMP (100 μM) and BN52021 (10 μM) each significantly reduced CC leakage in the presence of L-NAME

Rat intestinal mast cell amines are released during nitric oxide synthase inhibition in vitro

Inhibition of nitric oxide synthase increases microvascular permeability in rat small intestinal villi. To determine the mechanism(s) whereby this occurs we have perfused the vasculature of rat isolated small intestines with a gelatin-containing physiological salt solution. Inclusion of N-nitro-L-argintne methyl ester (L-NAME, 100 μM) or indomethacin (1 μM) in the perfusate increased leakage of injected colloidal carbon into microvessel walls. Pre-treatment with sodium nitroprusside (10 μM) significantly reduced the effects of both L-NAME and indomethacin, whereas carbacyclin (1 μM) only reduced the effects of indomethacin. PD151242 (1 μM) showed some antagonism towards the effects of L-NAME, but nordihydroguaiaretic acid (3 μM) was inactive. Pre-tment with cyproheptadine (10 μM) reduced the effects of both L-NAME and indomethacin, and also significantly reduced background (control) colloidal carbon leakage. Small intestines from polymixin B-treated rats showed significantly reduced colloidal carbon leakage in response to L-NAME. This suggests that the leakage-enhancing effects of both L-NAME and indomethacin in this preparation may be mediated by mast cell-derived amines

Microstructures of cast silver-copper alloy archaeological artefacts

Cast silver-copper alloys objects form a significant part of the archaeological record from the first millenium BC onwards. Although age hardening of annealed and quenched material has been well studied there has been little study of cast microstructures. This work used a combination of scanning electron microscopy (SEM), electron backscattered diffraction (EBSD) and optical microscopy (OM) to explore the range of microstructures in archaeological cast silver and compare them with those in modern cast silver. Modern cast silver showed extensive fine scale precipitation of copper within the grains which was unconnected to the copper-rich phase of the primary eutectic. Ancient cast objects displayed a wide range of microstructures, some very similar to the modern cast silver but some quite different. EBSD allowed interpretation of the microstructure close to some grain boundaries whose detail was obscured by the cast structure in OM and SEM images. Orientation images showed fine-scale interpenetration of the adjoining grains suggesting cellular growth, suggesting that over archaeological time boundary modification may take place in cast as well as wrought and annealed structures

Tainted ores and the rise of tin bronzes in Eurasia, c. 6500 years ago

The earliest tin bronze artefacts in Eurasia are generally believed to have appeared in the Near East in the early third millennium BC. Here we present tin bronze artefacts that occur far from the Near East, and in a significantly earlier period. Excavations at Pločnik, a Vinča culture site in Serbia, recovered a piece of tin bronze foil from an occupation layer dated to the mid fifth millennium BC. The discovery prompted a reassessment of 14 insufficiently contextualised early tin bronze artefacts from the Balkans. They too were found to derive from the smelting of copper-tin ores. These tin bronzes extend the record of bronze making by c. 1500 years, and challenge the conventional narrative of Eurasian metallurgical development