Radiocarbon dating is the most widely used dating technique in the world. Recent advances in Accelerator Mass Spectrometry (AMS) and sample preparation techniques have reduced the sample-size requirements by a factor of 1000 and decreased the measurement time from weeks to minutes. Today, it is estimated that more than 90 percent of all measurements made on accelerator mass spectrometers are for radiocarbon age dates. The production of 14 C in the atmosphere varies through time due to changes in the Earth's geomagnetic field intensity and in its concentration, which is regulated by the carbon cycle. As a result of these two variables, a radiocarbon age is not equivalent to a calendar age. Four decades of joint research by the dendrochronology and radiocarbon communities have produced a radiocarbon calibration data set of remarkable precision and accuracy extending from the present to approximately 12,000 calendar years before present. This paper presents high precision paired 230 Th/ 234 U/ 238 U and 14 C age determinations on pristine coral samples that enable us to extend the radiocarbon calibration curve from 12,000 to 50,000 years before present. We developed a statistical model to properly estimate sample age conversion from radiocarbon years to calendar years, taking full account of combined errors in input ages and calibration uncertainties. Our radiocarbon calibration program is publicly accessible at: http://www.radiocarbon.LDEO.columbia.edu/ along with full documentation of the samples, data, and our statistical calibration model. r

Simultaneous elucidation of the glycan structure and the glycosylation site are needed to reveal the biological function of protein glycosylation. In this study, we employed a recent type of fragmentation termed higher energy collisional dissociation (HCD) to examine fragmentation patterns of intact glycopeptides generated from a mixture of standard glycosylated proteins. The normalized collisional energy (NCE) value for HCD was varied from 30 to 60% to evaluate the optimal conditions for the fragmentation of peptide backbones and glycoconjugates. Our results indicated that HCD with lower NCE values preferentially fragmented the sugar chains attached to the peptides to generate a ladder of neutral loss of monosaccharides, thereby enabling the putative glycan structure characterization. In addition, detection of the oxonium ions enabled unambiguous differentiation of glycopeptides from non-glycopeptides. In contrast, HCD with higher NCE values preferentially fragmented the peptide backbone and, thus, provided information needed for confident peptide identification. We evaluated the HCD approach with alternating NCE parameters for confident characterization of intact N- and O-linked glycopeptides in a single liquid chromatography–tandem mass spectrometry (LC–MS/MS) analysis. In addition, we applied a novel data analysis pipeline, so-called GlycoFinder, to form a basis for automated data analysis. Overall, 38 unique intact glycopeptides corresponding to eight glycosylation sites (six N-linked and two O-linked sites) were confidently identified from a standard protein mixture. This approach provided concurrent characterization of both the peptide and the glycan, thereby enabling comprehensive structural characterization of glycoproteins in a single LC–MS/MS analysis.

Glycosylation is one of the most prominent and extensively studied protein post-translational modifications. However, traditional proteomic studies at the peptide level (bottom-up) rarely characterize intact glycopeptides (glycosylated peptides without removing glycans), so no glycoprotein heterogeneity information is retained. Intact glycopeptide characterization, on the other hand, provides opportunities to simultaneously elucidate the glycan structure and the glycosylation site needed to reveal the actual biological function of protein glycosylation. Recently, significant improvements have been made in the characterization of intact glycopeptides, ranging from enrichment and separation, mass spectroscopy (MS) detection, to bioinformatics analysis. In this review, we recapitulated currently available intact glycopeptide characterization methods with respect to their advantages and limitations as well as their potential applications.

Aim To investigate the population history and demographics of Jerdon's pitviper, Protobothrops jerdonii, and elucidate how the unique physical conditions and heterogeneous mountain environments resulting from the uplift of the Tibetan Plateau shaped the genetic diversity and evolutionary history of the species.Location China and Vietnam.Methods We sequenced and analysed a total of 1752 base pairs from two mitochondrial genes, cytochrome b (cyt b) and NADH dehydrogenase subunit (ND4), for 81 specimens sampled from 27 localities across the species' range, and a total of 464 base pairs from two nuclear genes for 28 representative samples from all mitochondrial DNA lineages. Based on these data, we constructed the genealogical relationships and estimated the divergence times of the mitochondrial DNA clades.Results The mitochondrial DNA results revealed the existence of five distinct, strongly supported and geographically structured DNA lineages within populations of P. jerdonii that are paraphyletic with respect to Protobothrops xiangchengensis. Estimation of divergence dates suggested that P. jerdonii possibly evolved in the western Hengduan Mountains region c. 6.6 Ma in the late Miocene. Nuclear DNA data did not provide sufficient resolution to distinguish the mitochondrial DNA lineages.Main conclusions Based on the present-day distribution and intraspecific genealogy, the evolutionary history of P. jerdonii can be explained by a pattern of dispersal followed by vicariance. All lines of evidence suggest that historical biogeographical factors, particularly the north-south orientation of the higher mountains, as well as low-elevation areas in western China, had the greatest influence on the population structure, lineage formation and species distribution of this snake. However, highly heterogeneous habitats and glacial cycles appear to have affected patterns of intraspecific differentiation. While our mitochondrial data provide evidence for clear phylogeographical structure, our small sampling of nuclear genes does not, suggesting that nuclear markers may not have had sufficient time to coalesce to match patterns observed in the mitochondrial data.