Localised and rapid Ca2+ micro-events in human neutrophils: conventional Ca2+ puffs and global waves without peripheral-restriction or wave cycling

Ultra-localised and peripherally restricted zones of elevated Ca2+ (z-waves) have been reported to cycle around the periphery of neutrophils at low frequency (1/20s) in the absence of conventional localised Ca2+ (puffs) and global Ca2+ (waves) signals. However, we report here that fast confocal laser scanning of human neutrophils loaded with either cytosolic fluo4 or its membrane associated analogue, MOMO reports both "conventional" stationary Ca2+ "puffs" (diameter c.3 microm) and global Ca2+ waves that sweep across the cell. The Ca2+ puff size and frequency of detection suggests that each neutrophil contained only a single release site and that its detection was limited by the location of the confocal plane relative to the event. Both formylated peptide receptor stimulation and cytosolic IP3 uncaging generated Ca2+ puffs (c.6% of cells) and global Ca2+ signals (c.75% of cells). The Ca2+ puffs peaked at approx. 250 nM and had a duration of approx. 235 msec and remained at a single locus. This was similar to other Ca2+ events in other cell types but in direct contrast to the reported z-waves. It was concluded that the micro-events which underlie Ca2+ signalling in neutrophils are conventional and that the existence of novel Ca2+z-waves is doubtful.     

Cloud-Enabled Microscopy and Droplet Microfluidic Platform for Specific Detection of Escherichia coli in Water

We report an all-in-one platform – ScanDrop – for the rapid and specific capture, detection, and identification of bacteria in drinking water. The ScanDrop platform integrates droplet microfluidics, a portable imaging system, and cloud-based control software and data storage. The cloud-based control software and data storage enables robotic image acquisition, remote image processing, and rapid data sharing. These features form a “cloud” network for water quality monitoring. We have demonstrated the capability of ScanDrop to perform water quality monitoring via the detection of an indicator coliform bacterium, Escherichia coli, in drinking water contaminated with feces. Magnetic beads conjugated with antibodies to E. coli antigen were used to selectively capture and isolate specific bacteria from water samples. The bead-captured bacteria were co-encapsulated in pico-liter droplets with fluorescently-labeled anti-E. coli antibodies, and imaged with an automated custom designed fluorescence microscope. The entire water quality diagnostic process required 8 hours from sample collection to online-accessible results compared with 2–4 days for other currently available standard detection methods.     

Application of the Bang and Ramm age at death estimation method to two known‐age archaeological assemblages

The Bang and Ramm method uses root dentine translucency (RDT) length in sectioned or unsectioned teeth as a sole indicator of chronological age at death in adult human remains. The formulae have been tested on modern remains of known age and on modern and archaeological remains of unknown age. This is the first published study of the method on known‐age archaeological specimens and tests whether RDT is a good indicator of chronological age in buried human remains. We applied the Bang and Ramm equations to two 18th and 19th century assemblages excavated from the crypt of Christ Church, Spitalfields, and the cemetery of All Hallows by the Tower. Translucency was defined by shining a light through the external unsectioned root surface and was measured from digital images of 583 and 83 nonmolar roots from 126 Spitalfields and 12 All Hallows individuals, respectively, aged 21–81 years. Average absolute difference between real age and estimated age was 10.7 years and 8.4 years for Spitalfields and All Hallows individuals, respectively, with 58% and 75% estimated within 10 years of known age, and 29% and 33% estimated within five years of known age. These estimations are comparable to results from other ageing methods applied to the Spitalfields collection. Ages from both populations were estimated largely to the middle ranges, with younger individuals overestimated and older individuals underestimated. This is a common occurrence when using inverse calibration, where age is treated as the dependent variable and the ageing feature as the independent variable. Am J Phys Anthropol 155:332–351, 2014. © 2014 Wiley Periodicals, Inc.     

Brief communication: The use of alternative dental measurements on deciduous teeth

The present study investigates the utility of cervical measurements in deciduous teeth and how they correlate with traditional measurements of the crown. First, this study establishes definitions by which these measurements could reliably be taken. Next, deciduous cervical and traditional crown diameters were taken on three distinct skeletal samples: a Neolithic assemblage from Central Anatolia (Çatalhöyük, n = 85), a precontact sample from Northern California (CA‐ALA‐329, n = 34), and a group of intrusive burials interred at Çatalhöyük that date between AD 60 and 1650 (n = 38). Across the dentition there are positive correlations between crown and cervical measurements, which tend to be higher in anterior teeth than in posterior teeth. Both measurements show low correlations with age; however, cervical measurements show fewer negative correlations with age. An intraobserver error study found low levels of error for both types of measurements. On a subset of the Çatalhöyük sample (n = 9), a principal components and biological distance analysis were conducted comparing the two types of measurements. Also, all three samples were subject to a canonical discriminant function analysis and the results from cervical and crown measurements were compared. All analyses produced slightly different results for each type of data suggesting that crown and cervical measurements capture different aspects of tooth shape. While cervical and crown measurements provide different statistical results, cervical measurements can provide information relevant to anthropological studies and may allow for larger datasets to be used by allowing the inclusion of teeth with modified crowns. Am J Phys Anthropol 149:299–306, 2012. © 2012 Wiley Periodicals, Inc.     

HYBRIDIZATION AND FLORAL VASCULARIZATION

Hillson, Charles J. (Pennsylvania State U., University Park.) Hybridization and floral vascularization. Amer. Jour. Bot. 50(10): 971–978. Illus. 1963.—The floral steles of 6 synthetic hybrid mints, representing the F₁ generation of a cross between Mentha spicata strain 199 and Mentha aquatica, were compared with each other and with the steles of the parents. Comparisons of the level of trace divergence, ovary wall vascularization, presence or absence of a nectary, and the nature of the gaps associated with dorsal carpellary bundles show that floral vascularization is an inherited character. Although slight male dominance is indicated in the hybrid steles, anatomical features of both parents are evident. Thus, while vascular tissue may be conservative when phylogenetic reduction of floral parts takes place through adnation and/or connation, stelar modification through hybridization is documented.     

Design, implementation and practice of JBEI-ICE: an open source biological part registry platform and tools

The Joint BioEnergy Institute Inventory of Composable Elements (JBEI-ICEs) is an open source registry platform for managing information about biological parts. It is capable of recording information about ‘legacy’ parts, such as plasmids, microbial host strains and Arabidopsis seeds, as well as DNA parts in various assembly standards. ICE is built on the idea of a web of registries and thus provides strong support for distributed interconnected use. The information deposited in an ICE installation instance is accessible both via a web browser and through the web application programming interfaces, which allows automated access to parts via third-party programs. JBEI-ICE includes several useful web browser-based graphical applications for sequence annotation, manipulation and analysis that are also open source. As with open source software, users are encouraged to install, use and customize JBEI-ICE and its components for their particular purposes. As a web application programming interface, ICE provides well-developed parts storage functionality for other synthetic biology software projects. A public instance is available at public-registry.jbei.org, where users can try out features, upload parts or simply use it for their projects. The ICE software suite is available via Google Code, a hosting site for community-driven open source projects.     

Detection of protein disulphide-isomerase in sheep pancreas fractions

Conclusions The use of diethylpyrocarbonate to inhibit endogenous ribonuclease in sheep pancreas allows the detection of protein-disulphide-isomerase activity in homogenates, at specific activities of up to 4 units/g. This is higher than the specific activity in sheep liver homogenates (about 2 units/g) or in homogenates of other sheep tissues (16). It is thus evident that high levels of protein-disulphide-isomerase activity are present in sheep pancreas. This is consistent both with the postulated general role of protein disulphide-isomerase in protein biosynthesis (10,11) and with the in vitro action of the enzyme on its conventional substrate scrambled ribonuclease, since pancreas is the major site of ribonuclease synthesis.