Cost‐effective and rapid lysis of Saccharomyces cerevisiae cells for quantitative western blot analysis of proteins,including phosphorylated eIF2α

The common method for liberating proteins from Saccharomyces cerevisiae cells involves mechanical cell disruption using glass beads and buffer containing inhibitors (protease, phosphatase and/or kinase inhibitors), followed by centrifugation to remove cell debris. This procedure requires the use of costly inhibitors and is laborious, in particular when many samples need to be processed. Also, enzymatic reactions can still occur during harvesting and cell breakage. As a result low‐abundance and labile proteins may be degraded, and enzymes such as kinases and phosphatases may still modify proteins during and after cell lysis. We believe that our rapid sample preparation method helps overcome the above issues and offers the following advantages: (a) it is cost‐effective, as no inhibitors and breaking buffer are needed; (b) cell breakage is fast (about 15 min) since it only involves a few steps; (c) the use of formaldehyde inactivates endogenous proteases prior to cell lysis, dramatically reducing the risk of protein degradation; (d) centrifugation steps only occur prior to cell lysis, circumventing the problem of losing protein complexes, in particular if cells were treated with formaldehyde intended to stabilize and capture large protein complexes; and (e) since formaldehyde has the potential to instantly terminate protein activity, this method also allows the study of enzymes in live cells, i.e. in their true physiological environment, such as the short‐term effect of a drug on enzyme activity. Taken together, the rapid sample preparation procedure provides a more accurate snapshot of the cell's protein content at the time of harvesting. Copyright © 2017 John Wiley & Sons, Ltd.     

Dried Tomato‐Flavored Probiotic Cream Cheese with Lactobacillus paracasei

Abstract: A dried tomato‐flavored probiotic cream cheese (P) containing Lactobacillus paracasei Lpc‐37 was developed for the purpose of this study. The same product, but without probiotic addition (C) was used as control. Lactococcus lactis subsp. lactis and Lactococcus lactis subsp. cremoris were used as lactic starter cultures. Chemical composition analyses and sensory tests were performed on days 1 and 7, respectively. Titratable acidity, pH value and L. paracasei population were determined every 7 d during the refrigerated storage (21 d) of the cream cheeses. The experiment and analyses were performed in triplicate, using standard methods. Probiotic population remained greater than 10⁷ CFU/g throughout the storage period, thereby characterizing the product as potentially probiotic. Cream cheeses C and P did not differ on the sensory tests, both obtaining good overall acceptance by the consumers, of which 82.6% stated that they certainly or probably would buy the product. Practical Application: Lactobacillus paracasei Lpc‐37 is a probiotic bacterium and clinical studies have shown that this microorganism beneficially affects its host. In general, dried tomato‐flavored products and cream cheese are products with good acceptance by the consumers. Thus, regular consumption of the probiotic cream cheese developed in this study may have positive effects on health and well being of people if incorporated into their diet.     

Atmospheric outflow of anthropogenic semivolatile organic compounds from East Asia in spring 2004

To estimate the emissions of anthropogenic semivolatile organic compounds (SOCs) from East Asia and to identify unique SOC molecular markers in Asian air masses, high-volume air samples were collected on the island of Okinawa, Japan between 22 March and 2 May 2004. Contributions from different source regions (China, Japan, the Koreas, Russia, and ocean/local) were estimated by use of source region impact factors (SRIFs). Elevated concentrations of hexachlorobenzene (HCB), hexachlorcyclohexanes (HCHs), dichlorodiphenyltrichloroethanes (DDTs), and particulate-phase polycyclic aromatic hydrocarbons (PAHs) were attributed to air masses from China. A large proportion of the variation in the current-use pesticides, gas-phase PAHs, and polychlorinated biphenyl (PCB) concentrations was explained by meteorology. Chlordanes showed a technical mixture profile and similar concentrations regardless of source region. alpha/gamma HCH and trans/cis chlordane ratios did not vary significantly with different source regions and had regional averages of 2.5 +/- 1.0 and 1.2 +/- 0.3, respectively. Particulate-phase PAH concentrations were significantly correlated (p value < 0.05) with other incomplete combustion byproduct concentrations, including elemental mercury (Hg0), CO, NOx, black carbon, submicrometer aerosols, and SO2. By use of measured PAH, CO, and black carbon concentrations and estimated CO and black carbon emission inventories, the emission of six carcinogenic particulate-phase PAHs was estimated to be 1518-4179 metric tons/year for Asia and 778-1728 metric tons/year for China, respectively. These results confirm that East Asian outflow contains significant emissions of carcinogenic particulate-phase PAHs.     

Extraction Behavior of Americium and Curium on Selected Extraction Chromatography Resins from Pure Acidic Matrices

Due to the similar chemical properties between the neighboring trivalent actinide elements americium and curium, their extraction behavior is often perceived as indistinguishable. In this work, the characterization of seven extraction chromatography resins (TEVA, TRU, DGA(N), Actinide, Ln, Ln2, and Ln3) for these trivalent actinides from acidic matrices (HNO₃, HCl, and HBr) has provided some evidence to the contrary. In most cases, Am(III) and Cm(III) exhibit identical extraction properties. However, separation is possible with TRU and DGA(N) resins as demonstrated in this study. The extraction shows a strong dependency on the specific anion in solution that follows the order NO₃^?>Br^?>Cl^?.     

Modeling of fouling in cross-flow microfiltration of suspensions

Cross-flow filtration of fine suspensions through microsieves occurs in microprocessing. The interaction of particles with surfaces in microenvironments has been extensively studied, but predominantly in monolayers and not with an eye to microfiltration. Here, we introduce a microfiltration model that pertains to particles that might be seen as fine in a macroscopic environment, but are large enough to intrude significantly into the shear layer of a microchannel. Thus, particle accumulation upon the sieve couples the steady-state filtrate flux and the suspension flow through the microchannel that feeds the sieve. We envision and create a stable, stationary multilayer of particles whose thickness is shear-limited and we identify and verify the structure and parameters that limit steady filtration in this environment. At first, a packed bed of particles forms, growing into and regulated by the micro channel's shear flow. A critical shear stress is shown to determine the thickness of the bed, seen as a stationary and stable multilayer of particles through which filtration may occur. As the bed thickens, at the expense of channel area for suspension flow, surface shear stress increases until no further particle adherence is possible. We built a simple example using hard noninteracting polymer microspheres and conducted cross-flow filtration experiments over Aquamarijn™ microsieves (uniform pore size of 0.8 μm). We observed a steady cake-layer thickness and because of the simple geometry afforded by uniform spheres, we could approximate the force balance, cake resistance, and filtration rate from first principles. The good fit of our data to the proposed mechanism lays a firm basis for the semiquantitative analysis of the behavior of more complex suspensions. © 2018 American Institute of Chemical Engineers AIChE J, 65: 207–213, 2019     

Distributed deadlock resolution in store-and-forward networks

We present a simple distributed algorithm that resolves store-and-forward deadlocks in data communication networks. The basic idea of the algorithm is to detect cycles of nodes that may cause store-and-forward deadlocks, and to rotate packets along these cycles. The algorithm uses a fixed amount of storage in each node for its execution, and, under reasonable assumptions upon the routing and packet handling, it ensures that packets that enter the network arrive at their destinations in finite time.     

SNA Networks of Small Systems

This paper discusses SNA/LEN, a possible extension of Systems Network Architecture intended to allow peer, dynamic, and easy to use networking functions for a variety of node sizes down to and including the new generation of personal computers. After reviewing the special requirements posed by small systems and the need for including them as equal partners in networks, the LEN architecture is described. This is done by following a sequence of steps that begin when a logical resource at some node that may not yet be part of the connectivity of an existing network requests a session with a remote logical resource of unknown location. After connectivity with the preexisiting network is established, directory services locates the remote object, route selection services determines the preferred path, a session is activated, and deadlock-free flow control assures a useful flow of data. Preliminary quantitative results from an LEN prototype are presented.     

Anthropogenic,Polyhalogenated, Organic Compounds in Sedentary Fish from Lake Huron and Lake Superior Tributaries and Embayments

Composite samples of fish taken from some of the major tributaries and embayments of Lake Superior and Lake Huron were analyzed for halogenated xenobiotics with the objective of detecting potential environmental problem areas, point source discharges of toxic materials, and previously unrecognized pollutants. The analyses were done by gas chromatographic, methane enhanced, negative ion, mass spectrometry. A variety of halogen-containing organic compounds and pesticides were identified. The highest concentration and variety of compounds were found in the fish from the highly industrialized Saginaw Bay drainage basin; fish from Lake Superior tributaries showed much lower concentrations.