Analysis of outer membrane proteins which are associated with growth of Bacteroides thetaiotaomicron on chondroitin sulfate.

By analyzing outer membrane proteins of Bacteroides thetaiotaomicron on two-dimensional polyacrylamide gels, we were able to identify 10 protein spots that were associated with growth on chondroitin sulfate but not with growth on glucuronic acid or other monosaccharides. These proteins were distinct from the outer membrane polypeptides that were associated with growth on two other negatively charged polysaccharides, polygalacturonic acid and heparin. Of the 10 protein spots that were associated with growth on chondroitin sulfate, 4 could be detected on immunoblots with antiserum that had been raised against outer membranes from bacteria grown on chondroitin sulfate and then cross-adsorbed with membranes from bacteria grown on glucose. Synthesis of these four proteins appeared to be regulated coordinately with synthesis of the two enzymes that degrade chondroitin sulfate, chondroitin lyase I and II. Although one of the four proteins (Mr 110,000) was similar in molecular weight to the chondroitin lyases, the cross-adsorbed antiserum which detected this outer membrane protein did not cross-react with either of these two enzymes.     

Vacuolar ion currents in the primitive green algaEremosphaera viridis: The electrical properties are suggestive of both the Characeae and higher plants

Summary The electrical properties of the vacuolar membrane of the primitive green algaEremosphaera virdis were investigated using the patch-clamp technique. In whole vacuole measurements two types of transport systems with long activation time-constants were identified. The first, showing marked outward rectification, was activated by an increase in the cytosolic calcium concentration. Furthermore, it displayed sensitivity to micromolar concentrations of the anion channel blocker Zn²⁺ and to acidification of the cytosol. In contrast, the second time-activated current component was almost insensitive to changes in cytosolic pH and was blocked by the potassium channel inhibitor TEA. In addition to these slowly activating current components, the vacuolar membrane contained at least two further transport systems, responsible for an instantaneous current. These two current components were distinguished by their different sensitivity to protons, cytosolic calcium, and TEA. Comparing these electrical properties to those observed in vacuoles of higher plants or in cytoplasmic droplets from characean algae, respectively, it seems thatEremosphaera is intermediate, corresponding to the systematic position of this simple green alga.Abbreviations [Ca²⁺]_cyt cytosolic free calcium concentration - EGTA ethyleneglycol-bis(-aminoethylether)N,N,N,N-tetraacetic acid - HEPES N-[2-hydroxyethyl]piperazine-N-[2-ethanesulfonic acid] - I electric current - IRC inward rectifying current - MES 2-[N-morpholino]ethanesulfonic acid - ORC outward rectifying current - pH_cyt cytosolic pH - pH_vac vacuolar pH - P_o open probability - P_x permeability coefficient of ion species X - TEA tetraethylammonium chloride - Tris tris[hydroxymethyl]aminomethane - V voltage     

Mycoplasma pneumoniae Pneumonia—Experience at a Referral Center

Mycoplasma pneumoniae pneumonia is usually a benign illness, and respiratory complications and extrapulmonary manifestations occur rarely. In this series, patients admitted to a referral hospital with this disorder had unusual symptoms, signs and findings on chest roentgenograms and laboratory studies. Pneumonia was often severe and extrapulmonary manifestations were frequent, resulting in prolonged hospital stays and illnesses. Although this extreme end of the spectrum of disease caused by M pneumoniae is not representative of this type of pneumonia as seen in outpatients, it is important to realize that patients admitted to hospital with severe, complicated pneumonia frequently have unusual manifestations of a common disease.     

Cytoplasmic filaments and cellular wound healing in amoeba proteus

The flexibility and self-healing properties of animal cell surface membranes are well known. These properties have been best exploited in various micrurgical studies on living cells (2, 3), especially in amoebae (7, 20). During nuclear transplantation in amoebae, the hole in the membrane through which a nucleus passes can have a diameter of 20-30 μm, and yet such holes are quickly sealed, although some cytoplasm usually escapes during the transfer. While enucleating amoebae in previous studies, we found that if a very small portion of a nucleus was pushed through the membrane and exposed to the external medium, the amoeba expelled such a nucleus on its own accord. When this happened, a new membrane appeared to form around the embedded portion of the nucleus and no visible loss of cytoplasm occurred during nuclear extrusion. In the present study, we examined amoebae that were at different stages of expelling partially exposed nuclei, to follow the sequence of events during the apparent new membrane formation. Unexpectedly, we found that a new membrane is not formed around the nucleus from inside but a hole is sealed primarily by a constriction of the existing membrane, and that cytoplasmic filaments are responsible for the prevention of the loss of cytoplasm.     

Clinical determinants of early spontaneous conversion to sinus rhythm in patients with atrial fibrillation

Netherlands Heart Journal - The current standard of care for acute atrial fibrillation (AF) focuses primarily on immediate restoration of sinus rhythm by cardioversion, although AF often terminates...     

Cherry Picking: A Characterization of the Temporal Hybridization Number for a Set of Phylogenies

Recently, we have shown that calculating the minimum–temporal-hybridization number for a set ${\mathcal{P}}$ of rooted binary phylogenetic trees is NP-hard and have characterized this minimum number when ${\mathcal{P}}$ consists of exactly two trees. In this paper, we give the first characterization of the problem for ${\mathcal{P}}$ being arbitrarily large. The characterization is in terms of cherries and the existence of a particular type of sequence. Furthermore, in an online appendix to the paper, we show that this new characterization can be used to show that computing the minimum–temporal hybridization number for two trees is fixed-parameter tractable.     

Saikosaponin-d,a novel SERCA inhibitor,induces autophagic cell death in apoptosis-defective cells

Autophagy is an important cellular process that controls cells in a normal homeostatic state by recycling nutrients to maintain cellular energy levels for cell survival via the turnover of proteins and damaged organelles. However, persistent activation of autophagy can lead to excessive depletion of cellular organelles and essential proteins, leading to caspase-independent autophagic cell death. As such, inducing cell death through this autophagic mechanism could be an alternative approach to the treatment of cancers. Recently, we have identified a novel autophagic inducer, saikosaponin-d (Ssd), from a medicinal plant that induces autophagy in various types of cancer cells through the formation of autophagosomes as measured by GFP-LC3 puncta formation. By computational virtual docking analysis, biochemical assays and advanced live-cell imaging techniques, Ssd was shown to increase cytosolic calcium level via direct inhibition of sarcoplasmic/endoplasmic reticulum Ca²⁺ ATPase pump, leading to autophagy induction through the activation of the Ca²⁺/calmodulin-dependent kinase kinase–AMP-activated protein kinase–mammalian target of rapamycin pathway. In addition, Ssd treatment causes the disruption of calcium homeostasis, which induces endoplasmic reticulum stress as well as the unfolded protein responses pathway. Ssd also proved to be a potent cytotoxic agent in apoptosis-defective or apoptosis-resistant mouse embryonic fibroblast cells, which either lack caspases 3, 7 or 8 or had the Bax-Bak double knockout. These results provide a detailed understanding of the mechanism of action of Ssd, as a novel autophagic inducer, which has the potential of being developed into an anti-cancer agent for targeting apoptosis-resistant cancer cells.