Phosphofructokinase isozymes from human organs and blood cells.

Phosphofructokinase (PFK) isozymes of blood cells and some human tissues were studied by starch gel electrophoresis and immunoprecipitation by anti-muscle and anti-erythrocyte PFK sera. PFK from muscle, heart, brain and placenta were totally precipitated by both antisera. PFK from blood cells (erythrocytes, lymphocytes, granulocytes, platelets) were precipitated more strongly by anti-erythrocyte PFK serum than by anti-muscle PFK serum. Liver, kidney and monoblast PFK were slightly precipitated by both antisera. From the electrophoretic patterns and the immunoprecipitation curves we may conclude that muscle contains the homotetrameric M4 forms; platelet, liver and kidney the homotetrameric E4 form, and blood cells the M-E hybrids. Monoblasts probably contain a E4 type PFK precursor, and heart, placenta and brain, a modified M4 type PFK. Other isozymes, unrelated with muscle and erythrocyte, were revealed in liver and kidney.     

Long-term anaesthesia with alfentanil and midazolam for lung transplantation in the dog

An anaesthetic regime was developed for lung transplantation in the dog using a continuous infusion of alfentanil and midazolam. This combination of agents provided excellent analgesia and also produced loss of consciousness. Cardiovascular stability was well maintained over a 24-h period of anaesthesia following lung transplantation. Although no animals were allowed to recover from anaesthesia in the present series, the regime described is likely to be suitable for recovery anaesthesia, particularly since both of the agents used can be reversed with specific antagonists.     

Immature red cells have ferritin receptors

Ferritin binds to immature red cells. The binding appears to be specific: (1) It is abolished by a large competing dose of nonradioactive ferritin. (2) There is little binding of ferritin to mature red cells. (3) Other high molecular weight proteins (gamma globulin and thyroglobulin) are not bound.     

Regional differences in foraging behaviour of invasive green crab (<Emphasis Type="Italic">Carcinus maenas</Emphasis>) populations in Atlantic Canada

Invasive green crab populations initially established in Canada within the Bay of Fundy, New Brunswick in the 1950s and were present in all five Atlantic provinces by 2007. Genetic evidence suggests that the Atlantic Canadian populations originated from two separate introductions with differences in time of establishment among regions and possible population-level behavioural differences. In this study, we examine intraspecific foraging behaviour among crabs from different populations, and interspecific foraging behaviour between genetically similar crabs and juvenile lobsters. Both sets of foraging experiments involved competition for a limited food source over a 1-h period. In intraspecific match-ups, recent invaders from Newfoundland (NL) were significantly superior foragers than long-established invaders from Nova Scotia (NS) and New Brunswick (NB) populations; however, we found no differences between NL and Prince Edward Island (PE) invaders. Crabs from PE were better competitors than those from NS and NB, but these differences were not significant. Interspecific competition experiments indicated that the feeding behaviour of recent invaders (NL) and genetically similar, but long-established invaders (NS), differed in the presence of juvenile lobsters. Our study documents striking behavioural differences among populations of green crab from a small geographic region, which may reflect a combination of both genetic differences and time since establishment. These differences may result in varying impacts on newly invaded habitats.     

Golgi complex beads and the transition region

Secretory proteins and membranes move in transfer vesicles from the rough endoplasmic reticulum through the transition region to the outer saccule of the Golgi complex. In both arthropod and vertebrate cells, the GC beads are a characteristic structural component of the transitional region. The beads are particles about half the size of ribosomes arranged equidistantly from one another and the smooth face of the ER. In an active GC, the beads are in rings through which the ER membrane emerges to form transfer vesicles. The beads may be part of the energy-dependent step required for the movement of proteins along eht secretory pathway, since they lose their ring arrangement under conditions that lower cellular ATP. The beads are organizers for Golgi complexes in the sense that they are the first recognizable components of new GCs as they arise from ER. Arthropod GC beads, but not those of vertebrates, can be visualized through their reaction with bismuth in vivo and in fixed tissue. Useful paradigms for traffic between the ER and the GC need to combine structural and biochemical information. Insect fat body, with its readily resolvable bismuth-strained beads and easily fractionated cell components may have particular value for this problem.     

A Screen for Genetic Suppressor Elements of Hepatitis C Virus Identifies a Supercharged Protein Inhibitor of Viral Replication

Genetic suppressor elements (GSEs) are biomolecules derived from a gene or genome of interest that act as transdominant inhibitors of biological functions presumably by disruption of critical biological interfaces. We exploited a cell death reporter cell line for hepatitis C virus (HCV) infection, n4mBid, to develop an iterative selection/enrichment strategy for the identification of anti-HCV GSEs. Using this approach, a library of fragments of an HCV genome was screened for sequences that suppress HCV infection. A 244 amino acid gene fragment, B1, was strongly enriched after 5 rounds of selection. B1 derives from a single-base frameshift of the enhanced green fluorescent protein (eGFP) which was used as a filler during fragment cloning. B1 has a very high net positive charge of 43 at neutral pH and a high charge-to-mass (kDa) ratio of 1.5. We show that B1 expression specifically inhibits HCV replication. In addition, five highly positively charged B1 fragments produced from progressive truncation at the C-terminus all retain the ability to inhibit HCV, suggesting that a high positive charge, rather than a particular motif in B1, likely accounts for B1’s anti-HCV activity. Another supercharged protein, +36GFP, was also found to strongly inhibit HCV replication when added to cells at the time of infection. This study reports a new methodology for HCV inhibitor screening and points to the anti-HCV potential of positively charged proteins/peptides.     

Heterogeneous biofilm activity in a segregated anaerobic fluidized bed bioreactor

Bed segregation in a fluidized bed bioreactor profoundly influenced biofilm thickness and microbial activities of the biofilm along the bed height. Bioparticles coated with a thin biofilm, observed at the bottom of the reactor, had a higher specific activity in propylene glycol and n-propanol degradation than in thick biofilms developed at the top of the reactor. Although no significant difference was observed in specific activity for propionate and acetate along the reactor flow axis, more total propionate and acetate conversion occurred in regions of thicker biofilm accumulation.