Murine C4-binding protein: a rapid purification method by affinity chromatography

A simple, 3-step method was described for purification of murine C4 binding protein (C4-bp), a recently recognized serum protein that functions as one of the regulatory proteins of the complement system. The method consists of 1) affinity chromatography using TNBS-BGG-conjugated Sepharose beads, 2) gel filtration on a Sepharose 6B column, and 3) heparin-Sepharose chromatography. By this method, milligram quantities of C4-bp can be easily purified by more than 500-fold from EDTA-serum of various mouse strains, and the whole purification process can be completed within 1 wk. The overall yield of C4-bp is about 15%. The C4-bp thus prepared is homogeneous as judged by SDS-polyacrylamide gel electrophoresis and immunelectrophoresis. The purified mouse C4-bp showed physicochemical properties very similar to those described for human C4-bp. Like human C4-bp, mouse C4-bp is composed of several apparently identical subunits of the m.w. of 80,000. However unlike the human counterpart, the subunits of mouse C4-bp are not linked by disulfide bonds but are connected by non-covalent forces that can be disrupted by SDS. The purified mouse C4-bp retained binding affinity for C4 and showed unaltered antigenicity. Immunization of rabbits with the purified mouse C4-bp resulted in the production of potent and monospecific antisera.     

Production of tetanolysin preparations and characteristics of their properties

As the result of the study of tetanolysin-producing Clostridium tetani strains, their populations have been found to be markedly heterogeneous with respect to the hemolytic activity of clone cultures. On the basis of normal and dialyzed cultures of selected variants with maximum activity the preparations of tetanolysin have been obtained, and their hemolytic activity and antigenic properties have been studied. Antihemolytic rabbit sera have also been obtained and characterized. Partially purified preparations of tetanolysin with high hemolytic activity have been obtained by the fractionation of C. tetani dialyzed cultures with ammonium sulfate.     

Isolation of mutants of the obligate methanotroph Methylomonas albus defective in growth on methane

A strain of Methylomonas albus BG8WM, a type 1 obligate methanotroph, which had been maintained for 2 ycars by serial passage on solid medium containing methanol as a carbon source was found to mutate at a frequency of 10^-5-10^-6 to resistance to dichloromethane (DCM^R), the parental strain BG8 did not give rise to DCM^R colonies. DCM^R strains were no longer capable of growth on methane as a carbon cource and exhibited greatly reduced or undetectable methane mono-oxygenase activity. The mutants fell into three groups on the basis of SDS-PAGE analysis of the polypeptide profiles of the particulate fraction of cell extracts. One or more of four polypeptides of Mr 70,000, 50,000, 25,000 and 23,000 were implicated as being components of the methane mono-oxygenase. Spontaneous reversion to growth on methane and sensitivity to dichloromethane occurred at a frequency of about 10^-8. The loss of methane mono-oxygenase activity was not associated with loss of the resident 55 kb plasmid.Abbreviations DCM^R dichloromethane-resistant - SDS-PAGE sodium dodecyl sulphate polyacrylamide gel electrophoresis - NMS nitrate minimal salts medium     

Increased near-ultraviolet induced DNA fragmentation in xeroderma pigmentosum variants.

Immediate fragmentation of parental DNA by near-ultraviolet irradiation at 313 nm was measured in cultured skin fibroblasts from normal individuals, patients with Xeroderma pigmentosum of complementation group A (XPA) and Xeroderma pigmentosum variants (XPV) by the alkaline elution procedure. For a dose of 2.25 KJm^?2 given at O^o fragmentation was comparable in all cell strains. However, fragmentation was strongly increased relative to O^o in XPV but not in normal fibroblasts and the XPA strains when irradiation was carried out at 37^o. From our results it appears that a step in the repair of $\text{parental}$ DNA is abnormal in XPV.     

Antibacterial activity of Turkish propolis and its qualitative and quantitative chemical composition

The antibacterial activity of propolis from different regions of Turkey was studied, accompanied by TLC and GC-MS analyses of its chemical composition and spectrophotometric quantification of the most important active principles. All six samples were active against the bacterial test strains used; however, samples 1 (Yozgat), 2 (Izmir) and 3 (Kayseri) were more active than samples 4 (Adana), 5 (Erzurum) and 6 (Artvin). By TLC comparison all samples were found to contain poplar taxonomic markers but in samples 4 (Adana), 5 (Erzurum) and 6 (Artvin), different substances were observed, which were not present in P. nigra L. bud exudate. The typical poplar samples 1 (Yozgat), 2 (Izmir) and 3 (Kayseri) displayed very similar phenolic and flavonoid content. Samples 4 (Adana), 5 (Erzurum) and 6 (Artvin) were characterized by low phenolic and very low flavonoid concentrations. Qualitative analysis by GC-MS revealed that sample 4 (Adana) contained diterpenic acids and high percent of cinnamyl cinnamate, sample 5 (Erzurum)-significant amounts of hydroxy fatty acids and triterpenic alcohoLs, and sample 6 (Artvin)-phenolic glycerides, characteristic for the bud exudate of Populus euphratica Oliv. The results confirm the importance of phenolics for propolis antibacterial activity, and the significance of P. nigra L. as a propolis source, which provides the hive with the best defense against microorganisms.     

Predator detection and avoidance by starlings under differing scenarios of predation risk

Practically all animals must find food while avoiding predators.An individual's perception of predation risk may depend on manyfactors, such as distance to refuge and group size, but it isunclear whether individuals respond to different factors ina similar manner. We tested whether flocks of foraging starlingsresponded in the same way to an increased perception of predationrisk by assessing three factors: (1) neighbor distances, (2)habitat obstruction, and (3) recent exposure to a predator.We found that in all three scenarios of increased risk, starlingsreduced their interscan intervals (food-searching bouts), whichincreased the frequency of their vigilance periods. We thenexamined how one of these factors, habitat obstruction, affectedescape speed by simulating an attack with a model predator.Starlings were slower to respond in visually obstructed habitats(long grass swards) and slower when they had their head downin obstructed habitats than when they had their head down inopen habitats. In addition, reaction times were quicker whenstarlings could employ their peripheral fields of vision. Ourresults demonstrate that different sources of increased riskcan generate similar behavioral responses within a species.The degree of visibility in the physical and social environmentaffects both the actual and perceived risk of predation.