Substrate-triggered recruitment of the TolC channel-tunnel during type I export of hemolysin by Escherichia coli.

A defining event in type I export of hemolysin by Escherichia coli is the substrate-triggered recruitment of the TolC channel-tunnel by an inner membrane complex. This complex comprises a traffic ATPase (HlyB) and the 478 residue adaptor protein (HlyD), which contacts TolC during recruitment. HlyD has a large periplasmic domain (amino acid residues 81-478) linked by a single transmembrane helix to a small N-terminal cytosolic domain (1-59). Export was disabled by deletion of the ca 60 amino acid residue cytosolic domain of HlyD, even though the truncated HlyD (HlyDDelta45) was, like the wild-type, able to trimerise in the cytosolic membrane, and interact with the traffic ATPase. The mutant HlyB/HlyDDelta45 inner membrane complex engaged the hemolysin substrate, but this substrate-engaged complex failed to trigger recruitment of TolC. Further analyses showed that HlyDDelta45 was specifically unable to bind the substrate. The result suggests that substrate engagement by the traffic ATPase alone is insufficient to trigger TolC recruitment, and that substrate binding to the HlyD cytosolic domain is essential. Analysis of three further N-terminal deletion variants, HlyDDelta26, HlyDDelta26-45 and HlyDDelta34-38, indicated that an extreme N-terminal amphipathic helix and a cytosolic cluster of charged residues are central to the cytosolic domain function. The cytosolic amphipathic helix was not essential for substrate engagement or TolC recruitment, but export was impaired without it. In contrast, when the charged amino acid residues were deleted, the substrate was still engaged by HlyD but engagement was unproductive, i.e. TolC recruitment was not triggered. Our results are compatible with the HlyD cytosolic domain mediating transduction of the substrate binding signal directly, presumably to the HlyD periplasmic domain, to trigger recruitment of TolC and assemble the type I export complex.     

Bayesian Insight into a Robust Test for Linear Contrast Based on Asymmetric Censored Samples

A robust test for linear contrast using modified maximum likelihood estimators based on symmetrically censored samples proposed by Tiku (1973, 1982a) is studied in this paper from the Bayesian point of view. The effects of asymmetric censoring on this testing procedure is investigated and a good approximation to its posterior distribution in this case is worked out. We also present an example which illustrates the method of obtaining the highest posterior density interval for the linear combination of the unknown location parameters.     

Chick embryonic cells as a source for generating in vitro model of muscle cell dystrophy

Chick embryonic cells can be used to develop an easy and economical in vitro model for conducting studies on the disease muscle dystrophy (MD). For this, the limb myoblasts from 11th day chick embryo were isolated and cultured. To this muscle cell culture, anti-dystroglycan antibody (IIH6) was added so as to target the α-dystroglycan and disrupt the connection between the cytoskeletal proteins and the extracellular matrix (which is a characteristic feature of MD). Cells were allowed to differentiate further and the morphometrics and mRNA expression were studied. The IIH6-treated muscle cells displayed changes in morphometry, contractibility, and also atrophy was observed when compared to the control cultures. Concomitant gene expression studies showed an upregulation in TGF-β expression, while the muscle sculpture genes MYOD1, MYF5, LAMA2 and MYOG were downregulated resembling the MD in vivo. This simple and cost-effective method can be useful in studies to further understand the disease mechanism and also in conducting initial studies on effect of novel pharmacological agents.     

Population ecology of the giant forest hog,Hylochoerus meinertzhageni in Chebera Churchura National Park,Ethiopia

Population, feeding habit, distribution and activity patterns of the giant forest hog (Hylochoerus meinertzhageni) were studied in Chebera Churchura National Park (CCNP), Ethiopia, using line transect sample counts. Population estimate of the hogs was 845 individuals. Among the counted individuals, 42.4% were males and 57.6% were females. Among them, 32.8% were adults, 26.9% were subadults, and 40.3% were piglets. Large sounders of up to 20 individuals were seen during the wet season. They were seen in all habitat types in CCNP with higher abundance in the riverine vegetation. Availability of food, green vegetation cover and water were factors governing distribution of forest hogs in the study area. Grass was the major food item of the hogs (90%), mainly of Panicum sp., Acalypha psilostachya, Cissapelos rigidifolia, Psilotrichum gnaphalobryum and Abutilon ceclii. Feeding was the major activity for which 49.7% of the active daytime was used. Resting (lying down, standing and wallowing) was the next major activity, for which they spent 38.8% of the daytime, followed by other activities (11.5%). Morning and late afternoon activity peaks were more intensive during the dry season. This female‐biased giant forest hog population is expected to sustain in CCNP.     

Seed Dispersal by the Sloth Bear (Melursus ursinus) in South India1

Previous studies have shown that sloth bears are mainly myrmecophagous, but in this study, 39 and 83 percent (wet and dry season, respectively) of scats surveyed in South India contained fruits. Seeds from three of six plant species collected from scats germinated faster than seeds not passing through bears; germination percentage was not higher in any of the six species. These results suggest that the sloth bear may play an important role in the population dynamics of fleshy‐fruited species of the region.     

Transfer and Expression of a Multiple Antibiotic Resistance Plasmid in Marine Bacteria

Conjugal transfer of a multiresistance plasmid from Pseudomonas fluorescens to halophilic and halotolerant bacteria was studied under in vitro and in situ conditions. Mating conducted in broth as well as on plates yielded a plasmid transfer frequency of as high as 10⁻³. Among these two, plate mating facilitated conjugal transfer of plasmid, because the cell-to-cell contact is more in plate mating. When P. fluorescens was incubated in seawater, the organism progressively lost its colony forming activity within 15 days. Microscopic examination revealed the presence of very short rods, indicating that the cells have become viable but nonculturable (VNC). Mating conducted in natural seawater without any added nutrients revealed that the conjugal transfer is influenced by the physical state of the donor and the recipients as well as the availability of nutrients. But a plasmid transfer frequency of 10⁻⁷ was obtained even after the donor cells have become VNC suggesting that the nonculturable state and nutrient deprived condition may not limit plasmid transfer. The results suggest that the terrestrial bacteria entering into the seawaters with antibiotic resistance plasmids may be responsible for the prevalence of resistance genes in the marine environment. Received: 4 May 1998 / Accepted: 18 June 1998     

Observations on the incidence and seasonal fluctuations of certain crustacean larvae in the plankton of the South-West Coast of India

The seasonal fluctuations in the incidence of planktonic organisms and the larval forms of certain crustaceans such as crabs, barnacles and post larvae of prawns in the plankton of the south-west coast of India have been followed for a period of three years from 1963. South-west monsoon period is the least productive period for zooplankton in this area. Brachyuran zoeae, post larvae of prawns and barnacle nauplii occur in the plankton throughout the year with distinct peaks for different groups. The zoeae ofUca annulipes occur in the plankton from September to May with a peak during November–December. The zoeae ofPortunus pelagicus are present in the plankton from September to June and their abundance is in February–March. The post larvae ofMetapenaeus affinis are found in the inshore plankton from November to June with the peak in March. The nauplii ofBalanus amphitrite communis occur in the plankton in all the months of the year, the peak incidence has been during November January. The zoeae ofU. annulipes are found to tolerate better the medium saline conditions. Of the ecological factors, salinity of the ambient water and the availability of planktonic food for the larvae seem to influence the seasonal fluctuations of these crustacean larvae in this locality.