Analysis ofDrosophila chromosome4 using pulsed field gel electrophoresis

Previous estimates of the size ofDrosophila melanogaster chromosome4 have indicated that it is 1% to 4% of the genome or 6 Mb. We have used pulsed field gel electrophoresis (PFGE) to separate megabase-sized molecules ofD. melanogaster chromosomal DNA. Southern blots of these gels were probed with DNA fragments from thecubitus interruptus andzfh-2 genes, which are located on chromosome4. They each identify the same-sized distinct band that migrates at approximately 5.2 Mb in DNA preparations from the Kc cell line. We interpret this band to be intact chromosome4. In DNA obtained from embryos of variousD. melanogaster wild-type strains, this chromosome band showed strain-specific size variation that ranged from 4.5 to 5.2 Mb. TheD. melanogaster chromosome4 probes also identified a single, 2.4 Mb band in embryonic DNA fromDrosophila simulans. We conclude thatD. simulans chromosome4 is substantially smaller than that ofD. melanogaster, presumably owing to diffirences in the amount of heterochromatic DNA sequences. Our simple DNA preparation from embryos and PFGE conditions should permit preparative isolation of chromosome4 DNA and will facilitate the molecular mapping of this chromosome.     

Coexisting mangrove-coral habitat use by reef fishes in the Caribbean

We conducted visual fish surveys in coexisting mangrove-coral (CMC) habitats in Panama to analyze the effect of coral presence in mangrove habitats on the fish assemblage. Our study revealed that CMC habitats harbor distinct fish assemblages compared to mangrove habitats without coral, with greater species richness and increased herbivore abundance. Abstract in Spanish is available with online material.     

Old,socially housed rhesus monkeys manipulate objects

Recent research has indicated that old, individually housed monkeys show little interest in novel objects. Yet unanswered is whether this effect is caused primarily by age or housing condition. The purpose of this study was to assess the role of social living in promoting responsiveness to objects. We measured the rates of object manipulation in older animals, assessed responsiveness over time to particular objects as a measure of habituation, and examined social influences on object use. Several social groups of rhesus monkeys that contained older adults were studied. These groups were housed in indoor pens or in an outdoor enclosure, and all monkeys had continuous access to a variety of objects in their home environment. In contrast to previous studies of individually housed monkeys, our group-housed monkeys showed sustained interest in objects. Old monkeys manipulated objects extensively, and this response was all the more significant, given that the objects were not novel. Monkeys housed in an outdoor enclosure showed object manipulation patterns that were not different from monkeys housed in indoor pens. However, females exhibited much higher object-related responses than males. Social facilitation played a role in the reactions of some monkeys to objects. Patterns of social facilitation as well as avoidance were present in two of the three indoor groups that were observed. Failure to manipulate objects in rhesus macaques appears to be more a function of individual housing than of old age. Factors such as environmental complexity, social needs, and early experience should be considered in order to understand why individually housed rhesus monkeys are unresponsive to objects. © 1993 Wiley-Liss, Inc.     

Mammal use of riparian corridors in semi-arid Sonora,Mexico

Wildlife populations in semi-arid regions are increasingly challenged by human activities and dependent on the connectivity of riparian corridors for access to surface water. The Madrean Archipelago is a biodiversity hotspot along the arid United States–Mexico borderlands that support both Neotropical and Nearctic wildlife. Infrastructure development (e.g., the border wall and the expansion of Mexican Federal Highway 2) in this region inhibits wildlife movement along the transnational mountain archipelago by disconnecting habitat. To explore the relationship between habitat variables and mammal use of riparian corridors in northern Sonora, Mexico, we collected data from 19 motion-sensitive cameras between October 2018 and April 2019 and used single-season occupancy models and Royle-Nichols abundance estimation models to analyze our data. We recorded 21 species of mammals, including the first sighting of jaguar (Panthera onca) in this region in 25 years. River characteristics (distance from river, riparian corridor width, water availability), remoteness (distance from highway, productivity, elevation), and topographic variety (vertical elevation difference) influenced patterns of occupancy probability and estimated abundance of mammals >1 kg, but the strength and direction of these relationships varied by species. Additionally, intermittently wet desert washes were comparable in species richness to the perennial system. These results highlight the importance of examining physical and biological aspects of habitat. This is especially true when identifying corridors where mitigation structures should be placed to improve wildlife connectivity in biodiversity hotspots like the Madrean Archipelago and semi-arid ecosystems worldwide.     

Double blind randomised controlled trial of effect of metoprolol on myocardial ischaemia during endoscopic cholangiopancreatography.

OBJECTIVE--To evaluate the effect of metoprolol, a beta adrenergic blocking drug, on the occurrence of myocardial ischaemia during endoscopic cholangiopancreatography. DESIGN--Double blind, randomised, controlled trial. SETTING--University Hospital. SUBJECTS--38 (two groups of 19) patients scheduled for endoscopic cholangiopancreatography. INTERVENTIONS--Metoprolol 100 mg or placebo as premedication two hours before endoscopy. MAIN OUTCOME MEASURES--Heart rate, arterial oxygen saturation by continuous pulse oximetry, ST segment changes during endoscopic cholangiopancreatography (an ST segment deviation > 1 mV was defined as myocardial ischaemia), electrocardiogram monitored continuously with a Holter tape recorder. RESULTS--All patients had increased heart rate during endoscopy compared with rate before endoscopy, but heart rate during endoscopy was significantly lower in the metoprolol group compared with the placebo group (P = 0.0002). Twenty one patients (16 placebo, 5 metoprolol; P = 0.0008) developed tachycardia (heart rate > 100/min) during the procedure, and 11 patients (10 placebo, 1 metoprolol; P = 0.003) developed myocardial ischaemia. One patient in the placebo group had an acute inferolateral myocardial infarction. In the 10 other patients with signs of myocardial ischaemia during endoscopy the ST deviation disappeared when the endoscope was retracted. In all patients myocardial ischaemia was related to increases in heart rate, and 10 of the 11 patients had tachycardia coherent with myocardial ischaemia. CONCLUSIONS--Metoprolol prevented myocardial ischaemia during endoscopic cholangiopancreatography, probably through lowering the heart rate. Thus, tachycardia seems to be a key pathogenic factor in the development of myocardial ischaemia during endoscopy.     

The hmu locus of Yersinia pestis is essential for utilization of free haemin and haem-protein complexes as iron sources

Yersinia pestis strains utilize haem and several haem-protein complexes as sole sources of iron. In this study, the haemin uptake locus (hmu) of Y. pestis KIM6+ was selected from a genomic library by trans-duction into an Escherichia coli siderophore synthesis (entC) mutant. Recombinant plasmids containing a common 16 kb BamHI insert were isolated that allowed E. coli entC to use haemin as an iron source. An 8.6 kb region of this insert was found to be essential for haemin utilization and encoded at least five proteins with molecular masses of 79/77, 44, 37, 35, and 30/27.5 kDa. A 10.9 kb Clal fragment containing the hmu locus showed varying degrees of homology to genomic DNA from Yersinia pseudotuberculosis, Yersinia enter-ocolitica, and other genera of Enterobacteriaceae. An E. coli hemA aroB strain harbouring cloned hmu genes used haemin as both an iron and porphyrin source but only on iron-poor medium, suggesting that haemin uptake is tightly iron regulated. Additionally, haemoglobin and myoglobin were used as iron sources by an E. coli entC (pHMU2.2) strain. Deletion of the hmu locus from Y. pestis KIM6+ chromosome generated a mutant that grew poorly on iron-depleted medium containing free haemin as well as mammalian haem-protein complexes including haemoglobin, haemoglobin-haptoglobin, myoglobin, haem-haemopexin, and haem-albumin unless it was complemented with cloned hmu genes.     

Site-directed mutagenesis of the katG gene of Mycobacterium tuberculosis: effects on catalase–peroxidase activities and isoniazid resistance

Recent studies examining the molecular mechanisms of isoniazid (INH) resistance in Mycobacterium tuberculosis have demonstrated that a significant percentage of drug-resistant strains are mutated in the katG gene which encodes a catalase–peroxidase, and the majority of these alterations are missense mutations which result in the substitution of a single amino acid. In previous reports, residues which may be critical for enzymatic activity and the drug-resistant phenotype have been identified by evaluating INH-resistant clinical isolates and in vitro mutants. In this study, site-directed mutagenesis techniques were utilized to alter the wild-type katG gene from M. tuberculosis at 13 of these codons. The effects of these mutations were determined using complementation assays in katG -defective, INH-resistant strains of Mycobacterium smegmatis and Mycobacterium bovis BCG. This mutational analysis revealed that point mutations in the katG gene at nine of the 13 codons can cause drug resistance, and that enzymatic activity and resistance to INH are inversely related. In addition, mutations in the mycobacterial catalase–peroxidase which reduce catalase activity also decrease peroxidase activity.     

Evaluation of a remotely operated vehicle (ROV) as a tool for studying the distribution and abundance of zooplankton

Conventional methods for measuring zooplankton distributionsare too laborious and time consuming to permit sufficient temporaland spatial resolution in many instances. An ability to makemore efficient and precise measurements would be useful. Weevaluated the potential for using the video system of a commerciallyavailable remotely operated vehicle (ROV) to measure the distributionand abundance of zooplankton by calibrating ROV counts withcounts based on a conventional sampling procedure (a Schindlertrap), and by using an ROV to measure the density of zooplanktonin a small lake. As configured here, this particular ROV wassuitable for measuring the density of the cladocerans Daphniaand Holopedium. It was also suitable for assessing the distribution,but not absolute densities, of Chaoborus and Leptodora. Imagequality was inadequate for quantitative estimates of copepod(Diaptomus minutus) abundance, and prevented us from studyingbehavioral responses of copepods to the vehicle. We concludethat the ROV has at least three useful features: it can be usedto locate patches of those species that are imaged effectively;a large number of samples (videotapes) can be collected almostsynoptically with high spatial resolution; the ROV enables insitu observation of zooplankton. The ROV also has three importantlimitations: the small image volume makes it difficult to studyrare organisms; inadequate image resolution precludes studiesof relatively small organisms (e.g. the calanoid copepod D.minutus);zooplankton respond to the presence of the ROV.