Bacteriophage lambda-E. coli K12 vector-host system for gene cloning and expression under lactose promoter control. II. DNA fragment insertion at the vicinity of the lac UV5 promoter.

Summary Recombinant plasmids containing the entire 16S RNA gene from the rrn B cistron of E. coli inserted in Col E1 and pBR322 plasmid vectors have been constructed. These plasmids have been mapped using several restriction endonucleases as well as by DNA-RNA hybridization. These maps reveal previously undetected restriction sites in the rrn B cistron and in Col E1 plasmid DNA.     

Semiautomation of preparation of fixed paraffin-embedded tissue for DNA analysis

The usual manual preparation of single-cell suspensions from fixed paraffin-embedded tissue sections for flow cytometric (FCM) DNA ploidy analysis is a time-consuming, labor-intensive technique that requires 70 minutes to deparaffinize and rehydrate 50 microns sections as the initial step. Manual deparaffinization was compared with two semiautomated methods using an automatic slide stainer with either a 70-minute or 35-minute schedule. Samples from 6 normal tissues and 21 tumors (13 diploid and 8 aneuploid) were prepared using all three methods and analyzed by FCM. The mean cell counts in all samples were over 10(6). The DNA indices for the three samples prepared from a given tissue showed no significant differences. Using the 70-minute automation schedule, no aneuploid peaks were lost, and the ratio of G0G1 normal cells to aneuploid tumor cells was maintained. The automation of deparaffinization can thus provide a significant reduction in the labor need to produce single-cell suspensions for FCM; it can be especially helpful when handling large numbers of tumors. At the same time, the automated procedure decreases the exposure to hazardous chemicals and lowers the chance of losing tissue.     

Ancient Cities of the Indus Valley Civilization

Ancient Cities of the Indus Valley Civilization. Jonathan Mark Kenoyer. Oxford: Oxford University Press; Karachi: American Institute of Pakistan Studies, 1998. 262 pp.     

A method for rapidly mass laser-marking individually coded ground beetles (Coleoptera: Carabidae) in the field

Abstract.  1. A laser-marking technique is described that may be used to rapidly mass-mark and individually code insects in the field. In this study, ground beetles were marked but the method is applicable to a wide range of organisms with a hard exoskeleton or shell.
2. In a field-scale capture–recapture study conducted in winter wheat, 8266 beetles were coded. Individuals marked in the first week of the trial and recaptured 2 months later retained their codes, which were therefore considered permanent.
3. Assemblages were dominated numerically by Pterostichus cupreus , which formed more than 98% of the total captures. Of the 8046 released P. cupreus , 2269 were recaptured. As an illustration of the use of these data, the POPAN parameterisation of the Jolly–Seber model was used to estimate a whole-field population of 17 237 individuals, equating to a density of 0.46 beetles m⁻².     

Reserpic acid as an inhibitor of norepinephrine transport into chromaffin vesicle ghosts

Reserpic acid, a derivative of the antihypertensive drug reserpine, inhibits catecholamine transport into adrenal medullary chromaffin vesicles. Since it does not affect the membrane potential generated by the H+-translocating adenosine triphosphatase but inhibits ATP-dependent norepinephrine uptake with a Ki of about 10 microM, reserpic acid must block the H+/monoamine translocator. Because reserpic acid is much more polar than reserpine, it does not permeate the chromaffin vesicle membrane, nor is it transported into chromaffin vesicle ghosts in the presence of Mg2+-ATP. Although it inhibits norepinephrine transport when added externally, reserpic acid does not inhibit when trapped inside chromaffin vesicle ghosts. Therefore, reserpic acid must bind to the external face of the monoamine translocator and should be a good probe of the translocator's structural asymmetry.     

Evaluation and use of a diffusion-controlled sampler for determining chemical and dissolved oxygen gradients at the sediment-water interface

Field and laboratory evaluations were made of a simple, inexpensive diffusion-controlled sampler with ports on two sides at each interval which incorporates 0.2-m polycarbonate membrane to filter samples in situ. Monovalent and divalent ions reached 90% of equilibrium between sampler contents and the external solution within 3 and 6 hours, respectively. Sediment interstitial water chemical gradients to depths of tens of centimeters were obtained within several days after placement. Gradients were consistent with those determined from interstitial water obtained by centrifugation of adjacent sediment. Ten milliliter sample volumes were collected at 1-cm intervals to determine chemical gradients and dissolved oxygen profiles at depth and at the interface between the sediment and water column. The flux of dissolved species, including oxygen, across the sediment-water interface can be assessed more accurately using this sampler than by using data collected from benthic cores.