Analysis of sequences and predicted structures required for viral satellite RNA accumulation by in vivo genetic selection.

In vivo genetic selection was used to study the sequences and structures required for accumulation of subviral sat-RNA C associated with turnip crinkle virus (TCV). This technique is advantageous over site-specific mutagenesis by allowing side-by-side selection from numerous sequence possibilities as well as sequence evolution. A 22 base hairpin and 6 base single-stranded tail located at the 3'-terminus of sat-RNA C were previously identified as the promoter for minus strand synthesis. Approximately 50% of plants co-inoculated with TCV and sat-RNA C containing randomized sequence in place of the 22 base hairpin accumulated sat-RNA in uninoculated leaves. The 22 base region differed in sat-RNA accumulating in all infected plants, but nearly all were predicted to fold into a hairpin structure that maintained the 6 base tail as a single-stranded sequence. Two additional rounds of sat-RNA amplification led to four sequence family 'winners', with three families containing multiple variants, indicating that evolution of these sequences was occurring in plants. Three of the four sequence family winners had the same 3 bp at the base of the stem as wild-type sat-RNA C. Two of the winners shared 15 of 22 identical bases, including the entire stem region and extending two bases into the loop. These results demonstrate the utility of the in vivo selection approach by showing that both sequence and structure contribute to a more active 3'-end region for accumulation of sat-RNA C.     

REITER PROTEIN COMPLEMENT FIXATION TEST—A Preliminary Comparison of the TPI Test with the Complement Fixation Test Employing a Soluble Protein Antigen Derived from the Reiter Strain

A comparative study of the specificity of the Reiter protein complement fixation (RPCF) test and the Treponema pallidum immobilization (TPI) test on 180 sera showed that the results in 178 instances or 98.9 per cent were in agreement.The sensitivity of the RPCF test, when compared to the TPI test, on 189 sera from patients known to have syphilis was in agreement in 182 or 96.3 per cent, assuming a correlation exists between positive TPI and both the “reactive” or “weakly reactive” RPCF test results.As to reproducibility of results, the RPCF test results agreed in 84 of the 87 sera tested (95.4 per cent). The sera were tested at least three times on different days.Anticomplementary reactions were observed in three of 91 normal sera.     

Stable antisense RNA expression neutralizes the activity of low-density lipoprotein receptor-related protein and promotes urokinase accumulation in the medium of an astrocytic tumor cell line.

Low-density lipoprotein receptor-related protein (LRP) binds and internalizes multiple ligands that are structurally and functionally diverse. However, the effects of LRP on cellular phenotype remain unclear. To study LRP in human astrocytic tumor cells, we designed LRP antisense RNA expression constructs in which the antisense cDNA fragment was expressed under the control of the cytomegalovirus (CMV) promoter. U-1242 MG astrocytic tumor cells were transfected with the antisense constructs and cloned from single cells to yield multiple cell lines with decreased LRP expression. Further studies were performed with two cell lines in which LRP antigen was completely eliminated (L(alpha)42) or substantially decreased (Lalpha47), as determined by Western blot analysis. Untransfected U-1242 MG cells and cells that were stably transfected with empty vector (pBK-CMV) bound activated alpha2-macroglobulin (alpha2M) in a specific and saturable manner. The Bmax was about 5000 receptors/cell. Lalpha42 cells did not bind alpha2M, and binding was decreased by >60% in Lalpha47 cells. Lalpha42 and Lalpha47 cells also demonstrated reduced susceptibility to the cytotoxin, Pseudomonas exotoxin A, and accumulated greatly increased levels of urokinase-type plasminogen activator (uPA) in conditioned medium. The accumulation of uPA demonstrates a major role for LRP in the catabolism of this protein in astrocytic tumor cells. The LRP-deficient cell lines, developed using antisense technology, represent a new model system for studying LRP function in astrocytes.     

Strategies for Obtaining Obsidian in Pre-European Contact Era New Zealand

Archaeological evidence of people''s choices regarding how they supply themselves with obsidian through direct access and different types of exchanges gives us insight in to mobility, social networks, and property rights in the distant past. Here we use collections of obsidian artefacts that date to a period of endemic warfare among Maori during New Zealand''s Late Period (1500–1769 A.D.) to determine what strategies people engaged in to obtain obsidian, namely (1) collecting raw material directly from a natural source, (2) informal trade and exchange, and (3) formal trade and exchange. These deposits represent a good cross-section of Late Period archaeology, including primary working of raw material at a natural source (Helena Bay), undefended sites where people discarded rubbish and worked obsidian (Bream Head), and a heavily fortified site (Mt. Wellington). We find that most of the obsidian described here was likely obtained directly from natural sources, especially those located on off-shore islands within about 60–70 km of sites. A smaller amount comes from blocks of material transported from an off-shore island a greater distance away, called Mayor Island, in a formal trade and exchange network. This study demonstrates the value of conducting tandem lithic technology and geochemical sourcing studies to understand how people create and maintain social networks during periods of warfare.     

Fibrinogen,Other Putative Markers of Inflammation,and Weight Gain in Middle‐aged Adults—The ARIC Study

Purpose: Weight gain is an important risk factor for the development of the metabolic syndrome, and inflammatory mediators are strongly associated with this syndrome. Our aim was to investigate whether inflammation predicts the development of weight gain in populations. Research Methods and Procedures: We investigated selected markers of inflammation in the prediction of weight gain over an approximately 3‐year period in a biethnic cohort of 13,017 men and women, 45 to 64 years of age, using multiple linear and logistic regression modeling. Results: In adjusted models, those in the highest quartile of fibrinogen gained, during the first 3 years of follow‐up, an estimated 0.23 kg/year more than those in the lowest quartile (p < 0.001). Adjusted odds of a large (greater than the 90th percentile) weight gain for those in the highest quartile of fibrinogen were 1.65 (95% confidence interval [CI], 1.38 to 1.97) times those in the lowest quartile. Similarly adjusted odds ratios for a large weight gain for those with high levels of white blood cell count, factor VIII, and von Willebrand factor were 1.38 (1.14 to 1.67), 1.28 (1.08 to 1.53), and 1.28 (1.08 to 1.51), respectively. Discussion: Fibrinogen and other putative markers of inflammation predict weight gain in middle‐aged adults. Given the known links between the inflammatory response and intermediary metabolism and the methodological strengths of the Atherosclerosis Risk in Communities (ARIC) cohort, these findings, though without immediate clinical applicability, suggest that inflammatory processes play a role in the development of the metabolic syndrome and cardiovascular disease in part through stimulation of weight gain.     

Molecular Phylogeography of Oncomelania Lindoensis (Gastropoda: Pomatiopsidae), the Intermediate Host of Schistosoma Japonicum in Sulawesi

Oncomelania lindoensis from Lake Lindu, Sulawesi, was characterizedfor genetic variation at 21 allozyme loci and compared withO. hupensis (China) and O. quadrasi (Philippines). Geneticdistances and interpopulation patterns of allele-sharing pointto a closer relationship between Sulawesi and the Philippines(Nei's unbiased genetic distances (D) averaged 0.50) than betweenSulawesi and China (D= 0.79). These data, coupled with a considerationof the geographic distribution of the genus, support the hypothesisthat the Sulawesi Oncomelaniaoriginated by avian-facilitated colonizationfrom the Philippines about two million years ago. Oncomelania from Sulawesi were originally described as subspecificallydistinct: Oncomelania hupensis lindoensis. However, the allopatricdistribution, unique alleles at five loci, and significant geneticdistances from congeners in Mindanao and elsewhere in the Philippinessuggest that this taxon should be distinguished as a full specieswithin the Oncomelania hupensis species group, namely: O. lindoensisDavis & Carney 1973. Comparison with published data on variationwithin quadrasi and in three Chinese subspecies of hupensisshowed that D values increase with taxonomic level in this speciesgroup. D averaged 0.15 (0–0.26) within Chinese subspeciesand 0.04 (0–0.13) within the Philippines, but was 0.30(0.20–0.45) between Chinese subspecies, and 0.48–0.80between the three species (hupensis, quadrasi and lindoensis).The genotypic cluster species concept and these multilocus geneticdistances can be used to help define species and subspeciesin these medically important snails. (Received 14 May 1997; accepted 20 April 1998)