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Byline: Adam Wollner Rep. John Delaney 's announcement that he would run for president left just about everyone in the political world scratching their heads. The Maryland Democrat's decision more [...]

This article revises the spatial and temporal boundaries of the Casas Grandes tradition associated with northwest Chihuahua, Mexico, based on new data collected in neighboring northeastern Sonora. The Casas Grandes tradition attained its greatest extent during the Medio period (AD 1200–1450/1500) followed by a dramatic demographic and political collapse. Hunter-gatherer groups subsequently occupied most of northwest Chihuahua. Data from the Fronteras Valley, Sonora, presents an alternative scenario, with a clear pattern of cultural continuity from the eleventh century to the colonial period in which sedentary farmers occupied the same landscapes and occasionally the same villages. These observations contribute to our understanding of the spread and subsequent demise of the Casas Grandes tradition in hinterland regions. For the Fronteras Valley, we infer that immigrant groups originally introduced Casas Grandes traditions and that uneven participation in a suite of shared religious beliefs and practices was common to all the hinterlands.

Br J Sports Med 2003;37:516-520 Objective: To examine venous partial pressure of oxygen (Pv[O.sub.2]), transcutaneous oxygen tension (tcP[O.sub.2]), and V[O.sub.2]MAX in a normobaric environment after a single hyperbaric oxygen (HB[O.sub.2]) [...]

It is well-established that at pH 7.4, intramolecular 1,3-N-alkylation reactions in isophosphoramide mustard (IPM) and phosphoramide mustard (PM) produce electrophilic alkylating agents with aziridinyl moieties. To investigate the role of 1,5-intramolecular cyclizations in the chemistry of IPM and PM, the five-membered ring phospholidine products of these reactions were independently synthesized and characterized by ³¹P NMR. In 0.33 M BisTris, pH 7.4, 37 °C, the intramolecular O-alkylation product of IPM [2-(2-chloroethylamino)-2-tetrahydro-2H-1,3,2-oxazaphospholidine-2-oxide (11)] had a chemical shift of δ 33.0 and a half-life of 3.3 h. The O-alkylation product of PM [2-amino-3-(2-chloroethyl)tetrahydro-2H-1,3,2-oxazaphospholidine-2-oxide (12)] displayed a chemical shift of δ 30.6 and a half-life of 26.9 h. For both IPM and PM, 1,5-N-alkylation provides the same product [1-(2-chloroethyl)-2-hydroxy-tetrahydro-2H-1,3,2-diazaphospholidine-2-oxide (13)]. Because of its instability, 13 was generated in situ and was not isolated; however, the chemical shift (δ 33.0) and reactivity (half-life 0.3 h at 25 °C) of the species attributed to 13 were consistent with the assigned structure. Resonances with ³¹P NMR chemical shifts indicative of 11 or 12 did not appear in reaction solutions of IPM or PM. The compound assigned as 13 gave hydrolysis products that were not found in reaction solutions of IPM or PM. The collective data supported the conclusion that intramolecular 1,5-alkylations do not contribute to the chemistry of IPM or PM in aqueous solutions at pH 7.4, 37 °C. Conversely, 11 and 12 were found to be the major if not exclusive products formed in DMSO solutions of the respective cyclohexylammonium salts of IPM and PM. Both 11 and 12 were relatively noncytotoxic against a series of cell lines, but there were differences in mutagenicities. Chinese hamster ovary cells were exposed to 11 or 12 for one half-life of each compound; 11 was nonmutagenic up to 500 μM, while 12 (500 μM) was mutagenic with 246 mutant colonies/10⁶ surviving cells.

We recently cloned and expressed a novel P2Y receptor (tp2y receptor) from a turkey cDNA library. Expression of this receptor in 1321N1 human astrocytoma cells confers nucleotide-dependent stimulation of phospholipase C activity; however, as we demonstrate here, it also confers nucleotide-dependent inhibition of adenylyl cyclase. Both the phospholipase C and adenylyl cyclase responses were promoted by receptor agonists over a similar range of concentrations. Moreover, not only did UTP and ATP activate the avian receptor but ITP, GTP, xanthosine 5'-triphosphate, and CTP were also agonists, with EC(50) values ranging between 0.1 and 1 microM. Similar potencies, rank-order, and selectivity of nucleotide agonists were also demonstrated for intracellular Ca(2+) mobilization measured during a 30-s stimulation under constant superfusion conditions. This observation indicates that receptor activation by nucleoside 5'-triphosphates is not produced by interconversion of these nucleotides into ATP or UTP. Pretreatment of cells with pertussis toxin completely abolished the inhibitory effect of nucleotide agonists on adenylyl cyclase, whereas the activation of phospholipase C was only partially inhibited. These results demonstrate that the avian P2Y receptor is a nucleoside triphosphate receptor of broad agonist selectivity that interacts with both pertussis toxin-insensitive and -sensitive G proteins to activate phospholipase C and to inhibit adenylyl cyclase. This is the first cloned P2Y receptor that is clearly Gi/adenylyl cyclase-linked.

1We investigated the effect of protein kinases and phosphatases on murine cystic fibrosis transmembrane conductance regulator (CFTR) Cl−channels, expressed in Chinese hamster ovary (CHO) cells, using iodide efflux and the excised inside‐out configuration of the patch‐clamp technique.2The protein kinase C (PKC) activator, phorbol dibutyrate, enhanced cAMP‐stimulated iodide efflux. However, PKC did not augment the single‐channel activity of either human or murine CFTR Cl−channels that had previously been activated by protein kinase A.3Fluoride, a non‐specific inhibitor of protein phosphatases, stimulated both human and murine CFTR Cl−channels. However, calyculin A, a potent inhibitor of protein phosphatases 1 and 2A, did not enhance cAMP‐stimulated iodide efflux.4The alkaline phosphatase inhibitor, (−)‐bromotetramisole augmented cAMP‐stimulated iodide efflux and, by itself, stimulated a larger efflux than that evoked by cAMP agonists. However, (+)‐bromotetramisole, the inactive enantiomer, had the same effect. For murine CFTR, neither enantiomer enhanced single‐channel activity. In contrast, both enantiomers increased the open probability (Po) of human CFTR, suggesting that bromotetramisole may promote the opening of human CFTR.5As murine CFTR had a low Poand was refractory to stimulation by activators of human CFTR, we investigated whether murine CFTR may open to a subconductance state. When single‐channel records were filtered at 50 Hz, a very small subconductance state of murine CFTR was observed that had a Pogreater than that of human CFTR. The occupancy of this subconductance state may explain the differences in channel regulation observed between human and murine CFTR.

Selective inhibitors of adenosine production, degradation and transport were used to potentiate in vivo levels of adenosine and to determine the source of both basal and N-methyl-D-aspartate (NMDA)-induced increases in levels of endogenous adenosine in vivo. Male Sprague-Dawley rats receiving unilateral intrastriatal injections of pharmacological agents were sacrificed 15 min postinjection by high-energy focused microwave irradiation (10 kW, 1.25 s). Ipsilateral and contralateral striata were dissected, and adenosine levels were measured by high-performance liquid chromatography. Inhibition of 5'-nucleotidase by alpha, beta-methylene ADP dose-dependently decreased adenosine levels under basal as well as NMDA-stimulated conditions. Inhibition of nucleoside transport by dilazep and adenosine deaminase by 2'-deoxycoformycin each dose-dependently increased basal adenosine levels. 2'-Deoxycoformycin potentiated NMDA-induced increases in adenosine levels. Inhibition of adenosine kinase by 5'-amino-5'-deoxyadenosine increased basal levels of adenosine, but did not significantly affect NMDA-induced increases in adenosine. 2'-Deoxycoformycin combined with 5'-amino-5'-deoxyadenosine produced a greater enhancement of NMDA-induced increases in levels of adenosine than when either drug was administered separately. Endogenous adenosine in vivo apparently originates from release of adenosine as well as from release and extracellular breakdown of a nucleotide under both basal and NMDA-stimulated conditions. Furthermore, inhibitors of adenosine kinase and adenosine deaminase work best to increase levels of endogenous adenosine under basal and NMDA-stimulated conditions, respectively.

Glutamate release after ischemia, hypoxia and seizure activity plays an important role in stimulating adenosine production and release. We characterized the ionotropic glutamate receptor subtype that regulates adenosine levels in vivo and investigated the role of nitric oxide and free radicals in mediating N-methyl-D-aspartate (NMDA)-induced increases in adenosine levels. Rats received unilateral intrastriatal injections and were sacrificed 15 min postinjection by high-energy focused microwave irradiation (10 kW, 1.25 s). Adenosine levels were measured by high-performance liquid chromatography in ipsilateral and contralateral striata. NMDA and kainic acid dose-dependently increased levels of adenosine whereas (+/-)-alpha-amino-3-hydroxy-5-methyl-4-isoxazol proprionic acid had no effect. The NMDA- and kainic acid-induced increases were blocked by dizocilpine, and the kainic acid response was decreased by 6-cyano-7-nitroquinoxaline-2,3-dione. The effects of NMDA and kainic acid on levels of adenosine were not additive. Intrastriatal L-arginine decreased, and the nitric oxide synthase inhibitor, NG-nitro-L-arginine methyl ester, increased basal adenosine levels. Coadministration of NMDA with L-arginine or NG-nitro-L-arginine methyl ester did not significantly affect NMDA-induced increases in levels of adenosine. N-Tert-butyl-phenylnitrone, a free radical scavenger, reversed L-arginine-induced decreases and NMDA-induced increases in levels of adenosine. Together, these results indicate that NMDA-type ionotropic receptors play an important role in regulating in vivo levels of adenosine in rat striatum and that free radicals, but not nitric oxide, apparently are involved in NMDA-induced increases in levels of adenosine. Conversely, nitric oxide, but not free radicals, apparently exert tonic control over basal levels of endogenous adenosine.

We have generated a mouse carrying the human G551D mutation in the cystic fibrosis transmembrane conductance regulator gene (CFTR) by a one‐step gene targeting procedure. These mutant mice show cystic fibrosis pathology but have a reduced risk of fatal intestinal blockage compared with ‘null’ mutants, in keeping with the reduced incidence of meconium ileus in G551D patients. The G551D mutant mice show greatly reduced CFTR‐related chloride transport, displaying activity intermediate between that of cftr(mlUNC) replacement (‘null’) and cftr(mlHGU) insertional (residual activity) mutants and equivalent to approximately 4% of wild‐type CFTR activity. The long‐term survival of these animals should provide an excellent model with which to study cystic fibrosis, and they illustrate the value of mouse models carrying relevant mutations for examining genotype‐phenotype correlations.

Certain gene mutations in Drosophila melanogaster cause sluggish motor activity. We have localized the transcription unit of the sluggish-A gene to a 14.7-kb region at the base of the X chromosome and have cloned corresponding cDNAs. The predicted protein product has significant sequence similarity to Saccharomyces cerevisiae proline oxidase (EC 1.5.99.8), a mitochondrial enzyme which catalyzes the first step in the conversion of proline to glutamate. In the mutant fly, mitochondrial proline oxidase activity is reduced and has kinetic properties different from those of the wild type, providing further evidence that the gene encodes proline oxidase. Indeed, the free proline level in mutant flies is elevated. When the mutant is rescued by transformation, the proline oxidase and free proline levels, as well as the motor and phototactic behavior, are restored to normal. During embryonic development the sluggish-A transcript is predominantly expressed in the nervous system. Significantly, it has previously been reported that a mouse mutant, PRO/Re, which has reduced proline oxidase activity and elevated free proline levels, also exhibits sluggish behavior.

Immunological evidence is presented which confirms that pp95, one of the major phosphoproteins accumulated in development of the cellular slime mould Dictyostelium discoideum, is identical with spore coat protein SP13. The site of phosphorylation is identified as phosphoserine. The second major phosphorylated component, pp74, corresponds to two co-migrating spore coat proteins known collectively as SP74.

Wnt genes have been implicated in a range of developmental processes in the mouse including the patterning of the central nervous system and limbs. Reported here for the first time is the expression of Wnt2 in the early heart field of 7.5-8.5 dpc (days post-coitum) mouse embryos, making Wnt2 a potentially useful gene marker for the early stages of heart development. Expression was also detected in the allantois from 8.0 dpc and at later stages in the placenta and umbilicus. Mice deficient in Wnt2, generated by gene targeting, displayed runting and approximately 50% died perinatally. Histological analysis revealed alterations in the size and structure of placentas from these mice from 14.5 dpc. The placental defects were associated primarily with the labyrinthine zone and included oedema and tissue disruption and accumulation of maternal blood in large pools. There was also an apparent decrease in the number of foetal capillaries and an increase in the amount of fibrinoid material in the Wnt2 mutant placentas. These results suggest that Wnt2 is required for the proper vascularisation of the mouse placenta and the placental defects in Wnt2-deficient mice result in a reduction in birthweight and perinatal lethality.

As a step towards constructing strains of Z. mobiliscapable of converting cellulose to ethanol, DNA fragments encoding endoglucanase (from Xanthomonas albilineans) and β-glucosidase (from either X.albilineansor Pseudomonassp.) were linked on the same vector and transferred to Z. mobilis. All clones expressed endoglucanase. β-Glucosidase was only produced by clones containing the Xanthomonasgene, and when two copies of this gene were present the β-glucosidase activity was higher.

1To investigate the function of the murine cystic fibrosis transmembrane conductance regulator (CFTR), a full‐length cDNA encoding wild‐type murine CFTR was assembled and stably expressed in Chinese hamster ovary (CHO) cells.2Like human CFTR, murine CFTR formed Cl−channels that were regulated by cAMP‐dependent phosphorylation and intracellular ATP. However, murine CFTR Cl−channels had a reduced single‐channel conductance and decreased open probability (Po) compared with those of human CFTR.3Analysis of the dwell time distributions of single channels suggested that the reduced Poof murine CFTR was caused by both decreased residence in the open state and transitions to a new closed state, described by an intermediate closed time constant.4For both human and murine CFTR, ATP and ADP regulated the rate of exit from the long‐lived closed state.55′‐Adenylylimidodiphosphate (AMP‐PNP) and pyrophosphate, two compounds that disrupt cycles of ATP hydrolysis, stabilized the open state of human CFTR. However, neither agent locked murine CFTR Cl−channels open, although AMP‐PNP increased the Poof murine CFTR.6The data indicate that although human and murine CFTR have many properties in common, some important differences in function are observed. These differences could be exploited in future studies to provide new understanding about CFTR.

Heparan sulphate by-products from the commercial manufacture of pig mucosal heparin were freed of chondroitin sulphate and fractionated according to anionic density. The fractions were treated with HNO2 at pH 1.5, and the resulting mixtures of oligosaccharides were reduced with NaB3H4 and analysed for their disaccharide composition by paper chromatography and by high-pressure liquid chromatography. The results show that the molar ratio of 2-O-sulpho-alpha-L-iduronosylanhydromannose to 6-O-sulpho-(2-O-sulpho-alpha-L-iduronosyl)anhydromannose decreased from 2.5 to 0.04 as the degree of sulphation of the fractions increased. In contrast, the molar ratio of 6-O-sulpho-(beta-D-glucuronosyl)anhydromannose to 6-O-sulpho-(alpha-L-iduronosyl)anhydromannose was approx. 2.4 in all heparan sulphate fractions and decreased to only half of this value in the most highly sulphated heparin fractions. These results are consistent with biosynthetic studies, which have shown that the N-sulpho-(2-O-sulpho-alpha-L-iduronosyl)D-glucosamine disaccharide is the metabolic precursor of the NO-disulpho-(2-O-sulpho-alpha-L-iduronosyl)-D-glucosamine disaccharide in heparin biosynthesis. The high-pressure liquid chromatography of the heparan sulphate oligosaccharides also revealed a number of unidentified oligosaccharides in the deamination mixtures.

Abstract: Genes encoding the mosquitocidal binary toxin of Bacillus sphaericus 2362 were introduced into Synechococcus PCC6301, a cyanobacterium that can tolerate a number of potential variations in the mosquito breeding environment, and can serve as a food source for mosquito larvae. The toxin genes, preceded by a Synechococcus rbcL promoter, were located on a mobilizable Escherichia coli Synechococcus shuttle vector, which was introduced into Synechococcus PCC6301 at frequencies of 10−5–10−7 exconjugants/recipient, depending on the selective conditions used. Recombinant Synechococcus exhibited significant toxicity against 2-day-old and 6-day-old Culex quinquefasciatus larvae, the concentration required to kill 50 % of larvae (LC50) being 2.1 × 105 and 1.3 × 105 cells/ml respectively. Mosquitocidal activity decreased tenfold after 20 generations of non-selective growth.

Mutations in the small optic lobes (sol) gene of Drosophila melanogaster cause specific cells to degenerate in the developing optic lobes, resulting in the absence of certain classes of columnar neurons. These neuronal defects lead to specific alterations in behavioral characteristics, particularly during flight and walking maneuvers. We have isolated the wild-type sol locus by microcloning and chromosomal walking and have established its genetic and molecular limits. Two major transcripts of 5.8 and 5.2 kilobases are produced from this locus by alternative splicing and are present throughout the entire life cycle. Sequence analyses of cDNAs corresponding to these two classes of transcripts predict two proteins of 1597 and 395 amino acids. The first shows similarity in its carboxyl-terminal region to the catalytic domain of a vertebrate calcium-activated neutral protease (calpain), whereas its amino-terminal region contains several zinc-finger-like repeats of the form WXCX2CX10-11CX2C. The second predicted protein contains only the first two of the zinc-finger-like repeats and is missing the calpain domain. By constructing transgenic flies carrying a single wild-type copy of the sol gene in a homozygous sol mutant background, we have restored the normal neuroanatomical phenotype to individuals that would have developed mutant brains.

KRE6 encodes a predicted type II membrane protein which, when disrupted, results in a slowly growing, killer toxin-resistant mutant possessing half the normal level of a structurally wild-type cell wall (1-->6)-beta-glucan (T. Roemer and H. Bussey, Proc. Natl. Acad. Sci. USA 88:11295-11299, 1991). The mutant phenotype and structure of the KRE6 gene product, Kre6p, suggest that it may be a beta-glucan synthase component, implying that (1-->6)-beta-glucan synthesis in Saccharomyces cerevisiae is functionally redundant. To examine this possibility, we screened a multicopy genomic library for suppression of both the slow-growth and killer resistance phenotypes of a kre6 mutant and identified SKN1, which encodes a protein sharing 66% overall identity to Kre6p. SKN1 suppresses kre6 null alleles in a dose-dependent manner, though disruption of the SKN1 locus has no effect on killer sensitivity, growth, or (1-->6)-beta-glucan levels. skn1 kre6 double disruptants, however, showed a dramatic reduction in both (1-->6)-beta-glucan levels and growth rate compared with either single disruptant. Moreover, the residual (1-->6)-beta-glucan polymer in skn1 kre6 double mutants is smaller in size and altered in structure. Since single disruptions of these genes lead to structurally wild-type (1-->6)-beta-glucan polymers, Kre6p and Skn1p appear to function independently, possibly in parallel, in (1-->6)-beta-glucan biosynthesis.

Previous attempts to isolate auxotrophic mutants of Anacystis nidulans produced only a limited range of phenotypes. The frequency of recovery of auxotrophic mutants has been quantified following different mutagenic and selective treatments, and their yield has been improved by using (1) a complete medium, (2) additional mutagens, (3) multiple cycles of penicillin enrichment and (4) altered pre-enrichment starvation conditions. These modified induction and selection conditions permitted the isolation of mutants defective in nitrate reductase, nitrite reductase or malate dehydrogenase, unable to reduce sulphate, or deficient in the synthesis of biotin, thiamine, paminobenzoate, serine, glutamate, adenine or uracil.

For biocontrol of mosquitoes, mosquitocidal toxin genes from Bacillus thuringiensissubsp. israelensisand Bacillus sphaericushave been cloned into a number of cyanobacteria. However, little is known about the persistence of such recombinant cyanobacteria in mosquito larval habitats. Four fresh water unicellular cyanobacteria, SynechococcusPCC6301, PCC7425, PCC7942 and SynechocystisPCC 6803, were evaluated under laboratory conditions related to mosquito breeding environments. Results indicated that SynechococcusPCC6301 was potentially the most suitable organism for use in the natural mosquito habitat as it could tolerate a wide range of temperatures, salinities, and biological and chemical insecticides. Moreover, strain PCC6301 could be ingested and digested by Culex quinquefasciatuslarvae and could support the development of larvae to full insect maturity.

The short story "The Doctor," by S.C. Delaney and Agnes Potier is presented.

The short story "The Wiper," by S.C. Delaney and Agnes Potier is presented.

Two unrelated South Australian families are presented who have a syndrome involving deafness, anal and digital abnormalities, with affected individuals in 2 generations. There is a single family reported in the literature with identical physical findings and a second family has been reported with digital and anal defects only.

The authors narrate their experience visiting their villages.

New research conducted by the University of Western Australia highlights the skin health concerns of urban-living Aboriginal children and the need for improved dermatology services. The study focused on two urban Aboriginal Community Controlled Health Organisations (ACCHOs) and found that skin conditions such as eczema, tinea, and acne were common among the children. The research emphasizes the importance of embedding dermatology practice within ACCHOs to ensure better access to care for Aboriginal children. The study was funded by various organizations, including the Western Australian Future Health and Innovation Fund and the National Health and Medical Research Council. [Extracted from the article]

A recent study conducted by the University of Washington examined the relationship between intimate partner violence (IPV) and health outcomes among people with HIV (PWH) in the United States. The study found that 9.3% of PWH reported experiencing IPV in the past year, with half of them reporting psychological violence without physical or sexual violence. The study revealed that IPV was associated with various negative health outcomes, including unstable housing, higher HIV viral load, depressive symptoms, substance use, and lower adherence to antiretroviral therapy. The findings highlight the importance of addressing psychological IPV in addition to physical and sexual IPV. [Extracted from the article]

V3 loop sequences were compared from 5 human immunodeficiency virus type 1(HIV-1)-infected patients over time. Three patients remained asymptomatic and 2 became symptomatic with large decreases in CD4 cell counts. The patient isolates were previously evaluated for phenotypic and antigenic properties and had different sensitivities to serum neutralization and changes in phenotype. This study showed a number of amino acid changes for the 2 symptomatic patients, each of whom progressed to AIDS during the study. The only amino acid substitution consistently associated with reduced CD4 cell counts, cytopathic effect, and progression to AIDS was Arg at position 11. Specific amino acid changes could not be correlated with increasing serum neutralization resistance or cytotropism changes. Increased loop charge was associated with a switch from macrophage to T cell tropism and a decrease in the number of CD4 cells. The study shows the importance of naturally occurring mutations in the V3 loop in controlling the biologic properties of HIV-1.

Progression to AIDS in patients harboring human immunodeficiency virus type 1 (HIV-1) isolates expressing a syncytium-inducing (SI) phenotype is faster than in those in whom the virus expresses a non-SI (NSI) phenotype. Zidovudine monotherapy does not appear to alter this outcome. To examine the role of didanosine (ddl) monotherapy in phenotype expression, HIV-1 isolates from 73 patients receiving ddl for up to 72 weeks were analyzed. After 12 weeks, the number of isolates expressing an NSI phenotype was 29% higher than at the start of treatment. Patients receiving high-dose ddI (375 mg twice daily) were significantly more likely to express the NSI phenotype at 12 weeks than patients who received low-dose ddI (100 mg twice daily), even after adjustment for phenotype and CD4 cell count at baseline, suggesting that ddI may be selective against the faster-replicating virus. ddI at 375 mg twice daily significantly increases the probability of an NSI phenotype over the short term in patients with advanced HIV disease.

The PMT2gene from Saccharomyces cerevisiaewas identified as FUN25, a transcribed open reading frame on the left arm of chromosome I (Ouellette, B. F. F., Clark, M. W. C., Keng, T., Storms, R. G., Zhong, W., Zeng, B., Fortin, N., Delaney, S., Barton, A., Kaback, D. B., and Bussey, H.(1993) Genome36, 32-42). The product encoded by the PMT2gene shows significant similarity with the dolichyl phosphate-D-mannose:protein O-D-mannosyltransferase, Pmt1p (EC 2.4.1.109), which is required for initiating the assembly of O-linked oligosaccharides in S. cerevisiae(Strahl-Bolsinger, S., Immervoll, T., Deutzmann, R., and Tanner, W.(1993) Proc. Natl. Acad. Sci. U. S. A.90, 8164-8168). The PMT2gene encodes a new protein O-D-mannosyltransferase. Yeast cells carrying a PMT2disruption show a diminished in vitroand in vivo O-mannosylation activity and resemble mutants with a nonfunctional PMT1gene. Strains bearing a pmt1 pmt2double disruption show a severe growth defect but retain residual O-mannosylation activity indicating the presence of at least one more protein-O-mannosyltransferase.