OBJECTIVES/GOALS: MiaA is a highly conserved prenyl transferase that catalyzes synthesis of the i6A37 tRNA modification in E. coli. While transcriptional regulation of MiaA is well characterized, there is no information on the MiaA post-transcriptional regulation. The aim of this study is to characterize the post-transcriptional regulation of the MiaA gene in E. coli. METHODS/STUDY POPULATION: To characterize the post-transcriptional regulation of miaA, we executed a targeted genetic screen of an E. coli small RNA library on a miaA-lacZ translational reporter fusion strain to identify small RNAs (sRNAs) that modulate MiaA translation or transcription termination. We also measured MiaA mRNA levels and miaA-lacZ activity in the absence or over-expression of candidate sRNA regulators of MiaA. We also measured MiaA mRNA levels in the absence of RNaseE and PNPase, two enzymes involved in mRNA turnover. Finally, we measured the ability of purified recombinant CsrA to bind to the MiaA mRNA transcript in vitro. RESULTS/ANTICIPATED RESULTS: We identified the carbon sensing sRNA CsrB and its cognate protein interaction partner CsrA, as potential post-transcriptional regulators of MiaA. Over-expression of CsrB fully repressed miaA-lacZ activity and MiaA mRNA levels. The absence of CsrA resulted in a defective miaA-lacZ activity and a 10-fold decrease in MiaA mRNA levels. We also identified an increase in the MiaA mRNA half-life particularly in the absence of RNaseE. Our results demonstrate an additional layer of regulation for the miaA operon by the CsrA/CsrB protein-sRNA system. DISCUSSION/SIGNIFICANCE: MiaA is a highly conserved bacterial protein. Our data may represent phenomena in an array of bacteria that could be targeted by novel antibiotics. The human MiaA homologue, TRIT1, plays a role in mitochondrial disorders. We anticipate that information garnered from MiaA studies will elucidate TRIT1 function and its role in mitochondrial disorders.

The Cincinnatian (Katian) of the Cincinnati Tri-State area is widely regarded as one of the most fossiliferous sections known (Meyer and Davis, 2009). Echinoderms from these strata include well-described asteroids, crinoids, cyclocystoids, edrioasteroids, glyptocystoids, mitrates, and ophiuroids. John Pope discovered a partially articulated echinoderm in float from the Fairview Formation that does not correspond to any known Cincinnatian echinoderm. Although mentioned in Ubaghs (1966, as a presumable personal communication from Pope, 1960), Haude and Langenstrassen (1976), Reich (2001), and Reich and Haude (2004), this specimen at the Cincinnati Museum Center (CMCPIP 51316) has neither been described nor illustrated; yet, these authors attributed it to Volchovia Hecker, 1938 in the Class Ophiocistioidea. Questions swirl around this fossil: what is its complete morphology; does it belong to Volchovia; whether or not it can be assigned to Volchovia, is it an ophiocistioid? The first step to understand this enigmatic echinoderm is to illustrate and describe the specimen, which is the objective of this note.

Herbicides have been a primary means of managing undesirable brush on grazing lands across the southwestern United States for decades. Continued encroachment of honey mesquite and huisache on grazing lands warrants evaluation of treatment life and economics of current and experimental treatments. Treatment life is defined as the time between treatment application and when canopy cover of undesirable brush returns to a competitive level with native forage grasses (i.e., 25% canopy cover for mesquite and 30% canopy cover for huisache). Treatment life of industry-standard herbicides was compared with that of aminocyclopyrachlor plus triclopyr amine (ACP+T) from 10 broadcast-applied honey mesquite and five broadcast-applied huisache trials established from 2007 through 2013 across Texas. On average, the treatment life of industry standard treatments (IST) for huisache was 3 yr. In comparison, huisache canopy cover was only 2.5% in plots treated with ACP+T 3 yr after treatment. The average treatment life of IST for honey mesquite was 8.6 yr, whereas plots treated with ACP+T had just 2% mesquite canopy cover at that time. Improved treatment life of ACP+T compared with IST life was due to higher mortality resulting in more consistent brush canopy reduction. The net present values (NPVs) of ACP+T and IST for both huisache and mesquite were similar until the treatment life of the IST application was reached (3 yr for huisache and 8.6 yr for honey mesquite). At that point, NPVs of the programs diverged as a result of brush competition with desirable forage grasses and additional input costs associated with theoretical follow-up IST necessary to maintain optimum livestock forage production. The ACP+T treatments did not warrant a sequential application over the 12-yr analysis for huisache or 20-yr analysis for honey mesquite that this research covered. These results indicate ACP+T provides cost-effective, long-term control of honey mesquite and huisache.

The target site of glyphosate [N-(phosphonomethyl)glycine] inhibition in plants and bacteria is 5-enolpyruvylshikimate 3-phosphate (EPSP) synthase. Our strategy for developing glyphosate-resistant crops has been to genetically engineer plants with a gene that codes for EPSP synthase with low sensitivity in glyphosate. We cloned such a gene from the aroA locus of a glyphosate-resistant mutagenized strain of Salmonella typhimurium. The enzyme encoded by this gene has a single amino acid change resulting in lower affinity for glyphosate and higher affinity for substrates than either plant or wild-type bacterial counterpart. A chimaeric gene containing the mutant aroA gene behind the octopine synthase promoter was constructed and integrated into Agrobacterium T-DNA vectors. Analysis of gall tissue from Brassica campestris L. (turnip rape) infected with A. tumefaciens K12 containing this chimaera showed mRNA and protein expressed from the bacterial gene; 50% of the total EPSP synthase activity present had kinetic properties of the mutant bacterial enzyme. Tobacco (Nicotiana tabacum L. ‘Xanthi′) plants have been regenerated from cocultivation with A. rhizogenes containing the same construct; analysis indicates expression of the gene and enhanced tolerance to glyphosate.

The final effort of the CLIMAP project was a study of the last interglaciation, a time of minimum ice volume some 122,000 yr ago coincident with the Substage 5e oxygen isotopic minimum. Based on detailed oxygen isotope analyses and biotic census counts in 52 cores across the world ocean, last interglacial sea-surface temperatures (SST) were compared with those today. There are small SST departures in the mid-latitude North Atlantic (warmer) and the Gulf of Mexico (cooler). The eastern boundary currents of the South Atlantic and Pacific oceans are marked by large SST anomalies in individual cores, but their interpretations are precluded by no-analog problems and by discordancies among estimates from different biotic groups. In general, the last interglacial ocean was not significantly different from the modern ocean. The relative sequencing of ice decay versus oceanic warming on the Stage 6/5 oxygen isotopic transition and of ice growth versus oceanic cooling on the Stage 5e/5d transition was also studied. In most of the Southern Hemisphere, the oceanic response marked by the biotic census counts preceded (led) the global ice-volume response marked by the oxygen-isotope signal by several thousand years. The reverse pattern is evident in the North Atlantic Ocean and the Gulf of Mexico, where the oceanic response lagged that of global ice volume by several thousand years. As a result, the very warm temperatures associated with the last interglaciation were regionally diachronous by several thousand years. These regional lead-lag relationships agree with those observed on other transitions and in long-term phase relationships; they cannot be explained simply as artifacts of bioturbational translations of the original signals.

Paleozoic echinoids are exceptionally rare, and little is known of their paleoenvironmental distribution. The echinoid fauna of the Fort Payne Formation (Late Osagean, Early Viséan) of south-central Kentucky is documented. Four genera, ?Archaeocidaris, Lepidocidaris, ?Lepidesthes, and an unidentified lepidocentrid, were recovered and represent three different families. This fauna, and their associated paleoenvironments, give important new insights into the facies distribution of Paleozoic echinoids and the taphonomic biases that affect this distribution. Lepidocidaris is known from the green shale facies, which comprises the core of Fort Payne’s carbonate buildups. ?Archaeocidaris and the lepidocentrid are known from the wackestone buildups and crinoidal packstone buildups. ?Lepidesthes is also known from crinoidal packstone and wackestone buildups, which argues against a semi-infaunal life mode for this taxon. All relatively semiarticulated echinoids were known from autochthonous facies, whereas the only echinoids from the allochthonous facies were disarticulated hemipyramids. Furthermore, deeper-water carbonate buildups were apparently capable of supporting diverse echinoid faunas during the Viséan.

Crinoid faunas from the Lower Devonian of South America are poorly known. Two new taxa are described from the Emsian Icla Formation at Cerro Kochis in the Cochabamba Department: the rhodocrinitid, Lutocrinus boliviaensis n. gen. n. sp. and Griphocrinus pirovanoi n. sp. Two additional camerates are reported from the Emsian Belèn Formation of the Altiplano region in the La Paz Department, including Ctenocrinus branisai n. sp. and Boliviacrinus isaacsoni McIntosh, 1988. In addition, a specimen of the blastoid, Pachyblastus dicki Bremier and Macurda, 1972, was recovered from the Altiplano region. These crinoids and material previously collected by Leonardo Branisa, including specimens of C. branisai n. sp., Apurocrinus sucrei McIntosh, 1981, B. isaacsoni McIntosh, 1988, and L. boliviaensis n. sp., are described and revised. The specimens give new insights into the Lower Devonian Malvinokaffric Realm of Bolivia, a relatively new frontier in crinoid paleontology.

Reports of bloodstream infections caused by methicillin-resistant Staphylococcus aureus among chronic hemodialysis patients to 2 Centers for Disease Control and Prevention surveillance systems (National Healthcare Safety Network Dialysis Event and Emerging Infections Program) were compared to evaluate completeness of reporting. Many methicillin-resistant S. aureus bloodstream infections identified in hospitals were not reported to National Healthcare Safety Network Dialysis Event.

Infect. Control Hosp. Epidemiol. 2016;37(2):205–207

Crinoids were relatively unaffected by the end-Devonian Hangenberg mass extinction event. Major clades of Devonian durophagous fishes suffered significant extinctions, however, and the dominant surviving clades were biting or nipping predators. In part as a response to the Hangenberg event, early Mississippian crinoids underwent an adaptive radiation, while fish clades with a shell-crushing durophagous strategy diversified. Durophagous predators are inferred to have been more effective predators on camerate crinoids; and it is hypothesized, following the predictions of escalation, that through the early Mississippian, camerate crinoids evolved more effective anti-predatory strategies in response. We test this hypothesis of escalation by examining the changes in spinosity and plate convexity among camerate crinoids throughout this interval. A new method was formulated to test for an increase in convexity of the tegmen plates. Traits in Agaricocrinus, Aorocrinus, and Dorycrinus (Family Coelocrinidae) were tested for congruence to the escalation hypothesis, and results were mixed. Convexity of tegmen plates in Agaricocrinus, spine length/calyx diameter in Aorocrinus, calyx size in Aorocrinus, central spine length in Dorycrinus, and spine width in Dorycrinus did not have size increase trends supporting escalation. Rather than an increase in convexity, the variance of convexity in Agaricocrinus tegmen plates narrowed, which could reflect an optimum. Alternatively, morphological change consistent with the escalation hypothesis occurred in calyx size of Agaricocrinus and in lateral spine length and calyx size in Dorycrinus. Furthermore, central and lateral spine length, parameters of the spine width, and size trends support escalation when Aorocrinus and Dorycrinus are treated as a lineage. Thus, inferred escalation acted on traits differently within a single lineage and was relevant for both speciation and the diversification of a new genus.