Cytomorphometry of large cell carcinoma of the lung

A computerized morphometry system was used to evaluate criteria for the cytologic diagnosis of large cell carcinoma (LCC) and poorly differentiated adenocarcinoma of the lung. There were 143 cells measured in six cases of LCC (five sputums and one bronchial washing) and 61 cells in four cases of adenocarcinoma (all sputum samples). Cellular and nuclear areas were significantly larger in adenocarcinoma whereas nucleolar area was greater in LCC, producing a higher nucleolar/nuclear area ratio in LCC. Cellular and nuclear form factors were smaller in LCC while the minor axis was longer in adenocarcinoma, resulting in a smaller axial ratio in adenocarcinoma. These data indicate that adenocarcinoma cells are larger and have a more rounded shape and less nucleolar material, as compared to the smaller, more ellipsoid and convoluted cells of LCC, which have more nucleolar area. A logistic regression identified cellular area, nucleolar/nuclear area ratio and cellular and nuclear form factors as significant contributors to the discrimination of LCC from adenocarcinoma, with a positive predictive value of 92%. Morphometry may therefore be helpful in the differential cytologic diagnosis of adenocarcinoma and LCC.     

The C region of human insulin-like growth factor (IGF) I is required for high affinity binding to the type 1 IGF receptor

We have produced and characterized the binding properties of three structural analogs of human insulin-like growth factor I (hIGF-I). These analogs are [1-62]hIGF-I, an analog lacking the carboxyl-terminal 8-amino acid D region of hIGF-I; [1-27, Gly4, 38-70]hIGF-I, an analog in which residues 28-37 of the C region of hIGF-I are replaced by a 4-reside glycine bridge; and [1-27,Gly4,38-62]hIGF-I, an analog with the C region glycine replacement and a D region deletion. The removal of the D region of hIGF-I has little effect on binding to the type 1 and type 2 insulin-like growth factor (IGF) receptors. [1-62]hIGF-I has 2-fold higher affinity for the insulin receptor and 4-fold higher affinity for IGF serum-binding proteins. The replacement of the C region of hIGF-I with a four-glycine span results in a 30-fold loss of affinity for the type 1 IGF receptor. However this analog has near normal affinity for the type 2 IGF receptor, the insulin receptor, and IGF serum-binding proteins. Incorporating the C region glycine replacement and the D region deletion into one analog does not affect binding to either the type 2 receptor or to IGF serum-binding proteins. As predicted from the single deletion analogs [1-27,Gly4,38-62]hIGF-I has reduced affinity for the type 1 IGF receptor (approximately 40-fold) and increased affinity for the insulin receptor (5-fold). These data indicate that determinants in the C region of hIGF-I are involved in maintaining high affinity binding to the type 1 IGF receptor and that neither the C region nor the D region are required for high affinity binding to the type 2 IGF receptor or to IGF serum-binding proteins.     

MOVEMENT MODALITIES AND RESPONSES TO ENVIRONMENTAL CHANGES OF THE MUDFLAT DIATOM CYLINDROTHECA CLOSTERIUM (BACILLARIOPHYCEAE)1

Cylindrotheca closterium (Ehrenberg) Reiman et Lewin is a raphid diatom widely distributed in mudflat assemblages. Video microscopy showed various movement modalities defined as smooth and corkscrew gliding, pirouette, pivot, rock and roll, rollover, and simultaneous pirouette and gliding. Z‐axis projection analysis of images revealed a unique gliding motif with corkscrew motions, which may have important ecological implications for C. closterium movement in muds. The general response to salinity alteration was a decrease in gliding movements with a concomitant increase in other modalities listed above. Short‐term responses to salinity change include dramatic alteration in modalities in hypo‐saline conditions and cessation of motility in extreme hyper‐saline environments. Modality changes were rapid and occurred within 5 s in response to hyper‐saline conditions. Hypo‐ or hyper‐saline conditions resulted in decreased gliding speed in standard media. Five‐ and 15‐day acclimation to salinity changes resulted in a progressive reduction in gliding movement, increased non‐gliding modalities and increased cell aggregation. Aggregation in hypo‐saline conditions was accompanied by a large increase in the polymer extracted by hot bicarbonate‐ and ethylenediamine tetraaceticacid‐ fractions of extracellular polymeric substance (EPS), the polymers of which have been implicated in cell attachment/motility phenomena. The monosaccharide profiles of these fractions were altered in response to hypo‐saline conditions. In general, monosaccharide profiles showed increased diversity upon cessation of motility and aggregation of cultures. The movement responses of C. closterium in response to environmental changes, accompanied by modifications in EPS, may form part of an adaptive strategy to survive in mudflats and could be useful as bioindicators of environmental changes.     

Neoselachian sharks and rays from the British Bathonian (Middle Jurassic)

Extensive sampling of Bathonian sediments from localities across southern and central England has produced over 8000 neoselachian teeth. These comprise diverse faunas, with over 25 species being represented in total, most of them previously undescribed. Seventeen new species and seven new genera are named: Palaeoscyllium tenuidens sp. nov., Praeproscyllium oxoniensis gen. et sp. nov., Eypea leesi gen. et sp. nov., Proheterodontus sylvestris gen. et sp. nov., Paracestracion bellis sp. nov., Palaeobrachaelurus mussetti sp. nov., Heterophorcynus microdon gen. et sp. nov., Dorsetoscyllium terraefullonicum gen. et sp. nov., Ornatoscyllium freemani gen. et sp. nov., Pseudonotidanus semirugosus gen. et sp. nov., Synechodus duffini sp. nov., Protospinax magnus sp. nov., P. bilobatus sp. nov., P. carvalhoi sp. nov., Belemnobatis kermacki sp. nov., B. stahli sp. nov. and Spathobatis delsatei sp. nov. In addition, a new family, the Pseudonotidanidae fam. nov., is defined and the status of Paranotidanus Ward and Thies, 1987, Hybodus levis Woodward, 1889 and Breviacanthus brevis (Phillips, 1871) are discussed. These taxa show strong facies specificity, with different species being restricted to different palaeoenvironments.     

Short-term fumigation of honey bee (Hymenoptera: Apidae) colonies with formic and acetic acids for the control of Varroa destructor (Acari: Varroidae)

Controlling populations of varroa mites is crucial for the survival of the beekeeping industry. Many treatments exist, and all are designed to kill mites on adult bees. Because the majority of mites are found under capped brood, most treatments are designed to deliver active ingredients over an extended period to control mites on adult bees, as developing bees and mites emerge. In this study, a 17-h application of 50% formic acid effectively killed mites in capped worker brood and on adult bees without harming queens or uncapped brood. Neither acetic acid nor a combined treatment of formic and acetic acids applied to the West Virginia formic acid fumigator was as effective as formic acid alone in controlling varroa mites. In addition, none of the treatments tested in late summer had an effect on the late-season prevalence of deformed wing virus. The short-term formic acid treatment killed > 60% of varroa mites in capped worker brood; thus, it is a promising tool for beekeepers, especially when such treatments are necessary during the nectar flow.     

Windows of opportunity: white-tailed deer and the dynamics of northern hardwood forests of the northeastern US

Herbivory, lighting regimes, and site conditions are among the most important determinants of forest regeneration success, but these are affected by a host of other factors such as weather, predation, human exploitation, pathogens, wind and fire. We draw together >50 years of research on the Huntington Wildlife Forest in the central Adirondack Mountains of New York to explore regeneration of northern hardwoods. A series of studies each of which focused on a single factor failed to identify the cause of regeneration failure. However, integration of these studies led to broader understanding of the process of forest stand development and identified at least three interacting factors: lighting regime, competing vegetation and selective browsing by white-tailed deer (Odocoileus virginianus). The diverse 100–200 year-old hardwood stands present today probably reflect regeneration during periods of low deer density (<2.0 deer/km²) and significant forest disturbance. If this hypothesis is correct, forest managers can mimic these “natural windows of opportunity” through manipulation of a few sensitive variables in the system. Further, these manipulations can be conducted on a relatively small geographic scale. Control of deer densities on a scale of 500 ha and understory American beech (Fagus grandifolia) on a scale of <100 ha in conjunction with an even-aged regeneration system consistently resulted in successful establishment of desirable hardwood regeneration.     

Nitrous Oxide Formation in the Colne Estuary, England: the Central Role of Nitrite

Nitrate and nitrite concentrations in the water and nitrous oxide and nitrite fluxes across the sediment-water interface were measured monthly in the River Colne estuary, England, from December 1996 to March 1998. Water column concentrations of N₂O in the Colne were supersaturated with respect to air, indicating that the estuary was a source of N₂O for the atmosphere. At the freshwater end of the estuary, nitrous oxide effluxes from the sediment were closely correlated with the nitrite concentrations in the overlying water and with the nitrite influx into the sediment. Increases in N₂O production from sediments were about 10 times greater with the addition of nitrite than with the addition of nitrate. Rates of denitrification were stimulated to a larger extent by enhanced nitrite than by nitrate concentrations. At 550 μM nitrite or nitrate (the highest concentration used), the rates of denitrification were 600 μmol N · m⁻² · h⁻¹ with nitrite but only 180 μmol N · m⁻² · h⁻¹ with nitrate. The ratios of rates of nitrous oxide production and denitrification (N₂O/N₂ × 100) were significantly higher with the addition of nitrite (7 to 13% of denitrification) than with nitrate (2 to 4% of denitrification). The results suggested that in addition to anaerobic bacteria, which possess the complete denitrification pathway for N₂ formation in the estuarine sediments, there may be two other groups of bacteria: nitrite denitrifiers, which reduce nitrite to N₂ via N₂O, and obligate nitrite-denitrifying bacteria, which reduce nitrite to N₂O as the end product. Consideration of free-energy changes during N₂O formation led to the conclusion that N₂O formation using nitrite as the electron acceptor is favored in the Colne estuary and may be a critical factor regulating the formation of N₂O in high-nutrient-load estuaries.     

Flux through Citrate Synthase Limits the Growth of Ethanologenic Escherichia coli KO11 during Xylose Fermentation

Previous studies have shown that high levels of complex nutrients (Luria broth or 5% corn steep liquor) were necessary for rapid ethanol production by the ethanologenic strain Escherichia coli KO11. Although this strain is prototrophic, cell density and ethanol production remained low in mineral salts media (10% xylose) unless complex nutrients were added. The basis for this nutrient requirement was identified as a regulatory problem created by metabolic engineering of an ethanol pathway. Cells must partition pyruvate between competing needs for biosynthesis and regeneration of NAD⁺. Expression of low-K_m Zymomonas mobilis pdc (pyruvate decarboxylase) in KO11 reduced the flow of pyruvate carbon into native fermentation pathways as desired, but it also restricted the flow of carbon skeletons into the 2-ketoglutarate arm of the tricarboxylic acid pathway (biosynthesis). In mineral salts medium containing 1% corn steep liquor and 10% xylose, the detrimental effect of metabolic engineering was substantially reduced by addition of pyruvate. A similar benefit was also observed when acetaldehyde, 2-ketoglutarate, or glutamate was added. In E. coli, citrate synthase links the cellular abundance of NADH to the supply of 2-ketoglutarate for glutamate biosynthesis. This enzyme is allosterically regulated and inhibited by high NADH concentrations. In addition, citrate synthase catalyzes the first committed step in 2-ketoglutarate synthesis. Oxidation of NADH by added acetaldehyde (or pyruvate) would be expected to increase the activity of E. coli citrate synthase and direct more carbon into 2-ketoglutarate, and this may explain the stimulation of growth. This hypothesis was tested, in part, by cloning the Bacillus subtilis citZ gene encoding an NADH-insensitive citrate synthase. Expression of recombinant citZ in KO11 was accompanied by increases in cell growth and ethanol production, which substantially reduced the need for complex nutrients.