An analog of ACTH/MSH (4–9), ORG-2766, reduces cerebral uptake of morphine

The uptake of morphine was significantly reduced in most regions of the brains of conscious, unrestrained rats within 10 minutes after treatment with an analog of ACTH/MSH (4–9), ORG-2766. The effect was most obvious in regions with significant densities of enkephalin receptors, namely basal ganglia, hippocampus and cortex. The results explain, in part, how some fragments and analogs of ACTH/MSH may antagonize behavioral actions of morphine, even though some of these peptides lack significant opiate receptor binding properties. We believe that this effect of ORG-2766 is related to an action on the permeability characteristics of the brain microvasculature. The underlying mechanism is unknown.     

The nature of regulation of hexose transport in cultured mammalian fibroblasts: Aerobic “repressive” control by d-glucosamine

Restrictive control (“repression”) of 3-O-methylglucose transport (or of galactose uptake) in confluent NIL hamster fibroblast cultures was found to be highly pronounced after preconditioning the cultures in medium containing d-glucosamine. The “repression” exerted by glucosamine developed slowly over several hours. The transport “repression” was counteracted by anaerobiosis, by 2,4-dinitrophenol (H. M. Kalckar, C. W. Christopher, and D. Ullrey, 1979, Proc. Nat. Acad. Sci. USA76, 6453–6455), and by fluoride as well as by malonate. In “de-repressed” cultures, i.e., in the absence of glucosamine in the medium or by using fructose during preconditioning, malonate did not affect regulation of the hexose transport system. In culture medium deprived of l-glutamine and serum, repressive control of the transport system by glucose as well as by glucosamine was greatly aggravated. However, the simultaneous addition of malonate abolished the severe “repression” by either of the hexoses. In all cases, preconditioning with fructose permitted high (“de-repressed”) transport activity. Unlike glucose, galactose, or glucosamine, fructose was not found to compete in the transport assay. The metabolic inhibitors which prevent the aerobic curtailment of the hexose transport system are all more or less directly interfering with the flow of metabolites through the tricarboxylate cycle, which may therefore play an important role in the “repressive” control of transport.     

Clusters of repeated sequences of chicken DNA are extensively methylated but contain specific undermethylated regions

In the chicken genome there are middle repetitive DNA sequences with a clustered organization. Each cluster is composed of members of different families of repeated DNA sequences and usually contains only one member of each family. Many clusters have the same assortment of repeated sequences but they are in scrambled order from cluster to cluster. These clusters usually exceed 20 × 10³ bases in length and comprise at least 10% of the repeated DNA of the chicken. The repeated sequences that are cluster components are extensively methylated. Methylation was detected by comparing HpaII and MspI digests of total DNA, where the occurrence of the sequence C-m⁵C-G-G is indicated when HpaII (cleaves C-C-G-G) fragments are larger than those generated by MspI (cleaves C-m⁵C-G-G or C-C-G-G). In hybridization experiments with Southern (1975) blots of total DNA digested with either HpaII or MspI, the cloned probes representing clustered repeated sequences showed a dramatic difference in the lengths of restriction fragments detected in the two digests. Many of the sequences that comprise these clusters are methylated in most of their genomic occurrences. There are patterns of methylation that are reproduced faithfully from copy to copy. The overall distribution of methylation within clusters seems to be regional, with long methylated DNA segments interrupted by specific undermethylated regions.     

The influence of the social environment on sexual maturation of female deermice (Peromyscus maniculatus bairdi)

Population Ecology - Female deermice housed from weaning with groups of five females, five males or five males plus five females had significantly smaller uteri at 35–38 days of age compared...     

Visualization of the 10-nm filament vimentin rings in vascular endothelial cells in situ : Close resemblance to vimentin cytoskeletons found in monolayers in vitro

Surface staining of the intact vascular endothelial cell layer lining the lumen of guinea pig thoracic aorta with antibodies to vimentin revealed that at least 70% of the cells contained intact perinuclear rings of 10-nm filaments. This correlated with the observations made on these cells in culture: 60–80% of the endothelial cells at confluence have complete perinuclear rings. By one- and two-dimensional gel electrophoresis and immunoprecipitation we confirmed that vimentin [17, 18] is the major constituent polypeptide of the 10-nm filaments in guinea pig endothelial cells. These results indicate that the vimentin [17] 10-nm filament cytoskeleton found in guinea pig endothelial cells in vitro is similar to the cytoskeleton found in situ.     

Timing and size of blooms of the ctenophore Mnemiopsis leidyi in relation to temperature in Narragansett Bay, RI

The ctenophore Mnemiopsis leidyi is at the northern extreme of its geographic range in Narragansett Bay, an estuary on the northeast coast of the United States. Blooms have typically been observed in late summer and fall according to records from 1950 to 1979. We document an expansion of the seasonal range of this important planktonic predator to include springtime blooms during the 1980s and 1990s. This shift to an earlier seasonal maximum is associated with increasing water temperature in Narragansett Bay. Temperatures in spring have risen, on average, 2 °C from 1950 to 1999 with warm years being associated with the positive phase of the North Atlantic Oscillation. During 1999, M. leidyi appeared earlier in spring and was more abundant than during any previous year for which records are available. Changes in the seasonal pattern and abundance of this predator are likely to have important effects on planktonic ecosystem dynamics of Narragansett Bay. These include reduction of zooplankton abundance in spring followed by increases in size and frequency of summer phytoplankton blooms. Earlier blooms of M. leidyi may also reduce survival of eggs and larvae of fish because, as in 1999, they coincide with the period of peak spawning.     

The Dehalococcoides Population in Sediment-Free Mixed Cultures Metabolically Dechlorinates the Commercial Polychlorinated Biphenyl Mixture Aroclor 1260

Microbial reductive dechlorination of commercial polychlorinated biphenyl (PCB) mixtures (e.g., Aroclors) in aquatic sediments is crucial to achieve detoxification. Despite extensive efforts over nearly two decades, the microorganisms responsible for Aroclor dechlorination remained elusive. Here we demonstrate that anaerobic bacteria of the Dehalococcoides group derived from sediment of the Housatonic River (Lenox, MA) simultaneously dechlorinate 64 PCB congeners carrying four to nine chlorines in Aroclor 1260 in the sediment-free JN cultures. Quantitative real-time PCR showed that the Dehalococcoides cell titer in JN cultures amended with acetate and hydrogen increased from 7.07 × 10⁶ ± 0.42 × 10⁶ to 1.67 × 10⁸ ± 0.04 × 10⁸ cells/ml, concomitant with a 64.2% decrease of the PCBs with six or more chlorines in Aroclor 1260. No Dehalococcoides growth occurred in parallel cultures without PCBs. Aroclor 1260 dechlorination supported the growth of 9.25 × 10⁸ ± 0.04 × 10⁸ Dehalococcoides cells per μmol of chlorine removed. 16S rRNA gene-targeted PCR analysis of known dechlorinators (i.e., Desulfitobacterium, Dehalobacter, Desulfuromonas, Sulfurospirillum, Anaeromyxobacter, Geobacter, and o-17/DF-1-type Chloroflexi organisms) ruled out any involvement of these bacterial groups in the dechlorination. Our results suggest that the Dehalococcoides population present in the JN cultures also catalyzes in situ dechlorination of Aroclor 1260 in the Housatonic River. The identification of Dehalococcoides organisms as catalysts of extensive Aroclor 1260 dechlorination and our ability to propagate the JN cultures under defined conditions offer opportunities to study the organisms' ecophysiology, elucidate nutritional requirements, identify reductive dehalogenase genes involved in PCB dechlorination, and design molecular tools required for bioremediation applications.     

Inter-specific differences in photosynthetic carbon uptake, photosynthate partitioning and extracellular organic carbon release by deep-water characean algae

1. The effect of light intensity on photosynthesis and the fate of newly fixed organic carbon was compared for three characean algae collected at the same depth (10 m) but differing in their depth distributions. For each species we determined photosynthesis–irradiance (P–E) responses, the partitioning of newly fixed carbon into four intracellular pools (low molecular‐weight compounds, polysaccharides, lipids and proteins) and the extracellular organic carbon (EOC) release at a range of photon flux densities (PFD) 0–60 μmol m^–2 s^–1. 2. The P–E responses differed between the three species, with the light compensation point (E_c) and dark respiration rate highest in the shallowest species (Chara fibrosa), intermediate in the mid‐range species (C. globularis) and lowest in the deepest species (C. corallina). Photosynthetic efficiency (α) and photosynthesis: respiration ratios were lowest in C. fibrosa and highest in C. corallina. 3. In all three species, the low molecular weight pool was the principal photosynthetic product (>60% of fixed C) at 3 μmol m^–2 s^–1 PFD, but its proportional contribution decreased rapidly with increasing irradiance. Polysaccharide rose to become the major product (>35% of fixed C) at saturating PFD (35 μmol m^–2 s^–1). 4. Protein synthesis was saturated at 5 μmol m^–2 s^–1 in all species and was consistently a lower proportion of the fixed carbon in C. corallina than the other species. The fraction incorporated in the lipid pool increased slightly with irradiance but was always less than 10% of fixed C, while the proportion lost as EOC was unaffected by light, being significantly higher in C. fibrosa than the other species. 5. A kinetic experiment with C. fibrosa at 35 μmol m^–2 s^–1 PFD revealed a continued increase in net polysaccharide, protein and lipid synthesis during a 22.5‐h light period, whereas the net size of the low molecular weight pool remained constant. In a subsequent dark period, protein and lipid synthesis continued at the expense of the polysaccharide and low‐molecular‐weight pools. The EOC release rose to a constant low release in the light, then peaked slightly immediately after the dark–light transition before returning to the same rate as in the light. Extrapolating these data over 24 h suggests that the proportion of fixed carbon lost as EOC may be as high as 10% in this species. 6. The interspecific differences in carbon acquisition between the three species reflected their depth distributions, with the deeper species having more efficient photosynthetic metabolism, lower P:R ratios and less EOC release, although no apparent differences in internal partitioning of photosynthate.     

Reductive Dehalogenation of Brominated Phenolic Compounds by Microorganisms Associated with the Marine Sponge Aplysina aerophoba

Marine sponges are natural sources of brominated organic compounds, including bromoindoles, bromophenols, and bromopyrroles, that may comprise up to 12% of the sponge dry weight. Aplysina aerophoba sponges harbor large numbers of bacteria that can amount to 40% of the biomass of the animal. We postulated that there might be mechanisms for microbially mediated degradation of these halogenated chemicals within the sponges. The capability of anaerobic microorganisms associated with the marine sponge to transform haloaromatic compounds was tested under different electron-accepting conditions (i.e., denitrifying, sulfidogenic, and methanogenic). We observed dehalogenation activity of sponge-associated microorganisms with various haloaromatics. 2-Bromo-, 3-bromo-, 4-bromo-, 2,6-dibromo-, and 2,4,6-tribromophenol, and 3,5-dibromo-4-hydroxybenzoate were reductively debrominated under methanogenic and sulfidogenic conditions with no activity observed in the presence of nitrate. Monochlorinated phenols were not transformed over a period of 1 year. Debromination of 2,4,6-tribromophenol, and 2,6-dibromophenol to 2-bromophenol was more rapid than the debromination of the monobrominated phenols. Ampicillin and chloramphenicol inhibited activity, suggesting that dehalogenation was mediated by bacteria. Characterization of the debrominating methanogenic consortia by using terminal restriction fragment length polymorphism (TRFLP) and denaturing gradient gel electrophoresis analysis indicated that different 16S ribosomal DNA (rDNA) phylotypes were enriched on the different halogenated substrates. Sponge-associated microorganisms enriched on organobromine compounds had distinct 16S rDNA TRFLP patterns and were most closely related to the δ subgroup of the proteobacteria. The presence of homologous reductive dehalogenase gene motifs in the sponge-associated microorganisms suggested that reductive dehalogenation might be coupled to dehalorespiration.     

Natural Transformation-Mediated Transfer of Erythromycin Resistance in Campylobacter coli Strains from Turkeys and Swine

Erythromycin resistance in Campylobacter coli from meat animals is frequently encountered and could represent a substantial barrier to antibiotic treatment of human infections. Erythromycin resistance in this organism has been associated with a point mutation (A2075G) in the 23S rRNA gene. However, the mechanisms responsible for possible dissemination of erythromycin resistance in C. coli remain poorly understood. In this study, we investigated transformation-mediated acquisition of erythromycin resistance by genotypically diverse C. coli strains from turkeys and swine, with total genomic DNA from erythromycin-resistant C. coli of either turkey or swine origin used as a donor. Overall, transformation to erythromycin resistance was significantly more frequent in C. coli strains from turkeys than in swine-derived strains (P < 0.01). The frequency of transformation to erythromycin resistance was 10⁻⁵ to 10⁻⁶ for turkey-derived strains but 10⁻⁷ or less for C. coli from swine. Transformants harbored the point mutation A2075G in the 23S rRNA gene, as did the erythromycin-resistant strains used as DNA donors. Erythromycin resistance was stable in transformants following serial transfers in the absence of the antibiotic, and most transformants had high MICs (>256 μg/ml), as did the C. coli donor strains. In contrast to the results obtained with transformation, spontaneous mutants had relatively low erythromycin MICs (32 to 64 μg/ml) and lacked the A2075G mutation in the 23S rRNA gene. These findings suggest that natural transformation has the potential to contribute to the dissemination of high-level resistance to erythromycin among C. coli strains colonizing meat animals.