Pathway-specific profiling identifies the NF-kappa B-dependent tumor necrosis factor alpha-regulated genes in epidermal keratinocytes

Identification of tumor necrosis factor alpha (TNF alpha) as the key agent in inflammatory disorders led to new therapies specifically targeting TNF alpha and avoiding many side effects of earlier anti-inflammatory drugs. However, because of the wide spectrum of systems affected by TNF alpha, drugs targeting TNF alpha have a potential risk of delaying wound healing, secondary infections, and cancer. Indeed, increased risks of tuberculosis and carcinogenesis have been reported as side effects after anti-TNF alpha therapy. TNF alpha regulates many processes (e.g. immune response, cell cycle, and apoptosis) through several signal transduction pathways that convey the TNF alpha signals to the nucleus. Hypothesizing that specific TNF alpha-dependent pathways control specific processes and that inhibition of a specific pathway may yield even more precisely targeted therapies, we used oligonucleotide microarrays and parthenolide, an NF-kappa B-specific inhibitor, to identify the NF-kappa B-dependent set of the TNF alpha-regulated genes in human epidermal keratinocytes. Expression of approximately 40% of all TNF alpha-regulated genes depends on NF-kappa B; 17% are regulated early (1-4 h post-treatment), and 23% are regulated late (24-48 h). Cytokines and apoptosis-related and cornification proteins belong to the "early" NF-kappa B-dependent group, and antigen presentation proteins belong to the "late" group, whereas most cell cycle, RNA-processing, and metabolic enzymes are not NF-kappa B-dependent. Therefore, inflammation, immunomodulation, apoptosis, and differentiation are on the NF-kappa B pathway, and cell cycle, metabolism, and RNA processing are not. Most early genes contain consensus NF-kappaB binding sites in their promoter DNA and are, presumably, directly regulated by NF-kappa B, except, curiously, the cornification markers. Using siRNA silencing, we identified cFLIP/CFLAR as an essential NF-kappa B-dependent antiapoptotic gene. The results confirm our hypothesis, suggesting that inhibiting a specific TNF alpha-dependent signaling pathway may inhibit a specific TNF alpha-regulated process, leaving others unaffected. This could lead to more specific anti-inflammatory agents that are both more effective and safer.     

Complementation of an Escherichia coli DnaK defect by Hsc70-DnaK chimeric proteins

Escherichia coli DnaK and rat Hsc70 are members of the highly conserved 70-kDa heat shock protein (Hsp70) family that show strong sequence and structure similarities and comparable functional properties in terms of interactions with peptides and unfolded proteins and cooperation with cochaperones. We show here that, while the DnaK protein is, as expected, able to complement an E. coli dnaK mutant strain for growth at high temperatures and lambda phage propagation, Hsc70 protein is not. However, an Hsc70 in which the peptide-binding domain has been replaced by that of DnaK is able to complement this strain for both phenotypes, suggesting that the peptide-binding domain of DnaK is essential to fulfill the specific functions of this protein necessary for growth at high temperatures and for lambda phage replication. The implications of these findings on the functional specificities of the Hsp70s and the role of protein-protein interactions in the DnaK chaperone system are discussed.     

New records of the rare North American endemic <Emphasis Type="Italic">Chara brittonii</Emphasis> (Characeae), with comments on its distribution

Characeae is a family of freshwater green algal macrophytes found on every continent except Antarctica. Although some species are thought to be cosmopolitan, others appear to be restricted to relatively small geographic areas. Chara brittonii is a North American endemic previously reported from eight scattered populations in Indiana, Michigan, New Jersey, and Ohio. Given that few extant populations were known, basic questions about its distribution, habitat preference, morphology, and phylogenetic placement remained unanswered. We have surveyed every reported locality for C. brittonii except the New Jersey locality, because the collection details are vague, and also surveyed numerous additional waterbodies in Indiana, Michigan, and Wisconsin. We have found extant populations in nine localities in three states. These include one newly reported site in Indiana and five newly reported sites in Wisconsin, the first known populations in that state. Chara brittonii seems to have been extirpated from several historical sites in recent decades. We expanded both the range and the number of known extant populations for C. brittonii and hypothesize that C. brittonii may be more widespread in the Midwest than previously thought. Factors contributing to the actual and apparent rarity of this species are discussed including its preferred habitat and small size. We observed that the number of antheridial scute cells of C. brittonii varied from 4–8 with both triangular and elongate scutes produced on the same thallus, an unusual condition for the family. Phylogenetic analyses using three plastid-encoded markers placed C. brittonii among a paraphyletic grade of C. foliolosa sensu lato strains in the monophyletic subsection Willdenowia. We propose that C. foliolosa, as currently recognized, represents a species complex of several phylogenetically distinct lineages, and conclude that C. brittonii is a structurally and phylogenetically distinct species worthy of conservation.     

Cophylogenetic assessment of New World warblers (Parulidae) and their symbiotic feather mites (Proctophyllodidae)

Host–symbiont relationships are ubiquitous in nature, yet evolutionary and ecological processes that shape these intricate associations are often poorly understood. All orders of birds engage in symbioses with feather mites, which are ectosymbiotic arthropods that spend their entire life on hosts. Due to their permanent obligatory association with hosts, limited dispersal and primarily vertical transmission, we hypothesized that the cospeciation between feather mites and hosts within one avian family (Parulidae) would be perfect (strict cospeciation). We assessed cophylogenetic patterns and tested for congruence between species in two confamiliar feather mite genera (Proctophyllodidae: Proctophyllodes, Amerodectes) found on 13 species of migratory warblers (and one other closely related migratory species) in the eastern United States. Based on COI sequence data, we found three Proctophyllodes lineages and six Amerodectes lineages. Distance‐ and event‐based cophylogenetic analyses suggested different cophylogenetic trajectories of the two mite genera, and although some associations were significant, there was little overall evidence supporting strict cospeciation. Host switching is likely responsible for incongruent phylogenies. In one case, we documented prairie warblers Setophaga discolor harboring two mite species of the same genus. Most interestingly, we found strong evidence that host ecology may influence the likelihood of host switching occurring. For example, we documented relatively distantly related ground‐nesting hosts (ovenbird Seiurus aurocapilla and Kentucky warbler Geothlypis formosa) sharing a single mite species, while other birds are shrub/canopy or cavity nesters. Overall, our results suggest that cospeciation is not the case for feather mites and parulid hosts at this fine phylogenetic scale, and raise the question if cospeciation applies for other symbiotic systems involving hosts that have complex life histories. We also provide preliminary evidence that incorporating host ecological traits into cophylogenetic analyses may be useful for understanding how symbiotic systems have evolved.     

Microbial and plant-derived compounds both contribute to persistent soil organic carbon in temperate soils

Our study tests the emerging paradigm that biochemical recalcitrance does not affect substantially long-term (50 years) SOC persistence. We analyzed the molecular composition of SOC in archived soils originating from four European long-term bare fallow experiments (Askov, Rothamsted, Versailles and Ultuna). The soils had been collected after various periods (up to 53 years) under bare fallow. With increasing duration of bare fallow without new organic inputs, the relative abundance of cutin- and suberin-derived compounds declined substantially, and the abundance of lignin-derived compounds was close to zero. Conversely, the relative abundance of plant-derived long-chain alkanes remained almost constant or increased during the bare fallow period. The relative abundance of N-containing compounds, considered to be abundant in SOC derived from microbial activity, increased consistently illustrating that microbial turnover of SOC continues even when plant inputs are stopped. The persistence of the different families of plant-derived compounds differed markedly over the scale of half a century, which may be ascribed to their contrasting chemical characteristics and recalcitrance, or to differences in their interactions with the soil mineral matrix, and likely some combination since chemical composition drives the degree of mineral association. Using soil from this unique set of long-term bare fallow experiments, we provide direct evidence that multi-decadal scale persistent SOC is enriched in microbe-derived compounds but also includes a substantial fraction of plant-derived compounds.     

Alcohol withdrawal-induced changes in brain biogenic amines in mice: Influence of the genotype

Alterations in striatal and hippocampal dopamine (DA) and serotonin (5HT) activities were investigated in two inbred strains of mice (C57B1 and Balb/c) after 3 withdrawal periods following 5 months chronic ethanol administration. Two groups of animals with different levels of ethanol administration (15% and 30%, v/v) were examined. A striking strain dependency has been noted. Striatal dopaminergic mechanisms of the Balb/c strain are profoundly disturbed in both groups. In contrast no changes were noted for either transmitter activities in C57B1 mice at any withdrawal time studied. Strain dependency has also been noted for hippocampal serotonin neurotransmission, since only Balb/c mice showed a progressive decrease in 5HT levels. These impairments observed in striatum and hippocampus could be involved in motor incoordinations and convulsions often associated with the withdrawal syndrome. The differences in withdrawal effects we noted between the two strains may be linked to the specific chemical neuroanatomy of the strains. Such specificities could be implied in the well known variability of withdrawal induced behavior in man.     

Demonstration of an interferon alpha receptor in human colonic cancer cells in culture

The IFN-alpha 2 receptor present in human cancerous colonic cells in culture has a capacity of 1,800 sites per cell with a kd of 0.5 nM. The molecular mass of the interferon alpha 2 (IFN-alpha 2) receptor complex is 134 kd. The IFN-alpha 2 induces the activity of the 2'5' oligoadenylate (2'5' A) synthetase activity in these cells and partly inhibits their growth.     

Transforming Global Health by Improving the Science of Scale-Up

In its report Global Health 2035, the Commission on Investing in Health proposed that health investments can reduce mortality in nearly all low- and middle-income countries to very low levels, thereby averting 10 million deaths per year from 2035 onward. Many of these gains could be achieved through scale-up of existing technologies and health services. A key instrument to close this gap is policy and implementation research (PIR) that aims to produce generalizable evidence on what works to implement successful interventions at scale. Rigorously designed PIR promotes global learning and local accountability. Much greater national and global investments in PIR capacity will be required to enable the scaling of effective approaches and to prevent the recycling of failed ideas. Sample questions for the PIR research agenda include how to close the gap in the delivery of essential services to the poor, which population interventions for non-communicable diseases are most applicable in different contexts, and how to engage non-state actors in equitable provision of health services in the context of universal health coverage.