Protein kinase C activation modulates pro- and anti-apoptotic signaling pathways

Activation of protein kinase C (PKC) by TPA in human U937 myeloid leukemia cells is associated with induction of adherence, differentiation, and G0/G1 cell cycle arrest. In this study, we demonstrate that in addition to these differentiating cells about 25% of U937 cells accumulated in the subG1 phase after TPA treatment. This effect proved to be phorbol ester-specific, since other compounds such as retinoic acid or vitamin D3 failed to induce apoptosis in conjunction with differentiation. Only a specific inhibitor of PKC, GF109203X, but not the broad-spectrum kinase inhibitor staurosporine or a tyrosine kinase inhibitor genistein could reverse the induction of apoptosis. Bryostatin-1, another specific PKC activator with distinct biochemical activity failed to induce apoptosis. Moreover, bryostatin-1 completely abolished the induction of apoptosis in U937 cells even if added 8 hours after TPA treatment. Apart from apoptosis induced by various chemotherapeutic drugs, TPA-related cell death is not mediated by an autocrine Fas-FasL loop and could not be prevented by a blocking antibody to the Fas receptor. However, a 75% reduction in the number of apoptotic cells after TPA stimulation was achieved by preincubation with a blocking antibody to the TNFalpha receptor. Tetrapeptide cleavage assays revealed a four-fold increase in the DEVD-cleavage activity in U937 cells compared to a three-fold increase in TUR cells. Immunoblotting demonstrated that TUR cells did not activate significant levels of caspase-3 or -7, whereas in U937 cells a 20-kDa cleavage product corresponding to activated caspase-3 was detectable after 3 d TPA exposure. Moreover, immunoblots revealed a strongly reduced expression of the adaptor molecule APAF-1, which is required for cytochrome c-dependent activation of caspase-9 and subsequently caspase-3. APAF-1 proved to be inducible after PKC activation with phorbol ester in U937, but not in TUR cells. Thus, APAF-1 expression may, at least in part, be regulated by PKC activity and reduced APAF-1 levels are associated with resistance to various inducers of apoptosis. Furthermore, TPA exposure of U937 cells is associated with increased levels of the pro-apoptotic proteins Bak and Bcl-xs, whereas simultaneously a decline in the Bcl-2 expression was noticable.     

The interleukin-1-related cytokine IL-1F8 is expressed in glial cells, but fails to induce IL-1beta signalling responses

The putative new interleukin (IL)-1 family member IL-1F8 (IL-1eta, IL-1H2) has been shown recently to activate mitogen activated protein kinases (MAPKs), extracellular signal-regulated protein kinase (ERK1/2) and c-Jun N-terminal kinase (JNK), and nuclear factor-kappa B (NFkappa B) via a mechanism that requires IL-1Rrp2 expression in cell lines. The aim of this study was to test the hypothesis that IL-1F8 contributes to brain inflammation and injury, by studying its expression and actions in the different cell types of the mouse brain in culture. Messenger RNA for IL-1F8 was detected in neurons and glia (microglial cells, oligodendrocytes progenitor cells and to a lesser extent astrocytes) by RT-PCR. Bacterial lipopolysaccharide (LPS) had no effect on IL-1F8 mRNA levels in mixed glial cultures. Recombinant mouse IL-1beta induced strong activation of ERK1/2, p38, JNK and NFkappa B, and significant release of IL-6 and PGE2, which was blocked by IL-1ra. In contrast, recombinant mouse IL-1F8 did not influence any of these parameters. These results demonstrate that CNS cells may be a source of IL-1F8, but the failure of LPS to modulate IL-1F8 mRNA expression, and of recombinant IL-1F8 to induce any of the classical IL-1 responses, suggest that this cytokine has restricted activities in the brain, or that it may act via alternative pathway(s).     

Phytotoxic chaetoglobosins are produced by the plant pathogen Calonectria morganii (anamorph Cylindrocladium scoparium)

By the application of a bioassay based on cresson seedlings, two phytotoxic compounds were isolated by thin-layer chromatography from the culture fluid of a Calonectria morganii isolate. The structure of both compounds was elucidated by ESI/MS and NMR spectroscopy. According to the Chemical Abstracts database, they were identified as chaetoglobosin A and 19-O-acetylchaetoglobosin A, mycotoxins originally described for Chaetomium globosum.     

Microbial linear plasmids

While plasmids were originally considered to be generally circular until almost two decades ago, linear elements were reported to exist as well. They are now known to be common genetic elements in both, pro- and eukaryotes. Two types of linear plasmids exist, the so-called hairpin plasmids with covalently closed ends and those with proteins bound to their 5′ termini. Hairpin plasmids are common in human-pathogenic Borrelia spirochetes, in which they are instrumental in escape from the immunological response; cryptic hairpin elements are present in mitochondria of the plant pathogenic fungus Rhizoctonia solani. Plasmids with 5′ attached proteins constitute the largest group. In actinomycetous bacteria they are conjugative and usually confer advantageous phenotypes, e.g. formation of antibiotics, degradation of xenobiotics, heavy-metal resistance and growth on hydrogen as the sole energy source. In contrast, the majority of linear plasmids from eukaryotes are cryptic, with only a few exceptions. In some yeasts a killer phenotype may be associated, the most thoroughly investigated elements being those from Kluyveromyces lactis killer strains. In Neurospora spp. and in Podospora anserina, senescence and longevity respectively are correlated with linear plasmids. This review focuses on the biology of linear plasmids, their environmental significance and their use as tools in molecular and applied microbiology. Received: 15 November 1996 / Received revision: 23 December 1996 / Accepted: 30 December 1996     

Effect of training in minimalist footwear on oxygen consumption during walking and running

The present study sought to examine the effect of 5 weeks of training with minimalist footwear on oxygen consumption during walking and running. Thirteen college-aged students (male n = 7, female n = 6, age: 21.7±1.4 years, height: 168.9±8.8 cm, weight: 70.4±15.8 kg, VO₂max: 46.6±6.6 ml·kg⁻¹·min⁻¹) participated in the present investigation. The participants did not have experience with minimalist footwear. Participants underwent metabolic testing during walking (5.6 km·hr⁻¹), light running (7.2 km·hr⁻¹), and moderate running (9.6 km·hr⁻¹). The participants completed this assessment barefoot, in running shoes, and in minimalist footwear in a randomized order. The participants underwent 5 weeks of training with the minimalist footwear. Afterwards, participants repeated the metabolic testing. Data was analyzed via repeated measures ANOVA. The analysis revealed a significant (F_4,32= 7.576, ηp2=0.408, p ≤ 0.001) interaction effect (time × treatment × speed). During the initial assessment, the minimalist footwear condition resulted in greater oxygen consumption at 9.6 km·hr⁻¹ (p ≤ 0.05) compared to the barefoot condition, while the running shoe condition resulted in greater oxygen consumption than both the barefoot and minimalist condition at 7.2 and 9.6 km·hr⁻¹. At post-testing the minimalist footwear was not different at any speed compared to the barefoot condition (p> 0.12). This study suggests that initially minimalist footwear results in greater oxygen consumption than running barefoot, however; with utilization the oxygen consumption becomes similar.     

Microsatellite-aided detection of genetic redundancy improves management of the International Cocoa Genebank, Trinidad

Cacao (Theobroma cacao L.), the tree from which cocoa butter and chocolate is derived, is conserved in field genebanks. The largest of these ex situ collections in the public domain is the International Cocoa Genebank, Trinidad (ICG,T). Reduction of genetic redundancy is essential to improve the accuracy and efficiency of genebank management. This study examined the pedigree and genetic diversity in a subset of 387 accessions in this collection. Sibship reconstruction of this subset revealed 56 full-sib families nested within 189 half-sib families. Sixteen centers of interconnectivity were identified, which suggested a high level of genetic redundancy in the collection. Generally, consistent phylogenetic trees were obtained using different genetic distance measures. However, a principal coordinate analysis of the D _est differentiation measure elicited the best representation of accession group clustering, and we recommend this approach when probing fine-scale genetic differentiation among cacao accessions. The composite genetic diversity of 414 cacao accessions was contained in a core set of 59 unique accessions. These results have significant implications in the conservation of genetic resources of the ICG,T and other cacao genebanks. The approach developed in this study is recommended as a strategy to curators in guiding conservation management practices of cacao and other similar ex situ genebanks.     

Drought-induced mortality of a foundation species (<Emphasis Type="Italic">Juniperus monosperma</Emphasis>) promotes positive afterlife effects in understory vegetation

Climate change-induced droughts have contributed to large-scale die-offs of dominant tree species throughout much of the southwestern United States. These mortality events provide ecologists with the opportunity to determine whether afterlife effects associated with the die-off occur and the potential implications for future ecosystem changes. We studied both the afterlife and interaction effects of condition (dead trees, living trees, and open areas) on understory vegetation in a Juniperus monosperma woodland of northern Arizona 7 years after a major mortality event. Five major findings resulted: (1) there was a positive afterlife effect on understory plants, in which vegetation under dead junipers contained almost double the amount of cover; (2) the competitive effect on understory plants was exemplified by a 1.3 times greater cover and 1.6 additional species in open areas compared to under living junipers; (3) plant community composition significantly differed by aspect and condition; (4) the highly invasive cheatgrass (Bromus tectorum) was 1.5 times greater under dead junipers compared to live junipers; and (5) litter depth and light availability were negatively and positively correlated with plant cover, respectively, but weakly correlated with afterlife effects. Our results indicate that mortality events can promote changes in understory vegetation through afterlife effects. In ecosystems where foundation species suffer high rates of mortality, changes in plant population dynamics and ecosystem function may promote an altered trajectory in community composition with the potential to increase the presence of invasive species. Continued species die-offs associated with climate change-induced drought may contribute to an increased occurrence and legacy of afterlife effects.