Influence of drainage basin topography and elevation on carbon dioxide and methane supersaturation of stream water

The partial pressures of CO₂ (pCO₂) andCH₄ (pCH₄) in streams are not only governed byinstream processes, but also by transformations occurring in soil andgroundwater ecosystems. As such, stream water pCO₂ andpCH₄ can provide a tool to assess ecosystem respiration andanaerobic metabolism throughout drainage basins. We conducted three surveyssampling the gas content of streams in eastern Tennessee and western NorthCarolina to assess factors regulating ecosystem metabolism in catchmentswith contrasting geomorphologies, elevations and soil organic matterstorage. In our first survey, the influence of drainage basin geomorphologyon ecosystem respiration was examined by sampling streams drainingcatchments underlain by either shale or dolomite. Geomorphology isinfluenced by geology with shale catchments having shallower soils, broader,unconstrained valley floors compared with dolomite catchments.pCO₂ varied little between catchment types but increased froman average of 3340 ppmv in spring to 9927 ppmv in summer or 9.3 and 28 timesatmospheric equilibrium (pCO_2(equilib)), respectively. Incontrast, pCH₄ was over twice as high in streams drainingshale catchments (306 ppmv; pCH_4(equilib) = 116) compared withmore steeply incised dolomite basins (130 ppmv; pCH_4(equilib)= 51). Using the ratio of pCH₄:pCO₂ as an indexof anaerobic metabolism, shale catchments had nearly twice as muchanaerobiosis (pCH₄:pCO₂ = 0.046) than dolomitedrainages (pCH₄:pCO₂ = 0.024). In our secondsurvey, streams were sampled along an elevational gradient (525 to 1700 m)in the Great Smoky Mountains National Park, USA where soil organic matterstorage increases with elevation. pCO₂ did not vary betweenstreams but increased from 5340 ppmv (pCO_2(equilib) = 15) to8565 ppmv (pCO_2(equilib) = 24) from spring to summer,respectively. During spring pCH₄ was low and constant acrossstreams, but during summer increased with elevation ranging from 17 to 2068ppmv (pCH_4(equilib) = 10 to 1216). The contribution ofanaerobiosis to total respiration was constant during spring(pCH₄:pCO₂ = 0.017) but during summer increasedwith elevation from 0.002 at 524 m to 0.289 at 1286 m. In our last survey,we examined how pCO₂ and pCH₄ changed withcatchment size along two rivers (ca. 60 km stretches in both riverscorresponding to increases in basin size from 1.7–477km² and 2.5–275 km²). pCO₂and pCH₄ showed opposite trends, with pCO₂decreasing ca. 50% along the rivers, whereas pCH₄roughly doubled in concentration downstream. These opposing shifts resultedin a nearly five-fold increase of pCH₄:pCO₂along the rivers from a low of 0.012 in headwaters to a high of 0.266 65-kmdownstream. pCO₂ likely declines moving downstream asgroundwater influences on stream chemistry decreases, whereaspCH₄ may increase as the prevalence of anoxia in riversexpands due to finer-grained sediments and reduced hydrologic exchange withoxygenated surface water.     

Long‐term successional forest dynamics: species and community responses to climatic variability

Question: Are trees sensitive to climatic variability, and do tree species differ in their responses to climatic variability? Does sensitivity of forest communities to climatic variability depend on stand composition? Location: Mixed young forest at Walker Branch Watershed near Oak Ridge, East Tennessee, USA. Methods: Using a long‐term dataset (1967–2006), we analyzed temporal forest dynamics at the tree and species level, and community dynamics for forest stands that differed in initial species composition (i.e., chestnut oak, oak–hickory, pine, and yellow poplar stands). Using summer drought and growing season temperature as defined climate drivers, we evaluated relationships between forest dynamics and climate across levels of organization. Results: Over the four‐decade study period, forest communities underwent successional change and substantially increased in biomass. Variation in summer drought and growing season temperature contributed to temporal biomass dynamics for some tree species, but not for others. Stand‐level responses to climatic variability were related to the responses of component species, except in pine stands. Pinus echinata, the dominant species in pine stands, decreased over time due to periodic outbreaks of pine bark beetle (Dendroctonus frontalis). These outbreaks at Walker Branch could not be directly related to climatic conditions. Conclusions: The results indicate that sensitivity of developing forests to climatic variability is stand type‐dependent, and hence is a function of species composition. However, in the long term, direct effects of climatic variability on forest dynamics may be small relative to autogenic successional processes or climate‐related insect outbreaks. Empirical studies testing for interactions between forest succession and climatic variability are needed.     

Genome-resolved viral ecology in a marine oxygen minimum zone

Oxygen minimum zones (OMZs) are critical to marine nitrogen cycling and global climate change. While OMZ microbial communities are relatively well-studied, little is known about their viruses. Here, we assess the viral community ecology of 22 deeply sequenced viral metagenomes along a gradient of oxygenated to anoxic waters (<0.02 μmol/l O₂) in the Eastern Tropical South Pacific (ETSP) OMZ. We identified 46 127 viral populations (≥5 kb), which augments the known viruses from ETSP by 10-fold. Viral communities clustered into six groups that correspond to oceanographic features. Oxygen concentration was the predominant environmental feature driving viral community structure. Alpha and beta diversity of viral communities in the anoxic zone were lower than in surface waters, which parallels the low microbial diversity seen in other studies. ETSP viruses were largely endemic, with the majority of shared viruses (87%) also present in other OMZ samples. We detected 543 putative viral-encoded auxiliary metabolic genes (AMGs), of which some have a distribution that reflects physico-chemical characteristics across depth. Together these findings provide an ecological baseline for viral community structure, drivers and population variability in OMZs that will help future studies assess the role of viruses in these climate-critical environments.     

Analysis of PTEN Complex Assembly and Identification of Heterogeneous Nuclear Ribonucleoprotein C as a Component of the PTEN-associated Complex

PTEN (phosphatase and tensin homolog deleted on chromosome 10) is well characterized for its role in antagonizing the phosphoinositide 3-kinase pathway. Previous studies using size-exclusion chromatography demonstrated PTEN recruitment into high molecular mass complexes and hypothesized that PTEN phosphorylation status and PDZ binding domain may be required for such complex formation. In this study, we set out to test the structural requirements for PTEN complex assembly and identify the component(s) of the PTEN complex(es). Our results demonstrated that the PTEN catalytic function and PDZ binding domain are not absolutely required for its complex formation. On the other hand, PTEN phosphorylation status has a significant impact on its complex assembly. Our results further demonstrate enrichment of the PTEN complex in nuclear lysates, suggesting a mechanism through which PTEN phosphorylation may regulate its complex assembly. These results prompted further characterization of other protein components within the PTEN complex(es). Using size-exclusion chromatography and two-dimensional difference gel electrophoresis followed by mass spectrometry analysis, we identified heterogeneous nuclear ribonucleoprotein C (hnRNP C) as a novel protein recruited to higher molecular mass fractions in the presence of PTEN. Further analysis indicates that endogenous hnRNP C and PTEN interact and co-localize within the nucleus, suggesting a potential role for PTEN, alongside hnRNP C, in RNA regulation.Phosphatase and tensin homolog deleted on chromosome 10 (PTEN)⁴ was cloned in 1997 (1–3) and has been well characterized for its tumor-suppressive role by dephosphorylating phosphatidylinositol 3,4,5-trisphosphate to phosphatidylinositol 4,5-bisphosphate and antagonizing the phosphoinositide 3-kinase pathway (4–7). PTEN also regulates cell migration, cell cycle progression, DNA damage response, and chromosome stability independently of its lipid phosphatase activity through its potential protein phosphatase activity and/or protein-protein interaction (8–11) (for recent reviews, see 12–14).PTEN is composed of an N-terminal catalytic domain and a C-terminal regulatory domain. The catalytic domain contains a conserved signature motif (HCXXGXXR) found in dual-specific protein phosphatases, and mutations within this catalytic domain, including the C124S mutation, are known to abrogate PTEN catalytic activity (4). The C terminus of PTEN contains a PDZ (PDS-95/Disc-large/Zo-1) binding domain, which interacts with PDZ-containing proteins such as MAGI-1b, MAGI-2, MAGI-3, hDLG, hMAST and NHERF (15–19). In addition to the PDZ binding domain, several key serine and threonine phosphorylation sites (Ser³⁸⁰, Thr³⁸², Thr³⁸³, and Ser³⁸⁵) at the PTEN C terminus are reported to play an important role in regulating its stability, localization, and activity (20–26).Recent studies suggest that PTEN may function within higher molecular mass complexes. Through size-exclusion chromatography, Vazquez et al. (27) demonstrated that PTEN can be separated into two populations: a monomeric hyperphosphorylated subpopulation and a higher molecular mass hypophosphorylated subpopulation. It was hypothesized that PTEN in its dephosphorylated form can interact with PDZ-containing proteins such as hDLG and be recruited into a higher molecular mass complex. Although the components within PTEN complex(es) have not been systematically studied and purified, MAGI-2, hDLG (27), NHERF2, PDGFR (19), NEP (28), and MVP (29) have been identified as potential components of the PTEN complex using the same size-exclusion chromatography methodology.In this paper, we aim to (i) investigate the essential elements of PTEN required for its complex formation and (ii) dissect the components of the PTEN-associated complex(es). Our results indicate that PTEN catalytic activity or its PDZ binding domain is not absolutely required for complex assembly. PTEN phosphorylation status on amino acids Ser³⁸⁰, Thr³⁸², Thr³⁸³, and Ser³⁸⁵, on the other hand, has a significant role in complex formation. In addition, we demonstrate that the PTEN complex is enriched in nuclear lysates, which suggests a mechanism through which phosphorylation can regulate complex assembly. Using two-dimensional difference gel electrophoresis (DIGE) analysis and comparing proteins present in higher molecular mass fractions in the presence and absence of PTEN followed by mass spectrometry analysis, we have identified heterogeneous nuclear ribonucleoprotein C (hnRNP C) as a potential component within the PTEN complex. PTEN and hnRNP C are shown here to interact and co-localize in the nucleus. We hypothesize that the PTEN and hnRNP C complex may play a role in RNA regulation.     

Bufadienolides from the southern African Drimia depressa (Hyacinthaceae: Urgineoideae)

Two bufadienolides, 3beta,16beta-dihydroxy-5beta-bufa-20,22-dienolide and 16beta-hydroxy-5beta-bufa-20,22-dienolide-3beta-O-beta-d-galactoside, have been isolated from bulbs of the poisonous South African geophyte Drimia depressa (Hyacinthaceae).     

The radial forearm flap: a biomechanical study of the osteotomized radius

An experimental study was undertaken to determine the effect of an osteotomy on radial strength and to compare two techniques used clinically to perform these osteotomies. Forty preserved human cadaveric radii were randomized into osteotomized (20) and nonosteotomized (20) groups. Osteotomized bones were further randomized into beveled-corner (10) and squared-corner (10) groups. A 9-cm-long, one-third thickness segment of bone was removed, similar to the defect resulting from a radial osteocutaneous transfer. All bones were tested to breaking using a four-point bending apparatus. Osteotomized radii were significantly weakened, with breaking strengths only 24 percent of the control group. Although the beveled osteotomy group appeared stronger than the squared osteotomy group, this finding was not significant with the numbers tested. In view of the weakness of the osteotomized radius, we recommend excising no more than one-third of the radial diameter and postoperative immobilization of the forearm for 8 weeks. A beveled osteotomy prevents overcutting at the corners and allows better visualization of the depth of cut. With these measures, the incidence of fracture may be reduced.     

Phenolic constituents from Drypetes armoracia

The methanol extract of the dried stem bark of Drypetes armoracia Pax & Hoffm. afforded two compounds named drypearmoracein A, (E)-4,5,6,7-tetrahydroxy-2-benzylhept-2-enoic acid and drypearmoracein B, 2,3-dihydroxy-9,10-tetrahydroanthra-1,4-quinone along with five known compounds: friedelan-3 beta-ol, friedelin, friedelane-3,7-dione, drypemolundein B and beta-stigmasterol. Their structures were established on the basis of spectroscopic analysis and chemical evidence.     

Regulated expression of heparin-binding EGF-like growth factor (HB-EGF) in the human endometrium: a potential paracrine role during implantation

Heparin-binding epidermal growth factor (HB-EGF) is a recently identified member of the EGF growth factor family found to be expressed in the uterus of both mouse and human at the time of implantation. In the present study, we investigated the expression patterns of HB-EGF in normal cycling endometrium and compared its expression with the fertility-associated endometrial epithelial biomarkers alpha(v)beta(3) integrin, leukemia inhibitory factor (LIF) and homeobox gene, HOXA-10. RNase protection assay (RPA) using RNA made from endometrium collected from different phases of the menstrual cycle demonstrated increased HB-EGF expression during the mid-secretory phase, a pattern similar to, but slightly preceding the expression of alpha(v)beta(3) integrin and HOXA-10. In vitro studies demonstrated stimulation of HB-EGF expression by estradiol-17beta (E(2)) and progesterone (P(4)) alone or in combination in stromal cells. Combined treatment with E(2) + P(4) was, however, required to stimulate epithelial HB-EGF expression. In vitro experiments demonstrated the ability of HB-EGF to stimulate epithelial expression of the key endometrial proteins including LIF, HOXA-10, and the beta(3) integrin subunit. Each has previously been demonstrated to be an important epithelial biomarker expressed during the implantation window. In addition, conditioned media from endometrial stromal cells treated with E(2) + P(4) + relaxin mimicked the stimulatory effect of HB-EGF on epithelial expression of the beta(3) integrin subunit. The stimulatory effect of the stromal-conditioned medium was blocked by antibodies that neutralize a known receptor for HB-EGF. These data suggest that uterine receptivity may be regulated in part by the stromal-derived HB-EGF.     

Testis fascin (FSCN3): a novel paralog of the actin-bundling protein fascin expressed specifically in the elongate spermatid head

During spermiogenesis, significant morphological changes occur as round spermatids are remodeled into the fusiform shape of mature spermatozoa. These changes are correlated with a reorganization of microfilaments and microtubules in the head and tail regions of elongating spermatids. There is also altered expression of specialized actin- and tubulin-associated proteins. We report the characterization of a novel, spermatid-specific murine paralog of the actin-bundling protein fascin (FSCN1); this paralog is designated testis fascin or FSCN3. Testis fascin is distantly related to fascins but retains its primary sequence organization. cDNA clones of mouse testis fascin predict a 498 amino acid protein of molecular mass 56 kD that shares 29% identity with mouse fascin. Mapping of murine and human FSCN3 genes shows localization to the 7q31.3 chromosome. Northern analysis indicates that FSCN3 expression is highly specific to testis and that in situ hybridization further restricts expression to elongating spermatids. Antibodies raised against recombinant FSCN3 protein identify a band at 56 kD in testis, epididymis, and epididymal spermatozoa, suggesting that testis fascin persists in mature spermatozoa. In accord with the in situ hybridization results, immunofluorescent microscopy localizes testis fascin protein to areas of the anterior spermatid head that match known distributions of F-actin in the dorsal and ventral subacrosomal spaces. It is possible that testis fascin may function in the terminal elongation of the spermatid head and in microfilament rearrangements that accompany fertilization.