Determination of oxygen saturation and hematocrit of flowing human blood using two different spectrally resolving sensors.

The blood parameters oxygen saturation and hematocrit were determined by two different spectral sensors using reflectance spectra from 550 to 900 nm and partial transmission spectra centered at 660 nm. The spectra were analyzed by the method of partial least squares. One sensor consists of a miniature integrating sphere, while the other was fiber-guided. The results show that the geometry of the sensors and different blood flows do not influence the spectral analysis significantly. Independent of the sensor geometry, both hematocrit and oxygen saturation could be determined with an absolute predicted root mean square error of less than 3%. Furthermore, the analysis showed that hematocrit prediction requires eight wavelength regions and oxygen saturation prediction requires four wavelength regions using reflectance spectroscopy. This implies that if the measurement is restricted to reflectance, a spectrometer is indispensable for determining both blood parameters. Hematocrit determination could be improved using reflectance measurements in combination with transmission.     

High-affinity binding of the staphylococcal HarA protein to haptoglobin and hemoglobin involves a domain with an antiparallel eight-stranded beta-barrel fold

Iron scavenging from the host is essential for the growth of pathogenic bacteria. In this study, we further characterized two staphylococcal cell wall proteins previously shown to bind hemoproteins. HarA and IsdB harbor homologous ligand binding domains, the so called NEAT domain (for "near transporter") present in several surface proteins of gram-positive pathogens. Surface plasmon resonance measurements using glutathione S-transferase (GST)-tagged HarAD1, one of the ligand binding domains of HarA, and GST-tagged full-length IsdB proteins confirmed high-affinity binding to hemoglobin and haptoglobin-hemoglobin complexes with equilibrium dissociation constants (K(D)) of 5 to 50 nM. Haptoglobin binding could be detected only with HarA and was in the low micromolar range. In order to determine the fold of this evolutionarily conserved ligand binding domain, the untagged HarAD1 protein was subjected to nuclear magnetic resonance spectroscopy, which revealed an eight-stranded, purely antiparallel beta-barrel with the strand order (-beta1 -beta2 -beta3 -beta6 -beta5 -beta4 -beta7 -beta8), forming two Greek key motifs. Based on structural-homology searches, the topology of the HarAD1 domain resembles that of the immunoglobulin (Ig) fold family, whose members are involved in protein-protein interactions, but with distinct structural features. Therefore, we consider that the HarAD1/NEAT domain fold is a novel variant of the Ig fold that has not yet been observed in other proteins.     

Whole plant response of Pongamia pinnata to drought stress tolerance revealed by morpho-physiological,biochemical and transcriptome analysis

Molecular Biology Reports - Pongamia is considered an important biofuel species worldwide. Drought stress in the early growth stages of Pongamia influences negatively on the germination and...     

Structure of yeast poly(A) polymerase in complex with a peptide from Fip1, an intrinsically disordered protein

In yeast, the mRNA processing enzyme poly(A) polymerase is tethered to the much larger 3'-end processing complex via Fip1, a 36 kDa protein of unknown structure. We report the 2.6 A crystal structure of yeast poly(A) polymerase in complex with a peptide containing residues 80-105 of Fip1. The Fip1 peptide binds to the outside surface of the C-terminal domain of the polymerase. On the basis of this structure, we designed a mutant of the polymerase (V498Y, C485R) that is lethal to yeast. The mutant is unable to bind Fip1 but retains full polymerase activity. Fip1 is found in all eukaryotes and serves to connect poly(A) polymerase to pre-mRNA processing complexes in yeast, plants, and mammals. However, the Fip1 sequence is highly divergent, and residues on both Pap1 and Fip1 at the observed interaction surface are poorly conserved. Herein we demonstrate using analytical ultracentrifugation, circular dichroism, proteolytic studies, and other techniques that, in the absence of Pap1, Fip1 is largely, if not completely, unfolded. We speculate that flexibility may be important for Fip1's function as a molecular scaffold.     

Highly functionalized 7-azaindoles as selective PPAR gamma modulators

A series of highly functionalized 3-aroyl and 3-phenoxy-2-methyl-7-azaindoles have been identified, which are potent selective PPARγ modulators (SPPARγMs). Addition of substituents at the 6-position of the 7-azaindoles improves in vitro potency and pharmacokinetics. 7-Azaindoles have significantly improved off-target profiles compared to the parent indole series.     

Identifying essential genes in Arabidopsis thaliana

Eight years after publication of the Arabidopsis genome sequence and two years before completing the first phase of an international effort to characterize the function of every Arabidopsis gene, plant biologists remain unable to provide a definitive answer to the following basic question: what is the minimal gene set required for normal growth and development? The purpose of this review is to summarize different strategies employed to identify essential genes in Arabidopsis, an important component of the minimal gene set in plants, to present an overview of the datasets and specific genes identified to date, and to discuss the prospects for future saturation of this important class of genes. The long-term goal of this collaborative effort is to facilitate basic research in plant biology and complement ongoing research with other model organisms.     

RBM6 splicing factor promotes homologous recombination repair of double-strand breaks and modulates sensitivity to chemotherapeutic drugs

RNA-binding proteins regulate mRNA processing and translation and are often aberrantly expressed in cancer. The RNA-binding motif protein 6, RBM6, is a known alternative splicing factor that harbors tumor suppressor activity and is frequently mutated in human cancer. Here, we identify RBM6 as a novel regulator of homologous recombination (HR) repair of DNA double-strand breaks (DSBs). Mechanistically, we show that RBM6 regulates alternative splicing-coupled nonstop-decay of a positive HR regulator, Fe65/APBB1. RBM6 knockdown leads to a severe reduction in Fe65 protein levels and consequently impairs HR of DSBs. Accordingly, RBM6-deficient cancer cells are vulnerable to ATM and PARP inhibition and show remarkable sensitivity to cisplatin. Concordantly, cisplatin administration inhibits the growth of breast tumor devoid of RBM6 in mouse xenograft model. Furthermore, we observe that RBM6 protein is significantly lost in metastatic breast tumors compared with primary tumors, thus suggesting RBM6 as a potential therapeutic target of advanced breast cancer. Collectively, our results elucidate the link between the multifaceted roles of RBM6 in regulating alternative splicing and HR of DSBs that may contribute to tumorigenesis, and pave the way for new avenues of therapy for RBM6-deficient tumors.     

Thermoregulation by Diabrotica undecimpunctata howardi and potential effects on overwintering biology

In North Carolina, southern corn rootworm (SCR), Diabrotica undecimpunctata howardi Barber, adults of both sexes (diapausing and nondiapausing) elevate body temperature (Tb) above ambient air temperature (Ta) by basking in direct sunlight on clear fall and winter days when Ta<13°C (Tb-Ta range: 0.8–13.3°C). On cloudy days, SCR adults did not exhibit basking behavior and Tb was more highly correlated with Ta, ground temperature (Tg), and substrate temperature (Ts) than on clear days. Ts was the best predictor of Tb regardless of Ta and the presence or absence of adult basking behavior (fall basking SCR, Ts vs. Tb: R²=0.94, Ta vs. Tb: R²=0.41; fall nonbasking SCR, Ts vs. Tb: R²=0.85, Ta vs. Tb: R²=0.55).These results suggest that SCR thermoregulation at low Ta is ectothermic regulation by microhabitat selection. Elevation of Tb by thermoregulation was often of sufficient magnitude to affect SCR behavior (flight, feeding, mating) and preovipositional development rate. This could explain why gravid SCR females have been found in North Carolina by 6 January when Ta's were too low for development. The relationship of Tb to Ta on cold sunny days would be important to include in any predictive day-degree model of SCR postdiapause development or SCR endoparasite development in the field.

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Résumé En Caroline du Nord, les adultes diapausants ou non des deux sexes de Diabrotica undecimpunctata howardi (SCR) élèvent la température de leurs corps (Tb) au-dessus de la température de l'air ambiant (Ta) en lézardant en plein soleil par jours clairs du printemps et de l'hiver, quand Ta<13°C (Tb-Ta de 0.8 à 13.3°C). Par temps nuageux, les adultes ne lézardent pas, et Tb dépend beaucoup plus de Ta, de la température du sol (Tg), et de la température du substrat (Ts), que par temps clair. Ts fournit la meilleure idée de Ta que les adultes lézardent ou non; quand SCR lézarde au printemps la corrélation de Ts avec Tb est R²=0.94, de Ta avec Tb:R²=0.41; au printemps, quand ils ne lézardent pas; Ts avec TbR²=0.85 et Ta avec Tb:R²=0.55. Ces résultats suggerent que la thermorégulation de SCR aux basses Ta est une régulation ectothermique par sélection du microhabitat. L'augmentation de Tb par thermorégulation est souvent suffisante pour modifier le comportement de SCR (vol, prise de nourriture, accouplement) et un développement préponte. Ceci pourrait expliquer pourquoi des femelles gravides ont été trouvées un 6 janvier en Caroline du Nord quand Ta était trop basse pour permettre le développement. II paraît important d'inclure la relation entre Tb et Ta par temps nuageux froid, dans tout modèle prédictif, basé sur la notion de degrés-jours, sur le développement de SCR après la diapause ou le dévelopment d'endoparasites de SCR dans la nature.