Crystal structure of outer membrane protein NMB0315 from Neisseria meningitidis

NMB0315 is an outer membrane protein of Neisseria meningitidis serogroup B (NMB) and a potential candidate for a broad-spectrum vaccine against meningococcal disease. The crystal structure of NMB0315 was solved by single-wavelength anomalous dispersion (SAD) at a resolution of 2.4 Å and revealed to be a lysostaphin-type peptidase of the M23 metallopeptidase family. The overall structure consists of three well-separated domains and has no similarity to any previously published structure. However, only the topology of the carboxyl-terminal domain is highly conserved among members of this family, and this domain is a zinc-dependent catalytic unit. The amino-terminal domain of the structure blocks the substrate binding pocket in the carboxyl-terminal domain, indicating that the wild-type full-length protein is in an inactive conformational state. Our studies improve the understanding of the catalytic mechanism of M23 metallopeptidases.     

Metal concentrations and mycorrhizal status of plants colonizing copper mine tailings: potential for revegetation

A field survey of metal concentrations and mycorrhizal status of plants growing on copper mine tailings was conducted in Anhui Province, China. Available phosphorus and organic matter in the tailings were very low. High concentrations of Pb, Zn, As and Cd as well as Cu were observed on some sites. The dominant plants growing on mine tailings belonged to the families Gramineae and Compositae, and the most widely distributed plant species wereImperata cylindrica, Cynodon dactylon andPaspalum distichum. Coreopsis drummondii also grew well on the arid sites but not on wet sites. Very low or zero arbuscular mycorrhizal (AM) fungal colonization was observed in most of the plants, but extensive mycorrhizal colonization was recorded in the roots ofC. drummondii andC. dactylon. Metal concentrations in plant tissues indicated that /.cylindrica andP. distichum utilized avoidance mechanisms to survive at high metal concentrations. The investigation suggests that remediation and revegetation of heavy metal contaminated sites might be facilitated by selection of tolerant plant species. Isolation of tolerant AM fungi may also be warranted.     

Hydrodynamics and mass transfer coefficient in activated sludge aerated stirred column reactor: experimental analysis and modeling

The aerated stirred reactor (ASR) has been widely used in biochemical and wastewater treatment processes. The information describing how the activated sludge properties and operation conditions affect the hydrodynamics and mass transfer coefficient is missing in the literature. The aim of this study was to investigate the influence of flow regime, superficial gas velocity (U(G)), power consumption unit (P/V(L)), sludge loading, and apparent viscosity (mu(ap)) of activated sludge fluid on the mixing time (t(m)), gas hold-up (epsilon), and volumetric mass transfer coefficient (k(L)a) in an activated sludge aerated stirred column reactor (ASCR). The activated sludge fluid performed a non-Newtonian rheological behavior. The sludge loading significantly affected the fluid hydrodynamics and mass transfer. With an increase in the U(G) and P/V(L), the epsilon and k(L)a increased, and the t(m), decreased. The epsilon, k(L)a, and t(m), were influenced dramatically as the flow regime changed from homogeneous to heterogeneous patterns. The proposed mathematical models predicted the experimental results well under experimental conditions, indicating that the U(G), P/V(L), and mu(ap) had significant impact on the t(m), epsilon, and k(L)a. These models were able to give the t(m), epsilon, and k(L)a values with an error around +/-8%, and always less than +/-10%.     

A truncated form of fibronectin is expressed in fish and mammals.

Fibronectin is a highly conserved and well-characterized extracellular matrix protein that plays a crucial role in many cellular processes. In amphibians, birds and mammals, multiple isoforms of fibronectin are generated by differential RNA splicing at three exons. Previously, we identified a novel form of fibronectin (FN2) in zebrafish that possesses a truncated structure and is generated by a unique RNA splicing pattern. Unlike other fibronectins, FN2 exists as a monomer and localizes on the cell surface. Here we show that this novel fibronectin is evolutionarily conserved, found in multiple species of fish and in mice and humans. The unusual structure, unique cellular distribution and conserved pattern of expression indicate that FN2 may have a novel function in normal cellular processes.     

Mitochondrial ND5 12338T>C variant is associated with maternally inherited hypertrophic cardiomyopathy in a Chinese pedigree

Hypertrophic cardiomyopathy is a primary disorder characterized by asymmetric thickening of the septum and left ventricular wall, which affects 1 in 500 individuals in the general population. Mutations in mitochondrial DNA have been found to be one of the most important causes of hypertrophic cardiomyopathy. Here we report the clinical, genetic and molecular characterization of a Han Chinese family with a likely maternally transmitted hypertrophic cardiomyopathy. Four (2 men/2 women) of 5 matrilineal relatives in this 3-generation family exhibited the variable severity and age at onset of 44 to 79years old. Sequence analysis of the entire mitochondrial DNA in this pedigree identified the known homoplasmic ND5 12338T>C variant. This mitochondrial DNA haplogroup belongs to the Eastern Asian F2a. The 12338T>C variant, highly evolutionarily conserved, resulted in the replacement of the translation initiating methionine with a threonine, shortening the ND5 polypeptide by 2 amino acids. The occurrence of ND5 12338T>C variant exclusively in maternal members of this Chinese family suggested that the 12338T>C variant is associated with maternally inherited hypertrophic cardiomyopathy. Our findings will provide theoretical basis for genetic counseling of maternally inherited hypertrophic cardiomyopathy.     

Functionally orthologous viral and cellular microRNAs studied by a novel dual-fluorescent reporter system

Recent research raised the possibility that some viral microRNAs (miRNAs) may function as orthologs of cellular miRNAs. In the present work, to study the functional orthologous relationships of viral and cellular miRNAs, we first constructed a dual-fluorescent protein reporter vector system for the easy determination of miRNA function. By expressing the miRNAs and the indicator and internal control fluorescent proteins individually from a single vector, this simple reporter system can be used for miRNA functional assays that include visualizing miRNA activity in live cells. Sequence alignments indicated that the simian virus 40 (SV40) encoded miRNA sv40-mir-S1-5p contains a seed region identical to that of the human miRNA hsa-miR423-5p. Using the new reporter system, it was found that sv40-mir-S1-5p and hsa-miR423-5p downregulate the expression of common artificial target mRNAs and some predicted biological targets of hsa-miR423-5p, demonstrating that they are functional orthologs. The human immunodeficiency virus 1 (HIV-1) encoded hiv1-miR-N367 also contains a seed sequence identical to that of the human miRNA hsa-miR192. Functional assays showed that hiv1-miR-N367 and hsa-miR192 could downregulate common artificial and predicted biological targets, suggesting that these miRNAs may also act as functional orthologs. Thus, this study presents a simple and universal system for testing miRNA function and identifies two new pairs of functional orthologs, sv40-mir-S1-5p and hsa-miR423-5p as well as hiv-1-miR-N367 and hsa-miR192. These findings also expand upon our current knowledge of functional homology and imply that a more general phenomenon of orthologous relationships exists between viral and cellular miRNAs.     

Targeted destruction of the orchestration of the pancreatic stroma and tumor cells in pancreatic cancer cases: Molecular basis for therapeutic implications

Pancreatic cancer is one of the most lethal malignancies, with a prominent desmoplastic reaction as its defining hallmark. The past several decades have seen dramatic progress in understanding of pancreatic cancer pathogenesis, including identification of precursor lesions, sequential transformation from normal pancreatic tissue to invasive pancreatic cancer and corresponding signature genetic events, and the biological impact of these events on malignant behavior. However, the currently used therapeutic strategies for epithelial tumor cells, which have exhibited potent antitumor activity in cell culture and animal models, have failed to produce significant effects in the clinic. The desmoplastic stroma surrounding pancreatic cancer cells, which accounts for about 90% of a tumor's mass, clearly is not a passive scaffold for cancer cells but an active contributor to carcinogenesis. Improved understanding of the dynamic interaction between cancer cells and the stroma will be important to designing effective therapeutic strategies for pancreatic cancer. This review focuses on the origin of stromal molecular and cellular components in pancreatic tumors, their biological effects on pancreatic cancer cells, and the orchestration of these two components.