An ITS-based phylogenetic framework for the genus Vorticella: finding the molecular and morphological gaps in a taxonomically difficult group

Vorticella includes more than 100 currently recognized species and represents one of the most taxonomically challenging genera of ciliates. Molecular phylogenetic analysis of Vorticella has been performed so far with only sequences coding for small subunit ribosomal RNA (SSU rRNA); only a few of its species have been investigated using other genetic markers owing to a lack of similar sequences for comparison. Consequently, phylogenetic relationships within the genus remain unclear, and molecular discrimination between morphospecies is often difficult because most regions of the SSU rRNA gene are too highly conserved to be helpful. In this paper, we move molecular systematics for this group of ciliates to the infrageneric level by sequencing additional molecular markers—fast-evolving internal transcribed spacer (ITS) regions—in a broad sample of 66 individual samples of 28 morphospecies of Vorticella collected from Asia, North America and Europe. Our phylogenies all featured two strongly supported, highly divergent, paraphyletic clades (I, II) comprising the morphologically defined genus Vorticella. Three major lineages made up clade I, with a relatively well-resolved branching order in each one. The marked divergence of clade II from clade I confirms that the former should be recognized as a separate taxonomic unit as indicated by SSU rRNA phylogenies. We made the first attempt to elucidate relationships between species in clade II using both morphological and multi-gene approaches, and our data supported a close relationship between some morphospecies of Vorticella and Opisthonecta, indicating that relationships between species in the clade are far more complex than would be expected from their morphology. Different patterns of helix III of ITS2 secondary structure were clearly specific to clades and subclades of Vorticella and, therefore, may prove useful for resolving phylogenetic relationships in other groups of ciliates.     

Protein molecular dynamics with electrostatic force entirely determined by a single Poisson-Boltzmann calculation

The electrostatic force including the intramolecular Coulombic interactions and the electrostatic contribution of solvation effect were entirely calculated by using the finite difference Poisson-Boltzmann method (FDPB), which was incorporated into the GROMOS96 force field to complete a new finite difference stochastic dynamics procedure (FDSD). Simulations were performed on an insulin dimer. Different relative dielectric constants were successively assigned to the protein interior; a value of 17 was selected as optimal for our system. The simulation data were analyzed and compared with those obtained from 500-ps molecular dynamics (MD) simulation with explicit water and a 500-ps conventional stochastic dynamics (SD) simulation without the mean solvent force. The results indicate that the FDSD method with GROMOS96 force field is suitable to study the dynamics and structure of proteins in solution if used with the optimal protein dielectric constant.     

An eukaryotic translation initiation factor,AteIF5A‐2, affects cadmium accumulation and sensitivity in Arabidopsis

Cadmium (Cd) is one of the most toxic elements and can be accumulated in plants easily; meanwhile, eIF5A is a highly conserved protein in all eukaryotic organisms. The present work tried to investigate whether eIF5A is involved in Cd accumulation and sensitivity in Arabidopsis (Arabidopsis thaliana L.) by comparing the wild‐type Columbia‐0 (Col‐0) with a knockdown mutant of AteIF5A‐2, fbr12‐3 under Cd stress conditions. The results showed that the mutant fbr12‐3 accumulated more Cd in roots and shoots and had significantly lower chlorophyll content, shorter root length, and smaller biomass, suggesting that downregulation of AteIF5A‐2 makes the mutant more Cd sensitive. Real‐time polymerase chain reaction revealed that the expressions of metal transporters involved in Cd uptake and translocation including IRT1, ZIP1, AtNramp3, and AtHMA4 were significantly increased but the expressions of PCS1 and PCS2 related to Cd detoxification were decreased notably in fbr12‐3 compared with Col‐0. As a result, an increase in MDA and H₂O₂ content but decrease in root trolox, glutathione and proline content under Cd stress was observed, indicating that a severer oxidative stress occurs in the mutant. All these results demonstrated for the first time that AteIF5A influences Cd sensitivity by affecting Cd uptake, accumulation, and detoxification in Arabidopsis.     

Optimal dispatching of multipriority jobs to two heterogeneous workstations

The general problem scenario of this paper is the following: Jobs of various priorities, stationed in a common storage area, are waiting to be dispatched to two non-identical workstations. Any of the waiting jobs can be accessed from the storage at any given time. Each job can be processed on either of the workstations, but once a job has been assigned it may not be preempted. By job priority it is meant that a higher priority job has disptach preference over a lower priority job. The processing time of a job on a given workstation is assumed to be random, the distribution being dependent on the job type and the configuration of the workstation. Specifically, the first problem studied considers only two classes of jobs: (1) “hot” jobs, whose processing is to be expedited and thus have the higher dispatch priority, and (2) “routine” jobs which may be assigned to an available workstation only if the workstation has been rejected by all “hot” jobs. The processing times are assumed to be exponentially distributed with means depending on the job class and workstation. We assume that, on the average, one workstation is faster than the other with regard to processing any job. The dispatching objective for each job class is to minimize its expected flowtime. It is shown that threshold dispatching policies are optimal for this problem. That is, the faster processor should be utilized whenever possible, and for each class there exists an explicit threshold such that when the number of jobs of that class in the buffer exceeds this threshold then a job of that class is dispatched to the slower processor, otherwise these jobs wait for the faster processor to become available. For the higher priority jobs, this threshold is shown to be a function only of the various processing rates of the two workstations. For the lower priority jobs, the threshold also depends on the number of higher priority jobs in the buffer. The results is extended to a system with n priority classes. Again, it is shown that when the processing times are exponentially distributed with different rates and the dispatching objective for each class is to minimize its expected flowtime, the optimal dispatching policies are of threshold type. Explicit thresholds are easily derived.     

Cloning and expression of a new rat procarboxypeptidase B gene in Escherichia coli and purification of recombination carboxypeptidase B

A new coding sequence of the procarboxypeptidase B gene was obtained from SD rat fresh pancreas by RT-PCR and highly expressed in Escherichia coli in inclusion bodies. The folded procarboxypeptidase B was subjected to trypsin enzymatic cleavage to produce active carboxypeptidase B, subsequently, carboxypeptidase B was effectively purified with anion exchange chromatography DEAE-FF and hydrophobic interaction chromatography Octyl FF, as a result, 40 mg carboxypeptidase B per litre cell culture with specific activity 7.42 u/mg was achieved. Further research showed that the obtained recombinant carboxypeptidase B could substitute carboxypeptidase B isolated from pancreas.     

Concepts and analysis of mass extinction with the late Ordovician event as an example

Twelve groups of fossils, including graptolites, brachiopods, nautiloids, trilobites, corals, crinoids, bryozoans, conodonts, ostracods, gastropods, chitinozoans, and acritarchs expired in different but substantial magnitude and global extent during the late Caradoc to latest Ashgill. It indicates a multiple‐episodic mass extinction containing the possible Prologue (late Caradoc), Climax episode (Rawtheyan) and Epilogue (late Hirnantian). The main causes of this mass extinction are recognized as a global sea‐level lowering in the climax and remarkable rapid rise at the final, and global cooling. The Chinese data, especially from the South China Paleoplate, are evaluated first. They are significant for explaining this global bioevent.     

Cs phytoremediation by Sorghum bicolor cultivated in soil and in hydroponic system

Cs accumulation characteristics by Sorghum bicolor were investigated in hydroponic system (Cs level at 50–1000 μmol/L) and in soil (Cs-spiked concentration was 100 and 400 mg/kg soil). Two varieties of S. bicolor Cowly and Nengsi 2# grown on pot soil during the entire growth period (100 days) did not show significant differences on the height, dry weight (DW), and Cs accumulation. S. bicolor showed the potential phytoextraction ability for Cs-contaminated soil with the bioaccumulation factor (BCF) and the translocation factor (TF) values usually higher than 1 in soil system and in hydroponic system. The aerial parts of S. bicolor contributed to 86–92% of the total removed amounts of Cs from soil. Cs level in solution at 100 μmol/L gave the highest BCF and TF values of S. bicolor. Cs at low level tended to transfer to the aerial parts, whereas Cs at high level decreased the transfer ratio from root to shoot. In soil, the plant grew well when Cs spiked level was 100 mg/kg soil, but was inhibited by Cs at 400 mg/kg soil with Cs content in sorghum reaching 1147 mg/kg (roots), 2473 mg/kg (stems), and 2939 mg/kg (leaves). In hydroponic system, average Cs level in sorghum reached 5270 mg/kg (roots) and 4513 mg/kg (aerial parts), without significant damages to its biomass at 30 days after starting Cs treatment. Cs accumulation in sorghum tissues was positively correlated with the metal concentration in medium.     

Alterations in microRNA expression linked to microcystin-LR-induced tumorigenicity in human WRL-68 Cells

Microcystin-LR (MC-LR) is a cyclic heptapeptide that acts as a potent hepatotoxin and carcinogen. However, the mechanism of its carcinogenic action remains undetermined. In this study, MC-LR was used to induce the malignant transformation of the WRL-68 cell line. Alterations in microRNA (miRNA) expression in the transformed cell were analyzed to determine the role of miRNAs in MC-LR-induced carcinogenesis. Cultured WRL-68 cells (labeled 25MC10) were continuously exposed to a low concentration (10 μg/L) of MC-LR for 25 passages. Compared with the mock-treated parental cells, the induced 25MC10 cells exhibited a higher growth rate, resistance to serum-induced terminal differentiation, and tumorigenicity in a nude mouse xenograft test. A pilot miRNA expression array analysis was conducted on the 25MC10 cells, followed by validation of select miRNAs by RT-PCR. We found that the onco-miRNAs miR-21 and miR-221 displayed upregulated expression while the liver-specific miR-122 was downregulated. These results suggest that chronic MC-LR exposure alters the miRNA expression profile of WRL-68 cells and causes phenotypic transformation. We propose that characteristic miRNA alterations could be used as molecular targets for the development of environmental water monitoring methods.