A new post-treatment system for anaerobic effluents containing a high Ca2+ content

A novel reactor concept has been developed that combines Ca²⁺ removal and BOD conversion under mesophilic (30 °C ) as well as under thermophilic (55 °C ) conditions. Soluble Ca²⁺, present in many industrial wastewaters, precipitates as a result of forced aeration in the aerobic bioreactor. The CaCO₃ precipitates, in turn, act as a carrier material for biofilm growth. This resulted in dense rapidly settling sludge granules covered with a biofilm on the surface. These sludge characteristics facilitate the application of hydraulic retention times of 1 h while maintaining a high biological activity in the reactor.     

Secondary structure as primary determinant of the efficiency of ribosomal binding sites in Escherichia coli.

Using a previously described vector (pKL203) we fused several heterologous ribosomal binding sites (RBSs) to the lacZ gene of E. coli and then studied the variation in expression of the fusions. The RBSs originated from bacteriophage Q beta and MS2 genes and the E. coli genes for elongation factor EF-Tu A and B and ribosomal protein L11 (rplK). The synthesis of the lacZ fusion proteins was measured by an immuno precipitation method and found to vary at least 100-fold. Lac-specific mRNA synthesis follows the variation in protein production. It appears that there is a correlation between the efficiency of an RBS to function in the expression of the fused gene and the lack of secondary structure, involving the Shine and Dalgarno nucleotides (SDnts) and/or the initiation codon. This efficiency is context dependent. The sequence of the SD nts and the length and sequence of the spacer region up to the initiation codon alone are not able to explain our results. Deletion mutations, created in the phage Q beta replicase RBS, reveal a complex pattern of control of expression, probably involving the use of a "false" initiation site.     

Mesophilic and thermophilic activated sludge post treatment of anaerobic effluent. Sludge and wastewater characterisation using batch experiments

Anaerobic pretreated paper process water was characterized interms of readily biodegradable, slowly biodegradable, very slowly biodegradable and inert wastewaterfractions under mesophilic and thermophilic conditions. The anaerobic pretreated paper process water containeda relatively high amount of slowly biodegradable components and few easily biodegradable componentsas indicated by the ratio of short term BOD over the BOD5. Wastewater readily biodegradable COD, determinedas short term BOD, was almost similar when measured under both temperature conditions. Fractions ofslowly biodegradable COD and inert COD of the same wastewater were found to depend on the type of biomassinvolved in the test. Thermophilic aerobic biomass was not able to degrade the wastewater to the sameextent as the mesophilic biomass resulting in higher apparent inert COD levels. Furthermore, wastewater colloidalCOD did not flocculate under thermophilic conditions and was thus not removed from the liquid phase.     

Rapid identification of lettuce seed germination mutants by bulked segregant analysis and whole genome sequencing

Lettuce (Lactuca sativa) seeds exhibit thermoinhibition, or failure to complete germination when imbibed at warm temperatures. Chemical mutagenesis was employed to develop lettuce lines that exhibit germination thermotolerance. Two independent thermotolerant lettuce seed mutant lines, TG01 and TG10, were generated through ethyl methanesulfonate mutagenesis. Genetic and physiological analyses indicated that these two mutations were allelic and recessive. To identify the causal gene(s), we applied bulked segregant analysis by whole genome sequencing. For each mutant, bulked DNA samples of segregating thermotolerant (mutant) seeds were sequenced and analyzed for homozygous single‐nucleotide polymorphisms. Two independent candidate mutations were identified at different physical positions in the zeaxanthin epoxidase gene (ABSCISIC ACID DEFICIENT 1/ZEAXANTHIN EPOXIDASE, or ABA1/ZEP) in TG01 and TG10. The mutation in TG01 caused an amino acid replacement, whereas the mutation in TG10 resulted in alternative mRNA splicing. Endogenous abscisic acid contents were reduced in both mutants, and expression of the ABA1 gene from wild‐type lettuce under its own promoter fully complemented the TG01 mutant. Conventional genetic mapping confirmed that the causal mutations were located near the ZEP/ABA1 gene, but the bulked segregant whole genome sequencing approach more efficiently identified the specific gene responsible for the phenotype.     

Assessment of effluent turbidity in mesophilic and thermophilic activated sludge reactors - origin of effluent colloidal material

Two lab-scale plug flow activated sludge reactors were run in parallel for 4 months at 30 and 55 degrees C. Research focussed on: (1) COD (chemical oxygen demand) removal, (2) effluent turbidity at both temperatures, (3) the origin of effluent colloidal material and (4) the possible role of protozoa on turbidity levels. Total COD removal percentages over the whole experimental period were 66+/-7% at 30 degrees C and 53+/-11% at 55 degrees C. Differences in total COD removal between both systems were due to less removal of soluble and colloidal COD at 55 degrees C compared to the reference system. Thermophilic effluent turbidity was caused by a combination of influent colloidal particles that were not effectively retained in the sludge flocs, and erosion of the thermophilic activated sludge itself, as shown by denaturing gradient gel electrophoresis (DGGE) profiles. DGGE analysis of PCR-amplified 16S rDNA fragments from mesophilic and thermophilic sludge differed, indicating that different microbial communities were present in the two reactor systems. The effects of protozoal grazing on the effluent turbidity of both reactors was negligible and thus could not account for the large turbidity differences observed.     

An open-channel blocker interacts with adjacent turns of alpha-helices in the nicotinic acetylcholine receptor.

The binding site for an open-channel blocker, QX-222, at mouse muscle nicotinic acetylcholine receptors was probed using site-directed mutagenesis, oocyte expression, and electrophysiological analysis. The proposed cytoplasmic end of the M2 transmembrane helix is termed position 1'. At position 10' (alpha S252, beta T263, gamma A261, delta A266), Ala residues yield stronger and longer binding of QX-222 than Ser or Thr residues. These effects are opposite and roughly equal (30%-50% per mutation) to previously reported effects at position 6'. The polar end of an anesthetic molecule seems to bind to the position 6' OH groups, which provide a water-like region; the nonpolar moiety is near position 10' and binds more strongly in a nonpolar environment. Interactions with adjacent OH-rich turns of an amphiphilic helix may explain the widespread blocking effects of local anesthetics at the conduction pore of ion channels.