Differential expression of a senescence-enhanced metallothionein gene in Arabidopsis in response to isolates of Peronospora parasitica and Pseudomonas syringae

The metallothionein gene, LSC54 , shows increased expression during leaf senescence in Brassica napus and Arabidopsis thaliana . A number of abiotic and biotic stresses have been shown to induce senescence-like symptoms in plants and, to investigate this further, the promoter of the LSC54 gene was cloned and fused to the GUS gene and transformed into Arabidopsis . The promoter was highly induced during leaf senescence and also in response to wounding; histochemical analysis indicated that this induction was localised to a few cells close to the wound site. The transgenic Arabidopsis tissue was infected with compatible and incompatible isolates of both the fungal biotroph, Peronospora parasitica and the bacterial necrotroph, Pseudomonas syringae. Incompatible isolates induced rapid cell death (the hypersensitive response) at the site of infection and, with both pathogens, early, localised expression of the GUS gene was observed. In contrast, relatively slow induction of the GUS gene was seen in the compatible interaction and this was correlated with the appearance of senescence-like symptoms in the biotrophic interaction and cell death by necrosis that occurred in response to the necrotrophic pathogen. These results suggest that there are common steps in the signalling pathways that lead to cell death in the hypersensitive response, pathogen induced necrosis and senescence.     

Synthesis, antioxidant and radical scavenging activities of novel benzimidazoles

The synthesis and antioxidant evaluation of some novel benzimidazole derivatives (10-24) are described. Antioxidant properties of the compounds were investigated employing various in vitro systems viz., microsomal NADPH-dependent inhibition of lipid peroxidation (LP), interaction of 2,2-diphenyl-1-picrylhydrazyl (DPPH) and scavenging of superoxide anion radical. Compounds 12 and 13 showed very good antioxidant capacity and were 17-18-fold more potent than BHT (IC50 2.3 x 10(-4) M) with 1.3 x 10(-5) M and 1.2 x 10(-5) M IC50 values, respectively, by interaction of the stable DPPH free radical.     

657del5 mutation of the Nijmegen breakage syndrome gene (NBS1) in the Turkish population

The 657del5 mutation of the NBS1 gene has been demonstrated in most patients with Nijmegen breakage syndrome (NBS). We identified four Turkish families in which probands were diagnosed as having NBS and found to be homozygous for the 657del5 mutation. The 657del5 allele in the four Turkish families had a single origin.     

Time-frequency analysis of visual evoked potentials for interhemispheric transfer time and proportion in callosal fibers of different diameters

This study is an extension of the experimental research of Nalçac et al., who presented 16 subjects with a reversal of checkerboard pattern as stimuli in the right visual field or left visual field and recorded EEG at O1, O2, P3, and P4. They applied the chosen bandpass filters (4–8, 8–15, 15–20, 20–32 Hz) to the VEPs of subjects and obtained four different components for each VEP. The first aim of this study is to improve the previous report using some methods in time-frequency domain to estimate interhemispheric delays and amplitudes in a time window. Using the improved estimates of interhemispheric delays, the second aim is to estimate the proportion of callosal fibers of different diameters that are activated by visual stimuli by comparing amplitudes of VEPs in different frequency bands. If the relation between frequency components of VEP and delays for callosal fibers of different dimension were reliable, it would give us an opportunity to deal with amplitude of bandpass-filtered VEPs in order to see approximately the proportion of these fibers activated by a certain stimulus. By using frequency-dependent shifts in time and maximizing the cross correlation of direct VEP (DVEP–VEP obtained from contralateral hemisphere)–indirect VEP (IVEP–VEP obtained from ipsilateral hemisphere) pairs in the time-frequency domain, we examined the delay not only at P100 and N160 peaks but along a meaningful time interval as well. Furthermore, by shifting back the IVEP according to the delay estimated at each time window, both the amplitudes and energies of the synchronized DVEP–IVEP pairs were compared at the chosen frequency bands. The percentages of IVEPs at each band was then examined further in conjunction with the distribution of axon diameters in the posterior pole of the CC, questioning the relation between the distributions of the axon diameters and activations at each band. We established an energy definition to express the activation in the fibers. When the energy percentages of IVEPs in theta and alpha were totaled, they were found to be between 76.2% and 81.6%, which is close to the value 74–77% for fibers of 0.4–1 m in diameter obtained from anatomical study of human CC. The sum of energy percentages in the beta1 and beta2 bands was between 20.1% and 24.2%, which probably reflects the proportion of activation of callosal fibers 1–3 m in diameter.     

Female reproductive pattern of Callinectes sapidus Rathbun, 1896 (Brachyura: Portunidae) in Iskenderun Bay,Eastern Mediterranean

We studied the female reproductive pattern of Callinectes sapidus, which was introduced to the Mediterranean in the 20^th century. We assessed female size at first maturity, fecundity, and fecundity relationship to size in Iskenderun Bay, eastern Mediterranean, Turkey. Samples were collected between July 2014 and June 2015 using bottom trawling at depths ranging from 1 to 50 m. A total of 322 crabs were caught of which 308 (95.7%) were females including 116 ovigerous ones. The minimum carapace width of the mature females was 39.1 mm and the mean carapace width 123.8 mm. The carapace width of ovigerous females varied between 95.1 and 144.5 mm, with a mean of 120.3 mm. The highest number of ovigerous females was observed in July and August. Mean fecundity was 1.91 million (667,950–4,669,853) eggs per female. A weak positive linear relationship between fecundity and carapace width was noted, as well as a high correlation with total egg weight. In the eastern Mediterranean, maturity sizes of females were smaller than those in the native region of the species.     

Normalization of TAM post-receptor signaling reveals a cell invasive signature for Axl tyrosine kinase

Background

Tyro3, Axl, and Mertk (TAMs) are a family of three conserved receptor tyrosine kinases that have pleiotropic roles in innate immunity and homeostasis and when overexpressed in cancer cells can drive tumorigenesis.

Methods

In the present study, we engineered EGFR/TAM chimeric receptors (EGFR/Tyro3, EGFR/Axl, and EGF/Mertk) with the goals to interrogate post-receptor functions of TAMs, and query whether TAMs have unique or overlapping post-receptor activation profiles. Stable expression of EGFR/TAMs in EGFR-deficient CHO cells afforded robust EGF inducible TAM receptor phosphorylation and activation of downstream signaling.

Results

Using a series of unbiased screening approaches, that include kinome-view analysis, phosphor-arrays, RNAseq/GSEA analysis, as well as cell biological and in vivo readouts, we provide evidence that each TAM has unique post-receptor signaling platforms and identify an intrinsic role for Axl that impinges on cell motility and invasion compared to Tyro3 and Mertk.

Conclusion

These studies demonstrate that TAM show unique post-receptor signatures that impinge on distinct gene expression profiles and tumorigenic outcomes.

     

Bypass of a primase requirement for bacteriophage T4 DNA replication in vivo by a recombination enzyme, endonuclease VII.

A primase, the product of phage T4 gene 61, is required to initiate synthesis of Okazaki pieces and to allow bidirectional replication from several T4 origins. However, primase-defective T4 gene 61 mutants are viable. In these mutants, leading-strand DNA synthesis starts at the same time as in wild type infections, but, in contrast to wild type, initiation is unidirectional and the first replicative intermediates are large displacement loops. Rapid double-strand DNA replication occurs later after infection, generating multiple branched concatemers, which are cut and packaged into viable progeny particles, as in wild-type T4. Evidence is presented that this late double-strand DNA replication requires functional endonuclease VII (endo VII), the product of the T4 gene 49. We propose that endo VII can provide a backup mechanism when primase is defective, because it cuts recombinational junctions, generating 3' ends. These ends can prime DNA synthesis to copy the DNA strands that had been displaced during the initial origin-dependent replication. We explain the DNA-delay phenotype and the commonly observed temperature dependence of DNA replication in primase-deficient gene 61 mutants as a consequence of temperature-dependent translational control of gene 49 expression. In the presence or absence of functional primase endo VII is essential for correct packaging of DNA. The powerful selection that keeps the function of endo VII and expression of its gene at levels that are optimal for T4 development determines both the efficiency and the limitations of the bypass mechanism.     

Microbial strain improvement for enhanced polygalacturonase production by Aspergillus sojae

Strain improvement is a powerful tool in commercial development of microbial fermentation processes. Strains of Aspergillus sojae which were previously identified as polygalacturonase producers were subjected to the cost-effective mutagenesis and selection method, the so-called random screening. Physical (ultraviolet irradiation at 254 nm) and chemical mutagens (N-methyl-N′-nitro-N-nitrosoguanidine) were used in the development and implementation of a classical mutation and selection strategy for the improved production of pectic acid-degrading enzymes. Three mutation cycles of both mutagenic treatments and also the combination of them were performed to generate mutants descending from A. sojae ATCC 20235 and mutants of A. sojae CBS 100928. Pectinolytic enzyme production of the mutants was compared to their wild types in submerged and solid-state fermentation. Comparing both strains, higher pectinase activity was obtained by A. sojae ATCC 20235 and mutants thereof. The highest polygalacturonase activity (1,087.2?±?151.9 U/g) in solid-state culture was obtained by mutant M3, which was 1.7 times increased in comparison to the wild strain, A. sojae ATCC 20235. Additional, further mutation of mutant M3 for two more cycles of treatment by UV irradiation generated mutant DH56 with the highest polygalacturonase activity (98.8?±?8.7 U/mL) in submerged culture. This corresponded to 2.4-fold enhanced polygalacturonase production in comparison to the wild strain. The results of this study indicated the development of a classical mutation and selection strategy as a promising tool to improve pectinolytic enzyme production by both fungal strains.     

Aldrin Epoxidation in Flathead Mullet (Mugil cephalus): Possible Involvement of CYP1A and CYP3A

The primary objective of this study was to determine specific cytochrome P450 isozyme(s) involved in the metabolism of aldrin to its toxic metabolite dieldrin in flathead mullet (Mugil cephalus) liver microsomes. To identify the cytochrome P450 isozyme responsible for the aldrin metabolism in mullet liver, the effects of mammalian‐specific cytochrome P450 inhibitors and substrates were determined in the epoxidation reaction of aldrin. CYP3A‐related inhibitors, ketoconazole, SKF‐525A, and cimetidine, inhibited the metabolism of aldrin. The contribution of CYP1A to the aldrin metabolism was shown by the inhibition of 7‐ethoxyresorufin‐O‐deethylase activity in the presence of aldrin. The results indicate that CY1A and CYP3A are the cytochrome P450s involved in aldrin epoxidase activity in mullet. In addition, the suitability of aldrin epoxidase activity for monitoring of environmental pollution was also assessed in the fish samples caught from four different locations of the West Black Sea coast of Turkey.