Characterization,Toxicity, and Optimization for the Growth and Production of Bacteriocin-like Substances by Lactobacillus curvatus

Probiotics and Antimicrobial Proteins - This study aimed to characterize, evaluate toxicity and optimize the conditions for the growth and production of bacteriocin-like substances by Lactobacillus...     

Theoretical computation of phase locking in embryonic atrial heart cell aggregates

The effects of periodic stimulation of spontaneously beating aggregates of chick atrial heart cells are considered. Provided the effects of a single stimulus do not change the properties of the oscillation, and that the oscillation is re-established rapidly following a stimulus, this system can be modeled by one-dimensional finite difference equations. These equations employ experimentally generated phase resetting data that describe the effects of a single isolated stimulus at different phases of the oscillation. A complete analysis of the predicted dynamics is given over a broad range of stimulation frequencies and amplitudes. Prominent features of the dynamics include phase locking, bistability, chaos, and disappearance of Arnold tongues at large stimulation amplitudes. The fine details of the bifurcations are sensitive to properties of the phase resetting curves, and consequently, the observed bifurcations are not expected to be "universal" for larger stimulation amplitudes. Experimental traces show many correspondences with theoretical computations.     

The GTP- and Phospholipid-Binding Protein TTD14 Regulates Trafficking of the TRPL Ion Channel in Drosophila Photoreceptor Cells

Recycling of signaling proteins is a common phenomenon in diverse signaling pathways. In photoreceptors of Drosophila, light absorption by rhodopsin triggers a phospholipase Cβ-mediated opening of the ion channels transient receptor potential (TRP) and TRP-like (TRPL) and generates the visual response. The signaling proteins are located in a plasma membrane compartment called rhabdomere. The major rhodopsin (Rh1) and TRP are predominantly localized in the rhabdomere in light and darkness. In contrast, TRPL translocates between the rhabdomeral plasma membrane in the dark and a storage compartment in the cell body in the light, from where it can be recycled to the plasma membrane upon subsequent dark adaptation. Here, we identified the gene mutated in trpl translocation defective 14 (ttd14), which is required for both TRPL internalization from the rhabdomere in the light and recycling of TRPL back to the rhabdomere in the dark. TTD14 is highly conserved in invertebrates and binds GTP in vitro. The ttd14 mutation alters a conserved proline residue (P75L) in the GTP-binding domain and abolishes binding to GTP. This indicates that GTP binding is essential for TTD14 function. TTD14 is a cytosolic protein and binds to PtdIns(3)P, a lipid enriched in early endosome membranes, and to phosphatidic acid. In contrast to TRPL, rhabdomeral localization of the membrane proteins Rh1 and TRP is not affected in the ttd14 ^P75L mutant. The ttd14 ^P75L mutation results in Rh1-independent photoreceptor degeneration and larval lethality suggesting that other processes are also affected by the ttd14 ^P75L mutation. In conclusion, TTD14 is a novel regulator of TRPL trafficking, involved in internalization and subsequent sorting of TRPL into the recycling pathway that enables this ion channel to return to the plasma membrane.     

Radiosynthesis of a ¹⁸F-labeled 2,3-diarylsubstituted indole via McMurry coupling for functional characterization of cyclooxygenase-2 (COX-2) in vitro and in vivo

The radiosynthesis of 3-(4-[(18)F]fluorophenyl)-2-(4-methylsulfonylphenyl)-1H-indole [(18)F]-3 as potential PET radiotracer for functional characterization of cyclooxygenase-2 (COX-2) in vitro and in vivo is described. [(18)F]-3 was prepared by McMurry cyclization of a (18)F-labeled intermediate with low valent titanium and zinc via a two-step procedure in a remote controlled synthesizer unit including HPLC purification and solid phase extraction. In this way [(18)F]-3 was synthesized in 80 min synthesis time in 10% total decay corrected yield from [(18)F]fluoride in radiochemical purity >98% and a specific activity of 74-91 GBq/μmol (EOS). [(18)F]-3 was evaluated in vitro using pro-inflammatory stimulated THP-1 and COX-2 expressing tumor cell lines (FaDu, A2058, HT-29), where the radiotracer uptake was shown to be consistent with up regulated COX-2 expression. The stability of [(18)F]-3 was determined by incubation in rat whole blood and plasma in vitro and by metabolite analysis of arterial blood samples in vivo, showing with 75% of original compound after 60 min an acceptable high metabolic stability. However, no substantial tumor accumulation of [(18)F]-3 could be observed by dynamic small animal PET studies on HT-29 tumor-bearing mice in vivo. This may be due to the only moderate COX-1/COX-2 selectivity of 3 as demonstrated by both cellular and enzymatic cyclooxygenase inhibition assay in vitro. Nevertheless, the new approach first using McMurry cyclization in (18)F-chemistry gives access to (18)F-labeled diarylsubstituted heterocycles that hold promise as radiolabeled COX-2 inhibitors.     

Testis differentiation in the glowworm, Lampyris noctiluca, with special reference to the apical tissue.

The gonads of Lampyris noctiluca are sexually undifferentiated during the first larval instars. They consist of many gonadal follicles that include the germ stem cells enclosed by the somatic cells of the follicle wall. Follicle wall cells are more numerous at the follicle apices than at the distal parts, but different cell types cannot be distinguished. In male larvae, the appearance of apical follicle tissue, derived from follicle wall cells, marks the onset of testis differentiation. When maximally expressed, the apical tissue occupies about the upper half of the testis follicles and can be observed in larvae of the fifth and sixth instar. The apical tissue is characterized by its "light" appearance (due to poor stainability) caused by the small number cellular organelles, especially a paucity of free ribosomes. Maximal expression of the apical tissue must be very brief, since in most examined fifth and sixth instar larvae the apical tissue is partly or mostly translocated into the center of the upper half of the follicles and spermatogonia then occupy the apical follicle tips. During and after translocation apical cells form projections that grow around clusters of spermatogonia (spermatocysts). Thus, the apical cells transform into spermatocyst envelope cells. They retain their "light" appearance but undergo dramatic subcellular differentiation: smooth ER becomes extremely prominent, forming stacks and whorls of parallel cisternae. Golgi complexes are also conspicuous. The cellular organization suggests secretory activity. The possibility of ecdysteroid production and its function is discussed. The spermatocyst envelope cells persist into the pupal stage. When spermiohistogenesis takes place in cysts, cyst envelope cells show signs of regression. At all stages of testis development apical cells and their derivatives, the spermatocyst envelope cells, phagocytize degenerating spermatogonia. Although this is an important task of these cells, the impressive formation of sER in the cyst envelope cells is indicative of an additional, as yet unknown, function.     

Detection of Shiga toxins,intimin and enterohemolysin in Escherichia coli strains isolated from children in eastern Slovakia

FiftyEscherichia coli strains isolated from stool samples of 51 healthy children, 143 strains isolated from stool samples of 327 children with diarrhea and 24 strains isolated from stool samples of 21 children with suspected hemolytic uremic syndrome were examined for the presence of Shiga toxin-producingE. coli virulence factors (shiga toxin 1 and 2, intimin and enterohemolysin) and their genes. Vero-cell assay and latex agglutination were used for detection of Shiga toxin 1 and 2, TSB agar with washed erythrocytes was used for detection of enterohemolysin; genes encoding shiga toxin 1 and 2, intimin and enterohemolysin were detected using multiplex PCR. The presence ofE. coli strains harboring genes encoding shiga toxin 1 and 2 (12 strains), intimin (34 strains) and enterohemolysin (12 strains) was demonstrated.