Main-chain length control of conformation, membrane activity, and antibiotic properties of lipopeptaibol sequential analogues

To investigate the effect of backbone length and amphiphilicity on the 3D structure, membrane permeability, and antibacterial properties of trichogins, a subclass of lipopeptaibols, we prepared, by the segment condensation approach in solution and chemically characterized, a set of N(alpha)-1-octanoylated -X-(GLUG)(n)-I-L- ( X=G or U where U=Aib; n=1-4) sequential peptide esters. In parallel, the 12-mer (UGGL)(3) aneurism peptide, an analogue of the 11-mer sequential peptide (n=2) with an amino acid insertion was also synthesized and studied. By FT-IR absorption technique, we clearly showed that, in CDCl(3) solution, all peptides essentially populate intramolecularly H-bonded, helical conformations. Moreover, CD spectroscopy indicates that all peptides, with the single exception of the shortest oligomer (the heptamer), adopt mixed 3(10)-/alpha-helical structures, to an extent approximately correlating with main-chain length, in MeOH solution and sodium dodecylsulfate (SDS) micelles. Significant membrane permeability properties were found only for the three longest GLUG-based peptides, with the 15-mer oligomer (n=4) resulting the most active. The lack of activity exhibited by the aneurism peptide in this experiment strongly suggests a relevant role for the sequence amphiphilicity. In addition, antibacterial activity and selectivity were highlighted and demonstrated to be dependent on peptide main-chain length and amphiphilicity, in the sense that the two shortest GLUG-based homologues are active against Gram-positive strains, whereas the two longest homologues are able to penetrate the membranes of the Gram-negative strains, and the UGGL-based aneurism peptide is inactive.     

Long term functional changes within the Oligochaeta communities within the Danube River Delta,Romania

Within the Danube River delta's lakes the Oligochaeta communities comprise between 7.9% and 36.2% of the total biomass of benthic fauna. Their importance in energy flow at the ecosystem level changed in relation to fast trophic transition of all shallow lakes to the hypertrophic state. The parameters of the energy budget of the dominant populations and the potential production of benthivorous fish species assessed during 1976–1994 interval support this conclusion. P/B ratio, K ₁ and K ₂ coefficients assessed for both the 1976–1980 and 1991–1994 intervals revealed different functional patterns of response of Potamothrix hammoniensis (Michaelsen, 1901) and Limnodrilus hoffmeisteri (Claparede, 1862) to varying trophic conditions. Changes in communities structure, size of the constituent populations and their age distribution, as well as the quantity and quality of food supply and level of hypoxia at the sediment/water interface were the main factors that affected the role of these populations as energy carrier from the huge energy pool represented by sedimented organic carbon to benthivorous fish species.

     

Ecological Significance of Nitrogen Cycling by Tubificid Communities in Shallow Eutrophic Lakes of the Danube Delta

Importance of tubificid populations on nitrogen cycle in two categories of shallow eutrophic lakes in the Danube Delta was quantitatively assessed for the 1992-1993 period. The structure of the primary producers in the studied lakes was used to discriminate between the two categories:(i) lakes dominated by macrophytes (A₁) and (ii) lakes dominated by phytoplankton (A₂). In both categories tubificid worms represented important fraction of the entire benthic community (35 and 32%, respectively, as number of individuals). They influence the sediment-water exchange of nutrients. The main processes involved are excretion of nutrients and their continuous release from sediments by molecular diffusion or through channels created by bioturbation. Inorganic nitrogen released from bottom sediments may regulate nitrogen load in the water body and thus, phytoplankton production. In 1992-1993, nitrogen stocks in tubificid biomass accounted for 5.3% in A₁ lakes and 15.6% in A₂ lakes of the amount stocked in phytoplankton, and only for 1.2 and 2.9% respectively, of the nitrogen load in water body. Nitrogen excretion rates ranged between 60.52 and 153.74 mg N m^–2 year^–1, and release rates from sediments between 378.26 and 960.87 mg N m^–2 year^–1, the lowest values being recorded for A₂ category. Differences are related to tubificid biomass, structure and abundance of primary producers and to nutrient load in different ecosystems. Ratios between release rate of inorganic nitrogen by tubificid worms and sedimentation rate of organic nitrogen in the two categories of lakes were 8.3 and 6.4% respectively. Contribution of nitrogen released daily from sediments to the dissolved inorganic nitrogen load in the water column was less than 0.5%. However, in A₁ and A₂ lakes, the released nitrogen had a potential to sustain 24.74 and 8.01%, respectively, of the annual phytoplankton production. These values suggest the significance of tubificids in keeping the eutrophication process at a high level, especially during the periods when nitrogen is the main limiting factor for phytoplankton production.     

Rapid deswelling response of poly(N-isopropylacrylamide)/poly(2-alkyl-2-oxazoline)/poly(2-hydroxyethyl methacrylate) hydrogels

Ternary poly( N-isopropylacrylamide)/poly(2-alkyl-2-oxazoline)/poly(2-hydroxyethyl methacrylate) (PNIPAAm/PROZO/PHEMA) hydrogels were prepared by the free-radical copolymerization of N-isopropylacrylamide (NIPAAm), 2-hydroxyethyl methacrylate (HEMA), and poly(2-alkyl-2-oxazoline) (PROZO) multifunctional macromonomers. The resulting polymeric materials were characterized by differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), and scanning electron microscopy (SEM), as well as by equilibrium swelling experiments. All synthesized hydrogels display temperature sensitivity in the 28-38 degrees C range. A high rate of response was registered as compared to that of materials based only on PNIPAAm. The swelling-deswelling peculiar behavior was related to the chemical composition (hydrophile/hydrophobe balance), the length of the inserted PROZO sequence, and inner morphology, an aspect which points on its possible control by synthesis. It was evidenced that the architecture of the resulting porous materials has a high order degree, emerging from the self-assembling of the microgel particles, which provided numerous, nearly uniform, large water release channels.     

Effect of Cross-Linking Methods on Structure and Properties of Poly(ε-caprolactone) Stabilized Hydrogels Containing Biopolymers

Different dense and porous biodegradable matrices based on solely atelocollagen, or with different atelocollagen and hyaluronic acid derivative ratios, were obtained by varying feeding formulations, cross-linking reaction parameters, and preparative protocols. The compositions and methods for forming hydrogels through a combination of physical and chemical cross-linking processes are provided. The chemical cross-linking was mainly mediated by a synthetic component, a poly(ε-caprolactone) reactive derivative, aiming the development of new hybrid hydrogels with tailored characteristics by an appropriate use of the advantages offered by the included natural and synthetic components and the selection of the preparative procedure. The structure and morphology of the 3D hybrid materials were comparatively investigated by means of Fourier-transform infrared spectroscopy (FTIR), differential scanning calorimetry (DSC), X-ray diffraction (XRD), and environmental scanning electron microscopy (ESEM). FTIR and XRD analysis showed no signs of collagen denaturation during the formation of 3D structures. The influence of various factors, such as the chemical composition of the resulted hydrogels and their morphology, on water uptake and water vapor sorption, mechanical behavior, as well as on in vitro degradation characteristics, was systematically investigated. The experimental results point on the advantage offered by the high and modular physicochemical stability of the ternary hydrogels cross-linked by combined approaches. All newly developed materials show no hemolytic effect, which recommends them for potential biomedical applications.     

Observations on the population dynamics of Potamothrix hammoniensis (Michaelsen, 1901) (Tubificidae,Oligochaeta) in Lake Isacova in the Danube Delta

The Danube Delta is one of the widest wetland systems in Europe and Lake Isacova is one of the biggest lakes in the Delta. The oligochaete community in the Lake comprised 52% of the total benthic fauna and Potamothrix hammoniensis (Michaelsen, 1901) (Tubificidae, Oligochaeta) represented more than 90% of the oligochaetes sampled. During September 1991 – August 1994, the proportion of ovigerous individuals of P. hammoniensis represented less than 20% of the total population, with localised differences in the Lake occurring over time. Breeding and cocoon production in the study period started in March of the year, when the water temperature was about 7 °C, and lasted until May 1992 and June 1993, respectively. The newly hatched individuals reached the highest proportion in May. Although large individuals were found again by the end of September (1991) and the beginning of October (1993), no spermatozeugmata in their spermathecae or cocoons were observed during November and October, respectively. Member of this cohort bred and started cocoon deposition during the following spring, after about 11 months of maturation (from April to March). A second, less intensive period of breeding was recorded during August 1992 and this lasted until the beginning of September 1993. However, cocoons from this latter period did not show embryonic growth and hatching until the autumn floods bring oxygenated water to the bottom level. This resulted in a period of rapid development followed by the mass hatching of cocoons. These specimens needed a maximum of 9 month (from October to July) to reach sexual maturity.     

Both sphingomyelin synthases SMS1 and SMS2 are required for sphingomyelin homeostasis and growth in human HeLa cells

Sphingomyelin (SM) is a vital component of cellular membranes in organisms ranging from mammals to protozoa. Its production involves the transfer of phosphocholine from phosphatidylcholine to ceramide, yielding diacylglycerol in the process. The mammalian genome encodes two known SM synthase (SMS) isoforms, SMS1 and SMS2. However, the relative contributions of these enzymes to SM production in mammalian cells remained to be established. Here we show that SMS1 and SMS2 are co-expressed in a variety of cell types and function as the key Golgi- and plasma membrane-associated SM synthases in human cervical carcinoma HeLa cells, respectively. RNA interference-mediated depletion of either SMS1 or SMS2 caused a substantial decrease in SM production levels, an accumulation of ceramides, and a block in cell growth. Although SMS-depleted cells displayed a reduced SM content, external addition of SM did not restore growth. These results indicate that the biological role of SM synthases goes beyond formation of SM.     

Nitrogen and Phosphorous Excretion Rates by Tubificids from the Prahova River (Romania)

Nitrogen and phosphorous exchange at the water–sediment interface is controlled both by complex physico-chemical factors and biological processes. Zoobenthos excretion is one of the most important processes in the mineralization of sedimented organic mater. In polluted freshwaters, tubificid worms are among the dominant components of the benthic community. Rates of ammonium and inorganic phosphate excretion by tubificids were experimentally assessed. They were related to the tubificid abundance in a stream ecosystem polluted with municipal and industrial wastewater. The relationship between these rates and temperature were investigated within the range of 4–23 °C. Relatively constant excretion rates were obtained for both nutrients in the first 8 h of excretion, ranging between 0.076 and 0.226 μg N mg d.w.⁻¹ h⁻¹ and 0.0065–0.01 μg P mg d.w.⁻¹ h⁻¹_, respectively. Q₁₀ values of 2.52 for ammonium and 1.31 for phosphate were calculated. If we presume that all excreta eventually enters the water column, then we can calculate that these invertebrates potentially add 39.17 mg N m⁻² day⁻¹ and 0.49 mg P m⁻² day⁻¹. These values accounts for 17.16 and 7.56% of the nutrient load in the river water, respectively.