Stability of a downflow anaerobic fixed-film reactor to feed change

Summary A stepped-loading start-up regime utilising volatile fatty acids as carbon and energy sources was applied to a downflow anaerobic fixed-film reactor operated at 37°C. A steadystate was attained with over 90% COD reduction at an organic loading rate of 6.1 Kg COD/m³·day. A complex wastewater coming from a citrus processing plant was then used to test the feasibility of feeding changes as well as the stability of the reactor performance. A final COD reduction of 60% was achieved with hydraulic retention times down to 1.5 days.     

The dynamics of (57)Fe nuclei in Fe(III)-DNA condensates.

The dynamics of iron nuclei in the condensates obtained by interaction of Fe(III) with DNA, Fe(III)(DNA monomer)(2), have been investigated by variable temperature (57)Fe M?ssbauer spectroscopy. Studies were effected on gel and freeze-dried samples, obtaining nearly coincident values of the parameters isomer shift and nuclear quadrupole splitting in T ranges 20-260 K. Functions ln(A(T)/A(77.3)) vs. T, here employed to investigate the dynamics of Fe nuclei, showed linear trends in the T ranges 20-150 and 150-260 K, respectively, the latter with larger slopes. Data coincided for gelled and freeze-dried specimens. No variation of delta or Delta E parameters occurred at the two T intervals, which suggests constancy of structure and bonding with the temperature changes. Functions (T) showed trends analogous to the corresponding functions determined for iron proteins, which were attributed to the occurrence of 'conformational substates'.     

Arcobacter cloacae sp. nov. and Arcobacter suis sp. nov., two new species isolated from food and sewage

Three strains recovered from mussels (F26), sewage (SW28-13^T) and pork meat (F41^T) were characterized as Arcobacter. They did not appear to resemble any known species on the basis of their 16S rDNA-RFLP patterns and the rpoB gene analyses. However, strains F26 and SW28-13^T appeared to be the same species. The 16S rRNA gene sequence similarity of strains SW28-13^T and F41^T to the type strains of all other Arcobacter species ranged from 94.1% to 99.6% and 93.4% to 98.8%, respectively. Phenotypic characteristics and the DNA–DNA hybridization (DDH) results showed that they belonged to 2 new Arcobacter species. A multilocus phylogenetic analysis (MLPA) with the concatenated sequences of 5 housekeeping genes (gyrA, atpA, rpoB, gyrB and hsp60) was used for the first time in the genus, showing concordance with the 16S rRNA gene phylogenetic analysis and DDH results. The MALDI-TOF mass spectra also discriminated these strains as two new species. The names proposed for them are Arcobacter cloacae with the type strain SW28-13^T (=CECT 7834^T = LMG 26153^T) and Arcobacter suis with the type strain F41^T (=CECT 7833^T = LMG 26152^T).     

Effect of temperature on the morphometric development of eggs in the prawn Macrobrachium americanum (Caridea: Palaemonidae) and larval success under experimental conditions

Abstract

This study evaluated the effect of temperature on morphometric features of the egg during the embryonic development of the prawn Macrobrachium americanum and the relationship with hatching and the survival of the larvae. Berried females were grouped (n = 3) and reared at three different temperatures, 26, 29, and 33 °C, for which seven developmental stages were recognized. At each stage, the apical and sagittal diameters of the eggs were measured, the volume was calculated, and the weights were recorded. Additionally, the duration of embryonic development, hatching percentage, and larval survival were determined. At 29 and 33 °C, the eggs’ volume increased by 50%, but at 26 °C, the increase was 25%. Larvae from eggs incubated at 33 °C died one day after hatching. At 29 °C, larvae survived until Zoea VII. Larvae from eggs incubated at 26 °C died at the end of Zoea I. The number of days of embryonic development was 20.5 ± 1.5 (26 °C), 15 ± 1 (29 °C), and 12 ± 1 (33 °C). A temperature of 29 °C was the most favorable for embryonic development in M. americanum.     

Tetanus Toxin Hc Fragment Induces the Formation of Ceramide Platforms and Protects Neuronal Cells against Oxidative Stress

Tetanus toxin (TeTx) is the protein, synthesized by the anaerobic bacteria Clostridium tetani, which causes tetanus disease. TeTx gains entry into target cells by means of its interaction with lipid rafts, which are membrane domains enriched in sphingomyelin and cholesterol. However, the exact mechanism of host membrane binding remains to be fully established. In the present study we used the recombinant carboxyl terminal fragment from TeTx (Hc-TeTx), the domain responsible for target neuron binding, showing that Hc-TeTx induces a moderate but rapid and sustained increase in the ceramide/sphingomyelin ratio in primary cultures of cerebellar granule neurons and in NGF-differentiated PC12 cells, as well as induces the formation of ceramide platforms in the plasma membrane. The mentioned increase is due to the promotion of neutral sphingomyelinase activity and not to the de novo synthesis, since GW4869, a specific neutral sphingomyelinase inhibitor, prevents neutral sphingomyelinase activity increase and formation of ceramide platforms. Moreover, neutral sphingomyelinase inhibition with GW4869 prevents Hc-TeTx-triggered signaling (Akt phosphorylation), as well as the protective effect of Hc-TeTx on PC12 cells subjected to oxidative stress, while siRNA directed against nSM2 prevents protection by Hc-TeTx of NSC-34 cells against oxidative insult. Finally, neutral sphingomyelinase activity seems not to be related with the internalization of Hc-TeTx into PC12 cells. Thus, the presented data shed light on the mechanisms triggered by TeTx after membrane binding, which could be related with the events leading to the neuroprotective action exerted by the Hc-TeTx fragment.     

Immunogenicity and efficacy of a chimpanzee adenovirus-vectored Rift Valley Fever vaccine in mice

Background

Rift Valley Fever (RVF) is a viral zoonosis that historically affects livestock production and human health in sub-Saharan Africa, though epizootics have also occurred in the Arabian Peninsula. Whilst an effective live-attenuated vaccine is available for livestock, there is currently no licensed human RVF vaccine. Replication-deficient chimpanzee adenovirus (ChAd) vectors are an ideal platform for development of a human RVF vaccine, given the low prevalence of neutralizing antibodies against them in the human population, and their excellent safety and immunogenicity profile in human clinical trials of vaccines against a wide range of pathogens.

Methods

Here, in BALB/c mice, we evaluated the immunogenicity and efficacy of a replication-deficient chimpanzee adenovirus vector, ChAdOx1, encoding the RVF virus envelope glycoproteins, Gn and Gc, which are targets of virus neutralizing antibodies. The ChAdOx1-GnGc vaccine was assessed in comparison to a replication-deficient human adenovirus type 5 vector encoding Gn and Gc (HAdV5-GnGc), a strategy previously shown to confer protective immunity against RVF in mice.

Results

A single immunization with either of the vaccines conferred protection against RVF virus challenge eight weeks post-immunization. Both vaccines elicited RVF virus neutralizing antibody and a robust CD8⁺ T cell response.

Conclusions

Together the results support further development of RVF vaccines based on replication-deficient adenovirus vectors, with ChAdOx1-GnGc being a potential candidate for use in future human clinical trials.

     

Studio Sul Ciclo Annuale del Fitoplancton del Lago di Piediluco (Umbria)

Abstract

Researches about the cycle of the phytoplankton in the lake of Piediluco (Umbria). — The Authors studied the phytoplanktonic productivity within an year (1960–1961) in the lake of Piediluco (Central Italy); they have tried also to bring into evidence eventual changes in the phytocoenosis of the lake, basing themselves on some phytoplanktonic analysis made by Pasquini (1924) and by Marchesoni (1940) on samples collected before that Nera River was put in the lake for hydroelectric purpose.

The lake of Piediluco, which is a relict of the ancient Pleistocenic lake (Velino lake), is prevàilingly alluvial in origin and it is the second lake of Umbria in extension, after Trasimeno lake, having a surface of Kmq. 1,52. It is situated in the plain of Rieti at 368 m. level s. l. m. and it is 19 m. deep (Fig. 1).

In 1932 this lake was exploited to have hydroelectric energy and the « Società Elettrica Terni » built the Stoney dike along Velino River and digged also a canal connecting the lake with the river; another derivation canal, which puts Nera River into the lake of Piediluco, was also digged.

Every month, from October 1960 to October 1961 in the eulimnic zone of the lake were collected qualitative (surface and vertical) and quantitative samples (at 1, 3, 5, 10, 15 and 19 m.) and some environmental factors as: temperature, contents in oxygen, in organic matter, in N-NH₄, in N-NO₃ and in P-PO₄ were studied.

After the putting of Nera River in the lake, the thermic regime changed: in summer there was a sinking and in winter a rise in temperature.

As we can see (Table I), the lake is of a tropical type and in winter it is in homothermal conditions; in summer there is a thermal stratification, which is of brief duration. The thermocline is evident only in June and July (Fig. 2).

The Table II, in which the quantities of oxygen dissolved in the water are reported, puts in evidence that the lake of Piediluco has an holomictic regime.

After the period of June, July and August, in which the oxygen contents of the deep waters is very low, there is in the late autumn the turnover which produces a complete mixing of superficial and deep waters (Fig. 3).

During the most part of the year phenomena of hypersaturation in O₂ take place in the surface (Table III): the highest amounts are reached in May, June, July and August. The highest percentage in O₂ is reached during the most part of the year in the surface waters (Fig. 4).

This high concentration is simply biologic in origin: it is due to the photosynthetic activity of phytoplankton which has numerically increased in this period.

Tables IV, V, VI and VII show the seasonal variations of the dissolved organic matter, of N-NH₄, of N-NO₃ and P-PO₄; they show also that the nutritive substances (mainly N-NO₃ and P-PO₄ which affect the productivity of the lake of Piediluco, undergo an annual cycle: a phase of consumption on the part of the algal flora living in the surface during spring and summer; a phase of production at the bottom during the summer-autumnal stratification and a winter phase of redistribution of these bottom reserves to all the mass of water owing to the late-autumnal turnover.

The qualitative (Table VIII and Table IX) and quantitative (Table 10/A and 10/B) analysis show an absolute prevalence of Diatoms during all the period in which the planktonic hauls have been carried out.

On a quantitative point of view they have a very large majority with regard to the other classes: Diatoms by themselves reaches 98%.

The Algae which tipically characterize the lake of Piediluco are: Cyclotella kuetzingiana, C. melosiroides, Fragilaria crotonensis, Synedra ulna, S. danica, S. acus, Asterionella formosa; these are followed by: Peridinium inconspicuum, P. cinctum, Dinobryon divergens, Rhizochrysis limnetica, Gloeococcus schroeteri and Closterium aciculare.

The prevalence of Diatoms during all the year is due not to one species, but to three species which characterize our lake (Table XI): in January and February there is a facies with Asterionella formosa, in April a facies with Synedra ulna and with Asterionella formosa, Cyclotella kuetzingiana, Synedra danica and Fragilaria crotonensis; in May and June a facies with Cyclotella kuetzingiana with Cyclotella melosiroides and Synedra ulna, while in August we have again a facies with Synedra ulna, and with Synedra acus and Cyclotella kuetzingiana. September and October are characterized by Synedra ulna, even though in a smaller amount.

The analysis of two qualitative hauls of surface carried out in the zone with a vegetation mainly constituted by Arundo phragmites (Table XII) put in evidence that the littoral vegetation affects the phytoplanktonic composition because it is on a qualitative point of view more rich of algal species than the eulimnic zone.

Before that Nera River was put in the lake, the phytoplankton of this lake was characterized by Peridinium cinctum and Ceratium hirundinella; now these species reduced themselves very much.

The Authors, in order to explain this modification in the algological composition, attach importance to the changing of the thermic conditions and perhaps to the increased turbulence of waters caused by the putting in of Nera River (on an average 16–17 mc/sec.).

Valuing the degree of trophism of the lake of Piediluco with the Diatom quozient (Nygaard et al.) and the Compound index (Nygaard, 1949), the lake must be considered eutrophic, but if we consider some environmental factors and the presence of some Algae, we conclude that the lake has a character tending to eutrophy.     

Ricerche istofisiologiche sullo spadice di arum

Riassunto

L'A. studia la struttura anatomica della clava dello spadice di Arum, indaga le intime modificazioni cellulari che avvengono durante il suo periodo di accrescimento, di maturità e di senescenza, e viene a rilevare alcuni segni della senescenza cellulare. Inoltre mette in evidenza le differenze morfologiche tra i procossi di necrobiosi in natura e in ambiente asfittico; segnala la vacuolizzazione delle masse cromatiniche come la causa principale della cariocinesi. Fa infine osservare che il parossismo termico (funzione prima di tale organo) rimane nei limiti biologici senza dirette azioni deleterie sulla struttura citologica della clava.     

The intracellular fate of zonula occludens 2 is regulated by the phosphorylation of SR repeats and the phosphorylation/O-GlcNAcylation of S257

Zona occludens 2 (ZO-2) has a dual localization. In confluent epithelia, ZO-2 is present at tight junctions (TJs), whereas in sparse proliferating cells it is also found at the nucleus. Previously we demonstrated that in sparse cultures, newly synthesized ZO-2 travels to the nucleus before reaching the plasma membrane. Now we find that in confluent cultures newly synthesized ZO-2 goes directly to the plasma membrane. Epidermal growth factor induces through AKT activation the phosphorylation of the kinase for SR repeats, serine arginine protein kinase 1, which in turn phosphorylates ZO-2, which contains 16 SR repeats. This phosphorylation induces ZO-2 entry into the nucleus and accumulation in speckles. ZO-2 departure from the nucleus requires intact S257, and stabilizing the β-O-linked N-acetylglucosylation (O-GlcNAc) of S257 with O-(2-acetamido-2-deoxy-d-glucopyranosylidene)amino-N-phenylcarbamate, an inhibitor of O-GlcNAcase, triggers nuclear exportation and proteosomal degradation of ZO-2. At the plasma membrane ZO-2 is not O-GlcNAc, and instead, as TJs mature, it becomes phosphorylated at S257 by protein kinase Cζ. This late phosphorylation of S257 is required for the correct cytoarchitecture to develop, as cells transfected with ZO-2 mutant S257A or S257E form aberrant cysts with multiple lumens. These results reveal novel posttranslational modifications of ZO-2 that regulate the intracellular fate of this protein.     

Numerical Analysis of Titanium Cast Devices for Dental Implantology

The present work pertains to a numerical investigation of the casting process of titanium devices adopted for dental implantology. The analysis of the titanium framework that connects different abutments, in a multi-implant configuration, is performed evaluating the characteristics of the material that depend on the manufacturing procedure. The connecting bar is obtained by a foundry process that is controlled through a numerical simulation by using a control volume technique. This analysis leads to the possibility to have detailed information on the process and control of the quality of the microstructure of the material produced that proves to be highly beneficial for defining mechanical properties. On the basis of the results obtained, a subsequent stress analysis can be performed, addressed to highlight critical conditions. Functional response of the whole bar-implant framework is studied by means of a numerical model, based on the geometric element method. This technique is particularly suited for describing complex morphology of the implant site. The present approach addresses a higher quality definition of the reliability of the device used in dental practice and represents a valuable tool in assisting optimisation procedures pertaining to manufacturing. In fact, the improvement of titanium devices is intended not only with regard to their mechanical performances, but also to requirements pertaining to manufacturing and clinical practice.