The role of coupled DNRA-Anammox during nitrate removal in a highly saline lake.

Nitrate (NO3-) removal from aquatic ecosystems involves several microbially mediated processes, including denitrification, dissimilatory nitrate reduction to ammonium (DNRA), and anaerobic ammonium oxidation (anammox), controlled by slight changes in environmental gradients. In addition, some of these processes (i.e. denitrification) may involve the production of undesirable compounds such as nitrous oxide (N2O), an important greenhouse gas. Saline lakes are prone to the accumulation of anthropogenic contaminants, making them highly vulnerable environments to NO3- pollution. The aim of this paper was to investigate the effect of light and oxygen on the different NO3- removal pathways under highly saline conditions. For this purpose, mesocosm experiments were performed using lacustrine, undisturbed, organic-rich sediments from the Pétrola Lake (Spain), a highly saline waterbody subject to anthropogenic NO3- pollution. The revised 15N-isotope pairing technique (15N-IPT) was used to determine NO3- sink processes. Our results demonstrate for the first time the coexistence of denitrification, DNRA, and anammox processes in a highly saline lake, and how their contribution was determined by environmental conditions (oxygen and light). DNRA, and especially denitrification to N2O, were the dominant nitrogen (N) removal pathways when oxygen and/or light were present (up to 82%). In contrast, anoxia and darkness promoted NO3- reduction by DNRA (52%), combined with N loss by anammox (28%). Our results highlight the role of coupled DNRA-anammox, which has not yet been investigated in lacustrine sediments. We conclude that anoxia and darkness favored DNRA and anammox processes over denitrification and therefore to restrict N2O emissions to the atmosphere.

Mode of fibroblast growth enhancement by human interleukin-1.

Previous studies have demonstrated that interleukin-1 (IL-1) stimulates fibroblast growth (Schmidt, J. A., S. B. Mizel, D. Cohn, and I. Green. 1982. J. Immunol. 128:2177-2182) and binds to specific, high affinity receptors of BALB/c3T3 cells (Bird, T. A., and J. Saklatval. 1986. Nature (Lond.). 324:263-265, 266-268). We have investigated the mechanism of fibroblast growth stimulation by IL-1. Addition of fibroblast growth factor derived from platelets (PDGF) to a quiescent culture of BALB/c3T3 cells produced 8-10-fold increase in DNA synthesis during 24-h incubation. The cellular action of PDGF was mediated through competence induction and required synergistic action of plasma-derived factors for full mitogenic activity. When tested at a wide range of concentrations (0.1-100 pM), natural IL-1 or recombinant IL-1 produced only a maximum of 5-10% of DNA synthesis elicited in response to PDGF or serum. Induction of DNA synthesis required continuous presence of IL-1 and did not exhibit synergism with plasma. Competence induction and mitogenic stimulation by PDGF was associated with early induction of proteins P32, P38, P46-48, P75, and changes in cytoskeletal organization. Examination of these early cellular changes showed that IL-1 did not produce similar induction of cellular proteins and the morphological changes associated with growth stimulation. These results suggest that the mode of IL-1 action on BALB/c3T3 was not through competence induction. When IL-1 was added to cells rendered competent by brief exposure to PDGF, 10-15% additional DNA synthesis occurred during the first 24 h. Extended incubation of PDGF-treated cells in the presence of IL-1 revealed that the stimulation by IL-1 occurred predominantly during the subsequent cycle of DNA replication, wherein DNA synthesis reached three- to fivefold higher than the untreated cultures. We conclude (a) IL-1 alone is not a potent mitogen for BALB/c3T3 cells, and does not bring cells out of the growth arrest Go phase, (b) treatment with PDGF renders the cells more responsive to IL-1, (c) part of the IL-1 action on competent cells may be characterized as progression inducing activity, further, (d) our results indicate that action of IL-1 on PDGF-treated cells produces sustained DNA synthesis for an extended period, perhaps by preventing the entry of cells into growth arrest Go phase.

[Study of 206 transfused sickle cell disease patients: immunization, transfusion safety and red blood cell supply]. / Etude d'une cohorte de 206 patients drépanocytaires adultes transfusés: immunisation, risque transfusionnel et ressources en concentrés globulaires.

BACKGROUND: Prevention of hemolytic transfusion reactions depends upon our capacity to prevent allo-immunization and conflicts between antigens of transfused red blood cells and antibodies produced by the recipient. In this study, we show that to secure transfusion of sickle cell disease patients, it is necessary to take into account their immunohematologic characteristics in the organization of transfusion. METHODS AND RESULTS: Immunohematological data of 206 chronically transfused patients have been collected as well as phenotypes of transfused units. In order to prevent allo-immunization against C and E antigens for patients typed D+C-E-c+e+ (56%), 26% of the transfused units were D-C-E-c+e+. We found that 47% of the patients had a history of allo-immunization, whereas only 15% produced an antibody the day of inclusion in the study. The non-detectable antibodies were frequently known as dangerous for transfusion. Finally, this study shows the frequency of anti-D in D+ patients and anti-C in C+ patients, pointing out the question of partial antigens. CONCLUSION: To insure optimal transfusion safety for sickle cell disease patients, three points have to be improved: blood donation within the Afro-Caribbean community living in France, access to history of immuno-hematological data, detection of variant antigens, especially within the RH blood system.

[ABO incompatibility and non myeloablative allogeneic stem cell transplantation]. / Incompatibilité ABO et greffe allogénique de cellules souches hématopoïétiques à l'ère des conditionnements non myéloablatifs.

ABO incompatibility is not a barrier for allogeneic hematopoietic stem cell transplantation but is associated with specific complications. Major ABO incompatibility is associated with delayed erythroid engraftment, increased transfusion requirement and cases of pure red cell aplasia. Minor ABO incompatibility may be responsible for acute haemolytic reactions in the first months following transplantation. The widely used non myeloablative conditioning regimens might modify the management of ABO incompatibility. They could favour pure red cell aplasia development in the setting of major ABO mismatch since they are associated with a prolonged persistence of host anti-donor isohemagglutinins after allogeneic hematopoietic stem cell transplantation. In the setting of minor ABO incompatibility, the use of peripheral blood stem cells and the nature of graft-versus-host disease prophylaxis regimen may have an impact on the incidence of haemolytic reactions. In that review, the clinical and therapeutic aspects of ABO incompatibility are studied, especially regarding the impact of the conditioning regimen intensity.

Salt requirements in the denitrifying bacterium Pseudomonas nautica 617.

Pseudomonas nautica 617, which was isolated from superficial marine sediment, was found to require sodium for growth. Growth also appeared to be sensitive to the divalent cation, Mg2+, the presence of which, together with that of Na+, was necessary for achieving maximal growth. We investigated cell capacity to resist lysis after washing with either 0.05 M MgCl2 or 0.5 M NaCl, by monitoring suspension optical density changes as well as the release of ultraviolet absorbing material. Mg2+ turned out to play a significant role in stabilizing the structure of the cell envelope. Respiratory activity was also sensitive to ionic environment. With cells washed with 0.05 M MgCl2 and suspended in 0.05 M Tris buffer, the respiration rate, assessed by N2O evolution, was 15% of that measured in artificial sea water. Upon addition of 0.5 M Na+, nitrous oxide production rose to 32% of the reference level. The dinitrification rate was fully restored by further addition of 0.05 M Mg2+. K+ alone had almost no effect, but when added with Na+, the rate of denitrification increased to 45%.

Metabolic differences between attached and free-living marine bacteria: inadequacy of liquid cultures for describing in situ bacterial activity.

Marinobacter sp. strain CAB was cultivated with or without porous glass beads as solid support. Two substrates were used: the hydrophilic sodium lactate and a hydrophobic C(18)-isoprenoid ketone (6,10,14-trimethylpentadecan-2-one (TMP)). The substrate adsorption onto the beads was measured. Bacterial adhesion was determined by a direct count technique and amounted to 70% of total cells. In the immobilised cell cultures (ICC), generation times were 1.5 and 1.8 times shorter than in the planktonic cultures (FCC) with sodium lactate and with TMP, respectively. In ICC, the growth yields were lower (15.3(FCC) x 10(9) and 0.8(ICC) x 10(9) bacteria mg(-1) of sodium lactate; 50(FCC) x 10(9) and 35(ICC) x 10(9) bacteria mg(-1) of TMP). The mineralisation of substrates was estimated after mass spectrometric determination of the CO2 production rates of both free and immobilised cell cultures. The results indicated a higher specific CO2 production rate in the ICC with sodium lactate (3.1(FCC)+/-0.2 and 3.5(ICC)+/-0.3 nmol CO2 mg(-1) protein min(-1)) but not in the ICC with TMP (1.9(FCC)+/-0.7 and 0.5(ICC)+/-0.3 nmol CO2 mg(-1) protein min(-1)). The affinities for the two substrates were lower in the presence of the solid support (K(m,ICC)=18.2+/-0.2 microM and 37.1+/-2.0 microM, for sodium lactate and TMP, respectively) than without support (K(m,FCC)=8.5+/-1.5 microM and 8.4+/-1.2 microM, for sodium lactate and TMP, respectively). Moreover, the presence of a solid support showed a lower inhibition by the TMP (K(i,FCC)=3.8+/-1.0 microM and K(i,ICC)=12.2+/-2.5 microM) which may explain why the immobilised cell cultures degraded hydrophobic TMP more efficiently than the planktonic cultures.

Inhibition of proliferation of fibroblasts by lazaroids (21-aminosteroids).

Lazaroids (21-aminosteroids) are a novel group of compounds that inhibit lipid peroxidation in biological systems and protect cells from oxidative damage during tissue injury. In vivo efficacy of lazaroids has been demonstrated in a variety of animal models of traumatic or ischemic injury of central nervous system (1-6). However, very little is known about the affects of lazaroids on cellular responses associated with tissue repair and remodeling eg cell proliferation and synthesis of extracellular matrix. In the present study, we show that lazaroids and certain related compounds inhibit cell proliferation in vitro. Light microscopic examination and determination of release of intracellular enzyme, lactate dehydrogenase suggested that cell growth inhibition by lazaroids was not due to cell death resulting from cytotoxic effects. Examination of several lazaroid related antioxidants and other known antioxidants eg vitamin E and Probucol showed that compounds with similar antioxidant potential did not exhibit identical antiproliferative activity, suggesting that cell growth inhibition by lazaroids may be unrelated to their antioxidant action. These results may have implication in the therapeutic actions of lazaroids.

Dentinal permeability of the dog canine after exposure of a cervical cavity to the beam of a CO2 laser.

The dentinal permeability of the axial wall of a class V cavities in the canine of the dog was analyzed before and after its laser CO2 irradiation at two energy levels: 285 J per cm2 and 570 J per cm2. The sealing of the exposed dentin was verified by means of a pellet of cotton wool placed in contact with the axial wall and on which 20 microliters of a 50 mg per ml solution of noradrenaline was injected with a micrometric syringe. The transdentinal passage of noradrenaline induces a vasoconstriction of the pulp which is attested by a drop in its blood pressure. The absence of any variation in the pulpal pressure is an indication of the sealing to noradrenaline of the dentinal wall irradiated with an energy level of 570 J per cm2. This experimentation illustrates that it is possible to obtain dentinal sealing by means of laser irradiation without affecting the underlying pulp. The quality of this reduction in permeability is dependent upon the different factors which define the energy level.

In vitro study of a Nd:YAP laser in endodontic retreatment.

Nd:YAP laser is a dental laser with a 1340 nm wavelength. The laser beam is carried by a 200 to 300 microns fiberoptic and is suitable for endodontic therapy. We used the Nd:YAP laser in an in vitro experiment to study its effectiveness in endodontic retreatment. Temperature measurements and irradiation parameters were first defined. Then Nd:YAP laser irradiation was used, alone or in combination with hand instruments, to remove various canal sealers and broken instruments. Clinical parameters were monitored and scanning electron microscopic observations were conducted. When used at 200 mJ--with a pulse duration of 150 ms, an exposure time of 1 s and a frequency of 10 Hz--Nd:YAP laser preserved the dentinal walls of the root canal and enabled root canal retreatment without thermic elevation harming periodontal tissue. It is concluded that, in combination with hand instruments, the Nd:YAP laser is an effective device for root canal preparation in endodontic retreatment.

In vitro evaluation of the antibacterial effects of intracanal micro plasma system treatment.

Forty freshly extracted single-rooted teeth were prepared to a size 25 master apical file, autoclaved, and inoculated with a known quantity of Actinomyces naeslundii. The teeth were divided into four groups (n = 10), including an untreated control group. The three treatment groups were exposed to Micro Plasma System (MPS), 0.5% NaOCl and 0.5% NaOCl + MPS respectively. The content of each root canal was absorbed by sterile paper points, diluted in 2 ml Schaedler Broth Medium, and incubated on blood agar. The number of CFU was determined. Data analysis, using an analysis of variance and Scheffe's test at the 1% level (Statview II software), indicated a significant reduction in CFU count for the three treatment groups compared to the control group. For the three treatment groups, no significant intergroup differences were observed.

A field trial of the Clinicult system for detection of asymptomatic gonorrhea in women.

Clinicult, a selective medium for culturing Neisseria gonorrhoeae, was field-tested in a gonorrhea screening program in Seattle, Wash., in 1973. The results with this medium and with the Transgrow and Thayer-Martin culture systems were compared as to sensitivity and specificity. A total of 5,141 women from three patient groups were included in the study. Group 1 consisted of 720 female patients of the venereal disease clinic of the Seattle-King County Health Department, who served as the control group. When this group was screened with the Clinicult and Thayer-Martin culture media, the Thayer-Martin medium proved superior in identifying positive carriers. Group 2 was composed of approximately 2,000 patients from five different facilities, including family planning clinics and hospital out patient services. No statistical difference in accuracy was found between the two culture systems used for this group-Clinicult and Transgrow. Group 3 was comprised of approximately 2,500 female patients who were screened with the Clinicult and Transgrow cultures by their own private physician or his staff. The Clinicult system proved significantly less effective than the Transgrow cultures in identifying infected females in group 3. The physicians varied greatly in their ability to use the Clinicult system successfully. Possible reasons for their errors may have been (a) lack of motivation and of care by their office personnel in conducting the necessary additional procedures required with Clinicult, (b) the inhibitory nature of the medium, and (c) the failure of the medium to produce colonies of adequate size. The staffs of communitywide screening programs for gonorrhea need to be highly selective in choosing the medical facilities in which to use the Clinicult culture system. When laboratory facilities are available for the full utilization of the Thayer-Martin medium, this system is preferable. When, however, standard culture procedures are not readily available, Clinicult, properly used, can reduce the central laboratory load by eliminating the need for processing negative cultures.

Masked form priming in writing words from pictures: evidence for direct retrieval of orthographic codes.

Three experiments used the masked priming paradigm to investigate the role of orthographic and phonological information in written picture naming. In all the experiments, participants had to write the names of pictures as quickly as possible under three different priming conditions. Nonword primes could be: (1) phonologically and orthographically related to the picture name; (2) orthographically related as in (1) but phonologically related to a lesser degree than in (1); (3) orthographically and phonologically unrelated except for the first consonant (or consonant cluster). Orthographic priming effects were observed with a prime exposure duration of 34 ms (Experiments 1 and 2) and of 51 ms (Experiment 3). In none of the experiments, did homophony between primes and picture names yield an additional advantage. Taken together, these findings support the view of the direct retrieval of orthographic information through lexical access in written picture naming, and thus argue against the traditional view that the retrieval of orthographic codes of obligatorily mediated by phonology.

Influence of oxygen supply on heptadecane mineralization by Pseudomonas nautica.

The influence of different states of oxygen supply on heptadecane mineralization has been investigated in resting cell suspensions of Pseudomonas nautica. The rate of heptadecane biodegraded was constant for oxygen concentrations between 21% and 10% (v/v) (about 100% and 50% of air saturation, 230 and 110 microM, respectively). A decline in biodegradation rates occurred for oxygen concentrations below 10% (about 50% of air saturation, 110 microM) and biodegradation stopped with 0.21% oxygen (2.3 microM). In the presence of 1% (11 microM) of oxygen, no CO2 was produced, showing that complete mineralisation of heptadecane was blocked.

[Transfusion and alloimmunization in sickle cell anemia patients]. / Transfusion et alloimmunisation chez les patients drépanocytaires.

Transfusion therapy for sickle cell anemia is limited by the development of antibodies to red cell antigens. The aim of this study was to evaluate whether transfusion of blood matched for antigens Rh and Kell would reduce the incidence of alloimmunization. We determined the transfusion history, red cell phenotype and development of alloantibodies in 173 patients with sickle all anemia who received transfusions. Forty nine patients were transfused exclusively with frozen red blood cells (RBL) matched for antigens Rh and Kell; the rate of alloimmunization was 8.2%; antibodies to the Jkb, Jka, Fya and S were developed; 1 patient developed 2 antibodies. In a control group of 124 patients who received standard red blood cells, the rate of alloimmunization was significantly increased to 30.6% (p < 0.05); antibodies against C, E, K, Fya were the most frequently developed and 19 patients (16%) developed antibodies reacting with different antigens. In the 2 groups, alloimmunization occurred after receiving a significantly different number of transfusions: mean 9 in the patients transfused with matched RBC and 32 in the control group. The influence of the kinetics of transfusion was not demonstrated. To assess the effect that racial differences might have on alloimmunization, comparison of the red cell phenotype of patients with that of a panel of unselected blood bank donors was performed: the patients had a significant decrease in the frequency of red cell antigens corresponding to most of the detected alloantibodies JkB, C, S. Fyb, Fya and Kell.(ABSTRACT TRUNCATED AT 250 WORDS)

Spatio-temporal diversity of free-living and particle-attached prokaryotes in the tropical lagoon of Ahe atoll (Tuamotu Archipelago) and its surrounding oceanic waters.

Spatio-temporal variability of prokaryotic water column communities inside and outside a Polynesian tropical lagoon subjected to pearl oysters farming was assessed in terms of abundance by quantitative PCR and diversity by DGGE. Communities and operational taxonomic units (OTUs) were analysed according to dry/rainy seasons and free-living/particle-attached state. Bacterial density was higher in the lagoon compared to ocean and a seasonal trend was observed. No influence of the localisation within lagoon or of the planktonic/attached states was noticed on bacterial abundance and diversity. The OTUs belonged to Cyanobacteria, to heterotrophic groups in Proteobacteria and Flavobacteria. Archaeal abundance showed seasonal tendency and particle-prevalence, but no effect of lagoon or oceanic location was observed. Lagoon and oceanic archaeal diversity were different and Euryarchaeota (MG-II, MBG, and Halobacteria) were detected. During the dry season, planktonic and particle-associated community differed, whereas at rainy season, both communities were similar and included members usually associated with coral.

Impact of nonmyeloablative conditioning regimens on the occurrence of pure red cell aplasia after ABO-incompatible allogeneic haematopoietic stem cell transplantation.

BACKGROUND AND OBJECTIVES: In the setting of major ABO-incompatible allogeneic haematopoietic stem cell transplantation (HSCT), pure red cell aplasia (PRCA) is linked to the persistence of host residual plasma cells secreting antidonor isohaemagglutinins (HA) after transplantation. There are conflicting results regarding the impact of the intensity of conditioning regimen on the occurrence of PRCA after major ABO-mismatched HSCT. MATERIAL AND METHODS: To address this question, we compared two cases occurring after nonmyeloablative (NMA) and myeloablative (MA) HSCT and reviewed previous cases reported in the NMA setting. RESULTS AND CONCLUSIONS: We observed a delayed disappearance of antidonor HAs in the NMA setting, associated to a more prolonged period of red blood cells transfusion dependence than in the MA setting. In our case as in several others, the disappearance of antidonor HAs and resolution of PRCA were observed after reinforcement of the graft-versus-host effect (i.e. immunosuppression removal or donor leukocytes infusion).

Molecular, biochemical, and physiological approaches for understanding the ecology of denitrification.

One of the major challenges in microbial ecology for the future is to establish links between structural and functional biodiversity. This is particularly difficult when one is interested in a phylogenetically diversified function such as denitrification. The data banks are very rich in functional gene sequences (nirS in this study), but most of them were obtained from not yet cultivated bacteria, and thus must be supplemented by sequences of organisms from the environment for which we could associate a taxonomic position and physiological characteristics. Combined analysis including molecular (16S-rRNA or nirS genes), physiological, and biochemical approaches was carried out on a bacterial set of 89 strains isolated from marine sediment. The denaturing gradient gel electrophoresis (DGGE) technique was successfully applied on unclamped polymerase chain reaction (PCR) products of nirS genes to compare the picture of the biodiversity obtained with 16S rRNA and nirS genes. The diversity of nirS genes and denitrifier characteristics were found within several of the 16S rDNA phylotypes. In contrast, the nirS phylotypes were no diverse both with respect to 16S rDNA and to physiology and biochemistry of denitrification. Sequences of the nirS PCR products were very close to marine environmental clones and were analyzed within the same phylogenetic tree.

Writing words from pictures: what representations are activated, and when?

In three experiments, the nature of the representations involved in written picture naming and the time course of their activation were investigated. French participants had to produce picture names while hearing distractors. In Experiment 1, distractors semantically related to the picture names yielded a semantic interference effect when a stimulus onset asynchrony (SOA) of--150 msec, but not when a SOA of 0 msec, was used, in both spoken and written picture naming. Experiment 2 showed that the semantic interference effect was not located at the conceptual level. In Experiment 3, participants wrote down picture names while hearing semantically related, phonologically related, both semantically and phonologically related, or unrelated distractors, presented at both SOAs. A semantic interference effect was obtained with phonologically unrelated distractors but was eliminated with phonologically related distractors. Facilitatory effects of phonologically related distractors were found at both SOAs. The implications of the findings for written picture naming are discussed.