Prediction of thermal hydrolysis pretreatment on anaerobic digestion of waste activated sludge.

Thermal hydrolysis is known for an efficient sludge disintegration capability to enhance biogas potential--but to which extent? Obviously, residual VSS concentration in digested sludge gives not sufficient information to predict additional biogas potential. In this paper, different types of waste activated sludge (WAS) were pre-hydrolysed by a full-scale Thermo-Pressure-Hydrolysis Process (Thermo-Druck-Hydrolyse, TDH) and break-down mechanisms on specific organic compounds were investigated. The IWA Anaerobic Digestion Model No.1 (ADM1) has been used for a systematic analysis of monitoring data gained from experimental work. The TDH process combined with anaerobic digestion can be well described by a modified ADM1 model that includes an X(P)-fraction (inactivated aerobic biomass and their decay products). More rapid and more complete degradation of TDH-treated sludge is represented by calibrated disintegration rate and disintegration factors, while biokinetic parameters of acetogenesis and methanogenesis show no sensitivity. TDH process impacts mainly biomass and decay products while inerts Xi already contained in the raw wastewater are hardly converted. Final concentration of soluble inerts in digestion effluent has been increased from 2% to 9% of influent COD due to thermal hydrolysis. An increase in biogas generation (ca. +80%) and in ammonia release (ca. +75%) can be explained by complete degradation of cell mass.

[Single extramedullary plasmacytoma of the nose]. / Plasmocitoma solitario extramedular de localización nasal.

A new solitary extramedullary plasmacytoma of nasal localization meeting the conditions for definition is reported. The patient was treated with surgery and irradiation, which controlled the disease and produced excellent cosmetic results. The bibliography is reviewed and the pathologic, clinical, and evolutive characteristics are discussed.

Basic residues of human group IIA phospholipase A2 are important for binding to factor Xa and prothrombinase inhibition comparison with other mammalian secreted phospholipases A2.

Human secreted group IIA phospholipase A2 (hGIIA) was reported to inhibit prothrombinase activity because of binding to factor Xa. This study further shows that hGIIA and its catalytically inactive H48Q mutant prolong the lag time of thrombin generation in human platelet-rich plasma with similar efficiency, indicating that hGIIA exerts an anticoagulant effect independently of phospholipid hydrolysis under ex vivo conditions. Charge reversal of basic residues on the interfacial binding surface (IBS) of hGIIA leads to decreased ability to inhibit prothrombinase activity, which correlates with a reduced affinity for factor Xa, as determined by surface plasmon resonance. Mutation of other surface-exposed basic residues, hydrophobic residues on the IBS, and His48, does not affect the ability of hGIIA to inhibit prothrombinase activity and bind to factor Xa. Other basic, but not neutral or acidic, mammalian secreted phospholipases A2 (sPLA2s) exert a phospholipid-independent inhibitory effect on prothrombinase activity, suggesting that these basic sPLA2s also bind to factor Xa. In conclusion, this study demonstrates that the anticoagulant effect of hGIIA is independent of phospholipid hydrolysis and is based on its interaction with factor Xa, leading to prothrombinase inhibition, even under ex vivo conditions. This study also shows that such an interaction involves basic residues located on the IBS of hGIIA, and suggests that other basic mammalian sPLA2s may also inhibit blood coagulation by a similar mechanism to that described for hGIIA.

[Analysis of hemopoietic progenitor cells in bone marrow and peripheral blood samples which have undergone different periods of cryopreservation]. / Analisis de progenitores hemopoyéticos en muestras de medula osea y de sangre periférica en periodos variables de criopreservación.

Bone marrow (BM) and peripheral blood stem cell (PBSC) samples from patients undergoing autologous transplant were tested to evaluate the effects of cryopreservation. Cell viability was assessed as well as the proliferative capability of CFU-GM and BFU-E (myeloid and erythroid progenitors respectively). Moreover, long term culture (LTC) of stromal cells was used to test their functionality. A total of 23 samples were studied: 5 from AML patients, 7 MM, 6 NHL, 3 ALL and 2 HL. Nine patients received autologous bone marrow transplant (ABMT) and the remaining 14 PBSC. The cells were frozen during 24 to 33 days before infusion and 16 to 40 months before culture. Forty percent of AML and MM samples gave rise to colonies in vitro while the other hematology diseases tested showed colony growth in almost 100% of the cases. Samples from patients with lymphoid diseases exhibited a good correlation between the percentage of CD34+ cells and the number of colonies developed in culture. Nevertheless, there was no correlation when ALL and MM were tested suggesting that the underlying disease may have affected the growth in culture. The stromal layer was fully developed on BM samples, but on PBSC samples it only generated macrophages and fibroblasts. Our results suggest that the efficacy of cryopreservation of hematopoietic cells can be measured by means of CFU-GM and BFU-E culture and that the period the samples remained frozen did not affect the growth capability of the cells.

Organization and expression of the two homologous genes encoding the NADP-malate dehydrogenase in Sorghum vulgare leaves.

We previously described the isolation and the nucleotide sequence of a nuclear gene from sorghum (NMDHI; 4.6 kb) encoding the NADP-malate dehydrogenase. Further analysis led us to identify a second homologous gene (NMDH II; 4.8 kb) located within the same 12.3 kb genomic clone (lambda LM17); these two genes are tandemly organized, in direct orientation. This second gene was entirely sequenced and comparison with the first gene showed that the positions on the 14 exons and 13 introns are conserved in both genes. The analysis of the genomic organization and copy number in the Sorghum vulgare genome revealed that there are no additional homologues and there is only one copy each of NMDH I and NMDH II. The isolation of two different cDNA clones in a previous work suggested that both genes were probably expressed. Analysis of specific mRNA accumulation during the greening process using synthetic oligonucleotide probes showed that the NMDH I gene is induced in the presence of light while the NMDH II gene seems to be constitutively expressed at low level.

Primary structure of sorghum malate dehydrogenase (NADP) deduced from cDNA sequence. Homology with malate dehydrogenase (NAD).

Malate dehydrogenase (NADP) (NADP-MDH) is an important enzyme of the photosynthetic CO2 fixation pathway of C4 plants. We have isolated two clones from a sorghum lambda gt11 cDNA library (CM3, 932 bp, and CM7, 1441 bp). Nucleotide sequence analysis of the cDNAs CM3 and CM7 showed the existence of two NADP-MDH mRNA species encoding different enzyme subunits. Microsequencing of the N-terminus of the mature protein indicated that a specific cleavage of 13 amino acids occurred during the purification steps of the enzyme. The full-length cDNA CM7 contains a large open reading frame encoding an NH2-terminal transit peptide of 40 amino acids and a mature protein of 389 amino acids (42.207 kDa). Alignment of the NADP-MDH sequence with those of several malate dehydrogenases revealed some similarities with NAD-MDHs.

Structure and characterization of the Sorghum vulgare gene encoding NADP-malate dehydrogenase.

A genomic clone encoding the NADP-malate dehydrogenase precursor has been isolated from a Sorghum vulgare lambda EMBL4 library using a cDNA probe. The entire structure of this nuclear gene (4.6 kb) encoding the 429-amino acid precursor is reported, as well as 602 bp of the 5'-flanking and 695 bp of the 3'-flanking regions. The gene is interrupted by 13 introns from 79 to 495 bp in length. S1 nuclease mapping showed the S. vulgare gene transcript to contain an untranslated leader sequence of 44 nucleotides. In the 5'-flanking region, several short sequences similar to the consensus motifs involved in the light-regulation process of other plant genes were found.

Identification of a cDNA clone for sorghum leaf malate dehydrogenase (NADP). Light-dependent mRNA accumulation.

The mechanisms underlying the photoregulation of the synthesis of sorghum leaf malate dehydrogenase (NADP) (EC 1.1.1.82) (NADP-MDH), a key enzyme in C4 photosynthesis, have been investigated. During the greening process a light-dependent increase in enzyme activity took place, accompanied by de novo synthesis of the protein. In vitro translation experiments showed that this chloroplastic protein is synthesized as a precursor (46 kDa) with a 'transit peptide' of about 2.5 kDa. A large increase in NADP-MDH-translatable RNAs was also observed during greening. We describe also the construction and characterization of a cDNA clone for NADP-MDH (pCM18A) in the expression vector lambda gt11. The use of this homologous probe demonstrated a light-dependent mRNA accumulation.