Evaluation of rice straw hemicellulose hydrolysate in the production of xylitol byCandida guilliermondii

Summary Rice straw was used as a lignocellulosic source to provide rich pentose media. By using a well characterized yeast strain,Candida guilliermondii FTI 20037, the hydrolysate obtained was converted to xylitol with an efficiency of 75% and production of 27 g of xylitol per liter in 48 hours. The satisfactory results reported here can be attributed to the low concentrations of toxic components generated throughout the chemical depolymerization of this raw material.     

A pre-formed Pyrogenic Factor Released by Lipopolysaccharide Stimulated Macrophages

The aim of this study was to investigate the pyrogenic activity of factor(s) released by rat peritoneal macrophages following a brief stimulation with LPS. The effect of this factor on the number of circulating leukocytes and serum Fe, Cu and Zn levels, was also evaluated. The possibility that the content of interleukin (IL)-1beta, IL-6 and tumour necrosis factor (TNF) in the supernatant could explain the observations was investigated. Supernatant produced over a period of 1 h by peritoneal macrophages, following a 30 min incubation with LPS at 37 degrees C, was ultrafiltered through a 10 000 MW cut-off Amicon membrane, sterilized, and concentrated 2.5, 5, 10 and 20 times. The intravenous (i.v.) injection of this supernatant induced a concentration-dependent fever in rats with a maximal response at 2 h. The pyrogenic activity was produced by macrophages elicited with thioglycollate and by resident cells. The supernatants also induced neutrophilia and reduction in Fe and Zn 6 h after the injection. Absence of activity in boiled supernatants, or supernatants from macrophages incubated at 4 degrees C with LPS, indicates that LPS was not responsible for the activity. In vitro treatment with indomethacin (Indo), dexamethasone (Dex), or cycloheximide (Chx) did not modify the release of pyrogenic activity into the supernatant or its effects on the reduction in serum metal levels. Although Chx abolished the production of mediator(s) inducing neutrophilia, and Dex reduced the induction of IL-1beta, TNF and IL-6, injection of the highest concentration of these cytokines detected in the supernatants did not induce fever. In vivo treatment with Dex, but not Indo, abolished the fever induced by the supernatant. These results suggest that macrophages contain pre-formed pyrogenic mediator(s), not related to IL-1beta, IL-6 or TNF, that acts indirectly and independently of prostaglandtn. It also seems likely that the pyrogenic activity is related to the factor responsible for the reduction of serum Fe and Zn levels, but not the neutrophilia.     

Tyrosine attack by free radicals derived from catalytic decomposition of carbon tetrachloride

The interaction between free radicals derived from the catalytic decomposition of carbon tetrachloride and tyrosine (the N-acetyl tyrosine ethyl ester, ATEE) under anaerobic and aerobic conditions was studied. The structure of the reaction products formed was desciphered by the GLC/MS analysis of their trimethylsilyl derivatives. Under anaerobic conditions the formation of the following products was found: (1) an unsaturated derivative of the amino acid; (2) the trimethylsilyl derivative of N-acetyl chloro tyrosine ethyl ester; (3) a hydroxyl adduct of ATEE ; (4) an ATEE adduct having a chlorine and a CCl₃ group in the molecule (it is suggested that CCl₃ is attached to the benzyl carbon and the chlorine located in the benzene ring); (5) an ATEE adduct having only a CCl₃ group tentatively assigned to be located on the benzyl carbon; and (6) and (7) were found to be two isomers of an ATEE having one CCl₃ on the aromatic ring. Under aerobic conditions the following reaction products were identified: Two products which were similar to those numbered (1) and (2) and formed anaerobically; (8) and (11) two isomeric dichlorinated adducts of ATEE; (9) and (10) two isomeric dichlorinated monohydroxylated derivatives of ATEE. Concerning the potential relevance of these findings, we consider that if similar interactions to those here reported occurred during CCl₄ poisoning, the activity of enzymes having tyrosine in their active center might result in impairment. Further, enzymes operating on tyrosine moieties in proteins might be perturbed in their action if tyrosine groups were attacked by the free radicals arising from catalytic decomposition of CCl₄ evidenced here.     

Effect of lipid supplements on the production and glycosylation of recombinant interferon-γ expressed in CHO cells

The effects of lipids on the glycosylation of recombinant human interferon- expressed in a Chinese Hamster Ovary cell line were investigated in batch culture. Lipids form an essential part of the N-glycosylation pathway, and have been shown to improve cell viability. In control (serum-free) medium the proportion of fully-glycosylated interferon- deteriorated reproducibly with time in batch culture, but the lipoprotein supplement ExCyte was shown to minimise this trend. Partially substituting the bovine serum albumin content of the medium with a fatty-acid free preparation also improved interferon- glycosylation, possibly indicating that oxidised lipids carried on Cohn fraction V albumin may damage the glycosylation process.Abbreviations BSA bovine serum albumin - CHO chinese hamster ovary - DHFR dihydrofolate reductase - FCS foetal calf serum - IFN- human interferon-gamma - q _IFN specific interferon production rate - specific growth rate - 2N doubly-gycosylated - 1N singly-glycosylated - ON non-glycosylated     

Coastal intrusion of copepods: mechanisms and consequences on the population biology of Rhincalanus nasutus

In contrast to its usual habitat as a copepod that may occurin shelf or slope waters with low oxygen content between depthsfrom the surface to >2000 m, Rhincalanus nasutus was foundin a shallow (<60 m) embayment, the Arauco Gulf in Chile.Here, we document the complex life history of this copepod wheredid and ontogenetic vertical migratory behavior was co-ordinatedwith the circulation pattern which helped to retain a summerresident population in the vicinity of the gulf. With the onsetof the upwelling season in the southern summer, the mid-depthEquatorial Sub Surface Waters intrude into the gulf, leadingto the formation of a strong thermocline at 10–20 m andthe development of a two-layer circulation pattern. CopepoditesI, C.II and C.III (first stage to exhibit migratory behavior)were found within the gulf in the layer where the net transportwas at a minimum while chlorophyll-a concentrations were ata maximum. Older stages (C.IV-C.VI females and males) migratefrom their daytime depth in the bottom, shoreward-moving, low-oxygenlayer, to their night-time depth in the shallow seaward-movinglayer. The population of copepods retained in the area thuslyreproduce, as reflected in the sequential pulses of differentdevelopmental stages. Because coastal intrusions such as thisof R.nasutus have been documented for several other speciesduring the seasons of maximum phytoplankton production (upwelling),they may form part of a more widespread reproductive strategyof the larger zooplankton of coastal upwelling systems thanpreviously suspected.     

Effect of copper sulfate on experimental atherosclerosis

Serum copper concentration increases significantly (p<0.01) in rats with experimental atherosclerosis compared to a control group. The serum zinc, the zinc, and copper concentration in abdominal aorta and in liver decreases significantly (p<0.05) compared to the control group. Administration of copper sulfate for 100 d in these animals induces a significant increase of serum copper (p<0.01), decrease of serum cholesterol (p<0.05) and increase of liver copper concentration as compared with the group fed only a high cholesterol diet. In the aorta of these animals the copper concentration increases and edema and lipid infiltration are considerably less than in the group of animals fed only a high lipid diet.     

Influence of oxygen on ethanol and xylitol production by xylose fermenting yeasts

The behaviour of Pichia stipitis, Pachysolen tannophilus, Candida shehatae and Candida parapsilosis was investigated to select the most suitable yeast to convert xylose either to ethanol or to xylitol, with little or no formation of by-products. The aeration rate was used as a variable parameter. P. stipitis and C. parapsilosis were the most effective producers or ethanol and xylitol, respectively, both reaching productivities at very low levels of oxygenation. With P. stipitis, better ethanol productivity was attained under microaerobic conditions (K_La = 4·8 h⁻¹) while with C. parapsilosis high yields and rates of xylitol production were detected at K_La values of about 16·3 h⁻¹. P. tannophilus and C. shehatae showed lower performances under all conditions used while changes in oxygenation modified the ratio of ethanol to xylitol produced by these yeasts, suggesting that they are more dependent on the oxygen power input than P. stipitis and C. parapsilosis. The influence of oxygen transfer rates on ethanol and xylitol formation with the best producers is discussed.     

K-Opioid Agonist Modulation of [3H]Thymidine Incorporation into DNA: Evidence for the Involvement of Pertussis Toxin-Sensitive G Protein-Coupled Phosphoinositide Turnover

Abstract: A body of evidence has indicated that μ-opioid agonists can inhibit DNA synthesis in developing brain. We now report that K -selective opioid agonists (U69593 and U50488) modulate [³H]thymidine incorporation into DNA in fetal rat brain cell aggregates in a dose- and developmental stage-dependent manner. K agonists decreased thymidine incorporation by 35% in cultures grown for 7 days, and this process was reversed by the K -selective antagonist, norbinaltorphimine, whereas in 21-day brain cell aggregates a 3,5-fold increase was evident. Cell labeling by [³H]thymidine was also inhibited by the K -opioid agonist as shown by autoradiography. In addition, U69593 reduced basal rates of phosphoinositide formation in 7-day cultures and elevated it in 21-day cultures. Control levels were restored by norbin-altorphimine. Pertussis toxin blocked U69593-mediated inhibition of DNA synthesis. The action of K agonists on thymidine incorporation in the presence of chelerythrine, a protein kinase C (PKC) inhibitor, or in combination with LiCl, a noncompetitive inhibitor of inositol phosphatase, was attenuated in both 7- and 21-day cultures. These results suggest that K agonists may inhibit DNA synthesis via the phosphoinositide system with a pertussis toxin-sensitive G protein as transducer. In mixed glial cell aggregates, U50488 increased thymidine incorporation into DNA 3.1-fold, and this stimulation was reversed by the opioid antagonist naltrexone.     

Aotus from the southwestern Amazon region is geographically and chromosomally intermediate betweenA. azarae boliviensis andA. infulatus

We described the karyotypes detected in 23Aotus specimens captured on Southwestern Amazon region (63°30'W, 9°00'S). This region should be occupied byA. nigriceps. However, the animals described here have a fur pattern and a karyotypes similar to those ofA. azarae boliviensis andA. infulatus. The females presented 50 chromosomes and the males 49 as a consequence of fusion between the chromosome Y and an autosome. This sample presented polymorphisms of G- and C-banding and NOR-staining. These animals are karyotypically intermediate betweenA. a. boliviensis andA. infulatus. The taxonomy and karyotypic evolution of these taxa are discussed.     

New method for peptide purification based on selective removal of truncation peptide impurities after SPPS with orthogonal capping

Peptide purification by high-performance liquid chromatography (HPLC) is associated with high solvent consumption, relatively large effort and lack of efficient parallelization. As an alternative, many catch-and-release (c&r) purification methods have been developed over the last decades to enable the efficient parallel purification of peptides originating from solid-phase peptide synthesis (SPPS). However, with one exception, none of the c&r systems has been widely established in industry and academia until today. Herein, we present an entirely new chromatography-free purification concept for peptides synthesized on a solid support, termed reactive capping purification (RCP). The RCP method relies on the capping of truncation peptides arising from incomplete coupling of amino acids during SPPS with a reactive tag. The reactive tag contains a masked functionality that, upon liberation during cleavage from the resin, enables straightforward purification of the peptide by incubation with a resin-bound reactive moiety. In this work, two different reactive tags based on masked thiols were developed. Capping with these reactive tags during SPPS led to effective modification of truncated sequences and subsequent removal of the latter by chemoselective reaction with a maleimide-functionalized solid support. By introducing a suitable protecting group strategy, the thiol-based RCP method described here could also be successfully applied to a thiol-containing peptide. Finally, the purification of a 15-meric peptide by the RCP method was demonstrated. The developed method has low solvent consumption, has the potential for efficient parallelization, uses readily available reagents, and is experimentally simple to perform.