DNA of some anaerobic rumen fungi: G + C content determination

The nuclear DNAs from five species of anaerobic rumen fungi have been isolated and purified by means of two extraction methods (with and without 8 M urea). Their G + C contents have been characterized by the thermal denaturation procedure of Marmur and Doty. As has already been shown in Neocallimastix frontalis, the results obtained by the two techniques demonstrated a very low G + C content (less than 20%) and the constant presence of satellite DNA.     

The biosynthesis of polyunsaturated fatty acids by rat sertoli cells.

1. The biosynthesis of polyunsaturated fatty acids (PUFA) of the n-6 and n-3 series was investigated in cultured Sertoli cells. 18:2n-6, 18:3n-6, 20:2n-6, 18:3n-3 and 20:3n-3 were added individually at a concentration of 20 mumol to culture media. 2. Maximum incorporation of 20- and 22-carbon PUFA into membrane lipids was observed after 72 hr of incubation with all the exogenous substrates used. 3. As reported in other cell systems, the delta 6 desaturation was the first rate-limiting step; the major factor regulating this activity was the concentration of linoleic acid or alpha-linolenic acid in the medium. 4. Our data show that the delta 5-desaturation represents a second regulatory step in PUFA biosynthesis. 5. The sum of n-6 and n-3 PUFA of the 22 carbon chain length constantly represented between 11 and 12% of total fatty acids, regardless of the exogenous substrate used. 6. Our kinetic studies of the incorporation of PUFA of the n-6 and n-3 series did not permit detection of a delta 8 desaturase activity.     

Genetic evidence in support of a shared Eurasian-North African dairying origin

The process by which pastoralism and agriculture spread from the Fertile Crescent over the past 10,000 years has been the subject of intense investigation by geneticists, linguists and archaeologists. However, no consensus has been reached as to whether this Neolithic transition is best characterized by a demic diffusion (with a significant genetic input from migrating farmers) or a cultural diffusion (without substantial migration of farmers). Milk consumption and thus lactose tolerance are assumed to have spread with pastoralism and we propose that by looking at the relevant mutations in and around the lactase gene in human populations, we can gain insight into the origin(s) and spread of dairying. We genotyped the putatively causal allele for lactose tolerance (–13910T) and constructed haplotypes from several polymorphisms in and around the lactase gene (LCT) in three North African Berber populations and compared our results with previously published data. We found that the frequency of the –13910T allele predicts the frequency of lactose tolerance in several Eurasian and North African Berber populations but not in most sub-Saharan African populations. Our analyses suggest that contemporary Berber populations possess the genetic signature of a past migration of pastoralists from the Middle East and that they share a dairying origin with Europeans and Asians, but not with sub-Saharan Africans.     

Charting the Irreversible Degradation Modes of Low Bandgap Pb-Sn Perovskite Compositions for De-Risking Practical Industrial Development

The commercialization of a solar technology necessitates the fulfillment of specific requirements both regarding efficiency and stability to enter and gain space in the photovoltaic market. These aims are heavily dependent on the selection of suitable materials, which is critical for suppressing any reliability risks arising from inherent instabilities. Focusing on the absorber material, herein the most suitable low bandgap lead-tin composition candidate for all-perovskite tandem applications is investigated by studying their degradation mechanisms with both widely available and advanced characterization techniques. Three irreversible degradation processes are identified in narrow bandgap Pb-Sn perovskite absorbers: 1) Tin (Sn) oxidation upon air exposure, 2) methylammonium (MA) loss upon heat exposure, and 3) formamidinium (FA) and cesium (Cs) segregation leading to impurity phase formation. From an industrial perspective, it is proposed to refocus attention on FASn_0.5Pb_0.5I₃ which minimizes all three effects while maintaining a suitable bandgap for a bottom cell and good performance. Moreover, a practical and highly sensitive characterization method is proposed to monitor the oxidation, which can be deployed both in laboratory and industrial environments and provide useful information for the technological development process, including, the effectiveness of encapsulation methods, and the acceptable time windows for air exposure.     

On the validity of time‐dependent AUC estimation in the presence of cure fraction

During the last decades, several approaches have been proposed to estimate the time‐dependent area under the receiver operating characteristic curve (AUC) of risk tools derived from survival data. The validity of these estimators relies on some regularity assumptions among which a survival function being proper. In practice, this assumption is not always satisfied because a fraction of the population may not be susceptible to experience the event of interest even for long follow‐up. Studying the sensitivity of the proposed estimators to the violation of this assumption is of substantial interest. In this paper, we investigate the performance of a nonparametric simple estimator, developed for classical survival data, in the case when the population exhibits a cure fraction. Motivated from the current practice of deriving risk tools in oncology and cardiovascular disease prevention, we also assess the loss, in terms of predictive performance, when deriving risk tools from survival models that do not acknowledge the presence of cure. The simulation results show that the investigated method is valid even under the presence of cure. They also show that risk tools derived from survival models that ignore the presence of cure have smaller AUC compared to those derived from survival models that acknowledge the presence of cure. This was also attested with a real data analysis from a breast cancer study.     

Characterization of the non-sulphated highly anionic kurloff cell glycoconjugates by lectin- and immunoblotting: Evidence for the presence of α(2,8)-linked polysialic acid-bearing molecules

Summary— Urea or guanidine hydrochloride-soluble extracts from highly purified Kurloff cells (KC) radiolabelled in vitro were subjected to DEAE-cellulose chromatography. Among the three anionic peaks obtained, a major and non-sulphated peak (designated as peak IV) strongly affected by glucosamine-labelling and eluted at about 0.3 M NaCl was analyzed. Gel filtration on Sepharose CL4B and 10% SDS-PAGE indicated its heterogeneous size. Peak IV consisted mainly of N-glycans as shown by its susceptibility to tunicamycin. Further insight into its chemical nature was obtained by examining its binding capacity to different lectins and by immunodot analysis. It strongly interacted with concanavalin A (Con A) after dot-blot or Western blotting. A large amount of these glycoproteins is not of the high-mannose type since Galanthus nivalis agglutinin reacted weakly with peak IV. Moreover, bindings to Phaseolus vulgaris and to wheat germ agglutinins suggest the presence of bisecting N-acetylglucosamine residues. Bindings to Sambucus nigra and to Ricinus communis agglutinins, dramatically lessened and increased respectively after desialylation, suggest the presence of Neu5Acα2,6Gal/GalNAc sequences. The absence of outer sialic acid residues linked α2,3 to galactose was demonstrated following Maackia amurensis agglutinin negativity. The use of poly(α2,8-sialyl) endo-N-acylneuraminidase combined with immunodot procedure with a monoclonal antibody that specifically recognizes α2,8-linked polysialic chains revealed that peak IV contains oligosac-charidic epitopes common to polysialyated neural cell adhesion molecules.     

Metabolic utilization of linoleate and alpha-linolenate in cultured Sertoli cells.

1. The capacity of cultured Sertoli cells to synthesize long-chain polyunsatured fatty acids (PUFA) from the essential fatty acid (EFA) precursors 18:2 n-6 and 18:3 n-3 was tested, and the concentrations of each EFA required to obtain maximal incorporation into membrane lipids were determined. 2. The two EFA were added to the culture medium as free fatty acids complexed to albumin in a molar ratio of 12:1. 3. When the substrates were added individually, the maximal levels of biosynthesis were obtained with 0.7 micrograms/ml of 18:2 n-6 and 2 micrograms/ml of 18:3 n-3. 4. When the two EFA were added together, clear alterations in the behavior of the desaturases with regard to the n-6 and n-3 fatty acids were observed. 5. It was found that a concentration of 0.35 micrograms/ml of each EFA represented the "ideal" required level in order to ensure optimal incorporation of 22-carbon PUFA into the membrane lipids. 6. These results provide the first data on the definition of EFA requirements for Sertoli cells in culture.     

The major Kurloff cell glycoproteins: lectin affinities, glycosidase susceptibilities and relationship with the sialylated acid phosphatases of the Kurloff body.

Urea-soluble fractions from purified Kurloff cells (KC) were analysed by affinoblotting. Lectin reactivities were quasi-exclusively confined to the 30-35 kDa major glycoproteins (mGPs) (responsible for the PAS positivity of the Kurloff body) with strong affinities for Canavalia ensiformis lectin, Phaseolus vulgaris erythroagglutinin and Sambucus nigra (SNA), Pisum sativum, Triticum vulgaris and Ulex europeus agglutinins. These data were consistent with the presence, among the KC mGPs, of large amounts of complex or hybrid N-glycosylproteins, in particular with Neu5Ac alpha 2,6Gal/GalNAc sequences, fucosyl residues and bisected residues. Their oligosaccharide sequences belong to more than one class, since some of these lectin reactivities had to be borne by distinct N-linked oligosaccharide chains. Before further analysis, KC mGPs were separated from other highly anionic glycoconjugates, by DEAE-cellulose chromatography. Their abundant potential RCA-binding sites masked by sialic acid were then revealed after neuraminidase (sialidase) or dilute acid pre-treatment. In remaining consistent with their lectin affinities, some KC mGPs were found to be PNGase F sensitive, while, either desialylated or not, they were all O-glycanase insensitive. Finally, by combined zymography and affinoblotting, the SNA-reactive fraction of KC mGPs was shown to correspond to denatured forms of the two zymographic size populations (190 kDa and 500 kDa) of KC acid phosphatases.     

Identification of the D2--Dopamine Receptor Binding Subunit in Several Mammalian Tissues and Species by Photoaffinity Labeling

Photoaffinity labeling of the D2-dopamine receptor in plasma membrane preparations of various tissues from several mammalian species was performed using the recently developed D2-dopaminergic antagonist probe [125I]N-(p-azidophenethyl)spiperone ([125I]N3-NAPS). In tissues containing D2-receptors such as the corpus striatum from rat, dog, calf, hamster, guinea pig, and rabbit as well as the anterior pituitary of rat, bovine, and hamster, the probe covalently labels a peptide of Mr = 94,000. Specificity of the labeling is typically D2-dopaminergic in character. The covalent labeling is blocked by (+)-butaclamol but not by the inactive (-)isomer. Agonists block incorporation with the order of potency: N-n-propylnorapomorphine greater than apomorphine greater than dopamine. The D2-selective antagonist spiperone blocks labeling of the Mr = 94,000 peptide whereas the D1-selective antagonist SCH-23390 is ineffective. Thus, these results indicate that the ligand binding subunit of the D2-dopamine receptor resides on a Mr = 94,000 peptide in these various tissues from several species. Under conditions where proteolysis is not stringently controlled, peptides of lower Mr (32-38,000) are labeled at the expense of the Mr = 94,000 peptide. The most efficient protease inhibitor tested in these systems was EDTA, suggesting that the generation of these lower Mr receptor fragments might be the result of a metal-dependent proteolysis in the membrane preparations. In the rat neurointermediate lobe, a tissue containing D2-receptors, [125I]N3-NAPS specifically labels a major peptide of Mr approximately equal to 120,000 in addition to the Mr = 94,000 peptide.(ABSTRACT TRUNCATED AT 250 WORDS)