Potentiation of GSK-3-catalyzed Alzheimer-like phosphorylation of human tau by cdk5

Tau protein from Alzheimer disease (AD) brain is hyperphosphorylated by both proline-dependent protein kinases (PDPKs) and non-PDPKs. It is presently unclear how PDPKs and non-PDPKs interact in tau hyperphosphorylation. Previously we have shown that non-PDPKs can positively modulate the activity of a PDPK (GSK-3) in tau phosphorylation (Singh et al. (1995) FEBS Lett. 358, 267-272). In this study we have investigated whether (A) non-PDPKs can also modulate the activity of the PDPK, cdk5, (B) a PDPK can modulate the activities of another PDPK, as well as non-PDPKs. We found that, like GSK-3, the activity of cdk5 is stimulated if tau were first prephosphorylated by any of several non-PDPKs (A-kinase, C-kinase, CK-1, CaM-kinase II). Prephosphorylation of tau by cdk5 stimulated both the rate and extent of a subsequent phosphorylation catalyzed by GSK-3. Under these conditions thr 231 phosphorylation was especially enhanced (9-fold). No significant stimulation of phosphorylation was obser ved when the order of these kinases was reversed (i.e. GSK-3 followed by cdk5). By contrast, prephosphorylation of tau by cdk5 served to inhibit subsequent phosphorylation catalyzed by C-kinase and CK-1, but not by A-kinase or CaM-kinase II. Our results suggest that in tau hyperphosphorylation in AD brain, cdk5-catalyzed phosphorylation may serve to up-regulate the activity of GSK-3 and down-regulate the activities of C-kinase and CK-1. (Mol Cell Biochem 167: 99-105, 1997)     

Involvement of Trihydroxyconjugated Bile Salts in Cholesterol Assimilation by Bifidobacteria

To determine the conditions of cholesterol assimilation, various strains of Bifidobacterium species were cultured in the presence of cholesterol and bile salts. During culturing, Bifidobacterium breve ATCC 15700 assimilates cholesterol in the presence of oxgall at pH values lower than 6. This strain was selected to study the influence of conjugated (taurocholic acid) and deconjugated (cholic acid) bile salts on cholesterol assimilation. B. breve ATCC 15700 assimilated cholesterol (up to 51%) when cultures were undertaken in the presence of taurocholic acid, whereas less than 13% of the initial amount of cholesterol was measured in the cells in the presence of cholic acid. Cultured in the presence of six individual di- or trihydroxyconjugated bile salts, bifidobacteria strains assimilated cholesterol. This assimilation appeared to be more important in the presence of trihydroxyconjugated bile salts (tauro- and glycocholic acids). It is concluded that trihydroxyconjugated bile salts are involved in the assimilation of cholesterol by bifidobacteria. Received: 20 June 1996 / Accepted: 19 July 1996     

Leaching of chalcopyrite byThiobacillus thiooxidans and oxidized copper ore byThiobacillus ferrooxidans isolated from local environments

Summary Feasibility for bacterial leaching of two different copper ores from Pakistan was investigated with locally isolated, acidophilic thiobacilli in shake flask culture at 28±2°C. After a lag period of 16 d,Thiobacillus thiooxidans solubilized up to 19% of copper present in chalcopyrite ore in 14 d before entering stationary phase. In oxidized copper ore amended with chalcopyrite ore and supplemented with sulphur to provide an energy source forT. ferrooxidans, the pH dropped as a result of bacterial growth and was accompanied by copper solubilization. The highest copper release, 61% of total, occurred in flasks having oxidized copper ore, sulphur and calcopyrite (111, by weight) and inoculated withT. ferrooxidans. Supplementation used in these tests resulted in nearly six times more solubilization as compared to tests conducted without supplementation.

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Resumen Se ha estudiado la posibilidad de realizar un lixiviado mediante bacterias de dos minerales de cobre del Pakistan. Las bacterias utilizadas son cepas, aísladas localmente, de tiobacilos acidófilos que se hacen crecer en un medio líquido en agitación a 28±2°C. Después de una fase lag de 16 dThiobacillus thiooxidans solubilizó hasta 19% del cobre presente en la calcopirita en 14 d, antes de alcanzar la fase estacionaria. El mineral de cobre oxidado se modificó con calcopirita y se suplementó con azufre afín de sumministrar una fuente de energía paraT. ferrooxidans, a resultas del crecimiento bacteriano el pH disminuyó solubilizándose cobre. La mayor cantidad de cobre sulubilizado se cencontró en frascos con mineral de cobre oxidado, azufre y calcopirita (111 en peso) e inoculados conT. ferrooxidans. La adición de suplementos incrementó 6 veces la solubilización comparada con la obtenida en ensayos sin adición de suplementos.

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Résumé On a testé la faisabilité de la lixiviation bactérienne de deux minerais différents de cuivre du Pakistan au moyen de thiobacilli acidophiles,isolés localement, en culture en flaçons agités à 28 ±2°C. Après une période de latence de 16 jours,Thiobacillus thiooxidans a solubilisé jusqu'à 19% du cuivre présent dans le minerai de chalcopyrite en 14 jours avant d'entrer en phase stationnaire de croissance. Dans un minerai de cuivre amendé par le minerai de chalcopyrite et additionné de soufre de manière à fournir une source d'énergie pourT. ferrooxidans, le pH a baissé comme suite de la croissance bactérienne, ce qui a entraîné la solubilisation du cuivre. La libération la plus élevée de cuivre, 61% au total, a eu lieu dans les flaçons qui avaient à oxyder le minerai de cuivre, le soufre et la chalcopyrite oans les proportions de 111 en poids, et qui avaient été inoculés parT. ferrooxydans. La supplémentation effectuée dans ces tests a eu pour effet de dissoudre six fois plus de cuivre que dans les tests conduits sans supplémentation.

     

Degradation of different hydrocarbons and production of biosurfactant byPseudomonas aeruginosa isolated from coastal waters

Summary A bacterium isolated from an oil spillage sample and identified asPseudomonas aeruginosa degraded hexadecane and heptadecane by 47% and 58% with 9% and 12% of total carbon from the respective substrates being liberated as CO₂. With octadecane and nonadecane as substrates, 73% and 60% were biodegraded while 27% and 25% of total carbon was evolved as CO², respectively. Production of biosurfactant by this bacterium was studied using hexadecane 5% (v/v) as substrate. The surface tension of spent culture medium, as well as the supernatant, was 30 mN/m compared to 71 mN/m for water. When the supernatant was mixed with hexadecane (15, v/v) a stable emulsion was formed which deteriorated only by 10% after one month.

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Dégradation de différents hydrocarbures et production de biosurfactants parPseudomonas aeruginosa isolé d'eaux côtières

Résumé Une bactérie, isolée d'un échantillon d'hydrocarbures épandus, et identifiée commePseudomonas aeruginosa, dégrade l'hexaet l'heptadécane à raison respective de 47 et de 58% du carbone total avec 9 et 12% des substrats respectifs libérés sous la forme de CO₂. Avec l'octa- et le nonadécane comme substrats, la biodégradation atteint respectivement 73 et 60% avec 27 et 25% du carbone total transformé en CO₂. On a étudié la production de biosurfactant par cette bactérie en utilisant l'hexadécane à 5% (v/v) comme substrat. La tension superficielle de la liqueur mixte, comme celle de son surnageant atteint 30 mN/m par comparaison à 71 mN/m pour l'eau. Lorsque le surnageant est mélangé à l'hexadécane (15, v/v), on forme une émulsion stable qui ne se détériore que de 10% au bout d'un mois.

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This work was carried out at the Biotechnology Laboratories of the Nuclear Institute for Agriculture & Biology (NIAB), Faisalabad.     

Patient and System-Related Delays of Emergency Medical Services Use in Acute ST-Elevation Myocardial Infarction: Results from the Third Gulf Registry of Acute Coronary Events (Gulf RACE-3Ps)

Background

Little is known about Emergency Medical Services (EMS) use and pre-hospital triage of patients with acute ST-elevation myocardial infarction (STEMI) in Arabian Gulf countries.

Methods

Clinical arrival and acute care within 24 h of STEMI symptom onset were compared between patients transferred by EMS (Red Crescent and Inter-Hospital) and those transferred by non-EMS means. Data were retrieved from a prospective registry of 36 hospitals in 6 Arabian Gulf countries, from January 2014 to January 2015.

Results

We enrolled 2,928 patients; mean age, 52.7 (SD ±11.8) years; 90% men; and 61.7% non-Arabian Gulf citizens. Only 753 patients (25.7%) used EMS; which was mostly via Inter-Hospital EMS (22%) rather than direct transfer from the scene to the hospital by the Red Crescent (3.7%). Compared to the non-EMS group, the EMS group was more likely to arrive initially at a primary or secondary health care facility; thus, they had longer median symptom-onset-to-emergency department arrival times (218 vs. 158 min; p˂.001); they were more likely to receive primary percutaneous coronary interventions (62% vs. 40.5%, p = 0.02); they had shorter door-to-needle times (38 vs. 42 min; p = .04); and shorter door-to-balloon times (47 vs. 83 min; p˂.001). High EMS use was independently predicted mostly by primary/secondary school educational levels and low or moderate socioeconomic status. Low EMS use was predicted by a history of angina and history of percutaneous coronary intervention. The groups had similar in-hospital deaths and outcomes.

Conclusion

Most acute STEMI patients in the Arabian Gulf region did not use EMS services. Improving Red Crescent infrastructure, establishing integrated STEMI networks, and launching educational public campaigns are top health care system priorities.     

Origin and Evolution of Rickettsial Plasmids

Background

Rickettsia species are strictly intracellular bacteria that have undergone a reductive genomic evolution. Despite their allopatric lifestyle, almost half of the 26 currently validated Rickettsia species have plasmids. In order to study the origin, evolutionary history and putative roles of rickettsial plasmids, we investigated the evolutionary processes that have shaped 20 plasmids belonging to 11 species, using comparative genomics and phylogenetic analysis between rickettsial, microbial and non-microbial genomes.

Results

Plasmids were differentially present among Rickettsia species. The 11 species had 1 to 4 plasmid (s) with a size ranging from 12 kb to 83 kb. We reconstructed pRICO, the last common ancestor of the current rickettsial plasmids. pRICO was vertically inherited mainly from Rickettsia/Orientia chromosomes and diverged vertically into a single or multiple plasmid(s) in each species. These plasmids also underwent a reductive evolution by progressive gene loss, similar to that observed in rickettsial chromosomes, possibly leading to cryptic plasmids or complete plasmid loss. Moreover, rickettsial plasmids exhibited ORFans, recent gene duplications and evidence of horizontal gene transfer events with rickettsial and non-rickettsial genomes mainly from the α/γ-proteobacteria lineages. Genes related to maintenance and plasticity of plasmids, and to adaptation and resistance to stress mostly evolved under vertical and/or horizontal processes. Those involved in nucleotide/carbohydrate transport and metabolism were under the influence of vertical evolution only, whereas genes involved in cell wall/membrane/envelope biogenesis, cycle control, amino acid/lipid/coenzyme and secondary metabolites biosynthesis, transport and metabolism underwent mainly horizontal transfer events.

Conclusion

Rickettsial plasmids had a complex evolution, starting with a vertical inheritance followed by a reductive evolution associated with increased complexity via horizontal gene transfer as well as gene duplication and genesis. The plasmids are plastic and mosaic structures that may play biological roles similar to or distinct from their co-residing chromosomes in an obligate intracellular lifestyle.     

A null mutation in TNIK defines a novel locus for intellectual disability

Intellectual disability (ID) is one of the most common disabilities and, although many genes have been implicated in its etiology, the genetic heterogeneity of ID continues to expand. The purpose of the study was to describe a novel autosomal recessive non-syndromic ID locus. Autozygome and linkage analysis, and exome sequencing followed by RNA and protein analysis of the candidate disease gene were performed. We describe two multiplex consanguineous families with non-syndromic ID phenotype, which maps to a critical linkage locus on 3q26. Exome sequencing of the index in each family revealed the same homozygous truncating mutation in TNIK that results in complete loss of the protein. TNIK is a kinase with a well-established role in dendrite development and synaptic transmission. The phenotype we observe in human patients who lack TNIK is consistent with the previously published Tnik ^?/? phenotype in the murine model. Our data strongly implicate TNIK deficiency in the causation of ID in humans.     

Detection and Characterization of Phytoplasma and Sugarcane Yellow Leaf Virus Associated with Leaf Yellowing of Sugarcane

Leaves from sugarcane were collected from Egyptian plantation fields and tested for phytoplasma (Sugarcane yellows phytoplasma, SCYP) and Sugarcane yellow leaf virus (SCYLV) using nested PCR (with different primers) and RT‐PCR, respectively. These results showed significant differences in the amplification of the PCR assays. The primer MLO‐X/MLO‐Y, which amplified the 16S‐23S rDNA spacer region, was the most precise to detect the phytoplasma in sugarcane plants. Sequencing and restriction fragment length polymorphism analysis revealed that all tested phytoplasmas belonged to the 16SrI (aster yellows phytoplasma) group, with the exception of cultivar G84‐47 belonged to the 16SrXI (Rice yellow dwarf phytoplasma) group. Three Egyptian sugarcane cultivars were phytoplasma free. Phylogenetic analyses of 34 screened accessions of 16S ribosomal DNA gene sequences of Candidatus phytoplasma including the ones collected from Egypt used in this study and those extracted from GenBank showed that they split into two distinct clusters. The phylogenetic analyses indicated that these phytoplasmas are closely related and share a common ancestor. All tested Egyptian sugarcane plants were infected by SCYLV with the exception of cultivar Phil‐8013 which was virus free.     

Diet of the Worm Lizard,Diplometopon zarudnyi (Nikolsky, 1907), in Riyadh province,Saudi Arabia (Reptilia: Trogonophidae)

We analysed the diet composition of 133 specimens of the Worm Lizard, Diplometopon zarudnyi, in Riyadh province, Saudi Arabia. Analysis of stomach contents revealed that 90 specimens (66.6%) had prey items in their gut. The stomach content consisted mainly of small invertebrate prey (beetles), mainly larvae of Dermestes sp., but also imagines of Dermestes maculatus and Rhynchophorus ferrugineus. Dermestes sp. constitutes over 99% of the total food items, showing a high selectivity in food intake. The high number of beetles in the stomach contents indicates that this lizard forages very frequently. Specimens collected during winter had empty stomachs. The high proportion of fossorial prey items and the occasional records of other stomach contents show that D. zarudnyi only occasionally forages on the surface.