WNT-LRP5 Signaling Induces Warburg Effect through mTORC2 Activation during Osteoblast Differentiation

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A randomized controlled phase Ib trial of the malaria vaccine candidate GMZ2 in African children

Background

GMZ2 is a fusion protein of Plasmodium falciparum merozoite surface protein 3 (MSP3) and glutamate rich protein (GLURP) that mediates an immune response against the blood stage of the parasite. Two previous phase I clinical trials, one in naïve European adults and one in malaria-exposed Gabonese adults showed that GMZ2 was well tolerated and immunogenic. Here, we present data on safety and immunogenicity of GMZ2 in one to five year old Gabonese children, a target population for future malaria vaccine efficacy trials.

Methodology/Principal Findings

Thirty children one to five years of age were randomized to receive three doses of either 30 µg or 100 µg of GMZ2, or rabies vaccine. GMZ2, adjuvanted in aluminum hydroxide, was administered on Days 0, 28 and 56. All participants received a full course of their respective vaccination and were followed up for one year. Both 30 µg and 100 µg GMZ2 vaccine doses were well tolerated and induced antibodies and memory B-cells against GMZ2 as well as its antigenic constituents MSP3 and GLURP. After three doses of vaccine, the geometric mean concentration of antibodies to GMZ2 was 19-fold (95%CI: 11,34) higher in the 30 µg GMZ2 group than in the rabies vaccine controls, and 16-fold (7,36) higher in the 100 µg GMZ2 group than the rabies group. Geometric mean concentration of antibodies to MSP3 was 2.7-fold (1.6,4.6) higher in the 30 µg group than in the rabies group and 3.8-fold (1.5,9.6) higher in the 100 µg group. Memory B-cells against GMZ2 developed in both GMZ2 vaccinated groups.

Conclusions/Significance

Both 30 µg as well as 100 µg intramuscular GMZ2 are immunogenic, well tolerated, and safe in young, malaria-exposed Gabonese children. This result confirms previous findings in naïve and malaria-exposed adults and supports further clinical development of GMZ2.

Trial Registration

ClinicalTrials.gov {"type":"clinical-trial","attrs":{"text":"NCT00703066","term_id":"NCT00703066"}}NCT00703066     

Maize genotypes classified as null at theglu locus have β-glucosidase activity and immunoreactive protein

Maize β-glucosidase (β-d-glucoside glucohydrolase; EC 3.2.1.21) was extracted from coleoptiles of 15 maize genotypes (3 normals, 10 nulls, and 2 hybrids) in two fractions, the soluble and the insoluble. The enzyme activity was measured spectrophotometrically in the soluble fraction and also studied on zymograms after native gel electrophoresis and isoelectric focusing. The enzyme was purified from a normal genotype by anion-exchange chromatography and preparative electrophoresis. Antisera were raised in four rabbits, and the soluble and the insoluble extracts of each genotype were analyzed for a cross-reacting material by ELISA and immunoblotting. The results showed that extracts from both the normal and the null genotypes had β-glucosidase activity, and the activity measured spectrophotometrically was 2- to 10-fold higher in normals than in nulls. Zymograms of the null genotypes were devoid of distinct bands that were present in those of normals and hybrids from crosses between normals and nulls. Zymograms of both the normal and the null genotypes had a diffuse, smeared zone of activity at the cathodic end of native gels. A cross-reacting antigen was present in extracts of both genotypes when assayed by ELISA and a 60-kD polypeptide (β-glucosidase monomer) was detected by four different monospecific β-glucosidase antisera on Western blots by immunostaining. Moreover, six of seven null genotypes had a larger amount of their 60-kD polypeptide in the insoluble fraction than in the soluble fraction. These data show that both the null and the normal genotypes have similar amounts of the enzyme protein, but the enzyme occurs mostly as insoluble or poorly soluble polymers in nulls, and the monogenic inheritance reported for the null alleles of theglu locus is likely to be for a factor encoded by another locus which affects directly or indirectly the solubility of the enzyme by increasing its polymerization into large quaternary structures.     

Screening expression libraries with nonradioactive immunological probes

An immunological screening method employing protein A-peroxidase which does not require radiolabelled antibodies for detection of Escherichia coli colonies synthesizing foreign proteins in a cDNA expression library is described. The technique is sensitive, simpler and more rapid than the procedures that rely on radiolabelled antibodies and autoradiography.     

Treatment of acute hepatitis C virus infection with interferon-α 2b and ribavirin: Case report and review of the literature

   

Hepatitis C virus (HCV) infection becomes chronic in about 85 % of individuals as demonstrated by the persistence of HCV. It is necesseray to treat acute hepatitis C infection. Interferon-α is generally used for the treatment of acute HCV infection.     

IMMUNOLOGICAL AFFINITIES AMONG SUBFAMILIES OF THE POACEAE

Immunological affinities were investigated among twenty taxa belonging to the grass subfamilies Pooideae, Chloridoideae, Panicoideae, Oryzoideae, and Bambusoideae. Antisera were raised to the prolamin fraction of seed proteins from species of eleven grass genera (Hordeum, Bromus, Festuca, Phleum, Elensine, Panicum, Pennisetum, Tripsacum, Dendrocalamus, and Oryza) and reacted with their homologous antigens and nineteen different heterologous antigens in Enzyme-Linked Immunosorbent Assay (ELISA). The immunological cross-reactivity among the eleven taxa whose prolamin was used for antisera production was analyzed quantitatively by generating matrices of averaged cross-reactivities, Q correlation and distance. The averaged cross-reactivity matrix was calculated from averaging reciprocal immunological reactions while the two other matrices were computed by considering each antiserum as a character and antigens as OTUs. The three matrices were subjected to clustering by the Unweighted Pair Group Method using Arithmetic Averaging (UPGMA). The phenogram based on the averaged similarity matrix showed some distortion, while the other two phenograms were similar in topology and were informative. A phenon line at r = 0.17 divided the phenogram based on Q correlation into four major groups: Pooideae, Oryzoideae, Bambusoideae, and Chloridoideae-Panicoideae. The two subfamilies in the Chloridoideae-Panicoideae group clustered at a correlation coefficient of 0.22. Within the Pooideae, the tribes Aveneae and Agrostideae were closely grouped together (r = 0.85), but they were quite distinct (r = 0.16) from the tightly clustered (r = 0.84–0.85) Bromeae, Poeae, and Triticeae. The Oryzoideae and Bambusoideae showed low immunological similarity (r = –0.07). The two tribes of the Panicoideae, Paniceae and Andropogoneae, displayed extensive immunological similarity clustering tightly at r = 0.84–0.86. The immunological data revealed a possible trend in grass evolution encompassing the chloridoid-panicoid groups and provided insights into the phylogenetic affinities of the bambusoid and oryzoid grasses. The results also underscored the heterogeneity of the taxa within the Pooideae.     

Mutational and structural analysis of aglycone specificity in maize and sorghum beta-glucosidases

Plant beta-glucosidases display varying substrate specificities. The maize beta-glucosidase isozyme Glu1 (ZmGlu1) hydrolyzes a broad spectrum of substrates in addition to its natural substrate DIMBOA-Glc (2-O-beta-d-glucopyranosyl-4-hydroxy-7-methoxy-1,4-benzoxaxin-3-one), whereas the sorghum beta-glucosidase isozyme Dhr1 (SbDhr1) hydrolyzes exclusively its natural substrate dhurrin (p-hydroxy-(S)-mandelonitrile-beta-d-glucoside). Structural data from cocrystals of enzyme-substrate and enzyme-aglycone complexes have shown that five amino acid residues (Phe198, Phe205, Trp378, Phe466, and Ala467) are located in the aglycone-binding site of ZmGlu1 and form the basis of aglycone recognition and binding, hence substrate specificity. To study the mechanism of substrate specificity further, mutant beta-glucosidases were generated by replacing Phe198, Phe205, Asp261, Met263, Phe377, Phe466, Ala467, and Phe473 of Glu1 by Dhr1 counterparts. The effects of mutations on enzyme activity and substrate specificity were studied using both natural and artificial substrates. The simple mutant replacing Phe198 by a valine had the most drastic effect on activity, because the capacity of this enzyme to hydrolyze beta-glucosides was almost completely abolished. The analysis of this mutation was completed by a structural study of the double mutant ZmGlu1-E191D,F198V in complex with the natural substrate. The structure reveals that the single mutation F198V causes a cascade of conformational changes, which are unpredictable by standard molecular modeling techniques. Some other mutations led to drastic effects: replacing Asp261 by an asparagine decreases the catalytic efficiency of this simple mutant by 75% although replacing Tyr473 by a phenylalanine increase its efficiency by 300% and also provides a new substrate specificity by hydrolyzing dhurrin.     

Could Fetuin-A Be a Biomarker for Autism Spectrum Disorder and Cognitive Developmental Delay?

Biochemistry (Moscow) - Early detection of cognitive developmental delay (CDD) and autism spectrum disorder (ASD) is challenging, despite the numerous scientific studies conducted and different...