Genetically modified L3,7 and L2 lipooligosaccharides from Neisseria meningitidis serogroup B confer a broad cross-bactericidal response.

Currently available Neisseria meningitidis serogroup B (MenB) vaccines are based on outer membrane vesicles (OMVs) that are obtained from wild-type strains. They are purified with the aim of decreasing the lipooligosaccharide (LOS) content and hence reduce the reactogenicity of the vaccine even though LOS is a potential protective antigen. In <2-year-old children, these MenB vaccines confer protection only against strains expressing homologous PorA, a major and variable outer membrane protein. Our objective was to develop a safe LOS-based vaccine against MenB. To this end, we used modified porA knockout strains expressing genetically detoxified (msbB gene-deleted) L2 and L3,7 LOSs, allowing the production of LOS-enriched OMVs. The vaccine-induced antibodies were found to be bactericidal against nearly all invasive strains, irrespective of capsular serogroup. In addition, we have also demonstrated that LOS lacking the terminal galactose (with a lgtB mutation; truncated L3 LOS), but not LOS produced without the galE gene, induced a bactericidal antibody response in mice similar to that seen for LOS containing the full lacto-N-neotetraose (L3,7 LOS). In conclusion, a bivalent detoxified LOS OMV-based vaccine demonstrated the potential to afford a broad cross-protection against meningococcal disease.

Cloning and expression of the TALE superclass homeobox Meis2 gene during zebrafish embryonic development.

Meis and Prep/Pknox (MEINOX family) proteins, together with Pbx (PBC family) proteins, belong to the TALE superfamily characterized by an atypical homeodomain containing three additional amino acids between helix 1 and helix 2. Members of the MEINOX and PBC families have been isolated in Caenorhabditis elegans, Drosophila, Xenopus, chick, mouse and human, and play crucial roles in many aspects of embryogenesis. Here, we report the isolation of meis2 in zebrafish. Expression of meis2 is first detected at the beginning of gastrulation. Later during embryogenesis, meis2 transcripts are found in distinct domains of the central nervous system with the strongest expression in the hindbrain. Expression was also detected in the isthmus, along the spinal cord and in the lateral mesoderm. As development proceeds, meis2 is also expressed in the developing retina, pharyngeal arches, and in the vicinity of the gut tube.

Functional analysis of ZNF85 KRAB zinc finger protein, a member of the highly homologous ZNF91 family.

We previously identified the ZNF85 (HPF4) KRAB zinc finger gene, a member of the human ZNF91 family. Here, we show that the ZNF85 gene is highly expressed in normal adult testis, in seminomas, and in the NT2/D1 teratocarcinoma cell line. Immunocytochemical localization of a panel of beta-Gal/ZNF85 fusion proteins revealed that ZNF85 contains at least one nuclear localization signal located in the spacer region connecting the KRAB domain with the zinc finger repeats. Bacterially expressed ZNF85 zinc finger domain bound strongly and exclusively to DNA in vitro in a zinc-dependent manner. The KRAB(A) domain of the ZNF85 protein and of several other members of the ZNF91 family exhibited repressing activity when tested in Gal4 fusion protein assays. The repression was significantly enhanced by the addition of the KRAB (B) domain, whereas further addition of other conserved regions had no effect. The ZNF85 KRAB(A) and (B) domains in vitro bound several nuclear proteins that might constitute critical cofactors for repression.

Inflammatory markers in younger vs elderly normal volunteers and in patients with Alzheimer's disease.

BACKGROUND: Recently it has been reported that activation of the inflammatory response system (IRS) may play a role in the aging process and in the pathogenesis of the degenerative changes associated with Alzheimer's disease (SDAT). The aims of the present study were to examine the peripheral IRS in normal aging and in SDAT patients. METHODS: Serum zinc (Zn), total serum protein (TSP), albumin (Alb), SP electrophoresis, and serum interleukin-6 (IL-6) and the stimulated production of tumor necrosis factor-alpha (TNFalpha) were determined in younger versus elderly healthy subjects and in SDAT patients vs. age-matched, healthy volunteers. RESULTS: Serum Zn and Alb were significantly lower in elderly than in younger healthy volunteers and were significantly and inversely correlated with age. The production of TNFalpha was significantly higher in elderly than in younger healthy volunteers and was significantly and positively correlated with age. In SDAT patients, no significant changes in serum Zn or TNFalpha production could be found. Serum Alb was significantly lower and serum IL-6 and the alpha1 and alpha2 globulin fractions significantly higher in SDAT patients than in controls. CONCLUSIONS: Activation of the IRS appears to accompany the normal aging process, i.e. lower serum Zn and Alb and increased TNFalpha production, as well as SDAT, i.e. lower serum Alb and increased serum IL-6 and alpha1 and alpha2 globulin fractions. The findings suggest that not all indicators of IRS activation in SDAT are related to those of the normal ageing process.

Objective human tooth colour measurements as a means of determining chronologic age in vivo and ex vivo.

Colour is a subjective sensation and as such is difficult to use in a quantitative study. However a number of clinical studies on extracted teeth have shown a good correlation between tooth colour and age. The purpose of this study was to examine the usefulness of a specific spectrophotometer in determining tooth colour on extracted and non- extracted teeth and to look for a possible age relationship. There were two parts in this study. An ex vivo study concentrated on collected tooth material. Single rooted teeth were selected out of each of the 5- year-age groups (ages ranged from 15-89 years). Colour measurements were performed on the mesial and vestibular aspects of the roots as well on the mid-vestibular aspects of the enamel crown. An in vivo study concentrated on the use of this specific shade taking system in living patients (n=70). Statistical analysis of the results revealed regression formulas for both ex vivo and in vivo situations displaying adjusted R-squares between 0.48 and 0.56. It may be concluded that age related trends were found. Having its shortcomings, the shade taking system was found to perform well as a convenient adjunct to dental age estimation in both the living and the deceased.

Protein adaptation to low temperatures: a comparative study of alpha-tubulin sequences in mesophilic and psychrophilic algae.

The alpha-tubulin genes from two psychrophilic algae belonging to the genus Chloromonas (here named ANT1 and ANT3) have been isolated and sequenced. The genes ant1 and ant3 contain 4 and 2 introns, respectively. The coding DNA sequences are 90% identical but the degree of isology is very high at the polypeptide level (more than 97% strict identities). The ANT1 and ANT3 alpha-tubulin amino acid sequences were compared to the corresponding sequence of the mesophilic alga Chlamydomonas reinhardtii. Of the 15 substitutions detected in ANT1 and/or ANT3, 5 are common to both psychrophilic algae. The recorded substitutions have been analyzed in terms of cold adaptation on the basis of the available three-dimensional structure of the alpha,beta-tubulin heterodimer from pig brain. Most of these are subtle changes, but two substitutions, M268V and A295V occurring in the region of interdimer contacts, could be of great significance for the cold stability of Antarctic algae microtubules due to the fact that the entropic control of microtubule assembly is particularly high in cold adaptes species.