Safety of cetirizine in infants 6 to 11 months of age: a randomized,double-blind,placebo-controlled study

BACKGROUND: H(1)-antihistamines are widely used for symptom relief in allergic disorders in infants and children; however, there are few prospective, randomized, double-blind, controlled studies of these medications in young children, and to date, no such studies have been conducted in infants. OBJECTIVE: This prospective, randomized, parallel-group, double-blind, placebo-controlled study was designed to evaluate the safety of the H(1)-antihistamine cetirizine, particularly with regard to central nervous system and cardiac effects, in infants age 6 to 11 months, inclusive. METHODS: Infants who met the entry criteria for age and had a history of treatment with an H(1)-antihistamine for an allergic or other disorder were randomized to receive 0.25 mg/kg cetirizine orally or matching placebo twice daily orally for 1 week. RESULTS: The mean daily dose in cetirizine-treated infants was 4.5 +/- 0.7 mg (SD). No differences in all-cause or treatment-related adverse events were observed between the cetirizine- and placebo-treated groups. A trend was observed toward fewer adverse events and sleep-related disturbances in the cetirizine group compared with the placebo group. No prolongation in the linear corrected QT interval was observed in cetirizine-treated infants compared with either baseline values or with values in placebo-treated infants. CONCLUSIONS: We have documented the safety of cetirizine in this short-term investigation, the first randomized, double-blind, placebo-controlled study of any H(1)-antihistamine in infants. Additional prospective, randomized, double-blind, placebo-controlled, long-term studies of cetirizine and other H(1)-antihistamines are needed in this population.     

The anti-apoptotic factor Bcl-2 can functionally substitute for the B cell survival but not for the marginal zone B cell differentiation activity of BAFF

The TNF family ligand B cell-activating factor (BAFF, BLyS, TALL-1) is an essential factor for B cell development. BAFF binds to three receptors, BAFF-R, transmembrane activator and CAML interactor (TACI), and B cell maturation antigen (BCMA), but only BAFF-R is required for successful survival and maturation of splenic B cells. To test whether the effect of BAFF is due to the up-regulation of anti-apoptotic factors, TACI-Ig-transgenic mice, in which BAFF function is inhibited, were crossed with transgenic mice expressing FLICE-inhibitory protein (FLIP) or Bcl-2 in the B cell compartment. FLIP expression did not rescue B cells, while enforced Bcl-2 expression restored peripheral B cells and the ability to mount T-dependent antibody responses. However, many B cells retained immaturity markers and failed to express normal amounts of CD21. Marginal zone B cells were not restored and the T-independent IgG3, but not IgM, response was impaired in the TACI-IgxBcl-2 mice. These results suggest that BAFF is required not only to inhibit apoptosis of maturating B cells, but also to promote differentiation events, in particular those leading to the generation of marginal zone B cells.     

Coatings of hydroxyapatite–bioactive glass microparticles for adhesion to biological tissues

Adsorption of particles across interfaces has been proposed as a way to create adhesion between hydrogels and biological tissues. Here, we explore how this particle bridging approach can be applied to attach a soft polymer substrate to biological tissues, using bioresorbable and nanostructured hydroxyapatite–bioactive glass microparticles. For this, microparticles of aggregated flower-like hydroxyapatite and bioactive glass (HA–BG) were synthesized via a bioinspired route. A deposition technique using suspension spreading was developed to tune the coverage of HA–BG coatings at the surface of weakly cross-linked poly(beta-thioester) films. By varying the concentration of the deposited suspensions, we produced coatings having surface coverages ranging from 4% to 100% and coating densities ranging from 0.02 to 1.0 mg cm⁻². The progressive dissolution of these coatings within 21 days in phosphate-buffered saline was followed by SEM. Ex vivo peeling experiments on pig liver capsules demonstrated that HA–BG coatings produce an up-to-two-fold increase in adhesion energy (9.8 ± 1.5 J m⁻²) as compared to the uncoated film (4.6 ± 0.8 J m⁻²). Adhesion energy was found to increase with increasing coating density until a maximum at 0.2 mg cm⁻², well below full surface coverage, and then it decreased for larger coating densities. Using microscopy observations during and after peeling, we show that this maximum in adhesion corresponds to the appearance of particle stacks, which are easily separated and transferred onto the tissue. Such bioresorbable HA–BG coatings give the possibility of combining particle bridging with the storage and release of active compounds, therefore offering opportunities to design functional bioadhesive surfaces.

Coatings of hydroxyapatite–bioactive glass microparticles are proposed as a way to create adhesion between hydrogels and biological tissues using adsorption of the microparticles across the interface.     

Efficient Method for Generating Point Mutations in the Vaccinia Virus Genome Using CRISPR/Cas9

The vaccinia virus (VACV) was previously used as a vaccine for smallpox eradication. Nowadays, recombinant VACVs are developed as vaccine platforms for infectious disease prevention and cancer treatment. The conventional method for genome editing of the VACV is based on homologous recombination, which is poorly efficient. Recently, the use of CRISPR/Cas9 technology was shown to greatly improve the speed and efficiency of the production of recombinant VACV expressing a heterologous gene. However, the ability to rapidly recover viruses bearing single nucleotide substitutions is still challenging. Notwithstanding, ongoing studies on the VACV and its interaction with the host cell could benefit from viral gene targeted mutagenesis. Here, we present a modified version of the CRISPR/Cas9 system for the rapid selection of mutant VACV carrying point mutations. For this purpose, we introduced a silent mutation into the donor gene (which will replace the wildtype gene) that serves a double function: it is located in the PAM (NGG) sequence, which is essential for Cas9 cleavage, and it alters a restriction site. This silent mutation, once introduced into the VACV genome, allows for rapid selection and screening of mutant viruses carrying a mutation of interest in the targeted gene. As a proof of concept, we produced several recombinant VACVs, with mutations in the E9L gene, upon which, phenotypic analysis was performed.