Establishment and Modification of Ninety-seven Pneumococcal Serotyping Assays Based on Quantitative Real-time Polymerase Chain Reaction.

Objective: To establish and modify quantitative real-time polymerase chain reaction (qPCR)-based serotyping assays to distinguish 97 pneumococcal serotypes. Methods: A database of capsular polysaccharide ( cps) loci sequences was generated, covering 97 pneumococcal serotypes. Bioinformatics analyses were performed to identify the cps loci structure and target genes related to different pneumococcal serotypes with specific SNPs. A total of 27 novel qPCR serotyping assay primers and probes were established based on qPCR, while 27 recombinant plasmids containing serotype-specific DNA sequence fragments were constructed as reference target sequences to examine the specificity and sensitivity of the qPCR assay. A panel of pneumococcal reference strains was employed to evaluate the capability of pneumococcal serotyping. Results: A total of 97 pneumococcal serotyping assays based on qPCR were established and modified, which included 64 serotypes previously reported as well as an additional 33 serotypes. Twenty-seven novel qPCR serotyping target sequences were implemented in the pneumococcal qPCR serotyping system. A total of 97 pneumococcal serotypes, which included 52 individual serotypes and 45 serotypes belonging to 20 serogroups, could not be identified as individual serotypes. The sensitivity of qPCR assays based on 27 target sequences was 1-100 copies/µL. The specificity of the qPCR assays was 100%, which were tested by a panel of 90 serotypes of the pneumococcal reference strains. Conclusion: A total of 27 novel qPCR assays were established and modified to analyze 97 pneumococcal serotypes.

Comparison of 0.025% FK-506, 0.05% Cyclosporin A, and 0.3% Sodium Hyaluronate Eye Drops for the Treatment of Botulinum Toxin B-Induced Mouse Dry Eye.

PURPOSE: To compare the effects of FK-506, cyclosporin A (CsA), and sodium hyaluronate (HA) eye drops for the treatment of botulinum toxin B (BTX-B)-induced mouse dry eye. METHODS: CBA/J mice were randomized into 5 groups. The groups received treatment with eye drops containing 0.025% FK-506 combined with 0.3% HA (FK-506+HA group), 0.025% FK-506 (FK-506 group), 0.05% CsA (CsA group), 0.3% HA (HA group), or 0.9% saline (saline group) 3 days after an intralacrimal gland injection with 20 mU of BTX-B. Tear production, corneal fluorescein staining, blink rate, and the mRNA and protein expression levels of inflammatory cytokines were measured. RESULTS: FK-506+HA eye drops increased tear production and reduced the corneal fluorescein staining scores at all time points after treatment compared with those in the saline group. Compared with those in the saline group, the tear production and severity of corneal epithelial defects in the FK-506 group were significantly improved at weeks 2 and 4. Compared with the saline eye drops, the CsA eye drops ameliorated only tear production and corneal fluorescein staining scores at week 4 after administration. The FK-506+HA, FK-506, and CsA eye drops downregulated the expression of inflammatory cytokines in both the keratoconjunctival tissues and lacrimal glands at week 4. CONCLUSIONS: The topical application of 0.025% FK-506 combined with 0.3% HA, 0.025% FK-506, or 0.05% CsA can suppress the expression of inflammatory cytokines and can alleviate the signs of dry eye. Topical application of 0.025% FK-506 combined with 0.3% HA showed the best therapeutic effect and may be a possible therapy for dry eye.

Self-assembly and anion sensing of metal-organic [M6L2] cages from fluorescent triphenylamine tri-pyrazoles with dipalladium(ii,ii) corners.

Three hexametal-organic cages [(N^N)6Pd6L2]6+ with syn, syn, syn conformation were synthesized via the coordination of tris(4-(1H-pyrazol-3-yl)phenyl)amine ligands and [(N^N)2Pd2(NO3)2](NO3)2 dimetallic corners (N^N = 2,2'-bipyridine for 1; 4,4'-dimethylbipyridine for 2; 1,10-phenanthroline for 3). These hexametal-organic cages exhibit anion sensing toward HSO3- in aqueous solution via fluorescent titration and NMR spectroscopy studies.

Programmable self-assembly of water-soluble organo-heterometallic cages [M12M'4L12] using 3-(3,5-dimethyl-1H-pyrazol-4-yl)pentane-2,4-dione (H2L).

A bifunctional ligand H2L featuring primary (pyrazole) and secondary (acetylacetone) coordination sites was preferentially reacted with dimetallic [M2(NO3)2](NO3)2 linkers at the pyrazolyl end of H2L, giving rise to dimetallic corners. Subsequently, the corners serve as the secondary site with M' to form water-soluble organo-heterometallic [M12M'4L12] cages in a stepwise mode.