BackgroundInflammation plays a key role in the pathophysiology of ischemic stroke. Some proinflammatory mediators, such as cytokines and chemokines, are produced in stroke. Chemokine-like factor 1 (CKLF1), as a novel C-C chemokine, displays chemotactic activities in a wide spectrum of leukocytes and plays an important role in brain development. In previous studies, we have found that the expression of CKLF1 increased in rats after focal cerebral ischemia and treatment with the CKLF1 antagonist C19 peptide decreased the infarct size and water content. However, the role of CKLF1 in stroke is still unclear. The objective of the present study was to ascertain the possible roles and mechanism of CKLF1 in ischemic brain injury by applying anti-CKLF1 antibody.MethodsMale Sprague–Dawley rats were subjected to one-hour middle cerebral artery occlusion. Antibody to CKLF1 was applied to the right cerebral ventricle immediately after reperfusion; infarct volume and neurological score were measured at 24 and 72 hours after cerebral ischemia. RT-PCR, Western blotting and ELISA were utilized to characterize the expression of adhesion molecules, inflammatory factors and MAPK signal pathways. Immunohistochemical staining and myeloperoxidase activity was used to determine the extent of neutrophil infiltration.ResultsTreatment with anti-CKLF1 antibody significantly decreased neurological score and infarct volume in a dose-dependent manner at 24 and 72 hours after cerebral ischemia. Administration with anti-CKLF1 antibody lowered the level of inflammatory factors TNF-α, IL-1β, MIP-2 and IL-8, the expression of adhesion molecules ICAM-1 and VCAM-1 in a dose-dependent manner. The results of immunohistochemical staining and detection of MPO activity indicated that anti-CKLF1 antibody inhibited neutrophil infiltration. Further studies suggested MAPK pathways associated with neutrophil infiltration in cerebral ischemia.ConclusionsSelective inhibition of CKLF1 activity significantly protects against ischemia/reperfusion injury by decreasing production of inflammatory mediators and expression of adhesion molecules, thereby reducing neutrophils recruitment to the ischemic area, possibly via inhibiting MAPK pathways. Therefore, CKLF1 may be a novel target for the treatment of stroke.

We have identified compound physical-chemical properties and structural criteria required to facilitate the adsorption of hydroxypyridines, quinolines, and pyrimidines onto aluminum(III), iron(III), and titanium(IV) oxides. We achieved this by measuring the adsorption behavior of a number of structurally related compounds, each with a nearly isolated variation in molecular structure, and systematically evaluating the effect of structural change on the nature and extent of adsorption. We find that the adsorption at the metal oxide-water interface is significant only when the oxo-hydroxy (keto-enol) tautomeric equilibrium in aqueous solution favors the hydroxy tautomer. Lack of adsorption of the oxo tautomer can be attributed to the absence of favorable electrostatic interactions between the compound and the surface, the absence of ligand groups capable of surface complexation, and the presence of strong intermolecular hydrogen bonding between ligand groups (carbonyl and amide) and water molecules. We find that adsorption of hydroxy tautomers with ortho-substituted cyclic N and -OH groups (such as trichloropyridinol) likely occurs via nonspecific electrostatic interactions, while non-ortho-substituted hydroxy tautomers (such as 8-hydroxyquinoline) and compounds such as (2-hydroxy-methylpyridine) likely adsorb via surface complexation. This study demonstrates that for compounds classified as tautomeric heterocycles it is imperative to accurately represent the predominant tautomer in aqueous solution so that the appropriate physical-chemical properties and reactivities are utilized in the evaluation of environmental fate.

In prior investigation, we discovered that (3'R,4'R)-3-cyanomethyl-4-methyl-3',4'-di-O-(S)-camphanoyl-(+)-cis-khellactone (4, 3-cyanomethyl-4-methyl-DCK) showed promising anti-HIV activity. In these current studies, we developed and optimized successfully a practical ten-step synthesis for scale-up preparation to increase the overall yield of 4 from 7.8% to 32%. Furthermore, compound 4 exhibited broad-spectrum anti-HIV activity against wild-type and drug-resistant viral infection of CD4+ T cell lines as well as peripheral blood mononuclear cells by both laboratoryadapted and primary HIV-1 isolates with distinct subtypes and tropisms. Compound 4 was further subjected to in vitro and in vivo pharmacokinetic studies. These studies indicated that 4 has moderate cell permeability, moderate oral bioavailability and low systemic clearance. These results suggest that 4 should be developed as a promising anti-HIV agent for development as a clinical trial candidate.As of February 2008, thirty-two anti-HIV drugs have been licensed by the US Food and Drug Administration (FDA) (http://www.fda.gov/oashi/aids/virals.html). These compounds include eleven HIV protease inhibitors, seventeen nucleoside and nonnucleoside reverse transcriptase (RT) inhibitors, one fusion inhibitor, one entry inhibitor (CCR5 co-receptor antagonist), one integrase inhibitor, and one multi-class combination product. Clinical combinations of these drugs, known as highly active antiretroviral therapy (HAART), have significantly reduced the morbidity and mortality of AIDS. However, increasing numbers of HIV/AIDS patients on HAART regimens fail to respond to current antiretroviral drugs due to the emergence of drugresistant HIV mutants. 1 Therefore, it is essential to develop additional potent anti-HIV drugs with novel mechanisms of action or resistance profiles different from those of current anti-HIV therapeutics.To whom correspondence should be addressed. Phone: 86-10-68181014. Fax: 86-10-6821-1656. E-mail: lanxieshi@yahoo.com; Phone: 919-962-0066. Fax: 919-966-3893. E-mail: khlee@unc.edu; Phone: 212-570-3058. Fax: 212-570-3099. E-mail: sjiang@nybloodcenter.org. NIH Public Access NIH-PA Author ManuscriptNIH-PA Author Manuscript NIH-PA Author ManuscriptIn our prior studies, 3',4'-di-O-(S)-camphanoyl-(+)-cis-khellactone (1, DCK, Figure 1) 2 and its analogs were identified as a novel class of anti-HIV agents with potent activity in H9 lymphocytes. Systematic modification of 1 provided more than 150 khellactone derivatives, including mono-, di-, and tri-substituted 1-analogs, and their SAR study results have been published. 3,4,5,6 Notably, mechanistic studies have demonstrated that 1 and its analogs do target HIV-1 RT; however, they do not interfere with its RNA-polymerase activity, but instead inhibit its DNA-dependent DNA polymerase activity. Thus, 1-analogs suppress the production of double-stranded viral DNA from a single-stranded DNA intermediate, 7 in stark contrast to current HIV-1 RT inhibitors that block the generation of single-stranded...

We previously reported that dinuclear copper(II) cryptate [Cu(2)L](4+) cleaves the C-C bond of acetonitrile at room temperature to produce a cyano-bridged dinuclear cryptate and methanol, whereby the reaction mechanism has not yet become clear. We have now systemically investigated this reaction, and four cryptates, [Cu(2)L](ClO(4))(4) (1), [Zn(2)L](ClO(4))(4) (2), [Cu(2)L(H(2)O)(2)](CF(3)SO(3))(4) (5), and [Cu(2)L(OH)(OH(2))](ClO(4))(3) (6) are reported here. Cryptates 1 and 2 can cleave the C--C bonds of acetonitrile, propionitrile, and benzonitrile at room temperature under open atmospheric conditions to give cyano-bridged cryptates [Cu(2)L(CN)](ClO(4))(3) (3) and [Zn(2)L(CN)](ClO(4))(3) (4), respectively, and the corresponding alcohol. In contrast, 5 and 6 do not show any C-C bond activation of nitriles, as the interior axial positions of Cu(II) in 5 and 6 are occupied by water/OH(-). The C-C bond cleavage of (S)-(+)-2-methylbutyronitrile by 2 produced (R)-(-)-2-butanol only; that is, the cleavage reaction proceeds through an S(N)2 pathway (Walden inversion).