4,6-Dimethoxy-2H-1-benzopyran-2-one

Human recombinant Fab fragment from combinatorial libraries of a B-cell lymphoma patient recognizes core protein of chondroitin sulphate proteoglycan 4.[Pubmed:29036679]

J Biochem. 2018 Jan 1;163(1):61-68.

CD antigens are well known as therapeutic targets of B-cell lymphoma. To isolate therapeutic antibodies that recognize novel targets other than CD antigens, we constructed a phage display combinatorial antibody Fab library from bone marrow lymphocytes of B-cell lymphoma patient. To eliminate antibodies reactive with known B-cell lymphoma antigen, non-hematopoietic and patient's sera reactive HeLaS3 cells was selected as a target of whole cell panning. Five rounds of panning against live HeLaS3 cells retrieved single Fab clone, termed AHSA (Antibody to HeLa Surface Antigen). Using phage display random peptide library, LSYLEP was identified as an epitope sequence of AHSA. LC-MS/MS analysis of AHSA-precipitated HeLaS3 cell lysates detected several fragments corresponding to the sequence of chondroitin sulphate proteoglycan 4 (CSPG4) core protein. Since LSYLEP sequence was at the position of 313-318 of CSPG4, we considered that CSPG4 was AHSA-associated antigen. Double staining of CSPG4-postive MDA-MB-435S cells with AHSA and anti-CSPG4 rabbit antibody showed identical staining position, and reduced AHSA reactivity was observed in CSPG4-siRNA treated MDA-MB-435S cells. In conclusion, we retrieved a human Fab from antibody library of B-cell lymphoma patient, and identified CSPG4 as a recognizing antigen. AHSA may have potential benefits for development of CSPG4-targeting theranostics for B-cell lymphoma.

Identifying Greener and Safer Plasticizers: A 4-Step Approach.[Pubmed:29036695]

Toxicol Sci. 2018 Feb 1;161(2):266-275.

The health and economic burden of endocrine disrupting chemicals, such as the plasticizer di(2-ethylhexyl) phthalate (DEHP), is prompting industry to develop alternatives. However, the absence of requirements for manufacturers to ensure the safety of these alternatives has led to the generation of replacements that may have similar or worse effects than the original chemicals. Consequently, there is increasing recognition by scientists, regulators and industry that proactive approaches are needed to develop safe chemical substitutes. We propose a 4-step approach for the design, characterization and toxicological testing of responsible alternative chemicals that we illustrate with our ongoing studies on DEHP replacements. Our approach is comprised of: (1) the design and characterization of alternative chemicals based on innovative chemical structures and environmental considerations; (2) large-scale in vitro cell-based high throughput and selective ex vivo studies to preselect the most innocuous alternatives; (3) an acute toxicity in vivo study to rule out overt toxicity of the selected candidates; and (4) an in utero and lactational exposure study comparing the effects of selected candidates to those currently in use, emphasizing commonly described phenotypes after exposure to the latter. Using this 4-step approach, we have identified 2 alternative chemicals displaying good plasticizing properties, better biodegradability, and less leaching than DEHP without any apparent toxicity in vivo. This process has thus far proven useful in the proactive identification of responsible chemical replacements for DEHP.

Fimbrins 4 and 5 Act Synergistically During Polarized Pollen Tube Growth to Ensure Fertility in Arabidopsis.[Pubmed:29036437]

Plant Cell Physiol. 2017 Nov 1;58(11):2006-2016.

The germination and polar growth of pollen are prerequisite for double fertilization in plants. The actin cytoskeleton and its binding proteins play pivotal roles in pollen germination and pollen tube growth. Two homologs of the actin-bundling protein fimbrin, AtFIM4 and AtFIM5, are highly expressed in pollen in Arabidopsis and can form distinct actin architectures in vitro, but how they co-operatively regulate pollen germination and pollen tube growth in vivo is largely unknown. In this study, we explored their functions during pollen germination and polar growth. Histochemical analysis demonstrated that AtFIM4 was expressed only after pollen grain hydration and, in the early stage of pollen tube growth, the expression level of AtFIM4 was low, indicating that it functions mainly during polarized tube growth, whereas AtFIM5 had high expression levels in both pollen grains and pollen tubes. Atfim4/atfim5 double mutant plants had fertility defects including reduced silique length and seed number, which were caused by severe defects in pollen germination and pollen tube growth. When the atfim4/atfim5 double mutant was complemented with the AtFIM5 protein, the polar growth of pollen tubes was fully rescued; however, AtFIM4 could only partially restore these defects. Fluorescence labeling showed that loss of function of both AtFIM4 and AtFIM5 decreased the extent of actin filament bundling throughout pollen tubes. Collectively, our results revealed that AtFIM4 acts co-ordinately with AtFIM5 to organize and maintain normal actin architecture in pollen grains and pollen tubes to fulfill double fertilization in plants.

Targeting the interleukin-4 and interleukin-13 pathways in severe asthma: current knowledge and future needs.[Pubmed:29036019]

Curr Opin Pulm Med. 2018 Jan;24(1):50-55.

PURPOSE OF REVIEW: Severe asthma is a heterogeneous disease that can be classified into phenotypes and endotypes based upon clinical or biological characteristics. Interleukin (IL)-4 and IL-13 play a key role in type 2 (T2) asthma. This article reviews the signaling pathway of IL-4 and IL-13 and highlights its targeted therapy in severe asthma. RECENT FINDINGS: Several clinical trials of biologics targeting the IL-4/IL-13 pathway have recently been completed. In patients with severe, uncontrolled asthma, targeting IL-13 alone with biologics including lebrikizumab and tralokinumab has not shown consistent reduction in asthma exacerbations. Simultaneous targeting of both IL-4 and IL-13 by blocking IL-4 receptor alpha using dupilumab has yielded more consistent results in reducing asthma exacerbations and improving lung function, especially in patients with increased blood eosinophils. Other biomarkers of T2 inflammation such as exhaled nitric oxide and serum periostin may also predict response to biologics targeting the IL-4/IL-13 pathway. SUMMARY: No biologic targeting the IL-4/IL-13 pathway is currently available for treatment of asthma, but emerging data suggest that biologics targeting IL-4 and IL-13 together may benefit patients with T2 high asthma. Additional data are needed about long-term efficacy and safety prior to incorporating these drugs into routine clinical practice.