Estrogen and Related Compounds: Biphasic Dose Responses

This article documents and quantitatively assesses the capacity of estrogen, phytoestrogens, and antiestrogens to affect biphasic dose-response relationships in animal/human models and across a broad range of cell types, affecting multiple endpoints. The range of endpoints displaying such biphasic dose responses includes plasminogen activation, oxytocin secretion, angiogenesis, cell proliferation, bone growth, monocyte chemotaxis, secretion of various cytokines, and other effects. The quantitative features of the dose response relationships revealed that the magnitude of the stimulatory responses was typically less than twofold, whereas the stimulatory responses were markedly variable ranging from about 5- to 10⁶fold. Mechanistic explanations of the biphasic responses are addressed.     

Androgens: Biphasic Dose Responses

Testosterone was reported to affect a variety of reproductive endpoints. More notable were the effects of dihydrotestosterone on cell proliferation in the prostate cancer cell model LNCaP and Sertoli cell function. Testosterone production was also biphasically affected by prolactin that was administered to adult testicular cells in vitro.     

Adenosine: Biphasic Dose Responses

This article characterizes the occurrence of biphasic dose responses by adenosine and its stable analogs in numerous systems, including the respiratory tract, kidney, cardiovascular system, and brain. Considerable mechanistic research on various systems explored and clarified the interactions of the A-1 and A-2 receptors and their interactive influence on the occurrence of the biphasic dose responses. In general, the maximum stimulatory response was 2.5-fold or less than controls, while the stimulatory concentration range was quite variable ranging from 10- to 10⁵-fold.     

Endothelial function and its assessment

Endothelial dysfunction is a characteristic aspect of most of the conditions associated with atherosclerosis and is commonly found as an early feature in atherothrombotic vascular disease. An appreciation of the underlying mechanisms of endothelial function, as well as dysfunction, is essential as this has critical influence on the different methods in the assessment of endothelial function and effects of various treatments on its quantification. Furthermore, endothelial dysfunction is recognised as a type of ‘target organ damage’ in common cardiovascular conditions (e.g., hypertension) and the area is of increasing interest for new drug development, as therapies that modulate the endothelium will have added advantages; thus, for the development of new/experimental drugs, an awareness of ways to assess the endothelium is necessary. In this review, an overview of different methods including biochemical markers, and invasive and non-invasive tools, to determine endothelial function is presented as well as their clinical relevance. Furthermore, the effects of various treatments on endothelial dysfunction and their underlying mechanisms are elucidated.     

Enantioselective uptake of fexofenadine by Caco‐2 cells as model intestinal epithelial cells

Objectives Fexofenadine contains a chiral carbon in its chemical structure and is orally administered as a racemic mixture. This study evaluated the selective uptake of fexofenadine enantiomers by Caco‐2 cells as a model of intestinal epithelial cells. Methods R(+)‐fexofenadine or S(?)‐fexofenadine was applied to Caco‐2 cells, followed by incubation. After incubation, the amounts of fexofenadine enantiomers in cells were determined. The kinetic parameters for the uptake of fexofenadine enantiomers by Caco‐2 cells were estimated using the Michaelis–Menten equation. Key findings The transporter‐mediated uptake rate of R(+)‐fexofenadine was 1.7‐fold higher than that of S(?)‐fexofenadine. The difference in transporter‐mediated R(+)‐fexofenadine and S(?)‐fexofenadine uptake was completely diminished under ATP‐depleted conditions and in the presence of organic anion transporter peptide (OATP) inhibitors. Also, a Dixon plot showed that each fexofenadine enantiomer was competitively inhibited by the other enantiomer. The ratio of R(+)‐fexofenadine uptake to S(?)‐fexofenadine uptake in the case of a racemic mixture was higher than that in the case of a single enantiomer. Conclusion This study suggested that the selective absorption of fexofenadine enantiomers by intestinal epithelial cells might have been due to the selective uptake mediated by OATPs and that the difference in intestinal absorption was enhanced with a racemic mixture.     

Bortezomib for Acute Antibody‐Mediated Rejection in Liver Transplantation

Antibody‐mediated rejection (AMR) is an uncommon, but challenging type of rejection after solid organ transplantation. We review three cases of AMR in ABO‐compatible liver transplant recipients. These cases were characterized by severe acute rejection resistant to steroids and antithymocyte globulin, histologic evidence of plasma cell infiltrates, C4d positivity and high serum anti‐HLA donor‐specific antibodies. All three patients were treated with bortezomib, a proteasome inhibitor effective in depleting plasma cells. After treatment, all patients had improved or normal liver function tests, resolution of C4d deposition and significant decline in their HLA donor‐specific antibodies.     

Lentiviral Vectors for Delivery of Gene-Editing Systems Based on CRISPR/Cas: Current State and Perspectives

CRISPR/Cas technology has revolutionized the fields of the genome- and epigenome-editing by supplying unparalleled control over genomic sequences and expression. Lentiviral vector (LV) systems are one of the main delivery vehicles for the CRISPR/Cas systems due to (i) its ability to carry bulky and complex transgenes and (ii) sustain robust and long-term expression in a broad range of dividing and non-dividing cells in vitro and in vivo. It is thus reasonable that substantial effort has been allocated towards the development of the improved and optimized LV systems for effective and accurate gene-to-cell transfer of CRISPR/Cas tools. The main effort on that end has been put towards the improvement and optimization of the vector’s expression, development of integrase-deficient lentiviral vector (IDLV), aiming to minimize the risk of oncogenicity, toxicity, and pathogenicity, and enhancing manufacturing protocols for clinical applications required large-scale production. In this review, we will devote attention to (i) the basic biology of lentiviruses, and (ii) recent advances in the development of safer and more efficient CRISPR/Cas vector systems towards their use in preclinical and clinical applications. In addition, we will discuss in detail the recent progress in the repurposing of CRISPR/Cas systems related to base-editing and prime-editing applications.     

What have we learned about how to prevent and treat antibody‐mediated rejection in kidney transplantation?

Antibody‐mediated rejection (ABMR) in kidney transplantation is a major cause of late graft loss, and despite all efforts to date the “standard of care” remains plasmapheresis, IVIg, and steroids, which itself is based on low quality evidence. This review focuses on the risk factors leading to memory and de novo donor‐specific antibody (DSA)‐associated ABMR, the optimal prevention strategies for ABMR, and advances in  adjunctive and emerging therapies for ABMR. Because new agents require regulatory approval via a Phase 3 randomized control trial (RCT), an overview of progress in innovative trial design for ABMR is provided. Finally, based on the insights gained in the biology of ABMR, current knowledge gaps are identified for future research that could significantly affect our understanding of how to optimally treat ABMR.