Helminth resistance is mediated by differential activation of recruited monocyte-derived alveolar macrophages and arginine depletion

Macrophages are known to mediate anti-helminth responses, but it remains uncertain which subsets are involved or how macrophages actually kill helminths. Here, we show rapid monocyte recruitment to the lung after infection with the nematode parasite Nippostrongylus brasiliensis. In this inflamed tissue microenvironment, these monocytes differentiate into an alveolar macrophage (AM)-like phenotype, expressing both SiglecF and CD11c, surround invading parasitic larvae, and preferentially kill parasites in vitro. Monocyte-derived AMs (Mo-AMs) express type 2-associated markers and show a distinct remodeling of the chromatin landscape relative to tissue-derived AMs (TD-AMs). In particular, they express high amounts of arginase-1 (Arg1), which we demonstrate mediates helminth killing through L-arginine depletion. These studies indicate that recruited monocytes are selectively programmed in the pulmonary environment to express AM markers and an anti-helminth phenotype

Molecular models of N-Benzyladriamycin-14-valerate (AD 198) in complex with the phorbol ester-binding C1b domain of protein kinase C-δ

N-Benzyladriamycin-14-valerate (AD 198) is a semisynthetic anthracycline with experimental antitumor activity superior to that of doxorubicin (DOX). AD 198, unlike DOX, only weakly binds DNA, is a poor inhibitor of topoisomerase II, and circumvents anthracycline-resistance mechanisms, suggesting a unique mechanism of action for this novel analogue. The phorbol ester receptors, protein kinase C (PKC) and β2-chimaerin, were recently identified as selective targets for AD 198 in vitro. In vitro, AD 198 competes with [3H]PDBu for binding to a peptide containing the isolated C1b domain of PKC-δ (δC1b domain). In the present study molecular modeling is used to investigate the interaction of AD 198 with the δC1b domain. Three models are identified wherein AD 198 binds into the groove formed between amino acid residues 6-13 and 21-27 of the δC1b domain in a manner similar to that reported for phorbol-13-acetate and other ligands of the C1 domain. Two of the identified models are consistent with previous experimental data demonstrating the importance of the 14-valerate side chain of AD 198 in binding to the C1 domain as well as current data demonstrating that translocation of PKC-α to the membrane requires the 14-valerate substituent. In this regard, the carbonyl of the 14-valerate participates in hydrogen bonding to the δC1b while the acyl chain is positioned for stabilization of the membrane-bound protein-ligand complex in a manner analogous to the acyl chains of the phorbol esters. These studies provide a structural basis for the interaction of AD 198 with the δC1b domain and a starting point for the rational design of potential new drugs targeting PKC and other proteins with C1 domains

Protection from Doxorubicin-Induced Cardiomyopathy Using the Modified Anthracycline N-Benzyladriamycin-14-valerate (AD 198)

The anthracycline doxorubicin (Dox) is an effective antitumor agent. However, its use is limited because of its toxicity in the heart. N-Benzyladriamycin-14-valerate (AD 198) is a modified anthracycline with antitumor efficacy similar to that of Dox, but with significantly less cardiotoxicity and potentially cardioprotective elements. In the present study, we investigated the possibility of in vivo protective effects of low-dose AD 198 against Dox-induced cardiomyopathy. To do this, rats were divided into four groups: vehicle, Dox (20 mg/kg; single injection day 1), AD 198 (0.3 mg/kg per injection; injections on days 1, 2, and 3), or a combination treatment of Dox + AD 198. Seventy-two hours after beginning treatment, hearts from the Dox group had decreased phosphorylation of AMP kinase and troponin I and reduced poly(ADP-ribose) polymerase, β-tubulin, and serum albumin expression. Dox also increased the phosphorylation of phospholamban and expression of inducible nitric-oxide synthase in hearts. Each of these Dox-induced molecular changes was attenuated in the Dox + AD 198 group. In addition, excised hearts from rats treated with Dox had a 25% decrease in left ventricular developed pressure (LVDP) and a higher than normal increase in LVDP when perfused with a high extracellular Ca2+ solution. The Dox-induced decrease in baseline LVDP and hyper-responsiveness to [Ca2+] was not observed in hearts from the Dox + AD 198 group. Thus Dox, with well established and efficient antitumor protocols, in combination with low levels of AD 198, to counter anthracycline cardiotoxicity, may be a promising next step in chemotherapy

Formulation, Development, and In Vitro Evaluation of a CD22 Targeted Liposomal System Containing a Non-Cardiotoxic Anthracycline for B Cell Malignancies

Doxorubicin cardiotoxicity has led to the development of superior chemotherapeutic agents such as AD 198. However, depletion of healthy neutrophils and thrombocytes from AD 198 therapy must be limited. This can be done by the development of a targeted drug delivery system that delivers AD 198 to the malignant cells. The current research highlights the development and in vitro analysis of targeted liposomes containing AD 198. The best lipids were identified and optimized for physicochemical effects on the liposomal system. Physiochemical characteristics such as size, ζ-potential, and dissolution were also studied. Active targeting to CD22 positive cells was achieved by conjugating anti-CD22 Fab’ to the liposomal surface. Size and ζ-potential of the liposomes was between 115 and 145 nm, and −8 to−15 mV. 30% drug was released over 72 h. Higher cytotoxicity was observed in CD22+ve Daudi cells compared to CD22−ve Jurkat cells. The route of uptake was a clathrin- and caveolin-independent pathway. Intracellular localization of the liposomes was in the endolysosomes. Upon drug release, apoptotic pathways were activated partly by the regulation of apoptotic and oncoproteins such as caspase-3 and c-myc. It was observed that the CD22 targeted drug delivery system was more potent and specific compared to other untargeted formulations

Circumvention of P-GP MDR as a function of anthracycline lipophilicity and charge

From a number of studies it has been suggested that positive charge and degree of lipophilicity dictate, or at least influence, whether anthracyclines are recognized by the apparently clinical important mechanism of tumor cell resistance, i.e., P-gp-mediated multidrug resistance. Using a selected series of analogs in which lipophilicity and or positive charge are altered we find the following: (1) Positively-charged anthracyclines as compared to their neutral counterparts are better recognized by MDR+ cells. (2) With increasing lipophilicity charge becomes less important for MDR recognition. (3) In MDR+ cells with a resistance index to Adriamycin (ADR) of 4534, as compared to an MDR- parental line, almost all of the resistance is circumvented (resistance index = 3) with an anthracycline which does not contain a protonatable nitrogen and is highly lipophilic (partition coefficient, log p = > 1.99). (4) As lipophilicity is increased to log p > 1.99 and nuclear binding is decreased, anthracycline localization switches from nuclear to cytoplasmic which most likely indicates a different cytotoxic target and mechanism of action. (5) Cytoplasmically localized anthracyclines appear to distribute also in mitochondria which suggests these organelles as possible new anthracycline targets. In contrast, ADR shows no mitochondrial localization. (6) Photoaffinity analysis suggests that the highly lipophilic analogs, regardless of charge, interfere with NASV-Vp binding to P-gp. This is consistent with the idea that highly lipophilic anthracyclines act as modulators of MDR which may contribute to their mechanism of overcoming this form of resistance. The possible clinical significance of these data is that highly lipophilic anthracyclines are shown to circumvent MDR which most likely reflects their ability to localize in the cytoplasm and affect targets other than nuclear DNA, i.e., mitochondria, and to act as self modulators of MDR. Thus, a new approach to circumventing MDR and other mechanisms of resistance which involve nuclear targets is the use of active anthracyclines which are highly lipophilic and localize in the cytoplasm/mitochondria

Deconstructing gender in trans-gender identities

In this theoretically informed clinical study the author draws upon the psychoanalytic and group therapeutic literature in addition to the works of Judith Butler and his own clinical group analytic work with trans-gender in order to discuss the author’s hypothesis that binary gender rigidity stands at the core of trans-gender states. The author suggests that the analytic task is to deconstruct gender and trans-gender constructions in working with these patients. In addition to working towards greater analytic understanding such an endeavour may also be considered as a social, political and cultural exercise in working towards shifts in our societal foundation matrix. Small group psychotherapy is used as a medium for these observations and as the basis for this study