Structure of the human κ-opioid receptor in complex with JDTic

Opioid receptors mediate the actions of endogenous and exogenous opioids on many physiological processes, including the regulation of pain, respiratory drive, mood, and—in the case of κ-opioid receptor (κ-OR)—dysphoria and psychotomimesis. Here we report the crystal structure of the human κ-OR in complex with the selective antagonist JDTic, arranged in parallel dimers, at 2.9 Å resolution. The structure reveals important features of the ligand-binding pocket that contribute to the high affinity and subtype selectivity of JDTic for the human κ-OR. Modelling of other important κ-OR-selective ligands, including the morphinan-derived antagonists norbinaltorphimine and 5′-guanidinonaltrindole, and the diterpene agonist salvinorin A analogue RB-64, reveals both common and distinct features for binding these diverse chemotypes. Analysis of site-directed mutagenesis and ligand structure–activity relationships confirms the interactions observed in the crystal structure, thereby providing a molecular explanation for κ-OR subtype selectivity, and essential insights for the design of compounds with new pharmacological properties targeting the human κ-OR

From small molecules to large proteins: Delivery through B family vitamin uptake pathways

Delivery of therapeutic molecules using the B family vitamin uptake pathways is presented. First a study examining targeting the folate receptor to deliver AZT was successfully conducted in vitro. Conjugation of AZT to folic acid resulted in a drug that showed an increase in cytotoxicity of ∼20-fold compared to free AZT, when tested in the A2780/AD drug resistant ovarian cell line. This compound was taken to pilot in vivo studies. Conjugates of the folic acid analogue methotrexate with nucleoside analogues AZT, IUdR and ddC were subsequently studied. The strategy was to conjugate two drugs that would work synergistically together. The individual conjugates did not show any synergism, but a cocktail of the AZT and IUdR conjugates showed a synergistic relationship with a lower IC 50 over 24 hours compared to cocktails of the free drugs. In an attempt to create an oral vaccine, conjugates of tetanus toxoid and B12 were explored. It was hypothesized that the B 12 uptake pathway could protect the toxoid in the digestive tract and facilitate crossing of the enterocyte. The pilot B 12 -tetanus toxoid conjugates were synthesized and characterized. Different ratios of B 12 /tetanus toxoid were synthesized. The conjugates were fluorescently tagged and intestinal uptake was modeled using BeWo placental carcinoma cells, followed by confocal microscopy

Assembly, Biochemical Characterization, Immunogenicity, Adjuvanticity, and Efficacy of Shigella Artificial Invaplex

ABSTRACT The native Invaplex (InvaplexNAT) vaccine and adjuvant is an ion exchange-purified product derived from the water extract of virulent Shigella species. The key component of InvaplexNAT is a high-molecular-mass complex (HMMC) consisting of the Shigella lipopolysaccharide (LPS) and the invasin proteins IpaB and IpaC. To improve product purity and immunogenicity, artificial Invaplex (InvaplexAR) was developed using recombinant IpaB and IpaC proteins and purified Shigella LPS to assemble an HMMC consisting of all three components. Characterization of InvaplexAR by various methods demonstrated similar characteristics as the previously reported HMMC in InvaplexNAT. The well-defined InvaplexAR vaccine consistently contained greater quantities of IpaB, IpaC, and LPS than InvaplexNAT. InvaplexAR and InvaplexNAT immunogenicities were compared in mouse and guinea pig dose escalation studies. In both models, immunization induced antibody responses specific for InvaplexNAT and LPS while InvaplexAR induced markedly higher anti-IpaB and -IpaC serum IgG and IgA endpoint titers. In the murine model, homologous protection was achieved with 10-fold less InvaplexAR than InvaplexNAT and mice receiving InvaplexAR lost significantly less weight than mice receiving the same amount of InvaplexNAT. Moreover, mice immunized with InvaplexAR were protected from challenge with both homologous and heterologous Shigella serotypes. Guinea pigs receiving approximately 5-fold less InvaplexAR compared to cohorts immunized with InvaplexNAT were protected from ocular challenge. Furthermore, adjuvanticity previously attributed to InvaplexNAT was retained with InvaplexAR. The second-generation Shigella Invaplex vaccine, InvaplexAR, offers significant advantages over InvaplexNAT in reproducibility, flexible yet defined composition, immunogenicity, and protective efficacy. IMPORTANCE Shigella species are bacteria that cause severe diarrheal disease worldwide, primarily in young children. Treatment of shigellosis includes oral fluids and antibiotics, but the high burden of disease, increasing prevalence of antibiotic resistance, and long-term health consequences clearly warrant the development of an effective vaccine. One Shigella vaccine under development is termed the invasin complex or Invaplex and is designed to drive an immune response to specific antigens of the bacteria in an effort to protect an individual from infection. The work presented here describes the production and evaluation of a new generation of Invaplex. The improved vaccine stimulates the production of antibodies in immunized mice and guinea pigs and protects these animals from Shigella infection. The next step in the product’s development will be to test the safety and immune response induced in humans immunized with Invaplex

China's Health Reform Update

China experienced both economic and epistemological transitions within the past few decades, greatly increasing demand for accessible and affordable health care. These shifts put significant pressure on the existing outdated, highly centralized bureaucratic system. Adjusting to growing demands, the government has pursued a new round of health reforms since the late 2000s; the main goals are to reform health care financing, essential drug policies, and public hospitals. Health care financing reform led to universal basic medical insurance, whereas the public hospital reform required more complex measures ranging from changes in regulatory, operational, and service delivery settings to personnel management. This article reviews these major policy changes and the literature-based evidence of the effects of reforms on cost, access, and quality of care. It then highlights the outlook for future reforms. We argue that a better understanding of the unintended consequences of reform policies and of how practitioners' and patients' interests can be better aligned is essential for reforms to succeed.SCI(E)SSCIREVIEW431-4483

PERCEPÇÕES SOBRE O TOQUE ENFERMEIRA E PACIENTE VISÃO DOS ALUNOS DE GRADUAÇÃO DE ENFERMAGEM

The touch is considered by the author a important sign nonverbal in the nursew — patient relationship. The authors are teaching in the nursing undergraduate a systematic approach about nonverbal communication, including the touch. The perception of the students after this knowledge are discussed in this paper and also when the touch may be a positive or negative experience to patients. The results of the 32 questionnaries administered to nursing students shows that the nurses are using the functional touch mainly.Por considerar o toque um dos sinais não verbais mais importantes na relação enfermeira-paciente, as autoras passaram a ensinar durante o curso de graduação de enfermagem, um bloco teórico, com aplicação teórico-prática sobre comunicação não verbal, incluindo conceitos gerais do tocar. Este trabalho teve como objetivos verificar como os alunos, após terem passado pelo bloco teórico citado, estavam percebendo a maneira das enfermeiras tocarem os pacientes durante a assistência de enfermagem, como eles tomam a decisão de tocar os pacientes que estão sob seus cuidados e, quando, segundo a percepção dos alunos, o toque pode ser considerado uma experiência positiva ou uma experiência negativa para o paciente. Dentre os dados obtidos, após questionário aplicado a 32 alunos, verificou-se que a maioria das vezes a enfermeira toca o paciente, segundo os alunos, para a execução do exame físico, ou seja, é um toque instrumental

Live-fibroblast IR imaging of a cytoprotective PhotoCORM Activated with Visible Light

Carbon monoxide releasing molecules (CORMs) are an emerging class of pharmaceutical compounds currently evaluated in several preclinical disease models. There is general consensus that the therapeutic effects elicited by the molecules may be directly ascribed to the biological function of the released CO. It remains unclear, however, if cellular internalization of CORMs is a critical event in their therapeutic action. To address the problem of cellular delivery, we have devised a general strategy which entails conjugation of a CO-releasing molecule (here a photoactivated CORM) to the 5'-OH ribose group of vitamin B12. Cyanocobalamin (B12) functions as the biocompatible water-soluble scaffold which actively transports the CORM against a concentration gradient into the cells. The uptake and cellular distribution of this B12-photoCORM conjugate is demonstrated via synchrotron FTIR spectromicroscopy measurements on living cells. Intracellular photoinduced CO release prevents fibroblasts from dying under conditions of hypoxia and metabolic depletion, conditions that may occur in vivo during insufficient blood supply to oxygen-sensitive tissues such as the heart or brain