Structure-based virtual screening for the discovery of natural inhibitors for human rhinovirus coat protein

Inhibitors of the human rhinovirus (HRV) coat protein are promising candidates to treat and prevent a number of upper respiratory diseases. The aim of this study was to find antiviral compounds from nature, focusing on the HRV coat protein. Through computational structure-based screening of an in-house 3D database containing 9676 individual plant metabolites from ancient herbal medicines, combined with knowledge from traditional use, we selected sesquiterpene coumarins from the gum resin asafetida as promising natural products. Chromatographic separation steps resulted in the isolation of microlobidene (1), farnesiferol C (2), farnesiferol B (3), and kellerin (4). Determination of the inhibition of the HRV-induced cytopathic effect for serotypes 1A, 2, 14, and 16 revealed a dose-dependent and selective antirhinoviral activity against serotype 2 for asafetida (IC50 = 11.0 microg/mL) and its virtually predicted constituents 2 (IC50 = 2.5 microM) and 3 (IC50 = 2.6 microM). Modeling studies helped to rationalize the retrieved results.     

Discovery of potent and selective agonists for the free fatty acid receptor 1 (FFA(1)/GPR40), a potential target for the treatment of type II diabetes

A series of 4-phenethynyldihydrocinnamic acid agonists of the free fatty acid receptor 1 (FFA(1)) has been discovered and explored. The preferred compound 20 (TUG-424, EC(50) = 32 nM) significantly increased glucose-stimulated insulin secretion at 100 nM and may serve to explore the role of FFA(1) in metabolic diseases such as diabetes or obesity.     

Seasonal changes and effect of harvest on glucosinolates in Isatis leaves

The seasonal fluctuation of glucosinolates in five defined Isatis tinctoria and one Isatis indigotica accessions (first year, rosette stage), grown on field plots under identical conditions, was investigated. Analysis of the intact glucosinolates was carried out with shock frozen, freeze dried leaf samples using a recently developed and validated PLE (pressurized liquid extraction) protocol and ion-pair HPLC coupled with ESI-MS in the negative mode. When comparing the two Isatis species, significant qualitative and quantitative differences in the glucosinolate patterns were observed. Differences among the various Isatis tinctoria accessions were much smaller. We studied the effects of repeated harvesting during the growth season on glucosinolate concentrations and found that repeated harvest did not have a major effect on glucosinolate concentrations of newly grown leaves. Glucosinolates could not be detected in woad leaves submitted to conventional drying.     

Transcutaneous immunization with CD40 ligation boosts cytotoxic T lymphocyte mediated antitumor immunity independent of CD4 helper cells in mice

Transcutaneous immunization (TCI) is a novel vaccination strategy that utilizes skin‐associated lymphatic tissue to induce immune responses. Employing T‐cell epitopes and the TLR7 agonist imiquimod onto intact skin mounts strong primary, but limited memory CTL responses. To overcome this limitation, we developed a novel imiquimod‐containing vaccination platform (IMI‐Sol) rendering superior primary CD8⁺ and CD4⁺ T‐cell responses. However, it has been unclear whether IMI‐Sol per se is restricted in terms of memory formation and tumor protection. In our present work, we demonstrate that the combined administration of IMI‐Sol and CD40 ligation unleashes fullblown specific T‐cell responses in the priming and memory phase, strongly enhancing antitumor protection in mice. Interestingly, these effects were entirely CD4⁺ T cell independent, bypassing the necessity of helper T cells. Moreover, blockade of CD70 in vivo abrogated the boosting effect of CD40 ligation, indicating that the adjuvant effect of CD40 in TCI is mediated via CD70 on professional APCs. Furthermore, this work highlights the so far underappreciated importance of the CD70/CD27 interaction as a promising adjuvant target in TCI. Summing up, we demonstrate that the novel formulation IMI‐Sol represents a powerful vaccination platform when applied in combination with sufficient adjuvant thereby overcoming current limitations of TCI.     

The in vitro effective antiviral action of povidone-iodine (PVP-I) may also have therapeutic potential by its intravenous administration diluted with Ringer's solution

The use of povidone-iodine (PVP-I) is well known in clinical medical practice. In vitro studies of cell cultures infected by HIV and H5N1 virus have shown that PVP-I has an antiviral action, while the cell hosts were not affected and survived. It is therefore worth investigating whether PVP-I, diluted with Ringer's solution, may have a therapeutic effect by parenteral administration. Specifically, the question is whether small concentrations of intravenous PVP-I could be well tolerated by the human organism, and in addition, if it would be possible to detect a beneficial activity. Its intravenous use may have a potential value against infections (by microbes, viruses, fungi and parasites), as well as an anti-inflammatory activity, especially in cases where antibiotics are ineffective. It could be used as a blood disinfectant, for treating burns, for the prevention of cancer, for the therapy of H5N1 influenza after its mutation, and other potential applications.     

Prevalence and course of sleep problems in childhood

STUDY OBJECTIVES: The Cologne Children's Sleep Study intended to provide information on prevalence and course of difficulties of initiating and maintaining sleep in childhood. DESIGN: Longitudinal study. SETTING: Children of the fourth grade of elementary schools in Cologne. PARTICIPANTS: 832 children and their parents; the mean age of the children was 9.4, 10.7, and 11.7 years at the 3 assessments. MEASUREMENTS AND RESULTS: Children and parents were surveyed using questionnaires 3 times on an annual basis. In self- and parental reports, about 30%-40% of the children of the longitudinal sample had problems falling asleep at the first assessment. One year later, about 30% to 40% of these children did not describe any difficulties initiating sleep, whereas about 60% did report continuing difficulties initiating sleep. Difficulties maintaining sleep are less common in childhood. The analysis of self- and parental reports revealed that in general children described significantly more difficulties initiating and maintaining sleep than their parents report. CONCLUSIONS: Difficulties initiating and maintaining sleep may be transient or persistent. In practice, children and adolescents should be included in the diagnostic and therapeutic process.     

Spinal muscular atrophy: SMN2 pre-mRNA splicing corrected by a U7 snRNA derivative carrying a splicing enhancer sequence.

Spinal muscular atrophy (SMA) is a lethal hereditary disease caused by homozygous deletion/inactivation of the survival of motoneuron 1 (SMN1) gene. The nearby SMN2 gene, despite its identical coding capacity, is only an incomplete substitute, because a single nucleotide difference impairs the inclusion of its seventh exon in the messenger RNA (mRNA). This splicing defect can be corrected (transiently) by specially designed oligonucleotides. Here we have developed a more permanent correction strategy based on bifunctional U7 small nuclear RNAs (snRNAs). These carry both an antisense sequence that allows specific binding to exon 7 and a splicing enhancer sequence that will improve the recognition of the targeted exon. When expression cassettes for these RNAs are stably introduced into cells, the U7 snRNAs become incorporated into small nuclear ribonucleoprotein (snRNP) particles that will induce a durable splicing correction. We have optimized this strategy to the point that virtually all SMN2 pre-mRNA becomes correctly spliced. In fibroblasts from an SMA patient, this approach induces a prolonged restoration of SMN protein and ensures its correct localization to discrete nuclear foci (gems).     

Training cardiothoracic surgeons of the future: The UK experience

Objective

The study aimed to assess targeted simulation courses, including live animal operating, as complementary training tools with regard to 2 key surgical skills in early cardiothoracic surgeon training.

Metastatic bone pain: treatment options with an emphasis on bisphosphonates

INTRODUCTION: One of the key targets for metastatic cancer cells is the skeleton. Once metastatic cells are established within the bone matrix, skeletal integrity becomes increasingly compromised. Bone lesions lead to various complications, including bone pain, fractures and spinal cord compression. MECHANISMS OF BONE PAIN: Bone pain is debilitating and affects quality of life of the patient. In addition, it increases the use of health care resources. Many patients with metastatic bone disease experience substantial bone pain despite state-of-the-art systemic analgesic treatment. Incident pain is the predominant pain syndrome. TREATMENT OPTIONS FOR BONE PAIN: Typically, this syndrome requires moderate baseline analgesia with increased on-demand doses. Other techniques for treating bone pain, including radiation therapy, neuraxial application of analgesics, nerve blocks and local stabilisation procedures, should be considered. In addition, therapy with bisphosphonates targeting bone-specific pain is an important strategy. This review discusses the various management options for bone pain arising from metastatic bone disease.