Therapeutic Responses to Two New SN-38 Derivatives in Colorectal Cancer Patient-Derived Xenografts and Respective 3D Cultures.

Background/aim: SN-38, an active metabolite of irinotecan, exhibits toxicity to all proliferating cells, causing dose-limiting and potentially life-threatening side effects. Newly established water-soluble derivatives of SN-38, 7-ethyl-9-(N-morpholinyl)methyl-10-hydroxycamptothecin (BN-MOA) and 7-ethyl-9-(N-methylamino)methyl-10-hydroxycamptothecin (BN-NMe), exhibit a unique mechanism of spontaneous alkylation of aromatic bases in DNA and show greater in vitro activity on cancer cell lines than SN-38. The aim of this study was to compare the therapeutic responses to irinotecan, BN-MOA and BN-NMe in vivo and in vitro in 3D cultures using colorectal cancer (CRC) patient derived xenografts (PDX).

Regulatory framework for genetically modified organisms in the Kingdom of Eswatini.

The Kingdom of Eswatini is a Party to the Convention on Biological Diversity and to the Cartagena Protocol on Biosafety. As Party, Eswatini has domesticated these agreements by passing the Biosafety Act, of 2012 to provide for the safe handling, transfer, and use of living modified organisms (LMOs) in the country. The Act regulates living modified organisms to be used for confined field trials, commercial release, import, export, and transit, and for food, feed, and processing.

High-dose cervical mobilization to improve central sensitization for a patient with post-fusion neck pain.

Background: Few studies have addressed the appropriate joint mobilization dosage for neck pain. Furthermore, the efficacy of manual therapy in patients with post-spinal fusion neck pain is unreported.

Case Description: A 63-year-old man with a 2-year history of unresolved neck pain post-cervical fusion presented to therapy with an exacerbation of neck and left-shoulder pain.

Use of orthopedic manual physical therapy to manage chronic orofacial pain and tension-type headache in an adolescent.

Temporomandibular dysfunction (TMD) is frequently associated with multiple headache types. While the efficacy of orthopedic manual physical therapy (OMPT) intervention for TMD with/without headache symptoms has been investigated, it has received less attention than other musculoskeletal conditions. This case describes the OMPT treatment and outcome of a 14 year-old female with a 2-year history of facial pain surrounding the right temporomandibular joint (TMJ), and bilateral occipital and temporal tension-type headaches (TTHs).

Preoperative mapping of fistula-in-ano: a new three-dimensional MRI-based modelling technique.

Aim: We aimed to develop an intuitive, interactive, three-dimensional (3D) MRI modelling technique to produce a 3D image of fistula-in-ano.

Method: The 3D model was created from standard two-dimensional (2D) MRI sequences to produce an image that is anatomically correct. Individual muscle and soft-tissue layers were extracted from T1-weighted sequences and fistula pathology from short TI inversion recovery (STIR) sequences, to produce two separate volumes.

Training requirements for point of care ultrasound in acute medicine.

Unlabelled: The aim of the study was to explore the ultrasound training requirements in acute medicine by comparing the ultrasound skill of an acute medicine trainee (AMT) with that of the radiology department following short ultrasound training.

Results: 43 participants (34 males and 9 females).The mean age was 58 (range 20-93 years).

Accurate whole human genome sequencing using reversible terminator chemistry.

DNA sequence information underpins genetic research, enabling discoveries of important biological or medical benefit. Sequencing projects have traditionally used long (400-800 base pair) reads, but the existence of reference sequences for the human and many other genomes makes it possible to develop new, fast approaches to re-sequencing, whereby shorter reads are compared to a reference to identify intraspecies genetic variation. Here we report an approach that generates several billion bases of accurate nucleotide sequence per experiment at low cost.

Psycho-educational group treatment for the severely and persistently mentally ill: how much leader training is necessary?

Psycho-educational groups (PEGs) have been shown to be a particularly effective form of treatment for patients classified under the rubric of severely and persistently mentally ill (SPMI). However, recent surveys suggest that certain professionals, such as nurses and clinical psychologists, called on to conduct these groups may have limited to no training. This study tested three methods (self-instructional, workshop, and workshop plus clinical supervision) designed to provide on-the-job training to practicing clinicians.

The kinetic model of the shikimate pathway as a tool to optimize enzyme assays for high-throughput screening.

Four-enzyme section of the shikimate pathway (Aro B, D, E, and K) of Streptococcus pneumoniae has been studied. Kinetic properties of the individual enzymes and three- and four-enzyme linked reactions have been characterized in vitro. On the basis of the data measured in spectrophotometric and LC-MS experiments, kinetic mechanisms of the enzymes have been suggested and all kinetic parameters have been identified.

Educational needs of inpatients with severe & persistent mental illness: a partial replication.

Although a great deal has been written about the importance of patient education, few replications of past research have been conducted to increase clinicians' confidence in the conclusions of previous studies. This is especially true of studies conducted in clinical settings where findings may or may not be generalizable to other institutions. This partial replication study tested whether previous findings about patient education in an outpatient setting were applicable to an inpatient setting using a modified version of the instrument.

Selection of outcome assessment instruments for inpatients with severe and persistent mental illness.

The purpose of this article is to describe a procedure to assist in selecting outcome measures for inpatients treated at a state psychiatric hospital. The procedure combines evidence-based criteria from the literature, instruments shown to be sensitive to change in clinical trials, and the perspectives of a multidisciplinary team of researchers, administrators, providers, and patient advocates. Recent efficacy and effectiveness studies were used to identify recurrently used outcome instruments.

Identification of small molecule inhibitors of the hepatitis C virus RNA-dependent RNA polymerase from a pyrrolidine combinatorial mixture.

HTS of the compound collection for inhibition of the HCV RNA dependent RNA polymerase identified two 168 member N-acyl pyrrolidine combinatorial mixture hits. Deconvolution and expansion of these mixtures by solid phase synthesis to establish initial SAR and identify a potent inhibitor is reported.

Enzymatic activities of the GB virus-B RNA-dependent RNA polymerase.

The GB virus-B (GBV-B) nonstructural protein 5B (NS5B) encodes an RNA-dependent RNA polymerase (RdRp) with greater than 50% sequence similarity to the hepatitis C virus (HCV) NS5B. Recombinant GBV-B NS5B was reported to possess RdRp activity (W. Zhong et al.

The peripheral benzodiazepine receptor ligand PK11195 binds with high affinity to the acute phase reactant alpha1-acid glycoprotein: implications for the use of the ligand as a CNS inflammatory marker.

The peripheral benzodiazepine receptor ligand PK11195 has been used as an in vivo marker of neuroinflammation in positron emission tomography studies in man. One of the methodological issues surrounding the use of the ligand in these studies is the highly variable kinetic behavior of [(11)C]PK11195 in plasma. We therefore undertook a study to measure the binding of [(3)H]PK11195 to whole human blood and found a low level of binding to blood cells but extensive binding to plasma proteins.

A systematic program to enhance clinician group skills in an inpatient psychiatric hospital.

This article describes the collaborative effort of a team of discipline directors, administrators, and academicians to create a systematic program to enhance the group competencies of a large clinical staff working at a state hospital. The effects of the program were tested by a quasi-experimental field study. Quantitative measures of group process provided limited support for program effectiveness.

Identification and biological characterization of heterocyclic inhibitors of the hepatitis C virus RNA-dependent RNA polymerase.

The hepatitis C virus (HCV) NS5B protein encodes an RNA-dependent RNA polymerase (RdRp), the primary catalytic enzyme of the HCV replicase complex. We established a biochemical RNA synthesis assay, using purified recombinant NS5B lacking the C-terminal 21 amino acid residues, to identify potential polymerase inhibitors from a high throughput screen of the GlaxoSmithKline proprietary compound collection. The benzo-1,2,4-thiadiazine compound 1 was found to be a potent, highly specific inhibitor of NS5B.

Novel spermine-amino acid conjugates and basic tripeptides enhance cleavage of the hairpin ribozyme at low magnesium ion concentration.

Combinations of the polyamine spermine and magnesium ions synergize to dramatically enhance cleavage of the hairpin ribozyme. Certain synthetic basic tripeptides stimulate hairpin cleavage significantly at limiting magnesium ion concentration, notably the tripeptide of L-diaminobutyric acid (Dab). Of a range of novel synthetic spermine-amino acid conjugates, L-Dab-spermine (but not D-Dab nor other amino acid conjugates) was more effective than spermine itself.

Real experiences of uHTS: a prototypic 1536-well fluorescence anisotropy-based uHTS screen and application of well-level quality control procedures.

This paper describes, for the first time, a true ultra-high throughput screen (uHTS) based upon fluorescence anisotropy and performed entirely in 1536-well assay plates. The assay is based upon binding and displacement of a BODIPY-FL-labeled antibiotic to a specific binding site on 70S ribosomes from Escherichia coli (Kd approximately 15 nM). The screen was performed at uHTS rates (i.

FlashPlate scintillation proximity assays for characterization and screening of DNA polymerase, primase, and helicase activities.

DNA replication proteins represent a class of extremely well-established anti-infective drug targets for which improvements in assay technology are required in order to support enzyme characterization, HTS, and structure-activity relationship studies. Replication proteins are conventionally assayed using precipitation/filtration or gel-based techniques, and are not yet all suitable for conversion into homogeneous fluorescence-based formats. We have therefore developed radiometric assays for these enzymes based upon FlashPlate technology that can be applied to a wide range of targets using a common set of reagents.

Time-Resolved Fluorescence Energy Transfer DNA Helicase Assays for High Throughput Screening.

DNA helicases are responsible for the unwinding of double-stranded DNA, facilitated by the binding and hydrolysis of 5'-nucleoside triphosphates. These enzymes represent an important class of targets for the development of novel anti-infective agents particularly because opportunity exists for synergy with existing therapies targeted at other enzymes involved in DNA replication. Unwinding reactions are conventionally monitored by low throughput, gel-based radiochemical assays; to overcome the limitations of low throughput to achieve comprehensive characterization of adenosine triphosphate (ATP)-dependent unwinding by viral and bacterial helicases and the screening for unwinding inhibitors, we have developed and validated homogeneous time-resolved fluorescence energy transfer (TRET) assays.

Investigation of the proposed interdomain ribose zipper in hairpin ribozyme cleavage using 2'-modified nucleosides.

The hairpin ribozyme achieves catalytic cleavage through interaction of essential nucleotides located in two distinct helical domains that include internal loops. Initial docking of the two domains is ion dependent and appears to be followed by a structural rearrangement that allows the ribozyme to achieve a catalytically active state that can undergo cleavage. The proposed structural rearrangement may also be ion dependent and is now of increased importance due to recent evidence that docking is not rate limiting and that metal ions are unlikely to be involved in the chemical cleavage step.

The harpin ribozyme: the roles of ions in RNA cleavage.

The cleavage reaction of the hairpin ribozyme is facilitated by divalent metal ions, such as Mg2+, or by non-metallic polycations, such as the polyamine spermine. We show substantial enhancement of cleavage with combinations of metallic and non-metallic ions. In addition we elucidate the locations of some ion binding sites by Fenton chemistry.