Genetic Relationships and Therapeutic Options for Relapsed Acute Lymphoblastic Leukemia

Acute lymphoblastic leukemia (ALL) is the most common form of cancer among children and can be lethal to the adult population. Though 80% of patients with ALL reach complete remission after treatment, about 20% of those diagnosed fail to remain cancer-free. Genetic rearrangements are the hallmark of relapsed ALL, but the mechanism by which these rearrangements occur is still unclear. Recent research suggests these mutations may be detectable during initial diagnosis. If researchers are able to accurately assess the probability of relapse during diagnosis by analyzing the genome of the leukemic cells, the likelihood of administering effective therapy would increase. Providing patients with a more appropriate therapy early on may prevent relapse altogether or increase survival rates after treatment for relapsed ALL patients

Zebrafish behavioural profiling identifies GABA and serotonin receptor ligands related to sedation and paradoxical excitation.

Anesthetics are generally associated with sedation, but some anesthetics can also increase brain and motor activity-a phenomenon known as paradoxical excitation. Previous studies have identified GABAA receptors as the primary targets of most anesthetic drugs, but how these compounds produce paradoxical excitation is poorly understood. To identify and understand such compounds, we applied a behavior-based drug profiling approach. Here, we show that a subset of central nervous system depressants cause paradoxical excitation in zebrafish. Using this behavior as a readout, we screened thousands of compounds and identified dozens of hits that caused paradoxical excitation. Many hit compounds modulated human GABAA receptors, while others appeared to modulate different neuronal targets, including the human serotonin-6 receptor. Ligands at these receptors generally decreased neuronal activity, but paradoxically increased activity in the caudal hindbrain. Together, these studies identify ligands, targets, and neurons affecting sedation and paradoxical excitation in vivo in zebrafish

Initial severity of depression and efficacy of cognitive-behavioural therapy: individual-participant data meta-analysis of pill-placebo-controlled trials

BACKGROUND: The influence of baseline severity has been examined for antidepressant medications but has not been studied properly for cognitive-behavioural therapy (CBT) in comparison with pill placebo. AIMS: To synthesise evidence regarding the influence of initial severity on efficacy of CBT from all randomised controlled trials (RCTs) in which CBT, in face-to-face individual or group format, was compared with pill-placebo control in adults with major depression. METHOD: A systematic review and an individual-participant data meta-analysis using mixed models that included trial effects as random effects. We used multiple imputation to handle missing data. RESULTS: We identified five RCTs, and we were given access to individual-level data (n = 509) for all five. The analyses revealed that the difference in changes in Hamilton Rating Scale for Depression between CBT and pill placebo was not influenced by baseline severity (interaction P = 0.43). Removing the non-significant interaction term from the model, the difference between CBT and pill placebo was a standardised mean difference of -0.22 (95% CI -0.42 to -0.02, P = 0.03, I2 = 0%). CONCLUSIONS: Patients suffering from major depression can expect as much benefit from CBT across the wide range of baseline severity. This finding can help inform individualised treatment decisions by patients and their clinicians.R01 MH060998 - NIMH NIH HHS; R34 MH086668 - NIMH NIH HHS; R01 AT007257 - NCCIH NIH HHS; R21 MH101567 - NIMH NIH HHS; K02 MH001697 - NIMH NIH HHS; R01 MH060713 - NIMH NIH HHS; R34 MH099311 - NIMH NIH HHS; R21 MH102646 - NIMH NIH HHS; K23 MH100259 - NIMH NIH HHS; R01 MH099021 - NIMH NIH HH

Clades of huge phages from across Earth's ecosystems.

Bacteriophages typically have small genomes1 and depend on their bacterial hosts for replication2. Here we sequenced DNA from diverse ecosystems and found hundreds of phage genomes with lengths of more than 200 kilobases (kb), including a genome of 735 kb, which is-to our knowledge-the largest phage genome to be described to date. Thirty-five genomes were manually curated to completion (circular and no gaps). Expanded genetic repertoires include diverse and previously undescribed CRISPR-Cas systems, transfer RNAs (tRNAs), tRNA synthetases, tRNA-modification enzymes, translation-initiation and elongation factors, and ribosomal proteins. The CRISPR-Cas systems of phages have the capacity to silence host transcription factors and translational genes, potentially as part of a larger interaction network that intercepts translation to redirect biosynthesis to phage-encoded functions. In addition, some phages may repurpose bacterial CRISPR-Cas systems to eliminate competing phages. We phylogenetically define the major clades of huge phages from human and other animal microbiomes, as well as from oceans, lakes, sediments, soils and the built environment. We conclude that the large gene inventories of huge phages reflect a conserved biological strategy, and that the phages are distributed across a broad bacterial host range and across Earth's ecosystems

Management of erectile dysfunction post-radical prostatectomy

© 2015 Saleh et al.Radical prostatectomy is a commonly performed procedure for the treatment of localized prostate cancer. One of the long-term complications is erectile dysfunction. There is little consensus on the optimal management; however, it is agreed that treatment must be prompt to prevent fibrosis and increase oxygenation of penile tissue. It is vital that patient expectations are discussed, a realistic time frame of treatment provided, and treatment started as close to the prostatectomy as possible. Current treatment regimens rely on phosphodiesterase 5 inhibitors as a first-line therapy, with vacuum erection devices and intraurethral suppositories of alprostadil as possible treatment combination options. With nonresponders to these therapies, intracavernosal injections are resorted to. As a final measure, patients undergo the highly invasive penile prosthesis implantation. There is no uniform, objective treatment program for erectile dysfunction post-radical prostatectomy. Management plans are based on poorly conducted and often underpowered studies in combination with physician and patient preferences. They involve the aforementioned drugs and treatment methods in different sequences and doses. Prospective treatments include dietary supplements and gene therapy, which have shown promise with there proposed mechanisms of improving erectile function but are yet to be applied successfully in human patients

Inhibition of Both HIV-1 Reverse Transcription and Gene Expression by a Cyclic Peptide that Binds the Tat-Transactivating Response Element (TAR) RNA

The RNA response element TAR plays a critical role in HIV replication by providing a binding site for the recruitment of the viral transactivator protein Tat. Using a structure-guided approach, we have developed a series of conformationally-constrained cyclic peptides that act as structural mimics of the Tat RNA binding region and block Tat-TAR interactions at nanomolar concentrations in vitro. Here we show that these compounds block Tat-dependent transcription in cell-free systems and in cell-based reporter assays. The compounds are also cell permeable, have low toxicity, and inhibit replication of diverse HIV-1 strains, including both CXCR4-tropic and CCR5-tropic primary HIV-1 isolates of the divergent subtypes A, B, C, D and CRF01_AE. In human peripheral blood mononuclear cells, the cyclic peptidomimetic L50 exhibited an IC50 ∼250 nM. Surprisingly, inhibition of LTR-driven HIV-1 transcription could not account for the full antiviral activity. Timed drug-addition experiments revealed that L-50 has a bi-phasic inhibition curve with the first phase occurring after HIV-1 entry into the host cell and during the initiation of HIV-1 reverse transcription. The second phase coincides with inhibition of HIV-1 transcription. Reconstituted reverse transcription assays confirm that HIV-1 (−) strand strong stop DNA synthesis is blocked by L50-TAR RNA interactions in-vitro. These findings are consistent with genetic evidence that TAR plays critical roles both during reverse transcription and during HIV gene expression. Our results suggest that antiviral drugs targeting TAR RNA might be highly effective due to a dual inhibitory mechanism

Part I: Design and Synthesis of Novel Drugs to Treat Asthma By Targeting GABAA Receptors in the Lung Part II: Design and Synthesis of Novel Α2/α3 Subtype Selective GABAAR Ligands for CNS Disorders

Part I. Asthma is a major healthcare challenge affecting an estimated 300 million people globally. Over $56 billion in asthma-related healthcare expenses occur in the United States annually. Moreover, asthma accounts for the majority of missed school/work days, Doctor and emergency room visits, and patient hospitalizations in young persons. Consequently, asthma continues to be a significant healthcare burden in terms of morbidity, productivity, and medical costs. Beta 2-adrenergic agonists and inhaled corticosteroids (ICs) are the most commonly prescribed treatments for the acute and chronic management of asthma. Both agents present efficacy, compliance, and adverse side effect concerns. Hence, there is an unmet need for asthma therapies with novel mechanisms of action to better control the disease with decreased adverse side effects. Previously, it was demonstrated that airway smooth muscle (ASM) cells express GABAA receptors (GABAAR\u27s) of the α4 and α5 subunits. Agonists of these GABAAR subtypes can relax ASM acutely. Targeting the limited and overlapping α subunits with subtype selective GABAAR agonists would effect both ASM relaxation and suppression of inflammation in the absence of any off-target CNS activity. Bz/GABAAergic agents have been proven to be safe and have a long clinical safety record. As a result, targeting Bz/GABAAR in the lung and the peripheral nervous system (PNS) would be a novel and effective strategy in a management of asthma in patients. In this vein, novel GABAAR positive allosteric modulators designed specifically for α4/α6 subunit selectivity were synthesized using iterative computational analyses. In addition, a series of deuterated analogs at key metabolic sites (C-3 and C-8 of the imidazobenzodiazepine scaffold) were synthesized to increase the drugs stability so that the drug stays in the body for a longer time to permit lower doses and still effect its anti-asthmatic properties for a longer duration, presumably with less side effects. Furthermore, a library of α4 subtype selective GABAAR ligands which were more hydrophilic to prevent blood brain barrier (BBB) penetration reduced CNS side effects. To obtain better in vitro and in vivo stability, bioisosteric moieties to replace the labile C-3 ester functional groups were designed and synthesized. Preclinical assays such as microsomal stability, cytotoxicity, and sensorimotor impairment have been studied on these novel analogs. Several ligands exhibited the desired properties required for better management of asthma. The results of studies in several models of asthma in vivo reinforces the novel hypothesis, which rests on relaxation of airway smooth muscle (ASM), a decrease in airway hyperresponsiveness (AHR), and a decrease in airway eosinophils, as well as modulating inflammatory cells. These ligands may be potential treatments for childhood asthma and also for the disease in adults. Part II. Nonselective ligands of the α1-3,5βγ 2 subtypes of GABAARs, such as diazepam have been used in the clinic for more than five decades for various central nervous system (CNS) disorders. These drugs exhibit various adverse CNS effects including sedation, ataxia, amnesia, tolerance, and addiction, which are believed to be mediated by α1 subtypes of GABAARs. As a result, these drugs are not applicable to all patients and have limited long-term applications. Despite their adverse CNS effects due to non-selective GABAAR efficacy, novel ligands with better subtype selectivity, efficacy and reduced adverse effects are now emerging to be suitable replacements for these benzos. The GABAAR agonists that possess superior α2/α3 subtype selectivity over the α1 and α5 subtypes are considered to be a promising avenue for development of novel GABAAR ligands to treat various CNS disorders including inflammatory pain, anxiety, neuropathic pain, and epilepsy, while avoiding side effects such as, ataxia, amnesia, tolerance and dependence. Previously, it had been shown the α2/α3 subtype selective Bz/GABAAR positive allosteric modulator (PAM) HZ-166 exhibited anticonvulsant, antihyperalgesic and anxiolytic properties while being devoid of sedation, ataxia, dependence and tolerance. However, the C-3 ester function in HZ-166 was too labile for studies of ADME toxicity. Consequently, research here was carried out to prepare new ligands with better efficacy and stability, which resulted in several new lead compounds including a 1,3-oxazole (KRM-II-81) and a 1,2,4-oxadiazole (MP-III-80). These bioisosteres were synthesized to overcome the problems with the metabolically labile ester functions. Among them KRM-II-81 exhibited prominent anxiolytic, anticonvulsant, antihyperalgesia, and antidepressant activity. An improved synthetic route was developed to better access the key ligand (HZ-166) in gram quantities for further optimization of this privileged scaffold. Synthesis of several new ester bioisosteres, importantly, those which contained deuterium in the scaffold at key metabolic sites, resulted in d1-MP-III-80, d3-MP-III-80, d5-MP-III-80, as well as a few 3-alkyl-1,2,4-oxadiazole derivatives. The in vitro and in vivo evaluation of these new ligands look promising and further investigations in vivo are underway. It is felt these new α2/α3 subtype selective ligands will result in novel compounds for development into effective treatments for anxiety disorders, for depression, for pain syndromes and for treatment of epilepsies with no tolerance nor dependence

Association of Superoxide Dismutase 2 (SOD2) Genotype with Gray Matter Volume Shrinkage in Chronic Alcohol Users: Replication and Further Evaluation of an Addiction Gene Panel.

BackgroundReduction in brain volume, especially gray matter volume, has been shown to be one of the many deleterious effects of prolonged alcohol consumption. High variance in the degree of gray matter tissue shrinkage among alcohol-dependent individuals and a previous neuroimaging genetics report suggest the involvement of environmental and/or genetic factors, such as superoxide dismutase 2 (SOD2). Identification of such underlying factors will help in the clinical management of alcohol dependence.MethodsWe analyzed quantitative magnetic resonance imaging and genotype data from 103 alcohol users, including both light drinkers and treatment-seeking alcohol-dependent individuals. Genotyping was performed using a custom gene array that included genes selected from 8 pathways relevant to chronic alcohol-related brain volume loss.ResultsWe replicated a significant association of a functional SOD2 single nucleotide polymorphism with normalized gray matter volume, which had been reported previously in an independent smaller sample of alcohol-dependent individuals. The SOD2-related genetic protection was observed only at the cohort's lower drinking range. Additional associations between normalized gray matter volume and other candidate genes such as alcohol dehydrogenase gene cluster (ADH), GCLC, NOS3, and SYT1 were observed across the entire sample but did not survive corrections for multiple comparisons.ConclusionConverging independent evidence for a SOD2 gene association with gray matter volume shrinkage in chronic alcohol users suggests that SOD2 genetic variants predict differential brain volume loss mediated by free radicals. This study also provides the first catalog of genetic variations relevant to gray matter loss in chronic alcohol users. The identified gene-brain structure relationships are functionally pertinent and merit replication

Osteopontin ablation ameliorates muscular dystrophy by shifting macrophages to a pro-regenerative phenotype.

In the degenerative disease Duchenne muscular dystrophy, inflammatory cells enter muscles in response to repetitive muscle damage. Immune factors are required for muscle regeneration, but chronic inflammation creates a profibrotic milieu that exacerbates disease progression. Osteopontin (OPN) is an immunomodulator highly expressed in dystrophic muscles. Ablation of OPN correlates with reduced fibrosis and improved muscle strength as well as reduced natural killer T (NKT) cell counts. Here, we demonstrate that the improved dystrophic phenotype observed with OPN ablation does not result from reductions in NKT cells. OPN ablation skews macrophage polarization toward a pro-regenerative phenotype by reducing M1 and M2a and increasing M2c subsets. These changes are associated with increased expression of pro-regenerative factors insulin-like growth factor 1, leukemia inhibitory factor, and urokinase-type plasminogen activator. Furthermore, altered macrophage polarization correlated with increases in muscle weight and muscle fiber diameter, resulting in long-term improvements in muscle strength and function in mdx mice. These findings suggest that OPN ablation promotes muscle repair via macrophage secretion of pro-myogenic growth factors

EFFECTS OF FAST FORWORD ON PHONOLOGICAL AWARENESS AND RAPID NAMING SKILLS OF AT-RISK STUDENTS

The current study examined the efficacy of Fast ForWord, a computer-based intervention designed to improve the auditory processing skills associated with language development and the subsequent acquisition of reading skills. The study used a randomized pre-test and post-test control design to examine the impact of Fast ForWord on the phonological awareness and rapid naming skills, of students who have failed to meet proficiency on the Connecticut Mastery Test in Reading as measured by the Comprehensive Test of Phonological Processing. The sample was recruited from a target population of 78 students from an urban school, between grades four and eight who were identified as at-risk students by scoring at the basic or below basic level on the 2008 Connecticut Mastery Test in Reading. A multiple regression and a two- group MANOVA were conducted as the methods in data analysis in this research. Results of the MANOVA indicated no significant differences in the levels of the independent variable, as defined by treatment and control group. Results of the multiple regression indicated that percentage of program completion predicted posttest phonological scores but not posttest rapid naming scores. Connecticut Mastery Test in Reading scores, when entered in the regression model, did not predict posttest phonological awareness or posttest rapid naming scores