Strategic targeting of the temporal lobe with orthogonal placement of responsive neurostimulation leads.

Responsive neurostimulation (RNS) is an effective treatment modality for refractory temporal lobe epilepsy (TLE). However, the optimal placement of RNS leads is not known. We use an orthogonal approach to lead placement instead of the more common longitudinal approach to target the entorhinal cortex (EC), given its potential for modulating activity entering and leaving the hippocampus. An orthogonal approach allows for coverage of the EC as well as the anterior lateral temporal cortex, which may be particularly beneficial for patients with mesial-lateral TLE and may also enable greater modulation of the limbic network. The objective of this study was to determine treatment outcomes for orthogonally placed RNS depth leads targeting the EC. We performed a retrospective analysis of prospectively collected data on a cohort of 13 patients. Mean follow-up duration was 57.3 months, and the 50% responder rate was 76.9%. These results show that orthogonally placed RNS leads are safe and effective for the treatment of refractory TLE. Larger cohorts are needed to further delineate the clinical utility of this novel targeting strategy.

Valorization of spent brewery yeast biosorbent with sonication-assisted adsorption for dye removal in wastewater treatment.

The effluent of textile industries containing synthetic dyes contributed to substantial pollution to water bodies. The biosorption process of Congo Red dye was successfully performed by integrating ultrasonication in the adsorption step with spent brewery yeast as a novel and renewable biosorbent. The adsorption process was hindered when ultrasonication was employed together with the biosorbent, indicating that desorption process had occurred. The adsorption process showed that 4 g/L of biosorbent was the optimum dosage for adsorption of 50 mg/L of Congo Red dye, and that the adsorption equilibrium fitted to the Langmuir model, with kinetics best fitted with pseudo-second order model. The maximum capacity of the adsorption was 52.6 mg/g, showing the potential of spent brewery yeast to aid in removing wastewater pollutants. Maximal Congo Red dye recovery (100%) was achieved in the sonication-assisted desorption studies using 0.01M NaOH as the eluting agent. The ultrasonication effects contributed to the efficient recovery of dye and good conversion of spent brewery yeast to biosorbent can be beneficial for treating pollution from textile wastewater.

GluA2 mRNA distribution and regulation by miR-124 in hippocampal neurons.

AMPA-type glutamate receptors mediate fast, excitatory neurotransmission in the brain, and their concentrations at synapses are important determinants of synaptic strength. We investigated the post-transcriptional regulation of GluA2, the calcium-impermeable AMPA receptor subunit, by examining the subcellular distribution of its mRNA and evaluating its translational regulation by microRNA in cultured mouse hippocampal neurons. Using computational approaches, we identified a conserved microRNA-124 (miR-124) binding site in the 3'UTR of GluA2 and demonstrated that miR-124 regulated the translation of GluA2 mRNA reporters in a sequence-specific manner in luciferase assays. While we hypothesized that this regulation might occur in dendrites, our biochemical and fluorescent in situ hybridization (FISH) data indicate that GluA2 mRNA does not localize to dendrites or synapses of mouse hippocampal neurons. In contrast, we detected significant concentrations of miR-124 in dendrites. Overexpression of miR-124 in dissociated neurons results in a 30% knockdown of GluA2 protein, as measured by immunoblot and quantitative immunocytochemistry, without producing any changes in GluA2 mRNA concentrations. While total GluA2 concentrations are reduced, we did not detect any changes in the concentration of synaptic GluA2. We conclude from these results that miR-124 interacts with GluA2 mRNA in the cell body to downregulate translation. Our data support a model in which GluA2 is translated in the cell body and subsequently transported to neuronal dendrites and synapses, and suggest that synaptic GluA2 concentrations are modified primarily by regulated protein trafficking rather than by regulated local translation.

Risk factors in idiopathic adhesive capsulitis: a case control study.

BACKGROUND: The etiology of idiopathic adhesive capsulitis (IAC) of the shoulder is poorly understood. In this case control study, we examine potential risk factors for the development of IAC. METHODS: Consecutive patients who presented to the senior author with IAC between 2000 and 2009 were included retrospectively in this case control study. Complete data were available for 87 patients. An age- and sex-matched group of 176 patients who presented to the same practice during the same time period with non-shoulder related orthopedic complaints were recruited as the control group. Health records and patient-completed questionnaires were utilized to identify comorbidities and other risk factors. RESULTS: Bivariate analyses demonstrated that diabetes, hypothyroidism, a lower body weight, a lower body mass index (BMI), and a positive family history of IAC were all risk factors for IAC. Diabetes, BMI, and positive family history of IAC remained independent variables with multivariate logistic regression analyses. There was a trend towards increased incidence of Dupuytren's disease in those with IAC, but this was not statistically significant. With regard to racial predilection, being born in the British Isles or having parents/grandparents born in the British Isles were risk factors for IAC. CONCLUSION: We confirm diabetes as an independent predictor of IAC. In addition, we identify a possible racial predilection for the development of IAC. Future research is needed to confirm whether a specific genetic component or environmental factors is responsible.

Cerebellar transcranial magnetic stimulation in psychotic disorders: intermittent, continuous, and sham theta-burst stimulation on time perception and symptom severity.

Background: The cerebellum contributes to the precise timing of non-motor and motor functions, and cerebellum abnormalities have been implicated in psychosis pathophysiology. In this study, we explored the effects of cerebellar theta burst stimulation (TBS), an efficient transcranial magnetic stimulation protocol, on temporal discrimination and self-reported mood and psychotic symptoms. Methods: We conducted a case-crossover study in which patients with psychosis (schizophrenias, schizoaffective disorders, or bipolar disorders with psychotic features) were assigned to three sessions of TBS to the cerebellar vermis: one session each of intermittent (iTBS), continuous (cTBS), and sham TBS. Of 28 enrolled patients, 26 underwent at least one TBS session, and 20 completed all three. Before and immediately following TBS, participants rated their mood and psychotic symptoms and performed a time interval discrimination task (IDT). We hypothesized that cerebellar iTBS and cTBS would modulate these measures in opposing directions, with iTBS being adaptive and cTBS maladaptive. Results: Reaction time (RT) in the IDT decreased significantly after iTBS vs. Sham (LS-mean difference = -73.3, p = 0.0001, Cohen's d = 1.62), after iTBS vs. cTBS (LS-mean difference = -137.6, p < 0.0001, d = 2.03), and after Sham vs. cTBS (LS-mean difference = -64.4, p < 0.0001, d = 1.33). We found no effect on IDT accuracy. We did not observe any effects on symptom severity after correcting for multiple comparisons. Conclusion: We observed a frequency-dependent dissociation between the effects of iTBS vs. cTBS to the cerebellar midline on the reaction time of interval discrimination in patients with psychosis. iTBS showed improved (adaptive) while cTBS led to worsening (maladaptive) speed of response. These results demonstrate behavioral target engagement in a cognitive dimension of relevance to patients with psychosis and generate testable hypotheses about the potential therapeutic role of cerebellar iTBS in this clinical population. Clinical Trial Registration: clinicaltrials.gov, identifier NCT02642029.

Cannabis use and social anxiety in young adulthood: A meta-analysis.

Young adulthood (ages 18 to 30 years old), a developmental age of exploration, is marked by new experiences and transitions. Cannabis use frequency is highest in young adulthood compared to other age periods. Social anxiety (characterized by fear, shyness, and inhibition in social situations where scrutiny and judgment is possible) is also prevalent during young adulthood. Social anxiety may be a complex predictor of cannabis use frequency and problems (e.g., any negative physical, emotional, or social outcome from use). Social anxiety may act as a risk factor as individuals may use cannabis frequently to manage their fear of negative evaluation and associated unpleasant affective states. The purpose of this meta-analysis was to quantify the magnitude of the associations between social anxiety and two cannabis variables (frequency of use and problems) in young adulthood. A comprehensive literature review was conducted to identify studies that included measures of social anxiety and at least one cannabis-related variable of interest among young adults. Eighteen studies were included in the meta-analysis. Results revealed a small, statistically significant positive association between social anxiety and cannabis problems (r = 0.197, k = 16, p = <0.001), and a nonsignificant association between social anxiety and cannabis use frequency (r = 0.002, k = 16, p = 0.929). The association between social anxiety and cannabis use frequency was moderated by the mean age such that samples with older mean ages exhibited a stronger correlation. Additionally, the association between social anxiety and cannabis problems was moderated by clinically significant levels of social anxiety, such that samples with fewer participants who met clinical levels of social anxiety exhibit a stronger correlation. This meta-analysis supports the idea that there is a complex relation between social anxiety and cannabis outcomes during young adulthood.

Predictors and outcome of aftercare participation of alcohol and drug users completing residential treatment.

This study attempts to identify factors associated with greater aftercare participation for 367 adults who completed abstinence-based residential addiction treatment between 2004 and 2007 at Bellwood Health Services in Toronto, Canada. Pre-treatment substance use, number of days spent in residential treatment, motivation, treatment satisfaction, and demographics were used to determine which characteristics predicted greater aftercare participation. The duration of residential treatment and treatment satisfaction emerged as significant predictors of aftercare attendance. Regular aftercare attendance was associated with lower levels of substance use at 6-month follow-up. Results suggest that a longer duration of residential treatment can influence continuing care engagement and highlight the importance of initial treatment retention for long-term recovery.

Activity-Dependent Regulation of Alternative Cleavage and Polyadenylation During Hippocampal Long-Term Potentiation.

Long-lasting forms of synaptic plasticity that underlie learning and memory require new transcription and translation for their persistence. The remarkable polarity and compartmentalization of neurons raises questions about the spatial and temporal regulation of gene expression within neurons. Alternative cleavage and polyadenylation (APA) generates mRNA isoforms with different 3' untranslated regions (3'UTRs) and/or coding sequences. Changes in the 3'UTR composition of mRNAs can alter gene expression by regulating transcript localization, stability and/or translation, while changes in the coding sequences lead to mRNAs encoding distinct proteins. Using specialized 3' end deep sequencing methods, we undertook a comprehensive analysis of APA following induction of long-term potentiation (LTP) of mouse hippocampal CA3-CA1 synapses. We identified extensive LTP-induced APA changes, including a general trend of 3'UTR shortening and activation of intronic APA isoforms. Comparison with transcriptome profiling indicated that most APA regulatory events were uncoupled from changes in transcript abundance. We further show that specific APA regulatory events can impact expression of two molecules with known functions during LTP, including 3'UTR APA of Notch1 and intronic APA of Creb1. Together, our results reveal that activity-dependent APA provides an important layer of gene regulation during learning and memory.

The cell biology of synaptic plasticity.

Synaptic plasticity is the experience-dependent change in connectivity between neurons that is believed to underlie learning and memory. Here, we discuss the cellular and molecular processes that are altered when a neuron responds to external stimuli, and how these alterations lead to an increase or decrease in synaptic connectivity. Modification of synaptic components and changes in gene expression are necessary for many forms of plasticity. We focus on excitatory neurons in the mammalian hippocampus, one of the best-studied model systems of learning-related plasticity.

Regulation of RB transcription in vivo by RB family members.

In cancer cells, the retinoblastoma tumor suppressor RB is directly inactivated by mutation in the RB gene or functionally inhibited by abnormal activation of cyclin-dependent kinase activity. While variations in RB levels may also provide an important means of controlling RB function in both normal and cancer cells, little is known about the mechanisms regulating RB transcription. Here we show that members of the RB and E2F families bind directly to the RB promoter. To investigate how the RB/E2F pathway may regulate Rb transcription, we generated reporter mice carrying an eGFP transgene inserted into a bacterial artificial chromosome containing most of the Rb gene. Expression of eGFP largely parallels that of Rb in transgenic embryos and adult mice. Using these reporter mice and mutant alleles for Rb, p107, and p130, we found that RB family members modulate Rb transcription in specific cell populations in vivo and in culture. Interestingly, while Rb is a target of the RB/E2F pathway in mouse and human cells, Rb expression does not strictly correlate with the cell cycle status of these cells. These experiments identify novel regulatory feedback mechanisms within the RB pathway in mammalian cells.

GFP reporter mice for the retinoblastoma-related cell cycle regulator p107.

The RB tumor suppressor gene is mutated in a broad range of human cancers, including pediatric retinoblastoma. Strikingly, however, Rb mutant mice develop tumors of the pituitary and thyroid glands, but not retinoblastoma. Mouse genetics experiments have demonstrated that p107, a protein related to pRB, is capable of preventing retinoblastoma, but not pituitary tumors, in Rb-deficient mice. Evidence suggests that the basis for this compensatory function of p107 is increased transcription of the p107 gene in response to Rb inactivation. To begin to address the context-dependency of this compensatory role of p107 and to follow p107 expression in vivo, we have generated transgenic mice carrying an enhanced GFP (eGFP) reporter inserted into a bacterial artificial chromosome (BAC) containing the mouse p107 gene. Expression of the eGFP transgene parallels that of p107 in these transgenic mice and identifies cells with a broad range of expression level for p107, even within particular organs or tissues. We also show that loss of Rb results in the upregulation of p107 transcription in specific cell populations in vivo, including subpopulations of hematopoietic cells. Thus, p107 BAC-eGFP transgenic mice serve as a useful tool to identify distinct cell types in which p107 is expressed and may have key functions in vivo, and to characterize changes in cellular networks accompanying Rb deficiency.

Expression and function of BMP3 during chick limb development.

Bone morphogenetic proteins (BMPs) play diverse roles in many aspects of skeletal development and bone homeostasis. During endochondral ossification, tight regulation of BMP activity is required to assure proper survival, proliferation and differentiation of skeletal progenitor cells into chondrocytes and osteoblasts. BMP3, a structurally divergent member of the BMP family, acts as a negative regulator of bone formation by limiting BMP signal transduction. In this study, we focus on the chick limb where we find BMP3 has a unique localization pattern with strong expression in the developing perichondrium. Overexpression of BMP3 in chick wing bud at the onset of chondrogenesis, using replication competent retrovirus, reduces BMP signaling leading to increased cell proliferation and delayed cell differentiation, resulting in expanded skeletal elements and joint fusions. Our results suggest that BMP3 expression in the perichondrium may serve to regulate cartilage cell proliferation by modulating the levels of BMP signaling, thus ensuring proper endochondral ossification.

Resistance surveillance in chronic hepatitis B patients treated with adefovir dipivoxil for up to 60 weeks.

Current therapies for chronic hepatitis B virus (HBV) infection do not provide adequate long-term control of viral replication in the majority of patients. Monotherapy with nucleoside analogs, such as lamivudine and famciclovir, is effective for short periods but results in the emergence of drug-resistant HBV in a substantial number of patients within 1 year of therapy. Adefovir dipivoxil (ADV) has demonstrated clinical activity against wild-type and lamivudine-resistant HBV, but it is unclear whether resistance mutations will emerge after long-term therapy with this drug. To determine whether extended treatment with ADV led to the emergence of drug-resistant populations of HBV, we analyzed virus isolated from patients currently enrolled in a long-term open-label study. The reverse transcriptase domain of HBV polymerase was amplified and sequenced from patients that had received a cumulative exposure of up to 60 weeks of ADV. During our analyses, several previously unreported amino acid substitutions were observed in the reverse transcriptase domain of HBV. Importantly, none of the observed mutations occurred in more than 1 patient, nor were they associated with an adefovir-resistant phenotype in vitro. Furthermore, none of the patients from whom these mutant viruses were isolated had evidence of virologic rebound. In conclusion, these results, although based on a limited number of patients, suggest that treatment with ADV does not lead to the emergence of resistant virus after up to 60 weeks of therapy.