A Novel Fusion of IL-10 Engineered to Traffic across Intestinal Epithelium to Treat Colitis.

IL-10 is a potent anti-inflammatory cytokine capable of suppressing a number of proinflammatory signals associated with intestinal inflammatory diseases, such as ulcerative colitis and Crohn's disease. Clinical use of human IL-10 (hIL-10) has been limited by anemia and thrombocytopenia following systemic injection, side effects that might be eliminated by a gut-restricted distribution. We have identified a transcytosis pathway used by cholix, an exotoxin secreted by nonpandemic forms of the intestinal pathogen Vibrio cholerae A nontoxic fragment of the first 386 aa of cholix was genetically fused to hIL-10 to produce recombinant AMT-101. In vitro and in vivo characterization of AMT-101 showed it to efficiently cross healthy human intestinal epithelium (SMI-100) by a vesicular transcytosis process, activate hIL-10 receptors in an engineered U2OS osteosarcoma cell line, and increase cellular phospho-STAT3 levels in J774.2 mouse macrophage cells. AMT-101 was taken up by inflamed intestinal mucosa and activated pSTAT3 in the lamina propria with limited systemic distribution. AMT-101 administered to healthy mice by oral gavage or to cynomolgus monkeys (nonhuman primates) by colonic spray increased circulating levels of IL-1R antagonist (IL-1Ra). Oral gavage of AMT-101 in two mouse models of induced colitis prevented associated pathological events and plasma cytokine changes. Overall, these studies suggest that AMT-101 can efficiently overcome the epithelial barrier to focus biologically active IL-10 to the intestinal lamina propria.

Characterizing Heart Rate Variability Response to Maximal Exercise Testing in People with Huntington's Disease.

Background: Huntington's disease (HD) is an autosomal dominant, neurodegenerative disease that involves dysfunction in the autonomic nervous system (ANS). Heart rate variability (HRV) is a valid and noninvasive measure for ANS dysfunction, yet no study has characterized HRV response to exercise in people with HD. Objective: Characterize HRV response to exercise in individuals with HD and explore its implications for exercise prescription and cardiac dysautonomia mechanisms. Methods: 19 participants with HD were recruited as part of a cohort of individuals enrolled in the Physical Activity and Exercise Outcomes in Huntington's Disease (PACE-HD) study at Teachers College, Columbia University (TC). 13 non-HD age- and gender-matched control participants were also recruited from TC. HRV was recorded with a Polar H10 heart rate (HR) monitor before, during, and after a ramp cycle-ergometer exercise test. Results: Participants with HD showed reduced HR peak (p < 0.01) and HR reserve (p < 0.001) compared with controls. Participants with HD demonstrated reduced root mean square of successive differences between normal-to-normal intervals (RMSSD) and successive differences of normal-to-normal intervals (SDSD) at rest (p < 0.001). Participants with HD also showed differences for low frequency (LF) power (p < 0.01), high frequency (HF) normalized units (nu) (p < 0.05), LF (nu) (p < 0.001), and HF/LF ratio (p < 0.05) compared with controls. Conclusions: We found reduced aerobic exercise capacity and sympathovagal dysautonomia both at rest and during post-exercise recovery in people with HD, suggesting modified exercise prescription may be required for people with HD. Further investigations focusing on cardiac dysautonomia and underlying mechanisms of sympathovagal dysautonomia in people with HD are warranted.

Validity and reliability of inertial measurement units for gait assessment within a post stroke population.

BACKGROUND: The ability to objectively measure spatiotemporal metrics within individuals post-stroke is integral to plan appropriate intervention, track recovery, and ultimately improve efficacy of rehabilitation programs. Inertial measurement units (IMUs) provide a means to systematically collect gait-specific metrics that could not otherwise be obtained from clinical outcomes. However, the use of IMUs to measure spatiotemporal parameters in stroke survivors has yet to be validated. The purpose of this study is to determine the validity and reliability of IMU-recorded spatiotemporal gait metrics as compared to a motion capture camera system (MCCS) in individuals post-stroke. METHODS: Participants (n = 23, M/F = 12/11, mean (SD) age = 50.2(11.1) spatiotemporal data were collected simultaneously from a MCCS and APDM Opal IMUs during a five-minute treadmill walking task at a self-selected speed. Criterion validity and test-retest reliability were assessed using Lin's concordance correlation coefficients (CCCs) and intraclass correlation coefficients (ICCs), respectively. Spatiotemporal values from MCCS and IMU were used to calculate gait asymmetry, and a t-test was used to assess the difference between asymmetry values. RESULTS: There were fair-to-excellent agreement between IMU and MCCS of temporal parameters (CCC 0.56-0.98), excellent agreement of spatial parameters (CCC >0.90), and excellent test-retest reliability for all parameters (ICC >0.90). CONCLUSIONS: Compared to motion capture, the APDM Opal IMUs produced accurate and reliable measures of spatiotemporal parameters. Findings support the use of IMUs to assess spatiotemporal parameters in individual's post-stroke.

Antitumor activity of an allosteric inhibitor of centromere-associated protein-E.

Centromere-associated protein-E (CENP-E) is a kinetochore-associated mitotic kinesin that is thought to function as the key receptor responsible for mitotic checkpoint signal transduction after interaction with spindle microtubules. We have identified GSK923295, an allosteric inhibitor of CENP-E kinesin motor ATPase activity, and mapped the inhibitor binding site to a region similar to that bound by loop-5 inhibitors of the kinesin KSP/Eg5. Unlike these KSP inhibitors, which block release of ADP and destabilize motor-microtubule interaction, GSK923295 inhibited release of inorganic phosphate and stabilized CENP-E motor domain interaction with microtubules. Inhibition of CENP-E motor activity in cultured cells and tumor xenografts caused failure of metaphase chromosome alignment and induced mitotic arrest, indicating that tight binding of CENP-E to microtubules is insufficient to satisfy the mitotic checkpoint. Consistent with genetic studies in mice suggesting that decreased CENP-E function can have a tumor-suppressive effect, inhibition of CENP-E induced tumor cell apoptosis and tumor regression.

A Wearable Vibratory Device (The Emma Watch) to Address Action Tremor in Parkinson Disease: Pilot Feasibility Study.

BACKGROUND: Parkinson disease (PD) is a neurodegenerative disease that has a wide range of motor symptoms, such as tremor. Tremors are involuntary movements that occur in rhythmic oscillations and are typically categorized into rest tremor or action tremor. Action tremor occurs during voluntary movements and is a debilitating symptom of PD. As noninvasive interventions are limited, there is an ever-increasing need for an effective intervention for individuals experiencing action tremors. The Microsoft Emma Watch, a wristband with 5 vibrating motors, is a noninvasive, nonpharmaceutical intervention for tremor attenuation. OBJECTIVE: This pilot study investigated the use of the Emma Watch device to attenuate action tremor in people with PD. METHODS: The sample included 9 people with PD who were assessed on handwriting and hand function tasks performed on a digitized tablet. Tasks included drawing horizontal or vertical lines, tracing a star, spiral, writing "elelelel" in cursive, and printing a standardized sentence. Each task was completed 3 times with the Emma Watch programmed at different vibration intensities, which were counterbalanced: high intensity, low intensity (sham), and no vibration. Digital analysis from the tablet captured kinematic, dynamic, and spatial attributes of drawing and writing samples to calculate mathematical indices that quantify upper limb motor function. APDM Opal sensors (APDM Wearable Technologies) placed on both wrists were used to calculate metrics of acceleration and jerk. A questionnaire was provided to each participant after using the Emma Watch to gain a better understanding of their perspectives of using the device. In addition, drawings were compared to determine whether there were any visual differences between intensities. RESULTS: In total, 9 people with PD were tested: 4 males and 5 females with a mean age of 67 (SD 9.4) years. There were no differences between conditions in the outcomes of interest measured with the tablet (duration, mean velocity, number of peaks, pause time, and number of pauses). Visual differences were observed within a small subset of participants, some of whom reported perceived improvement. The majority of participants (8/9) reported the Emma Watch was comfortable, and no problems with the device were reported. CONCLUSIONS: There were visually depicted and subjectively reported improvements in handwriting for a small subset of individuals. This pilot study was limited by a small sample size, and this should be taken into consideration with the interpretation of the quantitative results. Combining vibratory devices, such as the Emma Watch, with task specific training, or personalizing the frequency to one's individual tremor may be important steps to consider when evaluating the effect of vibratory devices on hand function or writing ability in future studies. While the Emma Watch may help attenuate action tremor, its efficacy in improving fine motor or handwriting skills as a stand-alone tool remains to be demonstrated.

Postural control and gait measures derived from wearable inertial measurement unit devices in Huntington's disease: Recommendations for clinical outcomes.

BACKGROUND: Postural control impairments begin early in Huntington's disease yet measures most sensitive to progression have not been identified. The aims of this study were to: 1) evaluate postural control and gait in people with and without Huntington's disease using wearable sensors; and 2) identify measures related to diagnosis and clinical severity. METHODS: 43 individuals with Huntington's disease and 15 age-matched peers performed standing with feet together and feet apart, sitting, and walking with wearable inertial sensors. One-way analysis of variance determined differences in measures of postural control and gait between early and mid-disease stage, and non-Huntington's disease peers. A random forest analysis identified feature importance for Huntington's disease diagnosis. Stepwise and ordinal regressions were used to determine predictors of clinical chorea and tandem walking scores respectively. FINDINGS: There was a significant main effect for all postural control and gait measures comparing early stage, mid stage and non-Huntington's disease peers, except for gait cycle duration and step duration. Total sway, root mean square and mean velocity during sitting, as well as gait speed had the greatest importance in classifying disease status. Stepwise regression showed that root mean square during standing with feet apart significantly predicted clinical measure of chorea, and ordinal regression model showed that root mean square and total sway standing feet together significantly predicted clinical measure of tandem walking. INTERPRETATIONS: Root mean square measures obtained in sitting and standing using wearable sensors have the potential to serve as biomarkers of postural control impairments in Huntington's disease.

Evaluation of gait initiation using inertial sensors in Huntington's Disease: insights into anticipatory postural adjustments and cognitive interference.

BACKGROUND: Understanding the contribution of anticipatory postural adjustments (APA) to walking ability in individuals with Huntington's disease (HD) may provide insight into motor planning and the functional consequences of HD-specific cortical-basal ganglia pathway dysfunctions. RESEARCH QUESTION: How do inertial measurement unit (IMU)-derived APAs and first step parameters differ between individuals with HD and non-HD peers under no load and cognitive load conditions, and what is their relationship to gait speed and clinical measures? METHODS: 33 individuals with manifest HD and 15 non-HD peers wore three Opal APDM IMUs during a 14-meter walk under no load and cognitive load conditions. APA acceleration amplitudes, APA durations, first step range of motion (ROM), and first step durations were compared, along with their relationship to gait speed. RESULTS: Individuals with HD had greater APA acceleration amplitudes, smaller first step ROM and longer first step durations compared to non-HD peers. No differences in APA durations were present between groups in both conditions. Cognitive loading influenced first step ROM but not other APA parameters. Mediolateral APA acceleration amplitudes were a significant predictor of gait speed and were related to disease-specific measures. SIGNIFICANCE: Larger acceleration amplitudes and smaller first step ROMs of greater duration, accompanied by the preservation of APA durations, reveal a discrepancy in movement scaling in HD. Additionally, the mediolateral component of the APA is likely a rate-limiting factor that drives a compensatory response in gait initiation. Further research is needed to explore the neural correlates of HD-related movement scaling.

Impact of diabetes and periodontal status on life quality.

OBJECTIVES: To investigate impact of periodontal status on quality of life (QoL) in type-1 (T1D) and type-2 (T2D) diabetes patients pre- and post-periodontal treatment using the Well-being Questionnaire 12 (W-BQ12) and Audit of Diabetes-Dependent Quality of Life-19 (ADDQoL-19). METHODS: W-BQ12 and ADDQoL-19 were self-completed by 56 T1D and 77 T2D patients at baseline and by those with periodontitis 3 and 6-months after therapy. RESULTS: At baseline, T1D patients had significantly higher general W-BQ12 [Median (IQR); 24.00 (20.25-27.75)] and positive well-being scores [8.00 (6.00-9.00)] (indicating better QoL) compared to T2D patients [22.00 (15.50-26.00) and 6.00 (3.50-9.00)], respectively (p < 0.05). Within both groups, general W-BQ12 scores did not differ significantly between patients with periodontal health, gingivitis, or periodontitis (p > 0.05). Significantly higher general W-BQ12 scores were observed in T1D patients at month 3 [28.00 (22.00-29.50)] compared to baseline [22.00 (17.00-24.50)] (p < 0.01), suggesting an initial improvement in QoL post-treatment. ADDQoL-19 identified that diabetes had greatest impact on the domain 'freedom to eat', with participants placing most importance on 'family life'. No significant changes in ADDQoL-19 scores were seen post-treatment (p > 0.05). CONCLUSIONS: Diabetes had impacts upon aspects of life quality in both T1D and T2D patients, though any additional impact based on periodontal status was not observed when using W-BQ12 and ADDQoL-19.

Metagenome sequencing of fingermillet-associated microbial consortia provides insights into structural and functional diversity of endophytes.

Endophytes confer unique ecological advantages to their host plants. In this study, we have characterized the diversity of endophytic consortia associated with the GPU-28 (GPU) and Udurumallige (UM) finger millet varieties, which are resistant and susceptible to the blast disease, respectively. Whole genome metagenome sequencing of GPU and UM helped to identify 1029 species (includes obligate endophytes) of microbiota. Among them, 385 and 357 species were unique to GPU and UM, respectively. Remaining 287 species were common to both the varieties. Actinobacteria and other plant-growth promoting bacteria were abundant in GPU as compared to UM. Functional annotation of genes predicted from genomes of endophytes associated with GPU variety showed that many genes had functional role in stress response, secondary metabolism, aromatic compounds, glutathione, and cysteine synthesis pathways as compared to UM. Based on in vitro and in planta studies, Bacillus cereus and Paenibacillus spp. were found to be effective in suppressing the growth of blast disease pathogen Magnaporthe grisea (strain MG03). In the future, these strains could serve as potential biocontrol agents to reduce the incidence of blast disease in finger millet crop.

ROCK-generated contractility regulates breast epithelial cell differentiation in response to the physical properties of a three-dimensional collagen matrix.

Breast epithelial cells differentiate into tubules when cultured in floating three-dimensional (3D) collagen gels, but not when the cells are cultured in the same collagen matrix that is attached to the culture dish. These observations suggest that the biophysical properties of collagenous matrices regulate epithelial differentiation, but the mechanism by which this occurs is unknown. Tubulogenesis required the contraction of floating collagen gels through Rho and ROCK-mediated contractility. ROCK-mediated contractility diminished Rho activity in a floating 3D collagen gel, and corresponded to a loss of FAK phosphorylated at Y397 localized to 3D matrix adhesions. Increasing the density of floating 3D collagen gels also disrupted tubulogenesis, promoted FAK phosphorylation, and sustained high Rho activity. These data demonstrate the novel finding that breast epithelial cells sense the rigidity or density of their environment via ROCK-mediated contractility and a subsequent down-regulation of Rho and FAK function, which is necessary for breast epithelial tubulogenesis to occur.

Effects of yoga on brain waves and structural activation: A review.

Previous research has shown the vast mental and physical health benefits associated with yoga. Yoga practice can be divided into subcategories that include posture-holding exercise (asana), breathing (pranayama, Kriya), and meditation (Sahaj) practice. Studies measuring mental health outcomes have shown decreases in anxiety, and increases in cognitive performance after yoga interventions. Similar studies have also shown cognitive advantages amongst yoga practitioners versus non-practitioners. The mental health and cognitive benefits of yoga are evident, but the physiological and structural changes in the brain that lead to this remain a topic that lacks consensus. Therefore, the purpose of this study was to examine and review existing literature on the effects of yoga on brain waves and structural changes and activation. After a narrowed search through a set of specific inclusion and exclusion criteria, 15 articles were used in this review. It was concluded that breathing, meditation, and posture-based yoga increased overall brain wave activity. Increases in graygray matter along with increases in amygdala and frontal cortex activation were evident after a yoga intervention. Yoga practice may be an effective adjunctive treatment for a clinical and healthy aging population. Further research can examine the effects of specific branches of yoga on a designated clinical population.

A DLL3-targeted antibody-drug conjugate eradicates high-grade pulmonary neuroendocrine tumor-initiating cells in vivo.

The high-grade pulmonary neuroendocrine tumors, small cell lung cancer (SCLC) and large cell neuroendocrine carcinoma (LCNEC), remain among the most deadly malignancies. Therapies that effectively target and kill tumor-initiating cells (TICs) in these cancers should translate to improved patient survival. Patient-derived xenograft (PDX) tumors serve as excellent models to study tumor biology and characterize TICs. Increased expression of delta-like 3 (DLL3) was discovered in SCLC and LCNEC PDX tumors and confirmed in primary SCLC and LCNEC tumors. DLL3 protein is expressed on the surface of tumor cells but not in normal adult tissues. A DLL3-targeted antibody-drug conjugate (ADC), SC16LD6.5, comprised of a humanized anti-DLL3 monoclonal antibody conjugated to a DNA-damaging pyrrolobenzodiazepine (PBD) dimer toxin, induced durable tumor regression in vivo across multiple PDX models. Serial transplantation experiments executed with limiting dilutions of cells provided functional evidence confirming that the lack of tumor recurrence after SC16LD6.5 exposure resulted from effective targeting of DLL3-expressing TICs. In vivo efficacy correlated with DLL3 expression, and responses were observed in PDX models initiated from patients with both limited and extensive-stage disease and were independent of their sensitivity to standard-of-care chemotherapy regimens. SC16LD6.5 effectively targets and eradicates DLL3-expressing TICs in SCLC and LCNEC PDX tumors and is a promising first-in-class ADC for the treatment of high-grade pulmonary neuroendocrine tumors.

A one-arm homologous recombination approach for developing nuclear receptor assays in somatic cells.

Drug discovery is in need of technologies that enable investigators to develop cell-based assays that accurately reflect the functional consequence of small molecule intervention on biological processes. Here, we describe a strategy that uses both one-arm homologous recombination and the beta-lactamase (BLA) reporter system, a sensitive and robust transcriptional reporter for gene activation. We demonstrate that this powerful approach can be utilized for developing cell-based assays for the glucocorticoid receptor (GR) and mineralocorticoid receptor (MR) in HEK293 somatic cells. Specifically, one-arm homologous recombination was used to introduce the GAL4 DNA-binding domain (GAL4DBD) in the GR and MR genomic loci such that a chimeric GAL4DBD-GR (ligand-binding domain) [GAL4DBD-GR(LBD)] and GAL4DBD-MR(LBD) transcript is produced from the strong CMV promoter in HEK293 cells previously stably transfected with the UAS(GAL4)-BLA reporter construct. Dexamethasone- and aldosterone-responding BLA-positive cells were isolated by fluorescence-activated cell sorting, and then further expanded into separate cell lines. The sensitivity and robustness of the resulting GR and MR assays are demonstrated by the fact that the addition of dexamethasone and aldosterone to the two transgenic clonal cell lines for 16 h results in high Z' values (>0.8) and EC(50) values of 1 and 0.3 nM, respectively. These assays illustrate the flexibility of this technology to generate high-performance cellular assays for nuclear receptor targets without the need for target-specific cDNA.

Does the mode of administration of the Oral Health Impact Profile-49 affect the outcome score?

OBJECTIVE: To determine if there are differences in outcome scores if the Oral Health Impact Profile-49 (OHIP-49) is delivered by two different modes of administration (manual-self complete versus telephone interview). METHODS: Patients with chronic periodontitis (n=83, 54% females and 46% males, mean age 49.1±9.5 years) completed the OHIP-49 using two modes of administration (manual self-complete and telephone interview) in a randomly assigned order, with a minimum washout period of 2 weeks between modes, both episodes occurring prior to any periodontal treatment being provided. To assess convergent validity, after each mode of administration, the patients were additionally asked a global question about their oral health-related quality of life (OHRQoL). RESULTS: Median OHIP-49 scores recorded by manual self-complete (median 36 [IQR=20-70]) were significantly higher than those recorded by telephone interview (median 27 [IQR=11-61]) (p<0.01). The global question was well correlated to the OHIP domains, but did not reveal any evidence of an order effect such as was seen with OHIP-49 itself (which showed a higher impact on OHRQoL during the first administration in either mode). CONCLUSIONS: The mode of administration (manual-self complete versus telephone interview) did substantially influence the OHIP-49 scores in patients with chronic periodontitis. The OHRQoL differed between the two modes of administration, with significantly higher scores (indicating poorer OHRQoL) when the questionnaire was manually self-completed. CLINICAL SIGNIFICANCE: The mode of administration of quality of life questionnaires such as OHIP-49 could potentially affect the outcome scores derived. This study investigated whether there is a difference in outcome scores if OHIP-49 is delivered via manual self-complete or by telephone interview in patients with chronic periodontitis. We found that there was a significant difference between the two modes: manual self-completion by the patients yielded significantly higher scores than completion by telephone interview. It is therefore important to be consistent in the mode of completion of OHIP-49, as mixing modes could introduce additional error into clinical studies that utilise this instrument.

Ack1 mediates Cdc42-dependent cell migration and signaling to p130Cas.

We previously showed that activation of the small GTPase Cdc42 promotes breast cell migration on a collagen matrix. Here we further define the signaling pathways that drive this response and show that Cdc42-mediated migration relies on the adaptor molecule p130(Cas). Activated Cdc42 enhanced p130(Cas) phosphorylation and its binding to Crk. Cdc42-driven migration and p130(Cas) phosphorylation were dependent on the Cdc42 effector Ack1 (activated Cdc42-associated kinase). Ack1 formed a signaling complex that also included Cdc42, p130(Cas), and Crk, formation of which was regulated by collagen stimulation. The interaction between Ack1 and p130(Cas) occurred through their respective SH3 domains, while the substrate domain of p130(Cas) was the major site of Ack1-dependent phosphorylation. Signaling through this complex is functionally relevant, because treatment with either p130(Cas) or Ack1 siRNA blocked Cdc42-induced migration. These results suggest that Cdc42 exerts its effects on cell migration in part through its effector Ack1, which regulates p130(Cas) signaling.