Scalable Measurements of Intrinsic Excitability in Human iPS Cell-Derived Excitatory Neurons Using All-Optical Electrophysiology.

Induced pluripotent stem (iPS) cells offer the exciting opportunity for modeling neurological disorders in vitro in the context of a human genetic background. While significant progress has been made in advancing the use of iPS cell-based disease models, there remains an unmet need to characterize the electrophysiological profile of individual neurons with sufficient throughput to enable statistically robust assessment of disease phenotypes and pharmacological modulation. Here, we describe the Optopatch platform technology that utilizes optogenetics to both stimulate and record action potentials (APs) from human iPS cell-derived excitatory neurons with similar information content to manual patch clamp electrophysiology, but with ~  3 orders of magnitude greater throughput. Cortical excitatory neurons were produced using the NGN2 transcriptional programming approach and cultured in the presence of rodent glial cells. Characterization of the neuronal preparations using immunocytochemistry and qRT-PCR assays reveals an enrichment of neuronal and glutamatergic markers as well as select ion channels. We demonstrate the scale of our intrinsic cellular excitability assay using pharmacological assessment with select ion channel modulators quinidine and retigabine, by measuring changes in both spike timing and waveform properties. The Optopatch platform in human iPS cell-derived cortical excitatory neurons has the potential for detailed phenotype and pharmacology evaluation, which can serve as the basis of cellular disease model exploration for drug discovery and phenotypic screening efforts.

Soft and hard tissue changes around laser microtexture single tooth implants--a clinical and radiographic evaluation.

PURPOSE: To investigate the periodontal parameters that affect the soft and hard tissues around Laser microtextured single tooth implants at 18 months after loading. METHODS: Twenty Laser Lok implants were placed in 20 single missing tooth sites using a 2-stage protocol. Clinical Parameters included Plaque Index (PI), Gingival Index (GI), Probing Pocket Depth (PPD), Bleeding on Probing (BOP), and Crestal Bone Loss (CBL). Clinical and radiographic evaluation was done at loading, 12 months and 18 months after loading. The data collected were analyzed statistically. RESULTS: The PI and GI during the entire follow-up period were well controlled. Eighty-six percent of implant sites were free of BOP at loading and 87% of sites were free of BOP at 18 months. A significant increase in PPD was not observed. The mean CBL was 0.59 mm at the time of loading, 0.80 mm at 12 months, and 1.06 mm at 18 months. CONCLUSION: The Laser Lok implants showed minimal CBL at 18 months than the commonly accepted 1.5 to 2.0 mm. The periimplant soft tissue stability was maintained throughout the study.

Piezoelectric energy extraction from a cylinder undergoing vortex-induced vibration using internal resonance.

A novel concept of utilizing the kinetic energy from ocean currents/wind by means of internal resonance is proposed to address the increasing global energy demand by generating clean and sustainable power. In this work, a non-linear rotative gravity pendulum is employed to autoparametrically excite the elastically mounted cylinder for a wide range of flow velocities. This concept is adopted to increase the oscillation amplitude of the cylinder due to vortex-induced vibration (VIV) in the de-synchronized region for energy harvesting. In this regard, a VIV-based energy harvesting device is proposed that consists of a cylinder with an attached pendulum, and energy is harvested with bottom-mounted piezoelectric transducers. The cylinder undergoes VIV when it is subjected to fluid flow and this excites the coupled fluid-multibody cylinder-pendulum system autoparametrically. In the de-synchronized region, when the vortex shedding frequency becomes two times the natural frequency of the pendulum, an internal resonance occurs. This helps in achieving a higher oscillation amplitude of the cylinder which does not happen otherwise. This study is focused on the two degree-of-freedom (2-DoF) cylinder-pendulum system where the cylinder is free to exhibit cross-flow vortex-induced vibrations subjected to the fluid. The objective of this work is to numerically investigate the effect of a non-linear rotative gravity pendulum (NRGP) on the VIV characteristics and piezoelectric efficiency of the system. The numerical model is based on the wake-oscillator model coupled with the piezoelectric constitutive equation. The influence of the frequency ratio, mass ratio, torsional damping ratio, and ratio of cylinder diameter to pendulum length of the NRGP device on response characteristics due to VIV is also investigated. A detailed comparative analysis in terms of electric tension and efficiency is performed numerically for flows with a wide range of reduced velocities for the cylinder with and without NRGP. A comprehensive study on the implications of internal resonance between the pendulum and a cylinder undergoing VIV on generated electric tension is also reported.

Developing antisense oligonucleotides for a TECPR2 mutation-induced, ultra-rare neurological disorder using patient-derived cellular models.

Mutations in the TECPR2 gene are the cause of an ultra-rare neurological disorder characterized by intellectual disability, impaired speech, motor delay, and hypotonia evolving to spasticity, central sleep apnea, and premature death (SPG49 or HSAN9; OMIM: 615031). Little is known about the biological function of TECPR2, and there are currently no available disease-modifying therapies for this disease. Here we describe implementation of an antisense oligonucleotide (ASO) exon-skipping strategy targeting TECPR2 c.1319delT (p.Leu440Argfs∗19), a pathogenic variant that results in a premature stop codon within TECPR2 exon 8. We used patient-derived fibroblasts and induced pluripotent stem cell (iPSC)-derived neurons homozygous for the p.Leu440Argfs∗19 mutation to model the disease in vitro. Both patient-derived fibroblasts and neurons showed lack of TECPR2 protein expression. We designed and screened ASOs targeting sequences across the TECPR2 exon 8 region to identify molecules that induce exon 8 skipping and thereby remove the premature stop signal. TECPR2 exon 8 skipping restored in-frame expression of a TECPR2 protein variant (TECPR2ΔEx8) containing 1,300 of 1,411 amino acids. Optimization of ASO sequences generated a lead candidate (ASO-005-02) with ∼27 nM potency in patient-derived fibroblasts. To examine potential functional rescue induced by ASO-005-02, we used iPSC-derived neurons to analyze the neuronal localization of TECPR2ΔEx8 and showed that this form of TECPR2 retains the distinct, punctate neuronal expression pattern of full-length TECPR2. Finally, ASO-005-02 had an acceptable tolerability profile in vivo following a single 20-mg intrathecal dose in cynomolgus monkeys, showing some transient non-adverse behavioral effects with no correlating histopathology. Broad distribution of ASO-005-02 and induction of TECPR2 exon 8 skipping was detected in multiple central nervous system (CNS) tissues, supporting the potential utility of this therapeutic strategy for a subset of patients suffering from this rare disease.

Evaluation of peri-implant soft tissue and bone levels around early loaded implant in restoring single missing tooth: A clinico-radiographic study.

BACKGROUND: One-stage nonsubmerged protocol which can achieve success rates comparable to implants placed in a two-staged submerged procedure also the preconditions for periimplant bone regeneration has lead to more refined concepts of implant loading. MATERIALS AND METHODS: Twenty sites with single missing tooth were included in this study. Clinical parameters included sulcus bleeding index (sBI), probing pocket depth (PD), and papilla index (PI) and radiographic parameters included crestal bone level were assessed for a period of 9 months. RESULTS: The crestal bone loss showed mean value ranging from baseline 0.25 ± 0.11 to 0.31 ± 0.08 at 3 weeks, to 0.67 ± 0.13 at 3 months, to 0.85 ± 0.09 at 6 months, and to 0.88 ± 0.12 at 9 months. Probing PD, the mean value for probing PD at 3 weeks 1.20 ± 0.83, 3 months 1.60 ± 1.1, at 6 months 1.40 ± 1.14, and at 9 months 1.20 ± 1.0. sBI, mean value for sBI at 3 weeks 0.00 ± 0.00, 3 months 0.3 ± 0.11, at 6 months 0.09 ± 0.25, and at 9 months 0.08 ± 0.24. PI, showed a significant difference among at different points of time with P = 0.000. CONCLUSION: The dental implants showed <1 mm of crestal bone loss at 9 months follow-up, clinically significant marginal bone loss occurred between the time of implant placement and 3 months. Subsequent to that, bone loss observed around the implant up to 9 months was minimal. The periimplant soft tissue maturity was maintained throughout the study.

Immediate Implant Placement in Anterior Aesthetic Region and Assessment using Cone-Beam Computed Tomography Scan Technology.

The esthetics and functional integrity of the periodontal tissues may be compromised by the dental loss. Dental implants have become an additional tool in the armamentarium of treatment options to offer the patient for the replacement of a missing tooth or teeth. Diagnosis and treatment planning is the key factors in achieving the successful outcome after placing and restoring implants placed immediately after tooth extraction. The introduction of cone-beam computed tomography (CBCT) for the maxillofacial region provides opportunities for dental practitioners to request multiplanar imaging. This case report demonstrates the use of CBCT scan technology in immediate implant placement in the maxillary anterior teeth region.

Gingival depigmentation: A split mouth comparative study between scalpel and cryosurgery.

Gingival hyperpigmentation is a major esthetic concern for many people. Although it is not a medical problem, many people complain of dark gums as unesthetic. Gingival depigmentation is a periodontal plastic surgical procedure, whereby the hyperpigmentation is removed or reduced by various techniques. For depigmentation of gingival, different treatment modalities have been reported, such as scalpel, cryosurgery, electrosurgery, lasers, etc., this article compares the management of three cases with scalpel and cryosurgery and also highlights the relevance of cryosurgery.