Protective effect of alpha-lipoic acid and epalrestat on oxaliplatin-induced peripheral neuropathy in zebrafish.

INTRODUCTION/AIMS: Oxaliplatin is a platinum-based anti-cancer drug widely used in colorectal cancer patients, but it may cause peripheral neuropathy. As one of the main causes of oxaliplatin-induced peripheral neuropathy (OPN) is oxidative stress, which is also a key factor causing diabetic peripheral neuropathy (DPN), the aim of this study was to evaluate the preventive effects of alpha-lipoic acid (ALA) and epalrestat (EP), which are used for the treatment of DPN, in an OPN zebrafish model. METHODS: Tg(nbt:dsred) transgenic zebrafish, with sensory nerves in the peripheral lateral line, were treated with oxaliplatin, oxaliplatin/EP, and oxaliplatin/ALA for 4 days. A confocal microscope was used to visualize and quantify the number of axon bifurcations in the distal nerve ending. To analyze the formation of synapses on sensory nerve terminals, quantification of membrane-associated guanylate kinase (MAGUK) puncta was performed using immunohistochemistry. RESULTS: The number of axon bifurcations and intensity of MAGUK puncta were significantly reduced in the oxaliplatin-treated group compared with those in the embryo medium-treated group. In both the oxaliplatin/EP and oxaliplatin/ALA-treated groups, the number of axon bifurcations and intensity of MAGUK puncta were greater than those in the oxaliplatin-treated group (p < .0001), and no significant difference was observed between larvae treated with oxaliplatin/ALA 1 µM and oxaliplatin/EP 1 µM (p = .4292). DISCUSSION: ALA and EP have protective effects against OPN in zebrafish. Our findings show that ALA and EP can facilitate more beneficial treatment for OPN.

Optimal placement for needle electromyography of the supinator muscle: Cadaveric studies.

INTRODUCTION/AIMS: The existing methods for needle electromyography are confusing as to which is the safest and most effective. Our aim was to identify the optimal and safest needle electromyographic insertion site in the supinator muscle. METHODS: We performed a two-step cadaveric dissection of the supinator muscle and related neurovascular structures. The study was performed using 18 upper limbs of 9 fresh adult cadavers (step 1) and 14 upper limbs of 7 fresh adult cadavers (step 2). In step 1, an imaginary line connecting the radial head (RH) and midpoint of the dorsal wrist (RW line) was drawn, and the distance from the RH to the point where the RW line and posterior interosseous nerve (PIN) intersect (L_CROSS) was measured on the RW line. In step 2, the needle was inserted 30 mm distal to the RH according to the results of step 1. After injection with India ink, dissection was performed to measure the distance between the needle insertion site and PIN (L_CROSS_Inj) on the RW line. RESULTS: The median L_CROSS was 51.4 (35.5-65.6) mm. Needle insertion spared the PIN in all cases during step 2, and the needle was inserted into the supinator muscle in all cases. The median L_CROSS_Inj was 27.4 (13.2-39.8) mm. DISCUSSION: A safe and accurate needle insertion site for the supinator muscle is approximately 30 to 40 mm distal to the RH along the RW line.

Safe Approach for Flexor Digitorum Profundus I and II Using the Palmaris Longus Tendon.

OBJECTIVE: To investigate a safe and accurate approach to achieve needle insertion for electromyography (EMG) of the flexor digitorum profundus (FDP) I and II muscles by identifying the anatomic relationship between the palmaris longus (PL) tendon, FDP muscle, and neurovascular bundle using ultrasonography. DESIGN: Descriptive study SETTING: Department of physical medicine and rehabilitation. PARTICIPANTS: Healthy individuals (age, 20-70y) without any diseases (N=29; 15 men, 14 women; 58 forearms). INTERVENTIONS: Ultrasonography. MAIN OUTCOME MEASURES: The FDP I and II muscles were transversely scanned on the volar aspect of the forearm at the junction of the middle and distal third between the medial epicondyle and ulnar styloid process. The distances and angles from the medial border of the PL tendon to FDP I, FDP II, and median nerve were measured. RESULTS: The probability of damage to the neurovascular structures and the accuracy of entering the FDP I and II muscles were calculated for 3 imaginary needle insertion angles (61.7°, 100.6°, and 90°). When the needle was inserted at an angle of 61.7°, it reached FDP I with an accuracy of 91.4%. Upon needle insertions at 90° and 100.6°, the needle reached FDP II with accuracies of 90% and 89.6%, respectively. In all 3 cases (61.7°, 90°, and 100.6°), there was no chance of penetrating the blood vessels or nerves. CONCLUSION: EMG of FDP I and II can be performed precisely and safely with the anterior approach at the distal one-third between the medial epicondyle and ulnar styloid process using the PL tendon.

Magnetic Control and Real-Time Monitoring of Stem Cell Differentiation by the Ligand Nanoassembly.

Native extracellular matrix (ECM) exhibits dynamic change in the ligand position. Herein, the ECM-emulating control and real-time monitoring of stem cell differentiation are demonstrated by ligand nanoassembly. The density of gold nanoassembly presenting cell-adhesive Arg-Gly-Asp (RGD) ligand on Fe3 O4 (magnetite) nanoparticle in nanostructures flexibly grafted to material is changed while keeping macroscale ligand density invariant. The ligand nanoassembly on the Fe3 O4 can be magnetically attracted to mediate rising and falling ligand movements via linker stretching and compression, respectively. High ligand nanoassembly density stimulates integrin ligation to activate the mechanosensing-assisted stem cell differentiation, which is monitored via in situ real-time electrochemical sensing. Magnetic control of rising and falling ligand movements hinders and promotes the adhesion-mediated mechanotransduction and differentiation of stem cells, respectively. These rising and falling ligand states yield the difference in the farthest distance (≈34.6 nm) of the RGD from material surface, thereby dynamically mimicking static long and short flexible linkers, which hinder and promote cell adhesion, respectively. Design of cytocompatible ligand nanoassemblies can be made with combinations of dimensions, shapes, and biomimetic ligands for remotely regulating stem cells for offering novel methodologies to advance regenerative therapies.

Optimal recording electrode placement for radial motor nerve conduction study using extensor indicis muscle: Cadaver and electrophysiological studies.

INTRODUCTION/AIMS: The aim of this study was to determine the optimal combination of active (E1) and reference (E2) recording electrode placements for the radial motor nerve conduction study recording over the extensor indicis muscle using surface electrodes. METHODS: Thirty-six upper limbs from 18 fresh frozen cadavers were dissected to determine the midpoint of the extensor indicis muscle. Radial nerve conduction study was performed in 112 arms of 56 healthy subjects. Six combinations of three E1 and two E2 sites were studied. The stimulation site was 8 cm proximal to the E1 electrode. RESULTS: The optimal combination of placement sites for the E1 and E2 electrodes to provide the largest amplitude is E1 electrode at the distal 1/4 point of the forearm length and E2 electrode at the extensor indicis tendon point in the wrist. DISCUSSION: Optimal recording electrode placement may increase the accuracy and reproducibility of radial motor nerve conduction studies.

Electrical stimulation accelerates neurite regeneration in axotomized dorsal root ganglion neurons by increasing MMP-2 expression.

Electrical stimulation (ES) can be useful for promoting the regeneration of injured axons, but the mechanism underlying its positive effects is largely unknown. The current study aimed to investigate whether ES could enhance the regeneration of injured neurites in dorsal root ganglion explants and regulate the MMP-2 expression level, which is correlated with regeneration. Significantly increased neurite regeneration and MMP-2 expression was observed in the ES group compared with the sham group. However, an MMP inhibitor significantly decreased this ES-induced neurite regeneration. Our data suggest that the positive effect of ES on neurite regeneration could likely be mediated by an increase in MMP-2 expression, thereby promoting the regeneration of injured neurites.

A cadaveric study for the volar needle approach to the pronator quadratus using the palmaris longus tendon landmark.

INTRODUCTION: Our aim in this work was to determine the safety and accuracy of the volar approach to the pronator quadratus (PQ) through cadaver dissection. METHODS: Twenty upper limbs from 10 fresh cadavers were investigated. At the level 3 cm proximal to the ulnar styloid process (USP), a needle was inserted just medial to the palmaris longus (PL) tendon. Distances of the median nerve (MN) and ulnar artery (UA) from the needle insertion point (IP) were measured using ultrasonography and cadaver dissection. RESULTS: The PQ was located at a depth of 10.8-19.9 mm from the skin and had a median thickness of 9.1 mm, measured 3 cm proximal to the USP. The median distances of the MN and UA from the IP were 7.6 and 13.4 mm, respectively. DISCUSSION: A needle insertion for the volar approach to the PQ was safe at 3 cm proximal to the USP, just medial to the PL tendon.

In Situ Blood Vessel Regeneration Using SP (Substance P) and SDF (Stromal Cell-Derived Factor)-1α Peptide Eluting Vascular Grafts.

OBJECTIVE: The objective of this study was to develop small-diameter vascular grafts capable of eluting SDF (stromal cell-derived factor)-1α-derived peptide and SP (substance P) for in situ vascular regeneration. APPROACH AND RESULTS: Polycaprolactone (PCL)/collagen grafts containing SP or SDF-1α-derived peptide were fabricated by electrospinning. SP and SDF-1α peptide-loaded grafts recruited significantly higher numbers of mesenchymal stem cells than that of the control group. The in vivo potential of PCL/collagen, SDF-1, and SP grafts was assessed by implanting them in a rat abdominal aorta for up to 4 weeks. All grafts remained patent as observed using color Doppler and stereomicroscope. Host cells infiltrated into the graft wall and the neointima was formed in peptides-eluting grafts. The lumen of the SP grafts was covered by the endothelial cells with cobblestone-like morphology, which were elongated in the direction of the blood flow, as discerned using scanning electron microscopy. Moreover, SDF-1α and SP grafts led to the formation of a confluent endothelium as evaluated using immunofluorescence staining with von Willebrand factor antibody. SP and SDF-1α grafts also promoted smooth muscle cell regeneration, endogenous stem cell recruitment, and blood vessel formation, which was the most prominent in the SP grafts. Evaluation of inflammatory response showed that 3 groups did not significantly differ in terms of the numbers of proinflammatory macrophages, whereas SP grafts showed significantly higher numbers of proremodeling macrophages than that of the control and SDF-1α grafts. CONCLUSIONS: SDF-1α and SP grafts can be potential candidates for in situ vascular regeneration and are worthy for future investigations.

Sonography-guided recording for superficial peroneal sensory nerve conduction study.

INTRODUCTION: We sought to establish the optimal recording position for antidromic conduction of the superficial peroneal nerve (SPN) by using ultrasonography (USG). METHODS: The sensory nerve action potentials (SNAPs) of the intermediate dorsal cutaneous nerve (IDCN) and medial dorsal cutaneous nerve (MDCN) in 64 limbs of 32 healthy participants were recorded (nerve conduction study [NCS]-1). Both nerves were identified by using USG, and the SNAPs were obtained from the USG-guided repositioned electrodes (NCS-2). RESULTS: The IDCN and MDCN were located at 29.3% ± 5.1% and 43.9% ± 4.9% of the intermalleolar distance from the lateral malleolus, respectively. Significantly greater amplitude was shown for SNAPs of both nerves in NCS-2 versus NCS-1. DISCUSSION: The optimal recording position is likely to be lateral, one-third from the lateral malleolus for the IDCN, and just lateral to the midpoint of the intermalleolar line for the MDCN. When the SPN response is unexpectedly attenuated, USG-guided repositioning of the electrodes should be considered. Muscle Nerve 57: 628-633, 2018.

Sonoanatomy of sensory branches of the ulnar nerve below the elbow in healthy subjects.

INTRODUCTION: We identify sensory branches of the ulnar nerve-palmar ulnar cutaneous nerve (PUCN), dorsal ulnar cutaneous nerve (DUCN), and superficial sensory branch-using ultrasonography. METHODS: In 60 forearms of 30 healthy adult volunteers, the origin and size of the PUCN, DUCN, and superficial sensory branch were measured by ultrasonography. The relative pathway of the DUCN to the ulnar styloid process was also investigated. RESULTS: The PUCN was observed in 47 forearms (78%), and the DUCN was observed in all forearms. Average distances from the pisiform to the origin of the PUCN and DUCN were 11.9 ± 1.4 and 7.0 ± 1.0 cm, respectively. Superficial and deep divisions split 0.9 ± 0.3 cm distal to the pisiform. Cross-sectional areas of the PUCN, DUCN, and superficial sensory branch were 0.3 ± 0.1, 1.5 ± 0.5, and 3.9 ± 1.0 mm2 , respectively. DISCUSSION: Sensory branches of the ulnar nerve can be visualized by ultrasonography, helping to differentiate ulnar nerve injury originating at either wrist or elbow. Muscle Nerve 57: 569-573, 2018.

Mechanics of the Cell Nucleus.

Nucleus is a specialized organelle that serves as a control tower of all the cell behavior. While traditional biochemical features of nuclear signaling have been unveiled, many of the physical aspects of nuclear system are still under question. Innovative biophysical studies have recently identified mechano-regulation pathways that turn out to be critical in cell migration, particularly in cancer invasion and metastasis. Moreover, to take a deeper look onto the oncologic relevance of the nucleus, there has been a shift in cell systems. That is, our understanding of nucleus does not stand alone but it is understood by the relationship between cell and its microenvironment in the in vivo-relevant 3D space.

Volume regulation and shape bifurcation in the cell nucleus.

Alterations in nuclear morphology are closely associated with essential cell functions, such as cell motility and polarization, and correlate with a wide range of human diseases, including cancer, muscular dystrophy, dilated cardiomyopathy and progeria. However, the mechanics and forces that shape the nucleus are not well understood. Here, we demonstrate that when an adherent cell is detached from its substratum, the nucleus undergoes a large volumetric reduction accompanied by a morphological transition from an almost smooth to a heavily folded surface. We develop a mathematical model that systematically analyzes the evolution of nuclear shape and volume. The analysis suggests that the pressure difference across the nuclear envelope, which is influenced by changes in cell volume and regulated by microtubules and actin filaments, is a major factor determining nuclear morphology. Our results show that physical and chemical properties of the extracellular microenvironment directly influence nuclear morphology and suggest that there is a direct link between the environment and gene regulation.

Safety Window for the Volar Needle Approach for Examination of the Pronator Quadratus Using Ultrasonography.

OBJECTIVES: To investigate pronator quadratus (PQ) anatomy and determine the proper volar needle insertion point based on landmarks, the tip of the ulnar styloid (an imaginary vertical line passing the tip of the ulnar styloid process [U line]), and the ulnar margin of the palmaris longus tendon (uPL) using ultrasonography. DESIGN: Descriptive study. SETTING: Department of physical medicine and rehabilitation. PARTICIPANTS: Participants between 20 and 60 years without any diseases. (N=25; 13 men, 12 women; 50 forearms). INTERVENTIONS: Ultrasonography. MAIN OUTCOME MEASURES: The proximal and distal volar surface points of origin, the proximal and distal insertion sites, and the midpoint of the PQ (PQ_M) were determined. The distance of each of the PQ surface indices from the U line was measured, and the probe was positioned at the level of PQ_M parallel to the U line. The relative distances from the vertical surface points of the median nerve and ulnar artery to the uPL were measured. RESULTS: The mean age and body mass index were 32.7±10.4 years and 21.98±2.83kg/m2. The PQ_M was located at a mean distance of 2.63±0.35cm proximal from the U line (men 2.79±0.37cm and women 2.45±0.21cm; P<.05). The mean safety window for the volar approach was 0.72±1.8cm toward the radial side and 1.51±0.30cm toward the ulnar side from the uPL. The PQ was at a mean depth of 1.30±0.19cm from the skin and had a mean thickness of 1.19±0.24cm at the level of PQ_M. The distance between the U line and the proximal edge of the PQ, as well as the PQ thickness, was greater in men than in women. CONCLUSIONS: The volar approach for needle electromyographic examination of the PQ can be performed precisely and safely.

Tight coupling between nucleus and cell migration through the perinuclear actin cap.

Although eukaryotic cells are known to alternate between 'advancing' episodes of fast and persistent movement and 'hesitation' episodes of low speed and low persistence, the molecular mechanism that controls the dynamic changes in morphology, speed and persistence of eukaryotic migratory cells remains unclear. Here, we show that the movement of the interphase nucleus during random cell migration switches intermittently between two distinct modes - rotation and translocation - that follow with high fidelity the sequential rounded and elongated morphologies of the nucleus and cell body, respectively. Nuclear rotation and translocation mediate the stop-and-go motion of the cell through the dynamic formation and dissolution, respectively, of the contractile perinuclear actin cap, which is dynamically coupled to the nuclear lamina and the nuclear envelope through LINC complexes. A persistent cell movement and nuclear translocation driven by the actin cap are halted following the disruption of the actin cap, which in turn allows the cell to repolarize for its next persistent move owing to nuclear rotation mediated by cytoplasmic dynein light intermediate chain 2.

A case of lateral calcaneal neuropathy: Lateral heel pain.

INTRODUCTION: Although isolated injury of the sural nerve is frequently reported, entrapment involving the lateral calcaneal branch of the sural nerve (LCSN) is very rare. METHODS: We report the case of a 29-year-old man with lateral calcaneal neuropathy of the sural nerve without any history of trauma. The diagnosis was based on clinical, electrophysiologic, and radiologic studies, including ultrasonography (US) and MRI. RESULTS: US revealed a spindle-shaped swelling in the posterior part of the sural nerve around the lateral malleolus. MRI showed a small lesion in the sural nerve close to the lesser saphenous vein around the level of the lateral malleolus. Two months after conservative management, the numbness and tingling in the lateral calcaneal region improved. CONCLUSIONS: Entrapment of the LCSN can arise from tight shoes. US is useful for evaluating the cutaneous nerve lesions that may be too small to detect by MRI. Muscle Nerve 54: 801-804, 2016.

Superficial radial sensory neuropathy: Medial and lateral branch injury.

INTRODUCTION: Superficial radial sensory nerve (SRN) injury may involve the main nerve or its distal medial or lateral branch. We investigated the utility of SRN conduction studies on the medial and lateral branches in patients with suspected SRN injury. METHODS: Fifteen patients with symptoms of SRN neuropathy were studied. Their clinical and electrophysiological findings were assessed according to sensory symptom areas. Three active recording electrode positions were used: snuff box (position A); medial branch (position B); and lateral branch (position C). RESULTS: In 7 patients with medial area symptoms, abnormal findings were seen in position B (100%) and position A (71%). In 3 patients with medial and lateral area symptoms, abnormal findings were seen in all positions. In 5 patients with lateral symptoms, abnormal findings were seen in position C only. CONCLUSIONS: Nerve conduction studies for each branch of the SRN are useful in patients with suspected SRN neuropathy.

Focal adhesion size uniquely predicts cell migration.

Focal adhesions are large protein complexes organized at the basal surface of cells, which physically connect the extracellular matrix to the cytoskeleton and have long been speculated to mediate cell migration. However, whether clustering of these molecular components into focal adhesions is actually required for these proteins to regulate cell motility is unclear. Here we use quantitative microscopy to characterize descriptors of focal adhesion and cell motility for mouse embryonic fibroblasts and human fibrosarcoma cells, across a wide range of matrix compliance and following genetic manipulations of focal adhesion proteins (vinculin, talin, zyxin, FAK, and paxilin). This analysis reveals a tight, biphasic gaussian relationship between mean size of focal adhesions (not their number, surface density, or shape) and cell speed. The predictive power of this relationship is comprehensively validated by disrupting nonfocal adhesion proteins (α-actinin, F-actin, and myosin II) and subcellular organelles (mitochondria, nuclear DNA, etc.) not known to affect either focal adhesions or cell migration. This study suggests that the mean size of focal adhesions robustly and precisely predicts cell speed independently of focal adhesion surface density and molecular composition.

Advanced Immunomodulatory Biomaterials for Therapeutic Applications.

The multifaceted biological defense system modulating complex immune responses against pathogens and foreign materials plays a critical role in tissue homeostasis and disease progression. Recently developed biomaterials that can specifically regulate immune responses, nanoparticles, graphene, and functional hydrogels have contributed to the advancement of tissue engineering as well as disease treatment. The interaction between innate and adaptive immunity, collectively determining immune responses, can be regulated by mechanobiological recognition and adaptation of immune cells to the extracellular microenvironment. Therefore, applying immunomodulation to tissue regeneration and cancer therapy involves manipulating the properties of biomaterials by tailoring their composition in the context of the immune system. This review provides a comprehensive overview of how the physicochemical attributes of biomaterials determine immune responses, focusing on the physical properties that influence innate and adaptive immunity. This review also underscores the critical aspect of biomaterial-based immune engineering for the development of novel therapeutics and emphasizes the importance of understanding the biomaterials-mediated immunological mechanisms and their role in modulating the immune system.

Transgenic zebrafish as a model for investigating diabetic peripheral neuropathy: investigation of the role of insulin signaling.

Diabetic peripheral neuropathy (DPN), a complication of diabetes mellitus (DM), is a neurodegenerative disorder that results from hyperglycemic damage and deficient insulin receptor (IR) signaling in peripheral nerves, triggered by failure of insulin production and insulin resistance. IR signaling plays an important role in nutrient metabolism and synaptic formation and maintenance in peripheral neurons. Although several animal models of DPN have been developed to identify new drug candidates using cytotoxic reagents, nutrient-rich diets, and genetic manipulations, a model showing beneficial effects remains to be established. In this study, we aimed to develop a DPN animal model using zebrafish to validate the effects of drug candidates on sensory neuropathy through in vivo imaging during the early larval stage. To achieve this, we generated Tg (ins:gal4p16);Tg (5uas:epNTR-p2a-mcherry) zebrafish using an enhanced potency nitroreductase (epNTR)-mediated chemogenetic ablation system, which showed highly efficient ablation of pancreatic ß-cells following treatment with low-dose metronidazole (MTZ). Using in vivo live imaging, we observed that sensory nerve endings and postsynaptic formation in the peripheral lateral line (PLL) were defective, followed by a disturbance in rheotaxis behavior without any locomotory behavioral changes. Despite defects in sensory nerves and elevated glucose levels, both reactive oxygen species (ROS) levels, a primary cause of DPN, and the number of ganglion cells, remained normal. Furthermore, we found that the activity of mTOR, a downstream target of IR signaling, was decreased in the PLL ganglion cells of the transgenic zebrafish. Our data indicates that peripheral neuropathy results from the loss of IR signaling due to insulin deficiency rather than hyperglycemia alone.

Electrodiagnostic Findings Using Radial Motor Segmental Conduction Study and Inching Test in Patients with Radial Neuropathy.

OBJECTIVE: The aim of this study was to characterize the electrodiagnostic findings of radial neuropathy using motor segmental conduction study (RMSCS), and to determine the utility of subsequent inching test in precise lesion localization. DESIGN: Twenty-three patients with radial neuropathy were evaluated using RMSCS with three-point stimulation. The pathomechanism of the lesions according to the RMSCS was classified into three groups: conduction block (CB), mixed lesion (combination of CB and axonal degeneration), and axonal degeneration. Inching test was performed in patients with CB to localize the lesion site, and needle EMG identified the most proximal radial nerve-innervated muscles affected. RESULTS: Out of twenty-three cases, the RMSCS demonstrated probable partial CB in 10, mixed lesions in 2, and axonal degeneration in 10. One case could not be categorized with RMSCS alone. As determined by RMSCS and inching test, the most common cause of CB was compression, while the most common cause of axonal degeneration was iatrogenic. In the CB group, the lesion locations identified by RMSCS and inching test were consistent with needle EMG localization. CONCLUSION: The combined RMSCS and inching test technique can precisely localize radial motor nerve injuries and provide detailed information on electrodiagnostic characteristics of radial mononeuropathy.