Changes In Peripheral Nerves Research Articles

Schwann cell stimulation induces functional and structural changes in peripheral nerves.

Signal propagation is the essential function of nerves. Lysophosphatidic acid 18:1 (LPA) allows the selective stimulation of calcium signaling in Schwann cells but not neurons. Here, the time course of slowing and amplitude reduction on compound action potentials due to LPA exposure was observed in myelinated and unmyelinated fibers of the mouse, indicating a clear change of axonal function. Teased nerve fiber imaging showed that Schwann cell activation is also present in axon-attached Schwann cells in freshly isolated peripheral rat nerves. The LPA receptor 1 was primarily localized at the cell extensions in isolated rat Schwann cells, suggesting a role in cell migration. Structural investigation of rat C-fibers demonstrated that LPA leads to an evagination of the axons from their Schwann cells. In A-fibers, the nodes of Ranvier appeared unchanged, but the Schmidt-Lanterman incisures were shortened and myelination reduced. The latter might increase leak current, reducing the potential spread to the next node of Ranvier and explain the changes in conduction velocity. The observed structural changes provide a plausible explanation for the functional changes in myelinated and unmyelinated axons of peripheral nerves and the reported sensory sensations such as itch and pain.

Exploratory Study of the Brain Response in Facial Synkinesis after Bell Palsy with Systematic Review and Meta-analysis of the Literature.

Facial synkinesis, characterized by unintentional facial movements paired with intentional movements, is a debilitating sequela of Bell palsy. Our aim was to determine whether persistent peripheral nerve changes arising from Bell palsy result in persistent altered brain function in motor pathways in synkinesis. A literature search using terms related to facial paralysis, Bell palsy, synkinesis, and fMRI through May 2021 was conducted in MEDLINE and EMBASE. Additionally, an fMRI study examined lip and eyeblink movements in 2 groups: individuals who fully recovered following Bell palsy and individuals who developed synkinesis. Task-based data of the whole brain that required lip movements in healthy controls were extracted from 7 publications. Three studies contributed similar whole-brain analyses in acute Bell palsy. The meta-analysis of fMRI in healthy control and Bell palsy groups determined common clusters of activation within each group using activation likelihood estimates. A separate fMRI study used multivariate general linear modeling to identify changes associated with synkinesis in smiling and blinking tasks. A region of the precentral gyrus contralateral to the paretic side of the face was hypoactive in synkinesis during lip movements compared with controls. This region was centered in a cluster of activation identified in the meta-analysis of the healthy controls but absent from individuals with Bell palsy. The meta-analysis relied on a small set of studies. The small sample of subjects with synkinesis limited the power of the fMRI analysis. Premotor pathways show persistent functional changes in synkinesis first identifiable in acute Bell palsy.

Peripheral Nerve Involvement at First Diagnosis of Multiple Sclerosis: A Prospective MR Neurography Study.

The aim of this study was to assess peripheral nerve involvement in patients with multiple sclerosis (MS) at first clinical presentation using quantitative magnetic resonance (MR) neurography in correlation with clinical, laboratory, electrophysiological, and central nervous MR imaging data. In this prospective monocentric study, 30 patients first diagnosed with MS according to the McDonald criteria (19 women; mean age, 32.4 ± 8.8 years) and 30 age- and sex-matched healthy volunteers were examined with high-resolution 3 T MR neurography using a dual-echo T2-relaxometry sequence covering the tibial and peroneal nerves from proximal thigh to distal calf. Magnetic resonance biomarkers of T2 relaxation time (T2 app ), proton spin density (PSD), and nerve cross-sectional area (CSA) were correlated with clinical symptoms, intrathecal immunoglobulin (Ig) synthesis, nerve conduction study, and lesion load on brain and spine MR imaging. The diagnostic accuracy of MR biomarkers was assessed using receiver-operating characteristic curves. Diffuse nerve changes were detected along the tibial and peroneal nerves in MS patients, who showed decreased PSD ( P < 0.001), increased T2 app ( P < 0.001), and smaller tibial nerve CSA ( P < 0.001) compared with healthy subjects. Tibial PSD was identified as best parameter separating patients from controls (area under the curve = 0.876). Intrathecal IgG and IgM synthesis correlated with PSD values ( r = -0.44, P = 0.016, and r = -0.42, P = 0.022). Contrast-enhancement of brain or spine lesions was related to larger tibial and peroneal CSA ( P < 0.001, P = 0.033). Abnormal electrophysiology correlated with higher tibial and peroneal T2 app ( P < 0.001 and P = 0.033), lower tibial and peroneal PSD ( P = 0.018 and P = 0.002), and smaller peroneal CSA ( P < 0.001). Quantitative MR neurography reveals peripheral nerve changes in patients with initial diagnosis of MS. Correlation of imaging findings with intrathecal immunoglobulin synthesis may indicate a primary coaffection of the peripheral nervous system in MS.

Can Diffusion Tensor Imaging Apparent Diffusion Coefficient Diagnose Carpal Tunnel Syndrome? A Systematic Review and Meta-Analysis.

Magnetic resonance diffusion tensor imaging (DTI) can detect microstructural changes in peripheral nerves. Studies have reported that the median nerve apparent diffusion coefficient (ADC), a quantification of water molecule diffusion direction, is sensitive in diagnosing carpal tunnel syndrome (CTS). Five databases were searched for studies using ADC to investigate CTS. Apparent diffusion coefficient (measured in mm2/s) were pooled in random-effects meta-analyses. Twenty-two studies met criteria yielding 592 patients with CTS and 414 controls. Median nerve ADC were measured at the level of the distal radioulnar joint (CTS ADC: 1.11, 95% CI: 1.07-1.15, I2 = 54%; control ADC: 1.04, 95% CI: 1.01-1.07, I2 = 57%), pisiform (CTS ADC: 1.39, 95% CI: 1.37-1.42, I2 = 0%; control ADC: 1.27, 95% CI: 1.23-1.31, I2 = 59%), hamate (CTS ADC: 1.40, 95% CI: 1.36-1.43, I2 = 58%; control ADC: 1.27, 95% CI: 1.25-1.28, I2 = 47%), and as an combination of several measurements (CTS ADC: 1.40, 95% CI: 1.37-1.47, I2 = 100%; control ADC: 1.39, 95% CI: 1.24-1.53, I2 = 100%). Median nerve ADC is decreased in individuals with CTS compared to controls at the levels of the hamate and pisiform. ADC cut-offs to diagnose CTS should be established according to these anatomic levels and can be improved through additional studies that include use of a wrist coil.

Expression of Protein Acetylation Regulators During Peripheral Nerve Development, Injury, and Regeneration.

Protein acetylation, regulated by acetyltransferases and deacetylases, is an important post-translational modification that is involved in numerous physiological and pathological changes in peripheral nerves. There is still no systematical analysis on the expression changes of protein acetylation regulators during sciatic nerve development, injury, and regeneration. Here, we sequenced and analyzed the transcriptome of mouse sciatic nerves during development and after injury. We found that the changes in the expression of most regulators followed the rule that “development is consistent with regeneration and opposite to injury.” Immunoblotting with pan-acetylated antibodies also revealed that development and regeneration are a process of increased acetylation, while injury is a process of decreased acetylation. Moreover, we used bioinformatics methods to analyze the possible downstream molecules of two key regulators, histone deacetylase 1 (Hdac1) and lysine acetyltransferase 2b (Kat2b), and found that they were associated with many genes that regulate the cell cycle. Our findings provide an insight into the association of sciatic nerve development, injury, and regeneration from the perspective of protein acetylation.

Fully Convolutional Neural Network Deep Learning Model Fully in Patients with Type 2 Diabetes Complicated with Peripheral Neuropathy by High-Frequency Ultrasound Image.

This study was aimed at exploring the diagnostic value of high-frequency ultrasound imaging based on a fully convolutional neural network (FCN) for peripheral neuropathy in patients with type 2 diabetes (T2D). A total of 70 patients with T2D mellitus were selected and divided into a lesion group (n = 31) and a nonlesion group (n = 39) according to the type of peripheral neuropathy. In addition, 30 healthy people were used as controls. Hypervoxel-based and FCN-based high-frequency ultrasound images were used to examine the three groups of patients to evaluate their diagnostic performance and to compare the changes of peripheral nerves and ultrasound characteristics. The results showed that the Dice coefficient (92.7) and mean intersection over union (mIOU) (82.6) of the proposed algorithm after image segmentation were the largest, and the Hausdorff distance (7.6) and absolute volume difference (AVD) (8.9) were the smallest. The high-frequency ultrasound based on the segmentation algorithm showed higher diagnostic accuracy (94.0% vs. 86.0%), sensitivity (87.1% vs. 67.7%), specificity (97.1% vs. 94.2%), positive predictive value (93.1% vs. 86.7%), and negative predictive value (94.4% vs. 84.0%) (P < 0.05). There were significant differences in the detection values of the three major nerve segments of the upper limbs in the control group, the lesion group, and the nonlesion group (P < 0.05). Compared with the nonlesion group, the patients in the lesion group were more likely to have reduced nerve bundle echo, blurred reticular structure, thickened epineurium, and unclear borders of adjacent tissues (P < 0.05). In summary, the high-frequency ultrasound processed by the algorithm proposed in this study showed a high diagnostic value for peripheral neuropathy in T2D patients, and high-frequency ultrasound can be used to evaluate the morphological changes of peripheral nerves in T2D patients.

LXR agonist improves peripheral neuropathy and modifies PNS immune cells in aged mice

BackgroundPeripheral neuropathy is a common and progressive disorder in the elderly that interferes with daily activities. It is of importance to find efficient treatments to treat or delay this age-related neurodegeneration. Silencing macrophages by reducing foamy macrophages showed significant improvement of age-related degenerative changes in peripheral nerves of aged mice. We previously demonstrated that activation of the cholesterol sensor Liver X receptor (LXR) with the potent agonist, GW3965, alleviates pain in a diet-induced obesity model. We sought to test whether LXR activation may improve neuropathy in aged mice.Methods21-month-old mice were treated with GW3965 (25 mg/Kg body weight) for 3 months while testing for mechanical allodynia and thermal hyperalgesia. At termination, flow cytometry was used to profile dorsal root ganglia and sciatic nerve cells. Immune cells were sorted and analyzed for cholesterol and gene expression. Nerve fibers of the skin from the paws were analyzed. Some human sural nerves were also evaluated. Comparisons were made using either t test or one-way ANOVA.ResultsTreatment with GW3965 prevented the development of mechanical hypersensitivity and thermal hyperalgesia over time in aged mice. We also observed change in polarization and cholesterol content of sciatic nerve macrophages accompanied by a significant increase in nerve fibers of the skin.ConclusionsThese results suggest that activation of the LXR may delay the PNS aging by modifying nerve-immune cell lipid content. Our study provides new potential targets to treat or delay neuropathy during aging.

MRI of the intraspinal nerve roots in patients with chronic inflammatory neuropathies: abnormalities correlate with clinical phenotypes.

Chronic inflammatory demyelinating polyneuropathy (CIDP) and multifocal motor neuropathy (MMN) are caused by inflammatory changes of peripheral nerves. It is unknown if the intra-spinal roots are also affected. This MRI study systematically visualized intra-spinal nerve roots, i.e., the ventral and dorsal roots, in patients with CIDP, MMN and motor neuron disease (MND). We performed a cross-sectional study in 40 patients with CIDP, 27 with MMN and 34 with MND. All patients underwent an MRI scan of the cervical intra-spinal roots. We systematically measured intra-spinal nerve root sizes bilaterally in the transversal plane at C5, C6 and C7 level. We calculated mean nerve root sizes and compared them between study groups and between different clinical phenotypes using a univariate general linear model. Patients with MMN and CIDP with a motor phenotype had thicker ventral roots compared to patients with CIDP with a sensorimotor phenotype (p = 0.012), while patients with CIDP with a sensory phenotype had thicker dorsal roots compared to patients with a sensorimotor phenotype (p = 0.001) and with MND (p = 0.004). We here show changes in the morphology of intra-spinal nerve roots in patients with chronic inflammatory neuropathies, compatible with their clinical phenotype.

In Vivo Cellular‐Level 3D Imaging of Peripheral Nerves Using a Dual‐Focusing Technique for Intra‐Neural Interface Implantation (Adv. Sci. 3/2022)

Neuroprosthesis The sprout dreams of becoming a huge tree. The sprout means our work (OCT-based imaging platform with a new DOF extension technique enabling to provide 3D microstructural changes in peripheral nerve with a metal wire inserted in vivo) and the tree means successful neuroprosthesis. This cover represents that our work will be the beginning of successful neuroprosthesis using intra-neural interface. More details can be found in article number 2102876 by Donghyun Hwang and co-workers.

EFFECT OF HYPERBARIC OXYGEN THERAPY ON NERVE REGENERATION IN RATS.

ABSTRACTObjective: To evaluate histological changes in peripheral nerves of rats after sciatic nerve neurorrhaphy, according to the time of exposure to hyperbaric oxygen chamber treatment. Methods: Twenty-five Wistar rats were divided into 5 groups according to the amount of exposure to hyperbaric oxygen chamber treatment. Group 1 was the control and there was no use of hyperbaric oxygen chamber; group 2 received one week of therapy; group 3, two weeks; group 4, three weeks; and group 5, four weeks. After the fourth postoperative week, the animals were submitted to euthanasia and a sciatic nerve sample sent for histological analysis. Axons proximal and distal to the neurorrhaphy were counted with axonal regeneration index measurement. Results: We observed that the number of axons distal to neurorrhaphy increases with the amount of hyperbaric oxygen chamber exposure, the results were more expressive from the third week of treatment. However, the statistical analysis found no significant difference between the groups. Conclusion: The descriptive analysis suggests benefit of using hyperbaric oxygen chamber directly proportional to the time of therapy. The study, however, did not present statistically relevant results, probably due to the reduced sample size. Subsequent studies with more significant sampling would be of great value. Level of Evidence II, Prospective Comparative Study.

In Vivo Cellular-Level 3D Imaging of Peripheral Nerves Using a Dual-Focusing Technique for Intra-Neural Interface Implantation.

In vivo volumetric imaging of the microstructural changes of peripheral nerves with an inserted electrode could be key for solving the chronic implantation failure of an intra‐neural interface necessary to provide amputated patients with natural motion and sensation. Thus far, no imaging devices can provide a cellular‐level three‐dimensional (3D) structural images of a peripheral nerve in vivo. In this study, an optical coherence tomography‐based peripheral nerve imaging platform that employs a newly proposed depth of focus extension technique is reported. A point spread function with the finest transverse resolution of 1.27 µm enables the cellular‐level volumetric visualization of the metal wire and microstructural changes in a rat sciatic nerve with the metal wire inserted in vivo. Further, the feasibility of applying the imaging platform to large animals for a preclinical study is confirmed through in vivo rabbit sciatic nerve imaging. It is expected that new possibilities for the successful chronic implantation of an intra‐neural interface will open up by providing the 3D microstructural changes of nerves around the inserted electrode.

Effect of vasodilator PGE1 on sciatic nerve function, VEGF level, vascular permeability and peripheral nerve in type 2 diabetic rats

Purpose: To investigate the effect of vasodilator prostaglandin E1 (PGE1) on sciatic nerve function, vascular endothelial growth factor (VEGF), vascular permeability and peripheral nerve in type 2 diabetes mellitus (T2DM) rats.&#x0D; Methods: Twenty-one rats in the study group were intraperitoneally injected with PGE1, while rats in T2DM untreated group (n = 21) were injected with an equivalent amount of normal saline. Seven rats were randomly selected from the study and the control groups every seven days for 21 days for determination of changes in sciatic nerve function, VEGF level, vascular permeability and peripheral nerves.&#x0D; Results: Sciatic nerve motor nerve conduction velocity (MNCV) was significantly higher in the study group than in T2DM untreated control group at the three time-points post-treatment (p &lt; 0.01). The level of VEGF in the study group decreased, relative to the T2DM untreated control group on the 7th 14th and 21st days post- treatment (p &lt; 0.05) while the water content of sciatic nerve tissue in study group was markedly decreased, relative to control value on day 21 (p &lt; 0.05). Rats in the study group showed decreased TTT, relative to those in T2DM untreated group on days 7, 14 and 21 post-treatment (p &lt; 0.01).&#x0D; Conclusion: PGE1 improves the sciatic nerve function of T2DM rats, reduces the level of serum VEGF and vascular permeability, and protects peripheral nerves. These findings provide a basis for the development of new T2DM drugs

The treatment of diabetes with new generation drugs

Introduction: Diabetes is a global problem, affecting nearly 422 million people around the world. Pathogenesis includes a defect in secretion or insulin activity. This results in an increase in the glucose level, which is associated with the development of complications. These include changes in peripheral vessels and nerves that lead to their damage. It is necessary to introduce appropriate treatment in the earliest stages of the disease to prevent these effects.Materials and methods: The aim of this work is to present knowledge about the treatment based on the recommendations of the Polish Diabetes Association, American Diabetes Association, as well as a literature review and analysis of publications published on PubMed and Google Scholar platforms.Results: Over the years, the recommendations and recommendations for treatment change. Recently, a greater role has been given to the latest antidiabetic drugs. This group includes SGLT-2 inhibitors as well as incremental drugs, the main representatives of which are GLP-1 analogues. These drugs affect the level of glycemia, but also have a beneficial effect on weight reduction and reduce the risk of cardiovascular events. Recently, Poland has introduced reimbursement of some new generation antidiabetic drugs such as dapaglyphosin, empaglyphosin and canaglyphosin.Conclusions: Dynamic development of diabetes treatment helps to slow down the course of the disease and postpones the introduction of insulin therapy as the final treatment method. Reimbursement of some antidiabetic drugs enables patients to have better access to drugs that have not been within their reach so far. The changes introduced are, in a way, groundbreaking in the treatment of diabetes.

Perioperative measures for prevention of phantom pain: an evidence-based approach to risk reduction

Prevention of phantom limb pain is one of the biggest and still largely unsolved challenges in perioperative medicine. Despite many study efforts and optimization of postoperative pain treatment over the last 30years, asignificant reduction in the incidence of phantom limb pain has not been achieved. Current studies have also shown that at least 50% of patients develop phantom pain after 6months. A possible approach could be to combine multiple synergistic interventions and implement them as aperioperative phantom pain management strategy bundle. In addition to regional anesthesia, NMDA antagonists, gabapentinoids, antidepressants and systemic lidocaine could play arelevant role. The aim of this pharmacological intervention was the modification of the pathophysiological changes in peripheral nerves and in the central nervous system after amputation.

Microstructural changes in the trigeminal nerve of patients with episodic migraine assessed using magnetic resonance imaging

BackgroundThere is histological evidence of microstructural changes in the zygomaticotemporal branch of the trigeminal nerve in migraineurs. This raises the possibility that altered trigeminal nerve properties contribute to migraine pathophysiology. Whilst it is not possible to explore the anatomy of small trigeminal nerve branches it is possible to explore the anatomy of the trigeminal root entry zone using magnetic resonance imaging in humans. The aim of this investigation is to assess the microstructure of the trigeminal nerve in vivo to determine if nerve alterations occur in individuals with episodic migraine.MethodsIn 39 migraineurs and 39 matched controls, T1-weighted anatomical images were used to calculate the volume (mm3) and maximal cross-sectional area of the trigeminal nerve root entry zone; diffusion tensor images were used to calculate fractional anisotropy, mean diffusion, axial diffusion and radial diffusion.ResultsThere were significant differences between the left and right nerve of controls and migraineurs with respect to volume and not cross-sectional area. Migraineurs displayed reduced axial diffusion in the right nerve compared to the left nerve, and reduced fractional anisotropy in the left nerve compared to left controls. Furthermore, although there were no differences in mean diffusion or radial diffusion, regional analysis of the nerve revealed significantly greater radial diffusion in the middle and rostral portion of the left trigeminal nerve in migraineurs compared with controls.ConclusionsMigraine pathophysiology is associated with microstructural abnormalities within the trigeminal nerve that are consistent with histological evidence of altered myelin and/or organization. These peripheral nerve changes may provide further insight into migraine pathophysiology and enable a greater understanding for targeted treatments of pain alleviation.

Protective mechanism against age-associated changes in the peripheral nerves

AimsSenescence is the normal decline in physiological functions due to aging that results in cell dysfunction. However, age-associated changes in peripheral nerves have not been elucidated. We observed histological changes in the sciatic nerves of young and older mice to investigate how peripheral nerves changed with age, and we evaluated protective mechanisms of peripheral nerves against aging. Main methodsSciatic nerves were collected from female C57BL/6 mice at the ages of 8 weeks (young group) and 78 weeks (aged group) and examined histologically. Using hematoxylin and eosin staining, the number and density of sciatic nerve axons were evaluated. Through immunofluorescence staining, the expression of nerve-specific proteins, oxidative stress markers, and a neuronal aging marker (REST/NRSF) were investigated, and the intensity of fluorescence was quantified. The differences between the groups were assessed, and age-associated peripheral nerve changes were evaluated. Statistical analysis was performed using the Mann–Whitney U test. Key findingsAlthough the number and density of axons did not differ significantly between the groups, they were lower in the aged group than in the young group. In addition, the fluorescence intensity of each marker did not differ significantly between the groups, but the expression of REST/NRSF alone was significantly higher in the aged group than in the young group (p < 0.05). SignificanceThis study suggested that peripheral nerve functions are preserved by the expression of REST/NRSF, which increases with age. Because oxidative stress did not change, the protective effects of REST/NRSF are considered to be related to oxidative stress.

Морфологическая характеристика изменений периферических нервов и тканей таргетных зон при соединении периферических нервов по типу «конец-в-бок» в эксперименте

Морфологическая характеристика изменений периферических нервов и тканей таргетных зон при соединении периферических нервов по типу «конец-в-бок» в эксперименте

Incidental Ultrastructural Findings in the Sural Nerve and Dorsal Root Ganglion of Aged Control Sprague Dawley Rats in a Nonclinical Carcinogenicity Study.

Xenobiotic-induced peripheral nerve damage is a growing concern. Identifying relative risks that a new drug may cause peripheral nerve injury over long periods of administration is gathering importance in the evaluation of animal models. Separating out age-related changes in peripheral nerves of rats caused by compression injury from drug-induced effects has been difficult. Biopsy of the sural nerve is utilized in humans for investigations of peripheral neuropathy, because it is largely removed from the effects of nerve compression. This study used transmission electron microscopy to identify incidental findings in the sural nerves and dorsal root ganglia of aged control rats over time. The goal was to establish a baseline understanding of the range of possible changes that could be noted in controls compared to rats treated with any new investigative drug. In this evaluation, most sural nerve fibers from aged control rats had few ultrastructural abnormalities of pathologic significance. However, glycogenosomes, polyglucosan bodies, swollen mitochondria, autolysosomes, split myelin, Schwann cell processes, and endoneural macrophages with phagocytosed debris (considered an indication of ongoing degenerative changes) were occasionally noted.

Analgesic Effect of Dizocine Combined with Ropivacaine on Recurrent Neuropathic Pain in Peripheral Nerve Compression Rats

To explore the analgesic effect of dizocine combined with ropivacaine on recurrent neuropathic pain in rat model of peripheral nerve compression. Rats were randomly divided into 5 groups: sham control group (S), peripheral nerve compression model group (M), dizocine group (D), ropivacaine group (R), and combined drug group (DR). Rat peripheral nerve compression model was constructed to observe the symptoms of the rats before and after surgery. Mechanical withdrawal threshold was measured on the 21st day after surgery. The electrophysiological changes of rat peripheral nerve were measured by biopotential recording system, including proximal latency, distal latency, and compound muscle action potential. The incubation period and nerve conduction velocity were further obtained. Histological changes were observed by HE staining and toluidine blue staining. Axon number and myelin damage grade were performed, and the ultrastructure was observed by transmission electron microscopy (TEM). The mechanical withdrawal threshold, nerve conduction velocity, and compound muscle action potential were effectively increased in combination group. However, the proximal latency, distal latency, and incubation period were significantly reduced. Furthermore, dizocine combined with ropivacaine can effectively reduce the degree of myelination. TEM shown that the DR group had the best therapeutic effect, and the histological appearance of the cross section was quite similar to that of the S group. Dizocine combined with ropivacaine has a significant analgesic effect in rat model of peripheral nerve compression.

Peripheral Nerve Microscopic Changes Related to Study Procedures: Two Nonclinical Case Studies.

Peripheral nerves are routinely examined microscopically during the nonclinical safety assessment of therapeutics. In addition to test article-related on- or off-target changes, microscopic changes in peripheral nerves may also be caused by study procedures, such as parenteral test article administration and blood or tissue sampling. We present 2 nonclinical case studies in which nonstandard peripheral nerves had study procedure-related histologic changes. The first case study describes mouse trigeminal nerve changes as a result of blood sampling via retro-orbital sinus puncture. These changes included minimal-to-mild nerve fiber (axonal) degeneration associated with macrophage infiltration. The second case study presents rat brachial plexus changes associated with animal handling and blood sampling. Brachial plexus changes included minimal-to-moderate inflammation, focal hemorrhage, and nerve fiber degeneration. In both cases, the histological changes were morphologically indistinguishable from those that might be due to test article. Therefore, careful consideration of the incidence and severity across groups and a review of study procedures to rule out handling-related nerve damage are essential before identifying a test article-related effect on peripheral nerves. Study design considerations to avoid such procedure-related changes will be discussed, as well as sampling strategies to help distinguish these from test article-related effects.