Progesterone may be a regulator and B12 could be an indicator of the proximal D4Z4 repeat methylation status on 4q35ter.

Facioscapulohumeral dystrophy (FSHD) has a hypomethylation-related epigenetic background and exhibits a different course in male and female patients. The differences between males and females have been linked to the levels of sex hormones. This study is the first to investigate the possible effect of these hormones on methylation status. We hypothesized that the levels of sex-related hormones, estradiol, testosterone, progesterone, and prolactin might be associated with the methylation status of the proximal part of the D4Z4. We also investigated the effect of fT3, folic acid, and vitamin B12 levels. We collected blood from 28 FSHD patients and 28 controls. DNA was extracted from each individual for bisulfite methylation analysis and serum was separated for biochemical analysis of estradiol, testosterone, progesterone, prolactin, fT3, folic acid, and B12 analysis. Methylation analysis was specified to the DR1, 5P regions and the proximal region covering both DR1 and 5P. Methylation levels were compared between FSHD patients and controls. The correlation of methylation levels with estradiol, testosterone, progesterone, prolactin, fT3, folic acid, and B12 was investigated. We found that the 5P region and the proximal region were significantly hypomethylated in FSHD patients compared to the controls, but not the DR1 region. Male patients exhibited a significant reduction in DNA methylation compared to male controls. Older FSHD patients exhibited a notable decrease in fT3 levels and hypomethylation of the 5P region. Analyses of each CpG revealed seven hypomethylated positions that were significantly different from the control group. Two of the positions demonstrated a correlation with progesterone in the control group. With the exception of one position, the methylation levels were inversely correlated with vitamin B12 in FSHD patients. The results of our study indicate that the methylation of the proximal D4Z4 region, particularly at specific positions, may be associated with progesterone. In addition, vitamin B12 may be an indicator of hypomethylation. We suggest that examining position-specific methylations may be a useful approach for the development of epigenetic treatment modalities.

Microglia cells treated with synthetic vasoactive intestinal peptide or transduced with LentiVIP protect neuronal cells against degeneration.

A common pathological hallmark of neurodegenerative disorders is neuronal cell death, accompanied by neuroinflammation and oxidative stress. The vasoactive intestinal peptide (VIP) is a pleiotropic peptide that combines neuroprotective and immunomodulatory actions. The gene therapy field shows long-term promise for treating a wide range of neurodegenerative diseases (ND). In this study, we aimed to investigate the in vitro efficacy of transduction of microglia using lentiviral gene therapy vectors encoding VIP (LentiVIP). Additionally, we tested the protective effects of the secretome derived from LentiVIP-infected "immortalized human" microglia HMC3 cells, and cells treated with Synthetic VIP (SynVIP), against toxin-induced neurodegeneration. First, LentiVIP, which stably expresses VIP, was generated and purified. VIP secretion in microglial conditioned media (MG CM) for LentiVIP-infected HMC3 microglia cells was confirmed. Microglia cells were activated with lipopolysaccharide, and groups were formed as follows: 1) Control, 2) SynVIP-treated, or 3) LentiVIP-transduced. These MG CM were applied on an in vitro neurodegenerative model formed by differentiated (d)-SH-SY5Y cells. Then, cell survival analysis and apoptotic nuclear staining, besides measurement of oxidative/inflammatory parameters in CM of cells were performed. Activated MG CM reduced survival rates of both control and toxin-applied (d)-SH-SY5Y cells, whereas LentiVIP-infected MG CM and SynVIP-treated ones exhibited better survival rates. These findings were supported by apoptotic nuclear evaluations of (d)-SH-SY5Y cells, alongside oxidative/inflammatory parameters in their CM. LentiVIP seems worthy of further studies for the treatment of ND because of the potential of gene therapy to treat diseases effectively with a single injection.

De Novo and Dominantly Inherited SPTAN1 Mutations Cause Spastic Paraplegia and Cerebellar Ataxia.

BACKGROUND: Pathogenic variants in SPTAN1 have been linked to a remarkably broad phenotypical spectrum. Clinical presentations include epileptic syndromes, intellectual disability, and hereditary motor neuropathy. OBJECTIVES: We investigated the role of SPTAN1 variants in rare neurological disorders such as ataxia and spastic paraplegia. METHODS: We screened 10,000 NGS datasets across two international consortia and one local database, indicative of the level of international collaboration currently required to identify genes causative for rare disease. We performed in silico modeling of the identified SPTAN1 variants. RESULTS: We describe 22 patients from 14 families with five novel SPTAN1 variants. Of six patients with cerebellar ataxia, four carry a de novo SPTAN1 variant and two show a sporadic inheritance. In this group, one variant (p.Lys2083del) is recurrent in four patients. Two patients have novel de novo missense mutations (p.Arg1098Cys, p.Arg1624Cys) associated with cerebellar ataxia, in one patient accompanied by intellectual disability and epilepsy. We furthermore report a recurrent missense mutation (p.Arg19Trp) in 15 patients with spastic paraplegia from seven families with a dominant inheritance pattern in four and a de novo origin in one case. One further patient carrying a de novo missense mutation (p.Gln2205Pro) has a complex spastic ataxic phenotype. Through protein modeling we show that mutated amino acids are located at crucial interlinking positions, interconnecting the three-helix bundle of a spectrin repeat. CONCLUSIONS: We show that SPTAN1 is a relevant candidate gene for ataxia and spastic paraplegia. We suggest that for the mutations identified in this study, disruption of the interlinking of spectrin helices could be a key feature of the pathomechanism. © 2022 International Parkinson and Movement Disorder Society.

Comparison of nerve conduction velocity distribution methods by cold exposure and ischemia.

PURPOSE: Non-invasive estimation of the conduction velocity distribution (CVD) of a peripheral nerve has the potential to both improve clinical diagnoses of pathology and to observe the progress of the disease or the efficacy of treatments. Comparisons were made of the performance of three non-invasive CVD estimation methods proposed by independent research groups on peripheral nerve bundles under different conditions. METHODS: The first method (Cummins) uses a nerve compound action potential (CAP) with temporal dispersion and a mathematical single fiber action potential (SFAP). The second method (Barker) uses two CAPs and a non-mathematical SFAP waveform. The third method (Hirose) uses two CAPs recorded from distal and proximal sites. The Cummins and Barker methods have iterative solutions in the time domain while the Hirose method is a deconvolution estimator in the frequency domain. In order to compare these methods, we used cold exposure to affect primarily motor fibers and ischemia to affect primarily sensory fibers on rat caudal nerve bundles. RESULTS: The Cummins method is sensitive to changes in motor and sensory fiber percentages in CVD if it is used with the volume conductor model. The Barker and Hirose methods are sensitive to motor fiber percentages in CVD but they cannot detect changes in sensory fiber percentages accurately. CONCLUSIONS: Estimation of the CVD using a priori SFAP created with a volume conductor model can non-invasively supply accurate and precise information about fiber groups in a peripheral nerve bundle.

Amplitude and velocity dependence of patellar pendulum triggered by T reflex in Parkinson's rigidity.

OBJECTIVES: The relation between increase of tonus and joint movement velocity is controversial in Parkinson's rigidity. It is accepted that the increase of tonus in rigidity is constant during joint movement, and does not change within all limits of movement. However, there is thoughtful evidence that the change in tonus in rigidity has a correlation with joint movement velocity and amplitude of movement. The pendulum movement that is formed by triggering of the patellar T reflex allows the examination of phasic stretching reflexes and physiological changes of passive stretching. Therefore, the velocity and amplitude properties of tonus in Parkinson's rigidity can be scanned together. MATERIALS AND METHODS: Patellar T reflex-triggered patellar pendulum was recorded in 40 Parkinson's patients. The velocity and amplitude changes in the pendulum were observed according to the rigidity scale. Muscle action potentials were recorded from the rectus femoris muscle and biceps femoris muscles simultaneously via superficial recording electrodes. Knee joint angle changes were recorded with a goniometer. The kinesiological and electromyographic features were compared with those of the control subjects. RESULTS: The number of pendulums decreased significantly, the angle of joint movement decreased, the peak time decreased and the angular velocity slowed down significantly in the Parkinson's group. While the latency of the patellar T reflex did not change significantly, its amplitude decreased, and the onset time of joint movement measured by accelerometer was prolonged. CONCLUSIONS: Parkinson's rigidity has a velocity-dependent component, and this correlates negatively with the rigidity scale.

Complementary Phenomena: Phantom Hand and Phantom Face.

After tissue or limb loss, the development of sensation and perception of the lost or deafferent tissue is defined as a phantom phenomenon. We investigated the presence of phantom phenomena in individuals who underwent a full face transplant as well as those who had a hand transplant. Specifically, we investigated sensory perception of the face on the fingers and sensory perception of the fingers on the face in three full face and four hand transplant patients. In all seven individuals, we used a brush to separately stimulate the right and left sides of the face or the palmar and dorsal faces of the hand. We then asked the individuals if they felt a sensation of touch on any other part of their body and, if so, to describe their perceptions. Changes in the regions of the primary sensory cortex representing the hand and face were defined using fMRI obtained via tactile sensory stimulation of the clinical examination areas. Two of the full face transplant patients reported sensory perceptions such as a prominent sensation of touch on their faces during sensory stimulation of their fingers. Three of the hand transplant patients reported sensory perceptions, which we referred to as finger patches, during sensory stimulation of the face area. In fMRI, overlaps were observed in the cortical hand and face representation areas. We consider the phantom hand and phantom face phenomena we observed to be complementary due to the neighborhood of the representations of the hand and face in the somatosensory cortex.

An Individualized Physiotherapy and Rehabilitation Program for Bilateral Hand Transplantations.

METHODS: This article describes a rehabilitation program and the long-term results after its application in 2 double-hand transplantation patients after 9- and 4-year follow-up periods. A personal rehabilitation program was planned to commence as early as possible after postoperative patient stabilization. Splinting, edema, positioning, passive and active joint movements, daily life activity modification and education, and occupational therapy were also emphasized. RESULTS: Positioning, edema, and passive joint movements were started in the acute phase. Strengthening and exercises were continued for 8 weeks and later for functional gains and adaptation to daily living activities. Subjective and overall results were quite satisfactory. The Hand Transplantation Score System, Disabilities of the Arm Shoulder and Hand, Semmes Weinstein Monofilaments, sensory tests, and modified Kapandji index improvements were all within good acceptable ranges. Although rapid recovery and functional development were observed in the first 4 years, these are still continuing at the time of writing. DISCUSSION: The most important determining factors in the success of rehabilitation are to return to daily life and the ability to acquire functional skills. Long-term follow-up of the patients in this study showed that they were able to return to their independent daily lives and that external assistance decreased to a minimum.

Replantation of forequarter amputation: Report of two cases with successful structural, motor and sensorial results.

Although there have been numerous reports of major replantation of upper extremity amputations, limited numbers of above-elbow amputation replantation have been reported. We present the technical details of two successful replantations of forequarter amputations in a nine-year-old girl and a three-year-old boy. In both cases, the forequarter was amputated due to avulsion traction injuries resulting in amputation including the entire upper limb, while the integrity of the scapula and parascapular muscles was maintained, with no injury to the glenohumeral joint. Replantation was performed, involving a shorter ischemia time with proper fixation, and vascular and neural repairs. Postoperative recovery was uneventful, and motor and sensorial acquisition were quite satisfactory during follow-up periods of 9 and 6 years, respectively. Proper fixation of the amputated part mimicking the original anatomy, radical debridement of avulsed vessels, and reconstruction of the defect using long vein grafts and neural repair while maintaining proper integrity are the most important factors in success. When the requirements are met, replantation of the forequarter in a child yields a superior outcome, from both the functional and esthetic perspectives. To the best of our knowledge, this is the first report in the English literature involving two sequential cases of such high-level replantation resulting in successful reacquisition of both viability and function.

Tibial nerve axonal excitability in type 1 diabetes mellitus.

INTRODUCTION: The aim of this study was to determine alterations in axonal excitability in tibial nerve as compared with median nerve axonal excitability in patients with diabetic polyneuropathy. METHODS: Six patients with diabetic polyneuropathy and 10 patients with diabetes mellitus without polyneuropathy were enrolled. RESULTS: Compared with diabetic patients without polyneuropathy, the tibial nerve strength-duration time constant was significantly longer and supernormality was lower in those with polyneuropathy. Threshold electrotonus studies showed abnormalities in patients with diabetic polyneuropathy, in which smaller threshold changes from long-depolarizing and hyperpolarizing conditioning, termed "fanning-in," were found. DISCUSSION: This study confirms that axonal excitability is significantly altered in the tibial nerve of patients with diabetic polyneuropathy. Evaluating the axonal excitability of the median and tibial nerves may reveal the presence of length-dependent polyneuropathy at an early stage. Muscle Nerve 59:76-81, 2019.

Medium latency excitatory reflex of soleus re-examined.

We aimed to study the receptor origin and postsynaptic potential profile of the medium latency reflex (MLR) response that develops in the soleus muscle when common peroneal nerve of antagonist tibialis anterior (TA) muscle is electrically stimulated. To achieve this aim, we electrically stimulated common peroneal nerve and recorded surface electromyography (SEMG) responses of soleus and TA muscles of informed volunteers. Additionally, we recorded intramuscular EMG from the soleus muscle. Stimulation of common peroneal nerve induced a direct motor response (M-response) in the TA and MLR in SEMG of the soleus. Using voluntarily-activated single motor units (SMUs) from the soleus muscle we noted that there were two distinct responses following the stimulus. The first response was a reciprocal inhibitory reflex probably originating from the antagonist muscle spindle primary (Ia) afferents. This was followed by an indirect reflex response activated by the contraction of the TA muscle during the M-response. This contraction generated a rapid acceleration in the direction of dorsiflexion hence inducing a stretch stimulus on soleus muscle. The response of soleus to this stimulus was a stretch reflex. We suggest that this stretch reflex is the main contributor to the so-called soleus MLR in the literature. This study illustrated the importance of using SMUs and also using discharge-rate based analysis for closely examining previously 'established' reflexes.

Functional lower lip reconstruction with the partial latissimus dorsi muscle free flap without nerve coaptation.

BACKGROUND: The repair of extensive lower lip defects is difficult due to the presence of both functional and esthetic requirements. This report describes functional lip repair using the partial latissimus dorsi free flap without nerve coaptation. METHODS: Reconstruction of the lower lip due to subtotal and total tissue defects was performed using latissimus dorsi free flap on twelve patients, between 2013 and 2017. The etiology was squamous cell carcinoma in six patients, malignant melanoma in one, firearm injury in three and microstomia in two. Following exposure of the lip defect, the partial latissumus dorsi muscle flap was harvested and transfered into the lower lip defect. The microvascular anastomoses was done and no nerve coaptation was performed. The muscle was covered with a skin graft taken from the thigh for mucosal and skin lining. Functional outcomes were assessed using physical examination, electromyography (EMG), electroneuromyography, cold/warm and pain sensation, two point discrimination (TPD), and Semmes Weinstein (SMW) tests. RESULTS: Postoperative course was uneventful for all of the flaps. Patients were followed for between fifteen months and four years (mean 28.2 months).Color match between the flap and the face and functional outcome was satisfactory. Reinnervation was detected at EMG in eleven patients undergoing surgery six months postoperatively.After 1 year, the patients demonstrated quite normal results of the test with SMW and TDP results. None of patients perceived both hot and cold sensation. CONCLUSION: The technique is an alternative option in lower lip reconstruction since it provides satisfactory functional and esthetic results despite absence of any neural coaptation.

Face Perception in Face Transplant Patients.

The authors tested face discrimination, face recognition, object discrimination, and object recognition in two face transplantation patients (FTPs) who had facial injury since infancy, a patient who had a facial surgery due to a recent wound, and two control subjects. In Experiment 1, the authors showed them original faces and morphed forms of those faces and asked them to rate the similarity between the two. In Experiment 2, they showed old, new, and implicit faces and asked whether they recognized them or not. In Experiment 3, they showed them original objects and morphed forms of those objects and asked them to rate the similarity between the two. In Experiment 4, they showed old, new, and implicit objects and asked whether they recognized them or not. Object discrimination and object recognition performance did not differ between the FTPs and the controls. However, the face discrimination performance of FTP2 and face recognition performance of the FTP1 were poorer than that of the controls were. Therefore, the authors concluded that the structure of the face might affect face processing.

Recovery of facial expressions using functional electrical stimulation after full-face transplantation.

BACKGROUND: We assessed the recovery of 2 face transplantation patients with measures of complexity during neuromuscular rehabilitation. Cognitive rehabilitation methods and functional electrical stimulation were used to improve facial emotional expressions of full-face transplantation patients for 5 months. Rehabilitation and analyses were conducted at approximately 3 years after full facial transplantation in the patient group. We report complexity analysis of surface electromyography signals of these two patients in comparison to the results of 10 healthy individuals. METHODS: Facial surface electromyography data were collected during 6 basic emotional expressions and 4 primary facial movements from 2 full-face transplantation patients and 10 healthy individuals to determine a strategy of functional electrical stimulation and understand the mechanisms of rehabilitation. A new personalized rehabilitation technique was developed using the wavelet packet method. Rehabilitation sessions were applied twice a month for 5 months. Subsequently, motor and functional progress was assessed by comparing the fuzzy entropy of surface electromyography data against the results obtained from patients before rehabilitation and the mean results obtained from 10 healthy subjects. RESULTS: At the end of personalized rehabilitation, the patient group showed improvements in their facial symmetry and their ability to perform basic facial expressions and primary facial movements. Similarity in the pattern of fuzzy entropy for facial expressions between the patient group and healthy individuals increased. Synkinesis was detected during primary facial movements in the patient group, and one patient showed synkinesis during the happiness expression. Synkinesis in the lower face region of one of the patients was eliminated for the lid tightening movement. CONCLUSIONS: The recovery of emotional expressions after personalized rehabilitation was satisfactory to the patients. The assessment with complexity analysis of sEMG data can be used for developing new neurorehabilitation techniques and detecting synkinesis after full-face transplantation.

Image-based Analysis of Emotional Facial Expressions in Full Face Transplants.

In this study, it is aimed to determine the degree of the development in emotional expression of full face transplant patients from photographs. Hence, a rehabilitation process can be planned according to the determination of degrees as a later work. As envisaged, in full face transplant cases, the determination of expressions can be confused or cannot be achieved as the healthy control group. In order to perform image-based analysis, a control group consist of 9 healthy males and 2 full-face transplant patients participated in the study. Appearance-based Gabor Wavelet Transform (GWT) and Local Binary Pattern (LBP) methods are adopted for recognizing neutral and 6 emotional expressions which consist of angry, scared, happy, hate, confused and sad. Feature extraction was carried out by using both methods and combination of these methods serially. In the performed expressions, the extracted features of the most distinct zones in the facial area where the eye and mouth region, have been used to classify the emotions. Also, the combination of these region features has been used to improve classifier performance. Control subjects and transplant patients' ability to perform emotional expressions have been determined with K-nearest neighbor (KNN) classifier with region-specific and method-specific decision stages. The results have been compared with healthy group. It has been observed that transplant patients don't reflect some emotional expressions. Also, there were confusions among expressions.

Assessment of Emotional Expressions after Full-Face Transplantation.

We assessed clinical features as well as sensory and motor recoveries in 3 full-face transplantation patients. A frequency analysis was performed on facial surface electromyography data collected during 6 basic emotional expressions and 4 primary facial movements. Motor progress was assessed using the wavelet packet method by comparison against the mean results obtained from 10 healthy subjects. Analyses were conducted on 1 patient at approximately 1 year after face transplantation and at 2 years after transplantation in the remaining 2 patients. Motor recovery was observed following sensory recovery in all 3 patients; however, the 3 cases had different backgrounds and exhibited different degrees and rates of sensory and motor improvements after transplant. Wavelet packet energy was detected in all patients during emotional expressions and primary movements; however, there were fewer active channels during expressions in transplant patients compared to healthy individuals, and patterns of wavelet packet energy were different for each patient. Finally, high-frequency components were typically detected in patients during emotional expressions, but fewer channels demonstrated these high-frequency components in patients compared to healthy individuals. Our data suggest that the posttransplantation recovery of emotional facial expression requires neural plasticity.

The effectiveness of TBL with real patients in neurology education in terms of knowledge retention, in-class engagement, and learner reactions.

In our medical school, we changed from a lecture-based method to a team-based learning (TBL) method to teach "polyneuropathies" in the neurology clerkship starting from the 2014 to 2015 academic year. Real patients were used instead of written scenarios in TBL sessions. This study aimed to compare former lecture-based and the current TBL methods in terms of knowledge retention, in-class learner engagement, and learner reactions. First, we determined in-class engagement and satisfaction of the students for the lectures given in the 2013-2014 academic year. The following year, besides the same criteria, we also determined individual (IRAT) and group readiness test (GRAT) scores in the TBL group. End-of-clerkship exam scores for both groups were recorded. Additionally, opinions of patients about their experiences throughout the TBL process were determined. One year later (2015 for lecture and 2016 for TBL), both groups sat for an MCQ test to determine their knowledge retention levels. We found no difference between groups regarding end-of-clerkship exam scores. The mean knowledge retention test score of the TBL group was significantly higher than that of the lecture group (5.85 ± 1.74 vs. 3.28 ± 1.70). The differences between IRAT, GRAT, and retention test scores in the TBL group were significant. The mean student satisfaction score on a five-point scale was 3.01 ± 0.9 (median = 3) in the lecture group and 4.11 ± 1.1 (median = 4) in the TBL group. Our results seem encouraging for use of TBL performed with real patients in neurology education to achieve better long-term knowledge retention and higher in-class engagement and student satisfaction.

Consideration of difficulties and exit strategies in a case of face allotransplantation resulting in failure.

We describe the first rescue procedure in a case of total face allotransplantation. The recipient was a 54-year-old man with severe disfigurement of the entire face following an accidental gunshot injury 5 years previously. The large defect included the maxilla, mandible, and mid-face. Full face procurement was performed from a multiorgan cadaveric donor and was allotransplanted to the recipient. The post-transplant induction immunosuppressive regimen included ATG combined with tacrolimus, mycophenolate mofetil, and prednisone, while maintenance was provided by the last three of these. Although the early postoperative period was uneventful, squamous cell carcinoma developed in the upper and lower extremities in the fifth postoperative month, and post-transplant lymphoproliferative disorder (PTLD) occurred in the sixth month postoperatively. Malignancies were treated, involving both surgical and medical approaches. The patient developed opportunistic pulmonary and cerebellar aspergillosis. In order to reduce the adverse affects and metabolic and immunological load, the transplanted face was removed and replaced with a free flap. Although the early postoperative period was promising, with the transferred flap surviving totally and all vital signs and general status appearing to be improving, the patient was eventually lost due to complicated infectious and metabolic events. Although this case was unsuccessful, we suggest that the immunological and metabolic load should be reduced as soon as stable medical conditions are established in case of diagnosis of a situation involving a high rate of mortality, such as PTLD and untreatable opportunistic infections. This should include withdrawal of all immunosuppressive drugs and removal of all allotransplanted tissues.

Late-onset Hirayama disease presenting with ulnar neuropathy: A case report.

Hirayama disease (HD) was first reported by Hirayama et al. in 1959. The disease is considered as a type of benign focal motor neuron disease that primarily affects upper limbs of young males. In this case report, we present a man aged 40-years with rapidly progressive weakness and atrophy in his left hand. The findings of nerve conduction studies were consistent with left ulnar neuropathy at the elbow. Flexion magnetic resonance imaging (MRI) revealed minimal enlargement of the posterior epidural space and anterior displacement of the spinal cord. After exclusion of relevant diseases the patient was diagnosed as having Hirayama disease with ulnar neuropathy. Mild ulnar entrapment at the elbow may be considered as a clinical feature of HD. Therefore, it is recommended that young male patients with wasting in upper extremities with findings of ulnar entrapment should not be judged to have ulnar neuropathy before HD has been carefully excluded.