Biochemical, biomechanical and imaging biomarkers of ischemic stroke: Time for integrative thinking.

Stroke is one of the leading causes of adult disability affecting millions of people worldwide. Post-stroke cognitive and motor impairments diminish quality of life and functional independence. There is an increased risk of having a second stroke and developing secondary conditions with long-term social and economic impacts. With increasing number of stroke incidents, shortage of medical professionals and limited budgets, health services are struggling to provide a care that can break the vicious cycle of stroke. Effective post-stroke recovery hinges on holistic, integrative and personalized care starting from improved diagnosis and treatment in clinics to continuous rehabilitation and support in the community. To improve stroke care pathways, there have been growing efforts in discovering biomarkers that can provide valuable insights into the neural, physiological and biomechanical consequences of stroke and how patients respond to new interventions. In this review paper, we aim to summarize recent biomarker discovery research focusing on three modalities (brain imaging, blood sampling and gait assessments), look at some established and forthcoming biomarkers, and discuss their usefulness and complementarity within the context of comprehensive stroke care. We also emphasize the importance of biomarker guided personalized interventions to enhance stroke treatment and post-stroke recovery.

Spectral Changes of EEG Following a 6-Week Low-Dose Oral Ketamine Treatment in Adults With Major Depressive Disorder and Chronic Suicidality.

BACKGROUND: Ketamine has considerable therapeutic potential in alleviating major depressive disorder and chronic suicidality. However, the clinical diagnosis of neuropsychiatric disorders requires more robust diagnostic criteria. Electroencephalography (EEG) has shown promise in classifying depressive and suicidal patients from healthy individuals. The present study aimed to identify changes in the spectral properties of EEG in patients with major depressive disorder and chronic suicidality after completing the 6-week Oral Ketamine Trial on Suicidality with follow-up occurring 4 weeks after final ketamine treatment and determine associations between EEG spectral output and clinical symptoms. METHODS: Participants (n = 25) had 4-minute eyes closed resting state EEG recorded at frontal, temporal, centro-parietal, and occipital regions. Spectral analysis was performed with Welch's power spectrum density method, and the power of 4 distinct frequency bands was analyzed: theta, alpha, low-beta, and high-beta. Correlation analyses between changes in clinical symptoms and spectral power were conducted using Spearman's ranked correlation. RESULTS: Between pre- and posttreatment, only centro-parietal alpha power decreased. Between posttreatment and follow-up, centro-parietal alpha increased again in addition to increases in temporal alpha, centro-parietal and temporal theta, and occipital low-beta and decreases in occipital theta and temporal low-beta. Additionally, the decrease of occipital theta positively correlated with clinical subscales for depression and stress. CONCLUSIONS: EEG spectral analysis revealed significant changes in theta, alpha, and low-beta frequency bands. Alpha band showed initial changes after treatment; however, this trended back toward baseline levels after the treatment cessation. In contrast, theta and low-beta showed significant power changes only after the treatment had ended.

Prophylactic Cerclage Cabling Decreases the Intraoperative Periprosthetic Fracture Rate during Cement-Less Total Hip Arthroplasty in Patients with Severe Developmental Hip Dysplasia.

PURPOSE OF THE STUDY Cementless stems in highly dysplastic hips are considered to increase the risk of femoral fracture and associated morbidity. Several authors speculated to use prophylactic cabling in this patient group to prevent intraoperative fractures. This study aims to reveal objective results regarding the perioperative complications in a large and consecutive patient group with respect to use of prophylactic cabling. MATERIAL AND METHODS A retrospective comparative study was planned. A total of 122 consecutive patients with dysplastic hips of Crowe type 3 or 4, operated on with total hip arthroplasty (THA) and shortening osteotomy using a rectangular femoral stem were included. Patients were stratified according to use of a diaphyseal prophylactic cerclage cable. Perioperative complications were recorded. Clinical outcome was measured in terms of Harris Hip Score and Visual Analog Scale (VAS) for pain. All results were compared between the groups. RESULTS The mean follow-up time was 27 months. Two (2%) versus five (14%) patients had a fracture at the diaphyseal level in cabled versus non-cabled groups. Difference between groups was statistically significant (p=0.01). Relative risk of fracture in case of a non-cabling was 5.8 (p=0.03). Eleven (9%) patients had a non-displaced fracture at the metaphyseal level. No significant differences were detected with respect to preoperative clinical outcome scores or change in these scores between groups. CONCLUSIONS Femoral diaphyseal fracture rates are low when cementless, rectangular stems are used in dysplastic high riding hips. Prophylactic cerclage cabling further decreases the fracture risk and eases treatment in case of a fracture without causing additional complications and therefore is recommended. Key words: intraoperative fracture, periprosthetic fracture, total hip arthroplasty, cerclage cabling, developmental hip dysplasia, transverse shortening osteotomy.

Effects of Kinesio Taping on Muscle Contractile Properties: Assessment Using Tensiomyography.

CONTEXT: Although functional effects of kinesio taping (KT) have been widely studied, its effects on contractile properties of the target muscle remain unclear. Tensiomyography is suitable for quantifying muscle stiffness and rate of force development upon imposed twitch contraction. OBJECTIVE: To test the hypothesis that KT has effects on contractile properties of targeted muscle using tensiomyography. DESIGN: Prospective cohort study. SETTINGS: Performance laboratory of a sports rehabilitation center. PARTICIPANTS: A total of 11 healthy volunteers. INTERVENTIONS: Tensiomyography measurements before KT facilitation technique applied (pre-KT), 45 minutes, and 24 hours after KT (post-KT1 and post-KT2, respectively) without removing the tape. MAIN OUTCOME MEASURES: Maximal radial displacement, contraction time, delay time, sustain time, relaxation time, and velocity of contraction. RESULTS: Significant effects were shown for maximal radial displacement (P = .004), contraction time (P = .013), relaxation time (P = .035), and velocity of contraction (P = .0033), but not for delay time (P = .060) and sustain time (P = .078). Post hoc testing indicated a significant decrease in maximal radial displacement for post-KT1 only (from 6.33 [1.46] to 4.87 [2.14] mm), and a significant increase in contraction time for both post-KT1 and post-KT2 (from 30.87 [11.39] to 39.71 [13.49] ms, and 37.41 [14.73] ms, respectively). Post hoc testing also showed a significant decrease in relaxation time for post-KT2 (from 65.97 [53.43] to 47.45 [38.12] ms), and a significant decrease in velocity of contraction for both post-KT1 and post-KT2 (from 0.22 [0.08] to 0.15 [0.09] mm/s, and 0.16 [0.07] mm/s), respectively. CONCLUSION: The findings indicate that KT leads to an increased muscle stiffness and a reduced muscle rate of force production despite the facilitation technique applied.

Bilioenteric Reconstruction Techniques in Pediatric Living Donor Liver Transplantation.

Biliary complications (BCs) are still a major cause of morbidity following liver transplantation despite the advancements in the surgical technique. Although Roux-en-Y (RY) hepaticojejunostomy has been the standard technique for years in pediatric patients, there is a limited number of reports on the feasibility of duct-to-duct (DD) anastomosis, and those reports have controversial outcomes. With the largest number of patients ever reported on the topic, this study aims to discuss the feasibility of the DD biliary reconstruction technique in pediatric living donor liver transplantation (LDLT). After the exclusion of the patients with biliary atresia, patients who received either deceased donor or right lobe grafts, and retransplantation patients, data from 154 pediatric LDLTs were retrospectively analyzed. Patients were grouped according to the applied biliary reconstruction technique, and the groups were compared using BCs as the outcome. The overall BC rate was 13% (n = 20), and the groups showed no significant difference (P = 0.6). Stricture was more frequent in the DD reconstruction group; however, this was not statistically significant (P = 0.6). The rate of bile leak was also similar in both groups (P = 0.6). The results show that the DD reconstruction technique can achieve similar outcomes when compared with RY anastomosis. Because DD reconstruction is a more physiological way of establishing bilioenteric integrity, it can safely be applied.

Live donor liver transplantation for type 1 tyrosinemia: An analysis of 15 patients.

Type 1 tyrosinemia is a rare metabolic disorder of the tyrosine degradation pathway. Due to the rarity of the disease, the best evidence literature offers is limited to guidelines based on expert opinions and optimal treatment is still a debate. LT serves as a definitive treatment of the defective metabolic pathway in the liver along with other serious disease manifestations such as LF and HCC. Nitisinone is a relatively new agent that is currently recommended for the medical management of the disease. Its mechanism of action is well understood, and efficacy is well established when started presymptomatically. This study aims to evaluate outcomes of 15 patients with type 1 tyrosinemia who underwent LT in nitisinone era and discuss its effect on prevention of HCC. A LT database of 1037 patients was reviewed. Data from 15 patients with type 1 tyrosinemia were retrospectively analyzed. All the patients except one were treated with nitisinone prior to LT. Most common indications for LT were LF and suspicious nodules. Seven patients had HCC. Mortality rate was 20% (n = 3). Nitisinone treatment has opened new horizons in the management of type 1 tyrosinemia, but LT still remains the only option for the patients developing LF and in the event of HCC. Neonatal screening programs utilizing blood succinyl acetone as the marker should be encouraged especially in the countries, such as Turkey, with high prevalence of consanguineous marriages.

Fascial nomenclature: Update on related consensus process.

The term fascia is increasingly used not only by anatomists but also by other professionals and authors in different health-oriented fields. This goes along with an inconsistent usage of the term, in which many different tissues are included by different authors causing an increasing amount of confusion. The Fascia Research Society acted to address this issue by establishing a Fascia Nomenclature Committee (FNC) with the purpose of clarifying the terminology relating to fascia. This committee conducted an elaborate Delphi process to foster a structured consensus debate among different experts in the field. This process led to two distinct terminology recommendations from the FNC, defining the terms "a fascia" and "the fascial system." This article reports on the process behind this proposed terminology as well as the implications for inclusion and exclusion of different tissue types to these definitions. Clin. Anat. 32:929-933, 2019. © 2019 The Authors. Clinical Anatomy published by Wiley Periodicals, Inc. on behalf of American Association of Clinical Anatomists.

Passive stiffness of monoarticular lower leg muscles is influenced by knee joint angle.

PURPOSE: While several studies demonstrated the occurrence of intermuscular mechanical interactions, the physiological significance of these interactions remains a matter of debate. The purpose of this study was to quantify the localized changes in the shear modulus of the gastrocnemius lateralis (GL), monoarticular dorsi- and plantar-flexor muscles induced by a change in knee angle. METHOD: Participants underwent slow passive ankle rotations at the following two knee positions: knee flexed at 90° and knee fully extended. Ultrasound shear wave elastography was used to assess the muscle shear modulus of the GL, soleus [both proximally (SOL-proximal) and distally (SOL distal)], peroneus longus (PERL), and tibialis anterior (TA). This was performed during two experimental sessions (experiment I: n = 11; experiment II: n = 10). The shear modulus of each muscle was compared between the two knee positions. RESULTS: The shear modulus was significantly higher when the knee was fully extended than when the knee was flexed (P < 0.001) for the GL (averaged increase on the whole range of motion: + 5.8 ± 1.3 kPa), SOL distal (+ 4.5 ± 1.5 kPa), PERL (+ 1.1 ± 0.7 kPa), and TA (+ 1.6 ± 1.0 kPa). In contrast, a lower SOL-proximal shear modulus (P < 0.001, - 5.9 ± 1.0 kPa) was observed. CONCLUSION: As the muscle shear modulus is linearly related to passive muscle force, these results provide evidence of a non-negligible intermuscular mechanical interaction between the human lower leg muscles during passive ankle rotations. The role of these interactions in the production of coordinated movements requires further investigation.

Behçet Disease serum is immunoreactive to neurofilament medium which share common epitopes to bacterial HSP-65, a putative trigger.

Autoimmune and dysimmune inflammatory mechanisms on a genetically susceptible background are implicated in the etiology of Behçet's Disease (BD). Heat-shock protein-65 (HSP-65) derived from Streptococcus sanguinis was proposed as a triggering factor based on its homology with human HSP-60. However, none of the autoantigens identified so far in sera from BD share common epitopes with bacterial HSP-65 or has a high prevalence. Here, we report that sera from BD patients are immunoreactive against filamentous neuronal processes in the mouse brain, retina and scrotal skin in great majority of patients. By using matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS) and peptide mass fingerprinting, Western blotting and peptide blocking experiments, we have identified neurofilament medium (NF-M) as the probable antigen for the serologic response observed. Clustal Omega analyses detected significant structural homology between the human NF-M and bacterial HSP-65 corresponding to amino acids 111-126, 213-232 and 304-363 of mycobacterial HSP-65, which were previously identified to induce proliferation of lymphocytes obtained from BD patients. We also found that sera immunoreactive against NF-M cross-reacted with bacterial HSP-65. These findings suggest that NF-M may be involved in autoimmunity in BD due to its molecular mimicry with bacterial HSP-65.

Intact muscle compartment exposed to botulinum toxin type A shows compromised intermuscular mechanical interaction.

INTRODUCTION: We tested the hypothesis that BTX-A diminishes epimuscular myofascial force transmission (EMFT) within an intact muscle compartment. METHODS: The tibialis anterior (TA) and extensor hallucis longus (EHL) muscles were kept at constant length, whereas the position of the extensor digitorum longus (EDL) muscle was changed exclusively. Two groups of Wistar rats were tested: a control group (no BTX-A injected) and a BTX group (0.1 unit of BTX-A injected into the mid-belly of TA). RESULTS: In controls, distally altered EDL position affected EDL distal and proximal forces and proximodistal force differences, indicating substantial EMFT. In the BTX group, EDL forces measured at the most proximal position did not change significantly with altered muscle position, and EDL proximodistal force differences became minimized. CONCLUSIONS: Use of BTX-A diminishes EMFT. It may be relevant clinically that BTX-A compromises intermuscular mechanical interaction, as recent studies have shown that such an interaction plays a role in the abnormal mechanics of spastic muscle.

The role of botulinum toxin type a-induced motor endplates after peripheral nerve repair.

INTRODUCTION: The aim of this study was to test the hypothesis that the increased number of new motor endplates induced by botulinum toxin type A (BTX-A) injection before nerve injury would be reinnervated after nerve repair, resulting in greater force generation. METHODS: Thirty male Wistar rats were divided randomly into 3 groups: (1) controls; (2) a group with saline solution injection; and (3) a group with BTX-A injection into gastrocnemius muscle (BTX group). Thirty-six days after the injections the left sciatic nerve was divided and coapted in all groups. Eight weeks later, muscle forces were measured, and histological samples were collected. RESULTS: No differences in the number of innervated endplates were found between the groups, but the number of denervated endplates was higher in the BTX group, as was the muscle tissue degeneration score. The BTX group showed distal muscle force measurements of up to 25.8% less compared with the control group. CONCLUSION: Although BTX-A injection increases the number of motor endplates, they are not functional.

Describing and classifying urban sound environments with a relevant set of physical indicators.

Categorization is a powerful method for describing urban sound environments. However, it has only been applied, until now, to discrete noise data collection, whereas sound environments vary continuously both in space and time. Therefore, a procedure is developed in this paper for describing the variations of urban sound environments. The procedure consists of mobile measurements, followed by a statistical clustering analysis that selects relevant noise indicators and classifies sound environments. Analysis are based on a 3 days + 1 night survey where geo-referenced noise measurements were collected over 19 1-h soundwalk periods in a district of Marseille, France. The clustering analysis showed that a limited subset of indicators is sufficient to discriminate sound environments. The three indicators that emerged from the clustering, namely, the Leq, A, the standard deviation σL eq, A, and the sound gravity spectrum SGC[50 Hz-10 kHz], are consistent with previous studies on sound environment classification. Moreover, the procedure proposed enables the description of the sound environment, which is classified into homogenous sound environment classes by means of the selected indicators. Thus, the procedure can be adapted to any urban environment, and can, for instance, favorably enhance perceptive studies by delimiting precisely the spatial extent of each typical sound environment.

Effects of botulinum toxin type A on non-injected bi-articular muscle include a narrower length range of force exertion and increased passive force.

INTRODUCTION: The goal of this study was to test the hypothesis that botulinum toxin type A (BTX-A) injection in rat tibialis anterior (TA) muscle affects the mechanics of its bi-articular synergist, both actively and passively. METHODS: Two groups of Wistar rats were tested: control (no BTX-A) and BTX (0.1 U of BTX-A) animals were injected exclusively to the mid-belly of TA. Extensor digitorum longus (EDL) muscle isometric forces were measured after proximal and distal lengthening. RESULTS: Five days after injection, BTX-A administration changed EDL mechanics: (1) active forces decreased (proximal muscle length dependently); (2) length range of active force exertion decreased both proximally and distally; and (3) passive muscle forces increased. CONCLUSIONS: Effects of BTX-A appear to not be limited to decreased active muscle tone, but may cause also a narrower active range of movement and increased passive resistance. Through spread of BTX-A to a bi-articular muscle, such effects are plausible for both joints spanned.

Magnetic resonance imaging assessment of mechanical interactions between human lower leg muscles in vivo.

Evidence on epimuscular myofascial force transmission (EMFT) was shown for undissected muscle in situ. We hypothesize that global length changes of gastrocnemius muscle-tendon complex in vivo will cause sizable and heterogeneous local strains within all muscles of the human lower leg. Our goal is to test this hypothesis. A method was developed and validated using high-resolution 3D magnetic resonance image sets and Demons nonrigid registration algorithm for performing large deformation analyses. Calculation of strain tensors per voxel in human muscles in vivo allowed quantifying local heterogeneous tissue deformations and volume changes. After hip and knee movement (Δ knee angle ≈ 25 deg) but without any ankle movement, local lengthening within m. gastrocnemius was shown to occur simultaneously with local shortening (maximally by +34.2% and -32.6%, respectively) at different locations. Moreover, similar local strains occur also within other muscles, despite being kept at constant muscle-tendon complex length. This is shown for synergistic m. soleus and deep flexors, as well as for antagonistic anterior crural and peroneal muscle groups: minimum peak lengthening and shortening equaled 23.3% and 25.54%, respectively despite global isometric conditions. These findings confirm our hypothesis and show that in vivo, muscles are in principle not independent mechanically.

The relationship of insulin resistance and metabolic syndrome with known breast cancer prognostic factors in postmenopausal breast cancer patients.

PURPOSE: The aim of this study was to investigate the effect of metabolic syndrome and insulin resistance at the time of diagnosis on the known prognostic factors of breast cancer in postmenopausal breast cancer patients. METHODS: The study included 71 patients with a recent diagnosis of postmenopausal breast cancer, admitted at the Medical Oncology outpatient clinic of the Izmir Ataturk Training and Research Hospital between June 2010 and June 2011. We determined whether the patients had metabolic syndrome and insulin resistance at diagnosis, and recorded known prognostic factors, such as tumor size, axillary lymph node involvement, presence of distant metastasis, tumor grade, estrogen receptor (ER), progesterone receptor (PR), and CerbB-2 status. RESULTS: Among 71 patients, 25 (35%) had metabolic syndrome at the time of diagnosis, and 33 (46%) had insulin resistance with Homeostasis Model of Assessment-Insulin Resistance (HOMA-IR)>2.7. No statistically significant difference was found in the prognostic values of breast cancer, i.e. tumor size, axillary lymph node involvement, distant metastasis, tumor grade, ER, PR, and CerbB-2 status between the patients with and without metabolic syndrome. There was no statistically significant difference in the prognostic factors of breast cancer at the time of diagnosis between 33 patients with insulin resistance and 38 patients without insulin resistance. CONCLUSION: Several previous studies showed a negative relationship between metabolic syndrome and insulin resistance and prognostic factors of breast cancer in postmenopausal breast cancer patients. However, our study failed to show such a relationship. The relationship between metabolic syndrome and insulin resistance and postmenopausal breast cancer was not well demonstrated due to the small number of patients, unknown duration of the metabolic syndrome and insulin resistance, and shorter follow-up period. Further studies are required to demonstrate the effect of metabolic syndrome and insulin resistance on the prognosis of breast cancer, including larger number of patients and longer follow-up periods.

The association of hematologic parameters on the prognosis of patients with metastatic renal cell carcinoma.

PURPOSE: Metastatic renal cell carcinoma (mRCC) bears a poor prognosis. We investigated the prognostic significance of some hematologic parameters of patients with mRCC. METHODS: We retrospectively reviewed the records of 53 patients with mRCC . The mean follow up time was 34 months (range 5-142).We assessed the prognostic value of hematologic parameters (leukocytes ,neutrophils, lymphocytes, platelets, neutrophil to lymphocyte ratio/NLR, platelet to lymphocyte ratio/PLR), and other clinical parameters with univariate and multivariate analysis. RESULTS: Memorial Sloan-Kettering Cancer Center (MSKCC) risk group , lung metastases, sunitinib treatment, lymphocyte count, NLR, and anemia significantly correlated with median overall survival (OS) on univariate analysis. The median OS in patients with a NLR < 3.4 was 32.2 months , significantly higher than the 13.9 months in patients with a ratio ≥ 3.4 (p = 0.006). Multivariate analysis revealed that MSKCC risk group and the NLR were independent predictors of OS. CONCLUSION: Hematologic parameters may be associated with OS in mRCC. However, further studies are needed to establish their routine use.

Evaluation of medication adherence of glaucoma patients during the COVID-19 pandemic.

PURPOSE: To evaluate changes in the medication adherence of glaucoma patients during the COVID-19 pandemic and the factors influencing medication adherence. MATERIALS AND METHODS: This cross-sectional study included a total of 197 glaucoma patients who were followed for at least six months in the Glaucoma Unit of the Ophthalmology Department of Inonu University, Faculty of Medicine. Patients were given a 28-item questionnaire, including the eight-item Morisky Medication Adherence Questionnaire, to evaluate medication adherence. Demographic and clinical data were recorded. P<0.05 was considered statistically significant. RESULTS: Interruption of glaucoma clinic visits during the pandemic was reported by 82 (41.6%) patients. Nonadherence was reported by 56 patients (28.4%) (95% confidence interval: 22.1-34.7). For these patients, the most common reasons for nonadherence were forgetfulness (50%), the inability to receive a prescription for the drug (10.7%) and being busy (10.7%). Factors influencing nonadherence were determined to be younger age, female gender, interruption of glaucoma clinic visits and high-income levels (P˂0.05). CONCLUSION: Interruption of glaucoma clinic visits during the COVID-19 pandemic and the resulting inability to have medications prescribed resulted in patient nonadherence with medication use.

Effects of elastic therapeutic taping on along-muscle fascicle local length changes: Magnetic resonance and diffusion tensor imaging based assessment.

Elastic therapeutic taping is utilized for prevention and treatment of various neuromusculoskeletal disorders and sports injuries. Kinesio taping (KT) is a popular version of this practice. Despite being widely used to improve muscular function, an understanding of KT effects on muscular mechanics are lacking. Considering the continuity of the fascial system and its mechanical interaction with muscle fascicles intramuscularly, the aim was to test the following hypothesis: mechanical loading induced on the skin by KT leads to along-muscle fascicle local length changes and shear strains in the targeted muscle. Magnetic resonance imaging (MRI)-based local tissue deformation analyses and diffusion tensor imaging (DTI)-based fiber tracking analyzes were combined. Anatomical MRI and DTI were acquired for 5 healthy female volunteers in 3 conditions: (1) without tape, (2) following sham application, and (3) after KT application. Local length changes and shear strains were calculated using image registration between conditions (1-2) and (2-3). Non-parametric Wilcoxon signed-rank test was performed to compare the two conditions. Data pooled from all subjects show that KT-imposed along-muscle fascicle lengthening (mean ± SD 0.026 ± 0.020), shortening (0.032 ± 0.027) and shearing (0.087 ± 0.049) occur and are significantly higher than those caused by sham application (0.012 ± 0.010; 0.013 ± 0.015; 0.029 ± 0.021, respectively) (p < 0.001). KT induced along-muscle fascicle length changes locally show heterogeneity. Our findings indicate that KT affects both along-muscle fascicle length changes and shear strains. This can be explained by KT imposed myofascial loads over the skin being transmitted via the fascial system, non-uniformly manipulating the mechanical equilibrium locally at different parts along the muscle fascicles.

In-vivo along muscle fascicle strain heterogeneity is not affected by image registration parameters: Robustness testing of combined magnetic resonance-diffusion tensor imaging method.

Coupled with diffusion tractography, non-rigid registration of high-resolution anatomical MR images allows the calculation of local strains along human skeletal muscle fascicles in-vivo. A reference study (passively imposed lengthening of gastrocnemius medial muscle) reported local shortening and lengthening occurring along the same muscle fascicles. However, the robustness of strain amplitudes and distribution patterns should be studied, as the heterogeneity of local length changes has major implications for muscle function. Using a previous image set of human medial gastrocnemius (GM) we aimed at testing: (1) the consistency of our MRI-DTI analysis workflow against changes made to the software environments, (2) the hypothesis that non-rigid demons algorithm tuning parameters (16 paired combinations were tested) are not a significant determinant of muscle fiber direction strain heterogeneity caused by passive knee extension. A profoundly altered analysis workflow did reproduce the original results well, showing a general pattern of proximally lengthened and distally shortened muscle fascicles (strain amplitude range: 21%-67%). Hierarchical shift function analyses and pairwise comparison of strain distributions between 10 equal parts of the tracked GM fascicles confirmed the hypothesis showing no significant effects of tuning parameters determining the in-vivo deformation field inhomogeneity. The findings show the robustness of the MRI-DTI method, and confirming the hypothesis, also the consistency of along muscle fascicle strain heterogeneity patterns against parameter selection. However, the strain amplitudes do vary with parameter choices. New studies are indicated to determine optimal tuning parameters to achieve accurate strain amplitudes compared to exact strains.