Healthy Adult Controls Research Articles

IMMU-46. MONITORING T-CELL DYNAMICS AND CHECKPOINT EXPRESSION IN THE IMMUNE MICROENVIRONMENT OF PEDIATRIC HIGH GRADE BRAIN TUMORS: THE MIMIC PROGRAM

Abstract BACKGROUND Pediatric high grade brain tumors (pHGBT) pose a significant challenge due to limited treatment options and poor prognosis. Understanding the immune landscape, in particular T cell profiles and functionality, is of high interest as numerous immunotherapeutic approaches focus on or utilize T-cells. This study aimed to characterize T-cell phenotypes and their potential for immunotherapeutic interventions in tumor fragments (obtained from cavitron ultrasonic surgical aspiration (CUSA)) and tumor tissue (biopsy), in comparison with peripheral blood, collected from pHGBT patients. METHODS Spectral flow cytometry was employed to characterize the presence of T-cell subsets and their functional phenotype. RESULTS T cells were detected in all materials with similar CD8/CD4 ratios. At diagnosis, T-cells in the tumor microenvironment exhibited significantly increased expression of PD1, LAG3 and TIM3 checkpoint receptors compared to peripheral blood. This difference was more pronounced in biopsy samples compared to CUSA fragments. T-cell composition and phenotype in peripheral blood were similar to healthy adult controls at diagnosis. However, during therapy, the T-cell compartment showed a shift towards a higher CD8/CD4 ratio, accompanied by increased activation markers (CD25, CD69) and checkpoint receptor expression . Notably, CD8+ T-cells in both peripheral blood and tumor microenvironment showed high production of Granzyme B. CONCLUSION Our findings demonstrate that despite the high expression of checkpoint receptors on CD8+ T-cells, these cells retain the capability to produce significant amounts of Granzyme B, indicating their potential for cytotoxic activity. Furthermore, variations in checkpoint receptor expression on T-cells were observed when comparing CUSA fragments to biopsy material. Functional assays will further validate these findings and elucidate the functional status of these cells. Understanding the function of T-cells within the tumor microenvironment is essential for designing novel combination therapies to enhance T-cell cytotoxicity against pHGBT.

Abstract 14833: Association of RV Hemodynamics From 4D Flow CMR With RV Restrictive Physiology From Pediatric to Adult Repaired Tetralogy of Fallot

Introduction: Right ventricular restrictive physiology (RVRP) is common after repair of Tetralogy of Fallot (rTOF), especially with residual pulmonary regurgitation. Four-dimensional (4D) flow cardiovascular magnetic resonance (CMR) quantitates biventricular blood flow components and kinetic energy (KE) parameters, but their association with RVRP is unclear. We aimed to study 4D flow CMR parameters in adult and pediatric rTOF patients vs healthy controls and associations of 4D flow parameters with RVRP in rTOF. Methods: 103 rTOF patients (40 pediatric, 63 adult; age 24±17 years; 53M:50F) and 171 healthy controls (14 pediatric, 157 adult; age 40±15 years; 97M:74F) were prospectively recruited from centers in Singapore (2) and China (1) to undergo 4D flow CMR on multi-vendor scanners. LV and RV flow components—direct flow, retained inflow, delayed ejection flow and residual volume—and KE parameters (normalized to end-diastolic volume, KEi EDV )—global, peak systolic, average systolic, average diastolic, peak E-wave and peak A-wave—were analyzed. KE discordance was calculated as the ratio of RV and LV average systolic KEi EDV . Results: See Table. RVRP was identified in 70 patients (68%). rTOF with RVRP exhibited higher median RV stroke volume index, RV ejection fraction, RV direct flow, RV peak E-wave and E-wave/A-wave, and lower RV residual volume compared to rTOF without RVRP (all P<0.05). Using binary multivariable logistic regression analysis—and after adjustment for age, pulmonary regurtitation, LV and RV ejection fraction, RV direct flow was found to be independently associated with RVRP in rTOF (P<0.001). Conclusion: Patients with RVRP had higher RV direct flow, RVEF and RV peak E-wave energy, indicating the potential compensatory adaptations in patients with RVRP.

Attentional Control in Subjective Cognitive Decline.

Attention is an essential cognitive ability that is necessary in other cognitive processes. Only few studies have focused on decline in specific functions of attention in older adults with cognitive decline. No research explores the difference in the proactive and reactive mode of control between the healthy control (HC) and older adults with subjective cognitive decline (SCD). The current work investigated whether there was any decline in alerting, orienting, and executive control in SCD. Particularly, the present study further explored the impairment of the proactive and reactive control in SCD. We recruited 25 HC and 26 SCD. All participants first finished a set of neuropsychological assessments. They then completed an Attention Network Test for measuring the alerting, orienting, and executive control, the List-wide and the Item-specific Proportion Congruency Effect task for measuring the proactive and reactive mode of control, respectively. No difference was found in alerting, orienting, and executive control measured by the ANT between SCD and HC. The results also indicated no difference in the reactive control between SCD and HC. However, older adults with SCD performed worse in the proactive control as compared to HC. Older adults with SCD showed an impairment in the proactive control. The current findings help us better understand objective decline in cognitive domains other than memory and shed light on early assessment and prevention of AD.

Characterization of Gla proteoforms and non-Gla peptides of gamma carboxylated proteins: Application to quantification of prothrombin proteoforms in human plasma

Gamma (γ) carboxylation is an essential post-translational modification in vitamin K-dependent proteins (VKDPs), involved in maintaining critical biological homeostasis. Alterations in the abundance or activity of these proteins have pharmacological and pathological consequences. Importantly, low levels of fully γ-carboxylated clotting factors increase plasma des-γ-carboxy precursors resulting in little or no biological activity. Therefore, it is important to characterize the levels of γ-carboxylation that reflect the active state of these proteins. The conventional enzyme-linked immunosorbent assay for protein induced by vitamin K absence or antagonist II (PIVKA-II) quantification uses an antibody that is not applicable to distinguish different γ-carboxylation states. Liquid chromatography-mass spectrometry (LC-MS) approaches have been utilized to distinguish different γ-carboxylated proteoforms, however, these attempts were impeded by poor sensitivity due to spontaneous neutral loss of CO2 and simultaneous cleavage of the backbone bond in the collision cell. In this study, we utilized an alkaline mobile phase in combination with polarity switching (positive and negative ionization modes) to simultaneously identify and quantify γ-carboxylated VKDPs. The method was applied to compare Gla proteomics of prothrombin (FII) in 10 μL plasma samples of healthy control and warfarin-treated adults. We also identified surrogate non-Gla peptides for seven other VKDPs to quantify total (active plus inactive) protein levels. The total protein approach (TPA) was used to quantify absolute levels of the VKDPs in human plasma.

Mediolateral foot placement control can be trained: Older adults learn to walk more stable, when ankle moments are constrained.

Falls are a problem, especially for older adults. Placing our feet accurately relative to the center-of-mass helps us to prevent falling during gait. The degree of foot placement control with respect to the center-of mass kinematic state is decreased in older as compared to young adults. Here, we attempted to train mediolateral foot placement control in healthy older adults. Ten older adults trained by walking on shoes with a narrow ridge underneath (LesSchuh), restricting mediolateral center-of-pressure shifts. As a training effect, we expected improved foot placement control during normal walking. A training session consisted of a normal walking condition, followed by a training condition on LesSchuh and finally an after-effect condition. Participants performed six of such training sessions, spread across three weeks. As a control, before the first training session, we included two similar sessions, but on normal shoes only. We evaluated whether a training effect was observed across sessions and weeks in a repeated-measures design. Whilst walking with LesSchuh, the magnitude of foot placement error reduced half-a-millimeter between sessions within a week (cohen's d = 0.394). As a training effect in normal walking, the magnitude of foot placement errors was significantly lower compared to the control week, by one millimeter in weeks 2 (cohen's d = 0.686) and 3 (cohen's d = 0.780) and by two millimeters in week 4 (cohen's d = 0.875). Local dynamic stability of normal walking also improved significantly. More precise foot placement may thus have led to improved stability. It remains to be determined whether the training effects were the result of walking on LesSchuh or from repeated treadmill walking itself. Moreover, enhancement of mechanisms beyond the scope of our outcome measures may have improved stability. At the retention test, gait stability returned to similar levels as in the control week. Yet, a reduction in foot placement error persisted.

GLUT-1 changes in paediatric Huntington disease brain cortex and fibroblasts: an observational case-control study

Paediatric Huntington disease with highly expanded mutations (HE-PHD; >80 CAG repeats) presents atypically, compared to adult-onset Huntington disease (AOHD), with neurodevelopmental delay, epilepsy, abnormal brain glucose metabolism, early striatal damage, and reduced lifespan. Since genetic GLUT-1 deficiency syndrome shows a symptom spectrum similar to HE-PHD, we investigated the potential role of the two main glucose transporters, GLUT-1 and GLUT-3, in HE-PHD. We compared GLUT-1 and GLUT-3 protein expression in HE-PHD, juvenile-onset (JOHD), and AOHD brains (n=2; n=3; n=6) and periphery (n=3; n=2; n=2) versus healthy adult controls (n=6; n=6). We also investigated mitochondrial complexes and hexokinase-II protein expression. GLUT-1 and GLUT-3 expression were significantly lower in HE-PHD frontal cortex (p=0.009, 95% [CI 13.4, 14.7]; p=0.017, 95% [CI 14.2, 14.5]) versus controls. In fibroblasts, GLUT-1 and GLUT-3 expression were lower compared to controls (p<0.0001, 95% [CI 0.91, 1.09]; p=0.046, 95% [CI 0.93, 1.07]). In the frontal cortex, this occurred without evidence of extensive neuronal degeneration. Patients with HE-PHD had deregulated mitochondrial complex expression, particularly complexes II-III, levels of which were lower in frontal cortex versus controls (p=0.027, 95% [CI 17.1, 17.6]; p=0.002, 95% CI [16.6, 16.9]) and patients with AOHD (p=0.052, 95% [CI 17.0, 17.6]; p=0.002, 95% [CI 16.6, 16.7]). Hexokinase-II expression was also lower in HE-PHD frontal cortex and striatum versus controls (p=0.010, 95% [CI 17.8, 18.2]; p=0.045, 95% [CI 18.6, 18.7]) and in frontal cortex versus patients with AOHD (p=0.013, 95% [CI 17.7, 18.1]). Expression JOHD levels were consistently different to those of HE-PHD but similar to those of AOHD. Our data suggest a dysfunctional hypometabolic state occurring specifically in paediatric Huntington disease brains. '5×1000' Personal Income Tax donation to LIRH Foundation; Italian Ministry of HealthRC2301MH04 and RF-2016-02364123 to CSS.

Sonographic evaluation of common carotid artery intima-media thickness in adults with coexisting type two (2) diabetes and hypertension: A case-controlled study

Diabetes mellitus (DM) and hypertension (HTN) are the two leading risk factors for atherosclerosis. Carotid intima-media thickness (CIMT) is considered a reliable surrogate measure for detecting subclinical atherosclerosis including its progression and regression. The study aim at evaluating the intima-media thickness of the common carotid artery in adults with coexisting diabetes mellitus and hypertension (DM+HTN) and compare with healthy controls.. A prospective case control study involving 115 adult subjects with coexisting DM+HTN and 144 healthy adult controls was carried out. Common carotid intima-media thickness (CCIMT) was measured at point approximately 1 cm distal to the right and left carotid bulb using the 7-12 MHz, multi-frequency linear array transducer of high resolution, touch Venue 50 ultrasound machine equipped with electronic callipers. The overall mean CCIMT was 1.13±0.40 mm and 0.82±0.23 mm for subjects with coexisting DM and HTN; and healthy controls, respectively. There was a statistically significant difference in CIMT of subjects with DM+HTN compared to healthy controls (p=0.000 each for both males and females, respectively). There were positive correlations between CCIMT and age in subjects with DM+HTN (r=0.260 and r=0.424 for males and females respectively); with healthy controls (r=0.619 and 0.626 for males and females, respectively). There was a statistically significant difference in CCIMT between subjects with DM+HTN and healthy controls. This implies that the increase in CCIMT is associated with both DM and HTN. In addition, age has a significant positive relationship with CCIMT in both subjects with DM+HTN and healthy controls.

No evidence for systemic low-grade inflammation in adult patients with early-treated phenylketonuria: The INGRAPH study.

The question of an increased cardiovascular risk has been recently raised in adults with phenylketonuria (PKU). As low-grade systemic inflammation increases cardiovascular risk, the INGRAPH study aimed to evaluate low-grade inflammation in adult PKU patients compared to healthy controls and to determine the potential influence of Phe-controlled diet on inflammation. Twenty early-treated adult PKU patients, including a subgroup of 15 classical PKU patients, and 20 healthy volunteers were included. PKU patients and healthy subjects were matched on age, sex and body mass index class. Plasma concentrations of CRP, IFNg, IL1a, IL1b, IL2, IL6, IL10, and TNFα were measured in PKU patients and compared to controls. Plasma CRP was not different in the PKU group as compared to controls. No significant differences were observed between the two groups concerning plasma cytokines concentrations. Plasma CRP and cytokine profile were not different between "on diet" and "off diet" PKU patients. All these results were similar considering only the classical PKU subgroup. No differences were shown in plasma CRP and pro-inflammatory cytokines between adult PKU patients and healthy controls. Further studies are needed, including more patients and extensive characterization of systemic low-grade inflammation, as cardiovascular risk appears to be a new concern in adult PKU population.

Evaluation of Serum Diamine Oxidase as a Diagnostic Test for Histamine Intolerance.

Histamine intolerance (HIT) is a clinical condition caused by decreased intestinal degradation of ingested histamine, primarily due to reduced enzyme diamine oxidase (DAO) activity, leading to histamine accumulation and causing various clinical manifestations. The measurement of serum DAO is commonly used as the main diagnostic test for HIT, although its diagnostic use is still uncertain. In this retrospective study, we aimed to assess the validity of DAO determination in patients with clinically suspected HIT. We measured DAO levels in 249 patients with suspected HIT and 50 healthy adult controls without HIT-related problems. Based on five clinical criteria, we divided patients into two groups: high (all five inclusion criteria; 41 patients) and low probability of HIT (≤4 inclusion criteria; 208 patients). Patients with a "high probability of HIT" had the lowest DAO (median: 8 U/mL, IQR: 6-10) in comparison to patients with a "low probability of HIT (median: 10 U/mL, IQR: 7-16, p = 0.0006) and healthy controls (median: 18 U/mL, IQR: 14-22, p < 0.0001). The specificity and sensitivity for DAO levels < 3/< 10 U/mL (manufacturer's set cut-off) to discriminate between patients with ''high probability of HIT'' and healthy controls were 100%/92% and 2%/71%. On the other hand, the specificity and sensitivity to discriminate between patients with ''high probability of HIT'' and ''low probability of HIT'' were 97%/61% and 2%/71%, respectively. Serum DAO determination represents an additional asset to the diagnosis of HIT based on clinical evaluation and assessment, but the diagnosis should not solely rely on DAO measurements.

Medication only improves limb movements while deep brain stimulation improves eye and limb movements during visually-guided reaching in Parkinson's disease.

Antiparkinson medication and subthalamic nucleus deep brain stimulation (STN-DBS), two common treatments of Parkinson's disease (PD), effectively improve skeletomotor movements. However, evidence suggests that these treatments may have differential effects on eye and limb movements, although both movement types are controlled through the parallel basal ganglia loops. Using a task that requires both eye and upper limb movements, we aimed to determine the effects of medication and STN-DBS on eye and upper limb movement performance. Participants performed a visually-guided reaching task. We collected eye and upper limb movement data from participants with PD who were tested both OFF and ON medication (n = 34) or both OFF and ON bilateral STN-DBS while OFF medication (n = 11). We also collected data from older adult healthy controls (n = 14). We found that medication increased saccade latency, while having no effect on reach reaction time (RT). Medication significantly decreased saccade peak velocity, while increasing reach peak velocity. We also found that bilateral STN-DBS significantly decreased saccade latency while having no effect on reach RT, and increased saccade and reach peak velocity. Finally, we found that there was a positive relationship between saccade latency and reach RT, which was unaffected by either treatment. These findings show that medication worsens saccade performance and benefits reaching performance, while STN-DBS benefits both saccade and reaching performance. We explore what the differential beneficial and detrimental effects on eye and limb movements suggest about the potential physiological changes occurring due to treatment.

Comparative Analysis of Red Blood Cells, White Blood Cells, Platelet Count, and Indices in Type 2 Diabetes Mellitus Patients and Normal Controls: Association and Clinical Implications.

Diabetes mellitus (DM) is a major health burden affecting 537 million adults worldwide, characterized by chronic metabolic disorder and various complications. This case control study aimed to assess the impact of type 2 diabetes mellitus (T2DM), including hyperglycemia levels, on hematological parameters and complete blood count (CBC) derived parameters. A total of 250 known diabetic patients from the Jazan Diabetic Center, Saudi Arabia, between January 2021 and December 2022, along with 175 healthy adult controls were recruited from Jazan Hospital's blood donation center. Demographic characteristics, medical histories, and relevant factors such as gender, age, BMI, treatment, disease duration, and comorbidities were collected with informed consent. The results of the red blood cell (RBC) count, RBC indices, and mean platelet volume showed significant differences between patients and controls, while the white cell (WBC) and platelet count were comparable between the two groups. CBC-derived parameters, especially neutrophil/lymphocyte ratio (NLR), and platelet/neutrophil ratio (PNR) exhibited significant differences. CBC and derived parameters serve as inexpensive tools for T2DM patients monitoring, indicating early blood cell alterations and potential development of anemia. Further studies are needed to explore their role in predicting T2DM pathogenesis and progression, aiming to reduce severe complications, mortality and morbidity.

The distribution of myeloid-derived suppressor cells subsets and up-regulation of programmed death-1/PD-L1 axis in peripheral blood of adult CAP patients.

Myeloid-derived suppressor cells (MDSCs) have been reported to expand and have a potent ability in the expansion of regulatory T cells in malignant and infectious disease. The current study was performed to investigate the role of MDSCs and possible immune mechanisms in dampening immune responses of community acquired pneumonia (CAP). This was a single-center cross-sectional study. The distribution of MDSCs subsets, the PD-1/PD-L1(L2) level of MDSCs subsets and Tregs in the peripheral blood of adult CAP patients and healthy control were measured by flow cytometry analysis. Peripheral blood mononuclear cells (PBMCs) from 63 adult CAP patients contained an elevated frequency of both G-MDSC (4.92±0.30 vs 2.25±0.21,p<0.0001) and M-MDSC (19.40±1.30 vs 9.64±0.57,p<0.001) compared to healthy controls. Treg in the peripheral blood of CAP patients exhibited increased expression of PD-1 and CTLA-4, accompanied by no difference of their frequency. Moreover, up-regulated expression of PD-L1 on MDSC subsets in the peripheral blood of CAP patients was also revealed. Of note, the frequency of circulating MDSCs subset displayed a positive correlation with neutrophil count percentage in blood in CAP patients. In summary, the significant expansion of circulating MDSCs subsets and the up-regulated expression of PD-1/PD-L1 level in CAP patients may suggest the possible involvement of PD-1/PD-L1axis in MDSCs mediated immune regulation on Treg at least partially in CAP patients.

Analysis of the serum bile acid profile to facilitate diagnosis and differential diagnosis of NA(+)-taurocholate cotransporting polypeptide deficiency

Objective: This study focuses on Na(+)-taurocholate cotransporting polypeptide (NTCP) deficiency to analyze and investigate the value of the serum bile acid profile for facilitating the diagnosis and differential diagnosis. Methods: Clinical data of 66 patients with cholestatic liver diseases (CLDs) diagnosed and treated in the Department of Pediatrics of the First Affiliated Hospital of Jinan University from early April 2015 to the end of December 2021 were collected, including 32 cases of NTCP deficiency (16 adults and 16 children), 16 cases of neonatal intrahepatic cholestasis caused by citrin deficiency (NICCD), 8 cases of Alagille syndrome, and 10 cases of biliary atresia. At the same time, adult and pediatric healthy control groups (15 cases each) were established. The serum bile acid components of the study subjects were qualitatively and quantitatively analyzed by ultra-high performance liquid chromatography-tandem mass spectrometry. The data were plotted and compared using statistical SPSS 19.0 and GraphPad Prism 5.0 software. The clinical and bile acid profiles of children with NTCP deficiency and corresponding healthy controls, as well as differences between NTCP deficiency and other CLDs, were compared using statistical methods such as t-tests, Wilcoxon rank sum tests, and Kruskal-Wallis H tests. Results: Compared with the healthy control, the levels of total conjugated bile acids, total primary bile acids, total secondary bile acids, glycocholic acid, taurocholic acid, and glycochenodeoxycholic acid were increased in NTCP deficiency patients (P < 0.05). Compared with adults with NTCP deficiency, the levels of total conjugated bile acids and total primary bile acids were significantly increased in children with NTCP deficiency (P < 0.05). The serum levels of taurochenodeoxycholic acid, glycolithocholate, taurohyocholate, and tauro-α-muricholic acid were significantly increased in children with NTCP deficiency, but the bile acid levels such as glycodeoxycholic acid, glycolithocholate, and lithocholic acid were decreased (P < 0.05). The serum levels of secondary bile acids such as lithocholic acid, deoxycholic acid, and hyodeoxycholic acid were significantly higher in children with NTCP deficiency than those in other CLD groups such as NICCD, Alagille syndrome, and biliary atresia (P < 0.05). Total primary bile acids/total secondary bile acids, total conjugated bile acids/total unconjugated bile acids, taurocholic acid, serum taurodeoxycholic acid, and glycodeoxycholic acid effectively distinguished children with NTCP deficiency from other non-NTCP deficiency CLDs. Conclusion: This study confirms that serum bile acid profile analysis has an important reference value for facilitating the diagnosis and differential diagnosis of NTCP deficiency. Furthermore, it deepens the scientific understanding of the changing characteristics of serum bile acid profiles in patients with CLDs such as NTCP deficiency, provides a metabolomic basis for in-depth understanding of its pathogenesis, and provides clues and ideas for subsequent in-depth research.

Immunological Characterization of Chronic Nonbacterial Osteomyelitis (CNO) in Adults: A Cross-Sectional Exploratory Study.

Chronic nonbacterial osteomyelitis (CNO) is a rare disease spectrum affecting children and adults. Adult CNO may occur as isolated bone inflammation, or with a broad range of extraskeletal features. CNO pathophysiology, including the key drivers of inflammation, remains largely unknown. For pediatric CNO, a role for pro-inflammatory cytokine dysregulation has been proposed, but studies in adults are scarce. We therefore provide immunological characterization of adult CNO. Cross-sectional study in our referral center including adult CNO patients (n = 172) and healthy controls (n = 65). Inflammation parameters and systemic inflammatory based scores(SIBS, including neutrophil/lymphocyte ratio [NLR] and systemic immune inflammation index [SII]) were compared between groups. Cytokine expression was explored with electrochemiluminescent immunoassays in 33 patients, eight healthy controls and 21 osteoporosis patients. Routine inflammation markers were higher in patients than in controls, but generally remained within reference range. Systemic inflammation was more pronounced in patients with additional vertebral involvement as compared to those osteitis in the anterior chest wall alone, in patients with comorbid pustulosis palmoplantaris or psoriasis, and in patients with strongly rather than moderately increased lesional uptake on nuclear imaging. SII was elevated in CNO patients too, but NLR was not. Cytokine expression was generally nondifferential between patients and both control groups, and patients displayed low absolute concentrations of pro-inflammatory cytokines. In this adult CNO cohort, systemic inflammation was generally subtle, but more pronounced in patients with vertebral lesions, associated skin disease, and strongly increased uptake on nuclear imaging. SII was increased in patients compared to healthy controls. Contrasting pediatric studies, we found no increased expression of the pro-inflammatory cytokines that have been proposed to drive the inflammatory cascade, like interleukin-6, -8, and -17 (IL-6, IL-8, and IL-17), and tumor necrosis α (TNF-α). Further studies are needed to evaluate the use of SII in diagnosis and monitoring of CNO, and elucidate the role of cytokine dysregulation in adult disease. © 2023 The Authors. JBMR Plus published by Wiley Periodicals LLC. on behalf of American Society for Bone and Mineral Research.

Temporal Dynamics of Serum Perforin and Granzymes in Three Different Clinical Stages of Virus-Induced Severe Fever with Thrombocytopenia Syndrome.

Virus-induced severe fever with thrombocytopenia syndrome (SFTS) induces a cell-mediated immune response that likely contributes to virus control in SFTS patients. To identify the temporal changes of the cell-mediated immune response, we investigated the changes in serum levels of perforin and granzymes at early periods after illness onset in SFTS patients. We analyzed 32 SFTS patients and compared the temporal patterns of serum perforin and granzyme A and B to that of 20 healthy control adults using the Mann-Whitney U test. Compared with healthy controls, the mean level of perforin was significantly reduced by 81% (P < 0.01) during the first week after illness onset, whereas granzyme B significantly increased by 4.6-fold (P = 0.02) in the first week after illness onset and decreased to normal afterward. During the study period, there was no significant difference in serum perforin and granzyme. These findings indicate that perforin and granzyme B in serum can be considered possible serologic markers that reflect the clinical stage of SFTS. Additional study is warranted for tracking circulating perforin and granzyme in different ages and for an extended period after illness onset.

Investigating the Association Between Muscular Ultrasonographic Alterations and Clinical Symptoms in Patients With Inflammatory Myopathy.

Muscle biopsy is commonly used to diagnose inflammatory myopathies. We evaluated the ability of muscle ultrasound, a non-invasive and simple tool, to distinguish between healthy subjects and patients with inflammatory myopathy. This study was conducted on 17 patients recently diagnosed with biopsy inflammatory myopathies (12 dermatomyositis, 5 polymyositis) compared with 17 age- and gender-matched healthy control adults. All patients underwent clinical assessments, including manual muscle testing, hand-held dynamometry, and muscle ultrasound evaluations, including thickness and echo intensity in predefined muscle groups. The disease duration was seven months (interquartile range: 3 to 11 months). Except for the biceps and gastrocnemius, patients' muscles had significantly higher echo intensity and lower thickness than the control group. The echo intensity sum-score manifested the highest area under the curve compared to the sum-scores of other variables (echo intensity vs manual muscle testing: Area under curves-difference=0.18, P<0.01; echo intensity vs dynamometry: Area under curves-difference=0.14, P=0.02; echo intensity vs thickness: Area under curves-differences-difference=0.25, P<0.01). The echo intensity of muscles differed significantly between healthy individuals and patients with inflammatory myopathies and may serve as a useful diagnostic biomarker.

Extrusion of the medial meniscus under a weight-loading condition in early knee osteoarthritis: an investigation using special upright magnetic resonance imaging

BackgroundWhether the medial meniscus morphology and movement occur under upright loading conditions in early knee osteoarthritis (OA) or medial meniscus posterior root tear (MMPRT) remains unknown. This study aimed to evaluate the medial and anteroposterior extrusion of the medial meniscus under unloaded and upright-loaded conditions in patients with early knee OA.MethodsTwelve patients with early knee OA and 18 healthy adult volunteers participated in this study. Magnetic resonance imaging using special equipment was performed with the participants in the unloaded and upright-loaded conditions. Medial, anterior, and posterior extrusions of the medial meniscus against the tibial edge were evaluated and compared between the early knee OA and healthy adult control groups. Additionally, 12 patients in the early knee OA group were divided into 2 subgroups based on whether MMPRT was observed, and the extrusion of the medial meniscus was compared.ResultsThe amount of medial extrusion of the medial meniscus in both the unloaded and upright-loaded conditions was significantly greater in the early knee OA group than in the control group (unloaded: 2.6 ± 1.0 mm vs 0.7 ± 0.5 mm; upright-loaded: 3.7 ± 0.9 mm vs 1.8 ± 0.8 mm). Similarly, the anterior and posterior extrusion of the medial meniscus in the upright-loaded condition was significantly larger in the early knee OA group (anterior: 4.6 ± 1.0 mm vs 3.7 ± 1.1 mm; posterior: -3.4 ± 1.1 mm vs -4.6 ± 1.6 mm). However, no difference was observed in meniscal extrusion between unloaded and upright-loaded conditions. The posterior extrusion of the medial meniscus in the upright-loaded condition was significantly greater in MMPRT cases than in non-MMPRT cases in the early knee OA group (MMPRT: -2.7 ± 1.1 mm; non-MMPRT -4.1 ± 1.5 mm).ConclusionsIn early knee OA, significantly large meniscal extrusions of the medial meniscus in both unloaded and upright-loaded conditions were found compared with healthy adults. Among patients with early knee OA, those with MMPRT showed a large posterior extrusion of the medial meniscus in the upright-loaded condition compared with those without MMPRT.Level of evidenceLevel IV.

Generation of two human iPSC lines from dermal fibroblasts of adult- and juvenile-onset Huntington's disease patients and two healthy donors

Huntington's disease (HD) is an autosomal dominant neurodegenerative disease caused by a mutation in the HTT gene. To generate human-induced pluripotent stem cells (hiPSCs), we used dermal fibroblasts from 1 healthy adult control (K-Pic2), 1 HD manifest patient (M-T2), 1 healthy juvenile control (jK-N1), and 1 juvenile HD patient (jHD-V1). HD stage of patients was assessed by neurological tests and donors were without comorbidities and were non-smokers. Characterization showed that the obtained hiPSCs have the same number of CAG repeats as the parental fibroblast lines, express pluripotency markers and have the ability to differentiate into all 3 germ layers.

Occipital transcranial direct current stimulation in episodic migraine patients: effect on cerebral perfusion

Cerebral blood flow differs between migraine patients and healthy controls during attack and the interictal period. This study compares the brain perfusion of episodic migraine patients and healthy controls and investigates the influence of anodal transcranial direct current stimulation (tDCS) over the occipital cortex. We included healthy adult controls and episodic migraineurs. After a 28-day baseline period and the baseline visit, migraine patients received daily active or sham anodal tDCS over the occipital lobe for 28 days. All participants underwent a MRI scan at baseline; migraineurs were also scanned shortly after the stimulation period and about five months later. At baseline, brain perfusion of migraine patients and controls differed in several areas; among the stimulated areas, perfusion was increased in the cuneus of healthy controls. At the first visit, the active tDCS group had an increased blood flow in regions processing visual stimuli and a decreased perfusion in other areas. Perfusion did not differ at the second follow-up visit. The lower perfusion level in migraineurs in the cuneus indicates a lower preactivation level. Anodal tDCS over the occipital cortex increases perfusion of several areas shortly after the stimulation period, but not 5 months later. An increase in the cortical preactivation level could mediate the transient reduction of the migraine frequency.Trial registration: NCT03237754 (registered at clincicaltrials.gov; full date of first trial registration: 03/08/2017).

Neuromelanin-sensitive MRI correlates of cognitive and motor function in Parkinson's disease with freezing of gait.

Substantia nigra pars compacta (SNc) and locus coeruleus (LC) are neuromelanin-rich nuclei implicated in diverse cognitive and motor processes in normal brain function and disease. However, their roles in aging and neurodegenerative disease mechanisms have remained unclear due to a lack of tools to study them in vivo. Preclinical and post-mortem human investigations indicate that the relationship between tissue neuromelanin content and neurodegeneration is complex. Neuromelanin exhibits both neuroprotective and cytotoxic characteristics, and tissue neuromelanin content varies across the lifespan, exhibiting an inverted U-shaped relationship with age. Neuromelanin-sensitive MRI (NM-MRI) is an emerging modality that allows measurement of neuromelanin-associated contrast in SNc and LC in humans. NM-MRI robustly detects disease effects in these structures in neurodegenerative conditions, including Parkinson's disease (PD). Previous NM-MRI studies of PD have largely focused on detecting disease group effects, but few studies have reported NM-MRI correlations with phenotype. Because neuromelanin dynamics are complex, we hypothesize that they are best interpreted in the context of both disease stage and aging, with neuromelanin loss correlating with symptoms most clearly in advanced stages where neuromelanin loss and neurodegeneration are coupled. We tested this hypothesis using NM-MRI to measure SNc and LC volumes in healthy older adult control individuals and in PD patients with and without freezing of gait (FOG), a severe and disabling PD symptom. We assessed for group differences and correlations between NM-MRI measures and aging, cognition and motor deficits. SNc volume was significantly decreased in PD with FOG compared to controls. SNc volume correlated significantly with motor symptoms and cognitive measures in PD with FOG, but not in PD without FOG. SNc volume correlated significantly with aging in PD. When PD patients were stratified by disease duration, SNc volume correlated with aging, cognition, and motor deficits only in PD with disease duration >5 years. We conclude that in severe or advanced PD, identified by either FOG or disease duration >5 years, the observed correlations between SNc volume and aging, cognition, and motor function may reflect the coupling of neuromelanin loss with neurodegeneration and the associated emergence of a linear relationship between NM-MRI measures and phenotype.