Normal Control Subjects Research Articles

Molecular Study on Single Nucleotide Polymorphism of Adiponectin Gene and its Association with Type II Diabetes in Egyptian Patients

Background: Type 2 diabetes mellitus is a prevalent hereditary condition that has multiple genetic risk factors as well as environmental influences. Aim: The Work aimed to find if there is an association between the presence of Single Nucleotide Polymorphism of the Adiponectin gene & its association with type II diabetes in Egyptian patients. Material & Techniques: This case-control research had been conducted at Molecular Biology and Biochemistry, Genetic Engineering & biotechnology institute, university of Sadat city. This research had been conducted on eighty patients; separated into 2 groups diabetic group and the control group. Results: significant reduction in adiponectin level in type II diabetes studied cases compared to normal control subjects (p <0.001). GG genotype is significantly more predominant in type 2 diabetes (54%) while the GT genotype is more predominant in control (60%) (p-value =0.04)., while no variation in allele distribution among both groups (p value=0.6). Conclusion: In studied cases with type 2 diabetes, there was a significant reduction in adiponectin levels in type II diabetes patients. GG genotype is significantly more predominant in type 2 diabetes. The genotype GG significantly rises the risk of occurrence of type 2 diabetes.

Macular Anatomy Differs in Dyslexic Subjects.

The macula, as the central part of the retina, plays an important role in the reading process. However, its morphology has not been previously studied in the context of dyslexia. In this research, we compared the thickness of the fovea, parafovea and perifovea between dyslexic subjects and normal controls, in 11 retinal segmentations obtained by optical coherence tomography (OCT). With this aim, we considered the nine sectors of the Early Treatment Diabetic Retinopathy Study (ETDRS) grid and also summarized data from sectors into inner ring subfield (parafovea) and outer ring subfield (perifovea). The thickness in all the four parafoveal sectors was significantly thicker in the complete retina, inner retina and middle retina of both eyes in the dyslexic group, as well as other macular sectors (fovea and perifovea) in the inner nuclear layer (INL), inner plexiform layer (IPL), IPL + INL and outer plexiform layer + outer nuclear layer (OPL + ONL). Additionally, the inner ring subfield (parafovea), but not the outer ring subfield (perifovea), was thicker in the complete retina, inner retina, middle retina (INL + OPL + ONL), OPL + ONL, IPL + INL and INL in the dyslexic group for both eyes. In contrast, no differences were found between the groups in any of the sectors or subfields of the outer retina, retinal nerve fiber layer, ganglion cell layer or ganglion cell complex in any eye. Thus, we conclude from this exploratory research that the macular morphology differs between dyslexic and normal control subjects, as measured by OCT, especially in the parafovea at middle retinal segmentations.

Photoacoustic Imaging of COVID-19 Vaccine Site Inflammation of Autoimmune Disease Patients.

Based on the observations made in rheumatology clinics, autoimmune disease (AD) patients on immunosuppressive (IS) medications have variable vaccine site inflammation responses, whose study may help predict the long-term efficacy of the vaccine in this at-risk population. However, the quantitative assessment of the inflammation of the vaccine site is technically challenging. In this study analyzing AD patients on IS medications and normal control subjects, we imaged the inflammation of the vaccine site 24 h after mRNA COVID-19 vaccinations were administered using both the emerging photoacoustic imaging (PAI) method and the established Doppler ultrasound (US) method. A total of 15 subjects were involved, including 6 AD patients on IS and 9 normal control subjects, and the results from the two groups were compared. Compared to the results obtained from the control subjects, the AD patients on IS medications showed statistically significant reductions in vaccine site inflammation, indicating that immunosuppressed AD patients also experience local inflammation after mRNA vaccination but not in as clinically apparent of a manner when compared to non-immunosuppressed non-AD individuals. Both PAI and Doppler US were able to detect mRNA COVID-19 vaccine-induced local inflammation. PAI, based on the optical absorption contrast, shows better sensitivity in assessing and quantifying the spatially distributed inflammation in soft tissues at the vaccine site.

Injury of the Spinothalamic Tract Following Whiplash Injury: A Diffusion Tensor Tractography Study

Using diffusion tensor tractography (DTT), we demonstrated the spinothalamic tract (STT) injury in patients with central pain following whiplash injury. Our primary hypothesis is that fractional anisotropy (FA) and tract volume (TV) of the STT in injured people differ from non-injured people. Our secondary hypothesis is that the direction of the collision results in a different type of injury. Nineteen central pain patients following whiplash injury and 19 normal control subjects were recruited. The STT was reconstructed by the DTT, the FA and TV of the STT were measured. In addition, different characteristics of the STT injury according to the collision direction were investigated. The FA value did not differ significantly between the patient and control groups (p > 0.05). However, the significantly lower value of the TV was observed in patient group than the control group (p < 0.05). The onset of central pain was significantly delayed (13.5 days) in patients who were involved in a frontal collision, compared to patients with rear-end collision (0.6 days) (p < 0.05). In contrast, the Visual Analogue Scale was higher in the patients with rear-end collision (p < 0.05). We found the STT injury mild traumatic brain injury (TBI) who suffered central pain after whiplash injury, using DTT. In addition, we demonstrated different characteristics of the STT injury according to the collision direction. We believe that injury of the STT would be usefully detected by DTT following whiplash injury.

Based on Tau PET Radiomics Analysis for the Classification of Alzheimer’s Disease and Mild Cognitive Impairment

Alzheimer's Disease (AD) and Mild Cognitive Impairment (MCI) are closely associated with Tau proteins accumulation. In this study, we aimed to implement radiomics analysis to discover high-order features from pathological biomarker and improve the classification accuracy based on Tau PET images. Two cross-racial independent cohorts from the ADNI database (121 AD patients, 197 MCI patients and 211 normal control (NC) subjects) and Huashan hospital (44 AD patients, 33 MCI patients and 36 NC subjects) were enrolled. The radiomics features of Tau PET imaging of AD related brain regions were computed for classification using a support vector machine (SVM) model. The radiomics model was trained and validated in the ADNI cohort and tested in the Huashan hospital cohort. The standard uptake value ratio (SUVR) and clinical scores model were also performed to compared with radiomics analysis. Additionally, we explored the possibility of using Tau PET radiomics features as a good biomarker to make binary identification of Tau-negative MCI versus Tau-positive MCI or apolipoprotein E (ApoE) ε4 carrier versus ApoE ε4 non-carrier. We found that the radiomics model demonstrated best classification performance in differentiating AD/MCI patients and NC in comparison to SUVR and clinical scores models, with an accuracy of 84.8 ± 4.5%, 73.1 ± 3.6% in the ANDI cohort. Moreover, the radiomics model also demonstrated greater performance in diagnosing AD than other methods in the Huashan hospital cohort, with an accuracy of 81.9 ± 6.1%. In addition, the radiomics model also showed the satisfactory classification performance in the MCI-tau subgroup experiment (72.3 ± 3.5%, 71.9 ± 3.6% and 63.7 ± 5.9%) and in the MCI-ApoE subgroup experiment (73.5 ± 4.3%, 70.1 ± 3.9% and 62.5 ± 5.4%). In conclusion, our study showed that based on Tau PET radiomics analysis has the potential to guide and facilitate clinical diagnosis, further providing evidence for identifying the risk factors in MCI patients.

Association of GABA receptor delta subunit gene variations with increased risk of methamphetamine dependence

ObjectiveEvidence reveals that γ-aminobutyric acid (GABA) receptors are involved in the development of methamphetamine (METH) dependence. The GABA receptor delta subunit gene (GABRD) might be a good candidate gene for METH dependence. In a case–control study, we investigated the association between the single nucleotide polymorphisms (SNPs) in GABRD and METH dependence in a Chinese Han population. MethodsA total of 300 METH dependent patients and 300 age and sex matched normal control subjects were recruited. Four SNPs (rs13303344, rs4481796, rs2376805, and rs2229110) in GABRD were determined with the TaqMan genotyping assay. The association of the SNPs with METH dependence was assessed. ResultsOnly the allele frequency of rs2376805 significantly differed between the patients and controls (P = 0.030). The G allele frequency of rs2376805 was higher in the METH dependent group than in the controls (odds ratio = 1.332, 95 % CI: 1.028–1.724). This association was found in females but not in males. In females, the frequencies of genotype and allele at rs2376805 significantly differed between the patients and controls (P = 0.025, 0.022, respectively); the rs2376805 G allele may also be a risk factor for METH dependence (odds ratio = 1.548, 95 % CI: 1.063–2.257). The haplotype ACGT frequency significantly differed between the patients and controls in total subjects (P = 0.008, odds ratio = 1.815, 95 % CI: 1.183–2.782), as well as in females (P = 0.005, odds ratio = 2.702, 95 % CI: 1.313–5.562). In females only, the METH craving score was significantly lower in patients harboring the G allele at rs2376805 than in those harboring the homozygous AA genotype (P = 0.044). ConclusionThe preliminary results indicate that GABRD rs2376805 is associated with METH dependence, especially in females.

Nuclear Magnetic Resonance-Based Metabolomics Approach Revealed the Intervention Effect of Using Complementary and Alternative Medicine (CAM) by CKD Patients.

Chronic kidney disease (CKD) is the end point of a number of systemic chronic diseases. The prevalence of CKD is increasing worldwide and recent epidemiological studies are showing the high prevalence of renal failure in CKD patients using complementary and alternative medicines (CAMs). Clinicians believe that biochemical profiles of CKD patients using CAM (referred here to as CAM-CKD) may be different compared to those on standard clinical treatment and should be managed differently. The present study aims to explore the potential of the NMR-based metabolomics approach to reveal the serum metabolic disparity between CKD and CAM-CKD patients with respect to normal control (NC) subjects and if the differential metabolic patterns can provide rationale for the efficacy and safety of standard and/or alternative therapies. Serum samples were obtained from 30 CKD patients, 43 CAM-CKD patients, and 47 NC subjects. The quantitative serum metabolic profiles were measured using 1D 1H CPMG NMR experiments performed at 800 MHz NMR spectrometer. The serum metabolic profiles were compared using various multivariate statistical analysis tools available on MetaboAnalyst (freely available web-based software) such as partial least-squares discriminant analysis (PLS-DA) and random forest (a machine learning) classification method. The discriminatory metabolites were identified based on variable importance in projection (VIP) statistics and further evaluated for statistical significance (i.e., p < 0.05) using either Student t-test or ANOVA statistics. PLS-DA models were capable of clustering CKD and CAM-CKD with considerably high values of Q 2 and R 2. Compared to CAM-CKD patients, the sera of CKD patients were characterized by (a) elevated levels of urea, creatinine, citrate, glucose, glycerol, and phenylalanine and phenylalanine-to-tyrosine ratio (PTR) and (b) decreased levels of various amino acids (such leucine, isoleucine, valine, and alanine), high-density lipoproteins, lactate, and acetate. These changes suggested that CKD patients manifest severe oxidative stress, hyperglycemia (with dampened glycolysis), increased protein energy wasting, and reduced lipid/membrane metabolism. Statistically significant and strong positive correlation of PTR with serum creatinine levels suggested the role of oxidative stress in the progression of kidney disease. Significant differences in metabolic patterns between CKD and CAM-CKD patients were observed. With respect to NC subjects, the serum metabolic changes were more aberrant in CKD patients compared to CAM-CKD patients. The aberrant metabolic changes in CKD patients with manifestations of higher oxidative stress compared to CAM-CKD patients could explain clinical discrepancies between CKD and CAM-CKD patients and further advocate the use of different treatment strategies for CKD and CAM-CKD patients.

Cranial Shape Changes in Nonsyndromic Unilateral Lambdoid Synostosis after Open Posterior Vault Remodeling.

The purpose of this study was to quantify change in cranial morphology in patients with nonsyndromic unilateral lambdoid craniosynostosis (ULC) from presentation (t0), after open posterior switch-cranioplasty (t1), and at 2-year follow-up (t2). Volumetric, linear, and angular analysis were performed on computed tomographic scans at the three time points and against normal control subjects. Significance was set at P < 0.05. Twenty-two patients were included. ULC cranial vault asymmetry index was higher than in control subjects before surgery (6.22 ± 3.55) but decreased after surgery (3.00 ± 2.53) to become comparable with the normal asymmetry range present in the controls. After surgery, both diagonals increased, but more on the fused side. In the 2 years after surgery, both diagonals in patients with ULC grew proportionately, but the fused diagonal remained slightly shorter than the patent side. Total cranial volume was higher in patients with ULC than in control subjects after surgery but became comparable at t2. Cranial base angulation improved by t2 but did not approach normal, and ear position remained unchanged. The facial twist was higher than in controls at t0 and t1 but was comparable at t2. Coronal asymmetry improved with surgery but remained undercorrected at t2, with the greatest residual asymmetry at opisthion. Open-switch cranioplasty normalizes cranial vault asymmetry index by increasing the fused cranial diagonal more than the patent side and is stable at 2 years. Skull base twist does not normalize, but facial twist approaches normal. Technique improvement should focus on residual coronal asymmetry present at opisthion. Therapeutic, IV.

Evaluation of Sleep Quality and Its Relationship With Pain, Mood, and Physical Function After Orthopaedic Trauma Surgery: A Prospective Cohort Study.

Sleep disturbances are associated with pain and mental health. We prospectively compared a cohort of patients with orthopaedic trauma with a control group to establish the prevalence and duration of sleep disturbance and associations between sleep disturbance and pain, mood scores, and functional outcome scores. Subjects were patients with orthopaedic trauma undergoing in-patient surgical procedures and healthy control subjects from the community. Questionnaires completed by all subjects included the VAS numerical pain rating scale, the abbreviated profile of mood states, Patient-Reported Outcomes Measurement Information System Physical Function, and Patient-Reported Outcomes Measurement Information System Sleep Disturbance. Control subjects completed the surveys once, and subjects with trauma completed them at 2, 6, and 24 weeks postoperatively. Healthy control subjects (28.6 ± 13.2) were significantly younger than patients with trauma (41.8 ± 18.9) ( P < 0.001). Compared with control subjects, at 2-week follow-up visit, patients with trauma had worse sleep ( P < 0.001) and worse mood ( P = 0.006). Across the study period, patients with trauma showed improvements in physical function ( P < 0.001) and pain (at rest [ P = 0.02], during activity [ P = 0.02], and at night [ P = 0.002]). In patients with trauma, better sleep disturbance scores were associated with better mood and less pain for all pain metrics ( P < 0.001). Patients with orthopaedic trauma have worse sleep disturbance scores at 2 weeks postoperatively compared with normal control subjects; this difference attenuated at 6 weeks. Sleep disturbance was found to markedly correlate with pain and mood, with worse sleep quality associated with higher pain and worse mood. Improvement in sleep quality across 24 weeks postoperatively was associated with improvement in mood scores. Patients should be counseled about the likely development of sleep disturbance and the possible association with worse emotional/mental health with worse sleep. Physicians should consider incorporating a multidisciplinary approach to the management of these select patients.

Humanin gene expression in subjects with Parkinson's disease.

Bradykinesia, tremor, rigidity and postural instability are the hallmark of Parkinson's disease (PD). Non-motor symptoms including cognitive, behavioral, and neuropsychiatric changes, sensory and sleep disturbances that may precede the motor symptoms by years. The peculiar pathological features of PD are decreased dopaminergic neurons and dopamine levels in the substantia nigra pars compacta and pontine locus coeruleus. Humanin is produced by a small gene peptide, which is located in the mitochondria genome. Inflammation, oxidative stress, mitochondrial dysfunction and altered transcription have been recognized as causative factors of PD. This evidence has prompted many researchers to focus on studying the functions of DNA and mitochondria. The purpose of the present study was to evaluate Humanin mRNA levels in peripheral blood mononuclear cells (PBMCs) of PD subjects, compared with those in PBMCs of normal control (NC) subjects. A total of 220 participants, including 154 PD patients (57 females and 97 males; mean age 71.54 years, SD 7.8) and 66 CN (28 females and 38 males; mean age 70.54 years, SD 9.45) were enrolled for the qRT-PCR analysis. Increased Humanin mRNA levels were found in PD samples, compared to controls. In conclusion, the present data confirm the tendency of mitochondria to overexpress mRNA in PD, which could be a cellular attempt to reduce apoptotic damage in PD subjects. Humanin might be useful as a marker for a better diagnosis of PD, and we cannot exclude that in the future it might also play a role on prognosis and in the possible therapies for PD.

Protective and Adverse Roles of DDX3X in Different Cell Types in Nonalcoholic Steatohepatitis Progression

Persistent hepatic cellular metabolic stress and liver inflammatory stimuli are key signatures of nonalcoholic steatohepatitis (NASH). DDX3X is a vital molecule involved in cell fate decisions in both pro-survival stress granule (SG) and pro-death NOD-like receptor family pyrin domain containing 3 (NLRP3) inflammasome assembly in response to stress signals. However, the role of DDX3X in NASH remains unclear. We characterized the cell type-specific roles of DDX3X in NASH. Human liver tissues from NASH patients and normal control subjects were collected to assess DDX3X expression and distribution. Nutritional steatohepatitis models were constructed by feeding macrophage-specific DDX3X knockout (DDX3X ΔMφ ), hepatocyte-specific DDX3X knockout (DDX3X Δhep ), and wild-type control (DDX3X fl/fl ) mice a high-fat and high-cholesterol (HFHC) diet, a methionine- and choline-deficient (MCD) diet, and a high-fat/high-iron/high-fructose/high-cholesterol, low-methionine, and choline-deficient (HFHIHFHC-MCD) diet. The study demonstrated that DDX3X was predominantly expressed in macrophages and hepatocytes in control liver tissues, and its expression was down-regulated in patients or mice with NASH. Compared to DDX3X fl/fl littermates, DDX3X ΔMφ mice showed improved liver histology in nutritional steatohepatitis models. Loss of macrophage DDX3X inhibited NLRP3 inflammasome-mediated pyroptosis, causing anti-inflammatory M2 polarization and alleviating hepatocyte steatohepatitic changes. DDX3X Δhep mice developed marked steatohepatitis in multiple nutritional steatohepatitis models compared to DDX3X fl/fl littermates. DDX3X-deleted hepatocytes showed impaired SG assembly, leading to increased sensitivity and intolerance to metabolic stimulation and resultant steatohepatitis. In conclusion, DDX3X plays opposite roles in different cell types during the progression of NASH. A better understanding of the cell-specific differences in the crosstalk between SG formation and NLRP3 activation is crucial for developing prospective targeted DDX3X inhibitors for the treatment of NASH.

Serum Cathepsin D was Elevated in Patients with Limb Lymphedema.

Lymphedema (LE) is a chronic disease that can lead to disability. Currently, the pathogenesis of LE remains unclear, and there is a lack of serum proteins applicable for diagnosis in clinical practice. This study aimed to screen and identify the differentially expressed proteins in serum samples of limb lymphedema and normal subjects and to further explore their value in the diagnosis of LE. Nano-flow reverse phase liquid chromatography-tandem mass spectrometry (Nano RPLC-MS/MS) was used to establish the serum protein profiles of primary lymphedema (PLE), secondary lymphedema (SLE), and normal controls (NC). Differentially expressed serum proteins were screened and identified. Subsequently, enrichment analysis was performed for proteins that were upregulated in the LE group compared to the NC group. The target protein was validated by western blot (WB) and enzyme-linked immunosorbent assay (ELISA). Both the receiver operating characteristic (ROC) curve and Spearman's correlation test were employed to evaluate the diagnostic performance of the protein and its relationship with disease severity. A total of 362 serum proteins were identified, among which 241 proteins were differentially expressed among PLE, SLE, and NC subjects (p < 0.05, fold change > 1.2). The enriched pathway correlated with cornified envelope formation was selected for further analysis. Cathepsin D (CTSD), a target protein involved in the selected pathway, was found to be up-regulated in the serum of PLE and SLE patients compared to NC. The AUCs of CTSD were 0.849 and 0.880 for patients with PLE and SLE, respectively. There was a significant positive correlation between the levels of serum CTSD and disease severity in the PLE group. Proteomic analysis found that the levels of serum proteins related to cornified envelope formation were elevated in patients with limb lymphedema. Serum CTSD was highly expressed in patients with limb lymphedema and showed good diagnostic value.

Sürnümerer Diş Gözlenen Çocuklarda Diş Gelişiminin Değerlendirilmesi: Olgu Kontrol Araştırması

Objective: The term supernumerary tooth is one of the important dental anomalies affecting children and is used to describe excess teeth in the dental arch. Knowledge of changes in tooth development accompanying supernumerary teeth provides information about both orthodontic problems and changes in bone and teeth. This study aims to evaluate the relationship between the presence of supernumerary teeth and dental development in children and to raise awareness about the conditions associated with the presence of supernumerary teeth. Material and Methods: 175 (114 males and 61 females) children with supernumerary teeth were examined and compared to 175 normal control subjects matched for sex and age. Dental development was evaluated from panoramic radiographs using the dental age estimation method of Demirjian et al. Data were statistically analyzed using the Shapiro-Wilk test. Results: Significant differences were found in the mean of dental age and chronological age difference between case controls and children with supernumerary teeth (0.4±0.7 and 0.8±0.7 years, respectively). The mean differences between dental age and chronological age in hyperdontic males and females were 0.77±0.7 years and 0.96±0.7 years, respectively, and the results of the controls were 0.39±0.7 for males and 0.47±0.8 years for females. The differences between the hyperdontic and control groups were significant for both genders (p<0.001). Conclusion: Children with supernumerary teeth appear to have advanced dental development compared to children with normal dentition. The presence of supernumerary teeth in children is a condition that affects dental development.

The Function of Retinal Thickness and Microvascular Alterations in the Diagnosis of Systemic Sclerosis.

In this study, we aim to investigate retinal thickness (RT) and superficial vascular density (SVD) differences between patients with systemic sclerosis (SSc) and healthy controls (HCs) by optical coherence tomography angiography (OCTA). Sixteen patients with a definitive SSc diagnosis without clinical signs of retinopathy and 16 normal control subjects were recruited. All individuals underwent OCTA scanning to assess macular RT and SVD. We divided each image into nine subregions as the early treatment diabetic retinopathy study (ETDRS). Visual acuity (VA) was considerably different between patients with SSc (32 eyes) and control subjects (32 eyes) (p < 0.001). Compared to the control group, individuals with SSc had decreased inner RT in inner superior, outer superior, outer temporal, inner temporal, center, and inner nasal regions (p < 0.05). Outer RT was decreased in the outer and inner temporal regions, and full RT was decreased in the regions of outer superior, inner superior, inner temporal, and outer temporal, in comparison to the control group (p < 0.05). Patients with SSc had significant reduction of SVD in the inner and outer of both superior and temporal, besides outer nasal regions than controls. (p < 0.05). Moreover, SVD was significantly associated with the outer temporal region of patients suffering from SSc (p < 0.05). Diagnostic Sensitivity of RT and SVD of Inner Superior Regions in SSc, as indicated by areas under curves of the Receiver Operating Characteristic (ROC), were 0.874 (95% CI: 0.786-0.962) and 0.827 (95% CI: 0.704-0.950), respectively. In conclusion, VA may be affected by RT variations inside the macula in patients with SSc. Measuring RT with OCTA could be a useful predictor of early diagnosis.

Evaluation and Comparison of Plasma miRNA-31 in Oral Squamous Cell Carcinoma

Background/Purpose: Oral Squamous cell carcinoma is sixth most common cancer with considerable morbidity and mortality. The microRNAs (miRNAs) are set of short RNAs involved in regulating the expression of protein coding genes. They are up or down-regulated in carcinogenesis and in oral cancer. The miRNA-31 (miR-31) is increased in oral cancer. Objective: To evaluate and compare the expression of miRNA-31 in plasma of Oral squamous cell carcinoma and control subjects. Materials and Methods: Case control study was carried out in 25 cases of oral squamous cell carcinoma subjects and 25 normal control subjects. The level of miRNA-31 in blood plasma was evaluated by miRNA easy kit (quagen) and miRNA-based qRT-PCR. The fold change was observed and compared between OSCC and controls. Results: The plasma level of miRNA-31 was significantly increased in OSCC patients compared to controls (p&lt;0.001). The patients with moderately differentiated, grade 4 OSCC patients showed significant increase in fold change compared to control, well differentiated and grade 3 OSCC (p&lt;0.001). Conclusion: Our results indicate that plasma miR-31 may be used as an adjuvant biomarker the detection of OSCC patient.

Therapeutic effect of ibrutinib, a selective Bruton's tyrosine kinase inhibitor, in orbital fibroblasts from patients with Graves' orbitopathy.

Bruton's tyrosine kinase (BTK) is an essential protein in B-cell antigen receptor (BCR) signaling pathway and is known to be related to pathogenetic effect on B-cell related malignancies and various autoimmune diseases. In this study, we investigated the therapeutic effect of ibrutinib, an orally bioavailable BTK inhibitor in the pathogenesis of Graves' orbitopathy (GO) in in vitro model. Expression of BTK in orbital tissues from GO and normal control subjects were evaluated by real-time polymerase chain reaction (PCR). Primary cultured orbital fibroblasts from each subject were exposed to ibrutinib and stimulated with interleukin (IL)-1β or insulin like growth factor (IGF)-1. Production of inflammatory cytokines was evaluated by real time PCR and enzyme-linked immunosorbent assays (ELISA). The downstream transcription factors were also determined by western blot assays. The expression of BTK in GO tissues were significantly higher than in healthy controls. After stimulation of GO orbital fibroblasts with IL-1β or IGF-1, BTK mRNA and phosphorylated (p)- BTK protein expression was also enhanced. Ibrutinib reduced the expression of BTK mRNA and proteins of p-BTK, and inhibited the IL-1β- and IGF-1-induced production of proinflammatory cytokines including IL-6, IL-8 and COX-2 in both GO and normal cells. Ibrutinib also significantly attenuated phosphorylation of Akt, p38, c-Jun N-terminal kinase (JNK), extracellular signal-regulated kinase (ERK), and nuclear factor kappa-light-chain-enhancer of activated B-cells (NF-κB) in IL-1β stimulated GO cells and Akt, JNK, and NF-κB in IL-1ß stimulated normal cells. BTK expression is enhanced in GO tissue and orbital fibroblasts. Ibrutinib, a BTK inhibitor suppresses proinflammatory cytokine production as well as phosphorylation of Akt and NF-κB protein. Our results suggest the potential role of BTK in GO inflammatory pathogenesis and possibility of a novel therapeutic target of GO.

High Expression of Inflammatory Cytokines and Chemokines in Human T-lymphotropic Virus 1-Associated Adult T-cell Leukemia/Lymphoma

Background: Human T-cell lymphotropic virus type 1 (HTLV-1) is the etiological agent of adult T-cell leukemia/lymphoma (ATLL) without any specific antiviral. Objectives: This study aimed to evaluate the expression level of inflammatory chemokines and pro-inflammatory cytokines in ATLL patients, asymptomatic carriers (ACs), and healthy individuals to assess the role of these inflammatory markers in ATLL pathogenicity. Methods: This study was conducted from May 2021 to August 2022. The ATLL blood samples were collected from the oncology wards of Imam Khomeini, Shariati, and Imam Hossein hospitals, in Tehran, Iran. The blood samples of ACs and normal control subjects were collected from blood donors referred to blood transfusion centers of Tehran and Alborz provinces, Iran. RNA extraction, complementary DNA (cDNA) synthesis, and real-time polymerase chain reaction (PCR) were done in targeted sample groups to investigate the correlation and expression rate of C-C motif chemokine ligand 3 (CCL3), C-C motif chemokine ligand 4 (CCL4), C-X-C motif chemokine ligand 8 (CXCL8), interleukin 23 subunit alpha (IL-23A), and interleukin 17 A (IL-17A). Results: A total of 30 samples were collected from 3 groups. The CCL3, CCL4, CXCL8, and IL-17A messenger RNA (mRNA) expression levels were significantly upregulated in the ATLL groups. There was a significant difference between CCL3 expression between the ACs and ATLL groups. In addition, CCL4 and CXCL8 expression levels were more significant in the ATLL group than in the normal control group. The IL-17A expression level significantly increased between groups. The IL-23A expression levels had no significant differences between the ATLL, ACs, and normal control groups. Conclusions: This study showed significant upregulation of pro-inflammatory cytokines and chemokines mRNAs in HTLV-1–associated ATLL compared to the ACs and normal control groups. Conducting more experiments to investigate the therapeutic effect of chemokines/cytokines in ATLL is essential.

Discriminative patterns of white matter changes in Alzheimer's

Changes in structural connectivity of the Alzheimer's brain have not been widely studied utilizing cutting-edge methodologies. This study develops an efficient structural connectome-based convolutional neural network (CNN) to classify the AD and uses explanations of CNNs' choices in classification to pinpoint the discriminative changes in white matter connectivity in AD. A CNN architecture has been developed to classify normal control (NC) and AD subjects from the weighted structural connectome. Then, the CNN classification decision is visually analyzed using gradient-based localization techniques to identify the discriminative changes in white matter connectivity in Alzheimer's. The cortical regions involved in the identified discriminative structural connectivity changes in AD are highly covered in the temporal/subcortical regions. A specific pattern is identified in the discriminative changes in structural connectivity of AD, where the white matter changes are revealed within the temporal/subcortical regions and from the temporal/subcortical regions to the frontal and parietal regions in both left and right hemispheres. The proposed approach has the potential to comprehensively analyze the discriminative structural connectivity differences in AD, change the way of detecting biomarkers, and help clinicians better understand the structural changes in AD and provide them with more confidence in automated diagnostic systems.

Association of serum klotho levels with different-staged vascular calcification status in patients with maintenance hemodialysis

BackgroundVascular calcification (VC) is suggested to be associated with serum klotho levels in patients with maintenance hemodialysis (MHD), whereas there is a lack of reports on the associations of VC status in whole arteries with serum klotho contents.MethodsOne hundred forty eligible patients with MHD and a total of age-and gender-matched normal controls (NCs) were recruited. We analyzed the VC statuses of large arteries and peripheral muscular arteries by calculating the sum of scores from each artery. The levels of serum klotho were determined by ELISA. In addition, the relationship between serum klotho and VC status was evaluated using correlation analysis and regression analysis.ResultsThe VC severity in MHD patients tended to be worse in comparison with NCs. Serum klotho level in patients with MHD was lower than that in the NC subjects (​P < 0.0001), which was correlated with VC scores as reflected by correlation analysis and regression analysis. Serum klotho concentrations exhibited a dynamic decline along with increased VC status stages. Subjects with higher levels of serum klotho had a higher prevalence of cardiovascular events.ConclusionOur study indicates serum klotho is strongly associated with VC status in a stage-dependent manner.

Morphologic, dynamic and high-resolution microscopy MRI in early-onset spondyloarthritis finger dactylitis

ObjectiveUp to now, the pathophysiology of SpA dactylitis has not been entirely clarified. It is not clear which are the involved tissues and which is the primary lesion of the “sausage-like” digit. The aim of our study was to examine the finger structures in early-onset finger dactylitis using high-resolution microscopy MRI together with morphologic and dynamic MRI.Subjects and methodsIn a 6-month period, 13 SpA patients (7 females and 6 males), mean age 54.07 years (range 37–73 years) and mean disease duration 7.07 years (range 1–44 years) with early-onset finger dactylitis (less than 3 months) were recruited. Nine patients had PsA, 3 HLA-B27-positive uSpA and 1 HLA-B27-negative uSpA. One patient had 2 dactylitis fingers. Ten healthy volunteers matched for age and sex with no personal and family history of SpA were enrolled. All dactylitis fingers and randomly selected fingers of the normal control subjects were imaged by morphologic, dynamic and high-resolution microscopy MRI.ResultsWe have found flexor tenosynovitis in all the 14 dactylitis fingers, joint synovitis in 5 and oedema in the finger soft tissue in 10. In 2 dactylitis fingers, there was oedema at the insertion of the joint capsule suggesting enthesitis. In 5 dactylitis fingers, there was only mild enhancement at the enthesis organ (collateral ligament, flexor and extensor tendons).ConclusionsOur MRI study on early-onset dactylitis demonstrates that flexor tenosynovitis, joint synovitis and oedema of the digit soft tissue are the predominant alterations visible in the early phase of evolution of dactylitis and that, therefore, enthesitis may not be considered the primary lesion of dactylitis.