Severe Infiltration Research Articles

Deletion of both anaerobic regulator genes fnr and narL compromises the colonization of Salmonella Typhimurium in mice model.

Salmonella Typhimurium (STM), a zoonotic pathogen, can adjust its metabolic pathway according to the variations in the partial pressure of atmospheric oxygen and nitrate via fumarate nitrate reductase regulator (Fnr) and NarL, the response regulator for nitrate reductase. Both Fnr and NarL have been individually reported to be the contributors of virulent phenotypes of STM. Hypoxia along with nitrate-rich environment are prevalent in macrophages and the Salmonella-induced inflammatory lumen of the host's large intestine activates both fnr and narL genes. In this study, the double (fnr and narL) knockout STM showed a synergistic reduction in the swimming (62%), swarming (84%) and biofilm density (86%) phenotypes anaerobically in association with its significant aerobic attenuation. The intracellular replication of the double mutant was reduced by 2.3 logs in chicken monocyte-derived macrophages. Furthermore, the competitive index of the double mutant in liver and spleen was found to be 0.3 and 0.44 respectively at 120h post-infection (PI) in mice. Surprisingly, no double mutant could be recovered from the infected mouse liver 3 days PI. Histopathological findings showed moderate infiltration of mononuclear cells in the large intestine of mice infected with double mutant, but severe infiltration was seen with the wild-type strain.

Clinical and laboratorial profile of patients with ATTR cardiomyopathy: comparison between wild type and p.Val142Ile forms in the Brazilian population

Abstract Background In Brazil, besides the wild type (wt) form, transthyretin amyloid cardiomyopathy (ATTR-CM) is predominantly caused by hereditary form with Val142Ile mutation. Considering that both forms occur in elderly people, the clinical presentation may be similar. We sought to compare the clinical presentation of patients with wt and Val142Ile mutation in a Brazilian cohort of ATTR-CM patients. Methods Among the 642 patients enrolled in REACT/SP, 283 presented ATTR-CM, being 85 wt and 90 Val142Ile patients. We compared the main clinical characteristics between groups. Results The wt in comparison to Val142Ile patients, respectively, presented: older age (78.4+/-8.5 vs 74.2+/-8.1 y.o., p = 0.0009); similar proportion of males (82% vs 81%, p=0.85); lower proportion of blacks (11% vs 39%, p=0.0001); similar prevalence of heart failure (HF) symptoms (85% vs 79%, p=0.33); higher prevalence of syncope (13% vs 2%, p=0.008) and Pacemakers (PM) implantation (8% vs 1%, p=0.027); similar prevalence of neuropathy manifestations (38% vs 51%, p=0.17); lower creatinine (1.5+/-0.8 vs 2.1+/-2.1 mg/dL, p=0.02) and NT-ProBNP levels (2860.0+/-2843.3 vs. 5488.3+/-5455.6 pg/ml, p=0.0001); reduced interventricular septal thickness (15.6+/-3.3 vs 17.0+/-3.3 mm, p= 0.006), posterior left ventricular (LV) posterior wall thickness (14.2+/-2.4 vs 16.2+/-4.4 mm, p=0.0003), higher LV ejection fraction (52.0+/-10.1 vs 48.2+/-13.6%, p=0.038), higher global LV longitudinal strain (8.6+/-8.7 vs 3.4+/-9.8, p=0.0003), smaller LV diastolic diameter (45.9+/-6.1 vs 43.0+/-7.3 mm, p=0.005) at 2D-Echocardiogram. Conclusions In the Brazilian population wt and Val142Ile patients had similar clinical presentation regarding HF and neuropathy symptoms, but higher prevalence of syncope and PM in wt patients. Conversely, Val142Ile patients presented more severe amyloid cardiac infiltration.

Fabry cardiomyopathy: myocardial fibrosis, inflammation and reduced mannose6phosphate receptors cause low accessibility to enzyme therapy

Abstract Background Clinical impact of enzyme replacement therapy (ERT) on advanced Fabry disease cardiomyopathy (FDCM) appears limited. Purpose Clarify the pathologic mechanisms responsible of ERT inefficacy in the advanced FDCM. Methods Ten male patients with advanced FDCM (echocardiographic maximal wall thickness 19.3 ± 2.1 mm) underwent left ventricular endomyocardial biopsy before and 4 hours after beta-agalsidase infusion. Comparative studies between pre and post infusion samples included: histology and electron microscopy; assessment of myocardial alpha-galactosidase A activity; immunohistochemistry for alpha-galactosidase A and semiquantitative evaluation (from 0 to 3) of its cardiomyocyte content; Ultrastructural immunogold analysis with anti-alpha-galactosidase A ab. Western Blot (WB) quantification of mannose-6-phosphate receptors (M6Pr). Controls were surgical left ventricular biopsies from patients with mitral stenosis. Results Histologic and Ultrastructural evaluation showed no removal of storage material while myocardial fibrosis was 9.8 ± 6.8 vs 3.8 ± 2.0 of controls and virus-negative lymphocytic inflammation was observed in 7 out of 10 patients. At Ultrastructural immunogold analysis, Myocardial alpha-galactosidase A activity increased in post infusion samples by overall 1.89-fold. Alpha-galactosidase A immunostaining in cardiomyocytes was absent at baseline in all patients and did not significantly improve in post-infusion samples. Immunogold particles increased by 1.33-fold remaining far from normal controls (86.9 ± 6.6). Protein analysis showed M6Pr in advanced FDCM to be 81% lower than in normal heart. Conclusions Our study shows a low accessibility to ERT of cardiomyocytes affected by advanced FDCM. It is sustained by myocardial fibrosis, inflammation and severe down-regulation of M6Pr. Figure Legend Histologic and ultrastructural and mannose6phosphate receptors characteristics of patients with severe Fabry Disease Cardiomyopathy. Panel A shows severe cellular glycolipid infiltration associated with diffuse interstitial fibrosis (Masson trichrome, 100x). Panel B reveals an overlapping lymphocytic CD45Ro+ myocardial inflammation with focal necrosis of adjacent cardiomyocytes. Insert(i), Ultrastructural detail showing vacuoles (black arrows) to consist of myelin bodies,( scale bar=3μm). Panel C (upper) Western blot analysis of all 10 patients of Mannose-6-phosphate Receptor, molecular weight 275 Kd. Alpha sarcomeric actin (43 kDa) was used as a loading control. (Panel C -lower) Panel D: Graphs document western blot of Mannose-6-phosphate Receptor, in all 10 patients with severe FDCM showing 3-fold lower of Mannose-6-phosphate Receptor values in patients versus normal heart (4.830 ± 1116,724 vs 14.737± 1145,386; p < 0.001).

Quantitative ultrasound assessment of fatty infiltration of the rotator cuff muscles using backscatter coefficient

BackgroundTo prospectively evaluate ultrasound backscatter coefficients (BSCs) of the supraspinatus and infraspinatus muscles and compare with Goutallier classification on magnetic resonance imaging (MRI).MethodsFifty-six participants had shoulder MRI exams and ultrasound exams of the supraspinatus and infraspinatus muscles. Goutallier MRI grades were determined and BSCs were measured. Group means were compared and the strength of relationships between the measures were determined. Using binarized Goutallier groups (0–2 versus 3–4), areas under the receiver operating characteristic curves (AUROCs) were calculated. The nearest integer cutoff value was determined using Youden’s index.ResultsBSC values were significantly different among most Goutallier grades for the supraspinatus and infraspinatus muscles (both p < 0.001). Strong correlations were found between the BSC values and Goutallier grades for the supraspinatus (τb = 0.72, p < 0.001) and infraspinatus (τb = 0.79, p < 0.001) muscles. BSC showed excellent performance for classification of the binarized groups (0–2 versus 3–4) for both supraspinatus (AUROC = 0.98, p < 0.0001) and infraspinatus (AUROC = 0.98, p < 0.0001) muscles. Using a cutoff BSC value of −17 dB, sensitivity, specificity, and accuracy for severe fatty infiltration were 87.0%, 90.0%, and 87.5% for the supraspinatus muscle, and 93.6%, 87.5%, and 92.7% for the infraspinatus muscle.ConclusionBSC can be applied to the rotator cuff muscles for assessment of fatty infiltration. For both the supraspinatus and infraspinatus muscles, BSC values significantly increased with higher Goutallier grades and showed strong performance in distinguishing low versus high Goutallier grades.Relevance statementFatty infiltration of the rotator cuff muscles can be quantified using BSC values, which are higher with increasing Goutallier grades.Key PointsUltrasound BSC measurements are reliable for the quantification of muscle fatty infiltration.BCS values increased with higher Goutallier MRI grades.BCS values demonstrated high performance for distinguishing muscle fatty infiltration groups.Graphical

Influence of Natural Dentin Biomodification Agent on Push-Out Bond Strength and Nanoleakage of Self-Adhesive Resin Cement Luting of Glass-Fiber Posts.

To evaluate the plant-derived compound lignin (LIG) as a pretreatment of intraradicular dentin in combination with EDTA on push-out bond strength (PBS) and nanoleakage of the glass fiber posts (GFPs) cemented using adhesive resin cement. Twenty-eight human incisor roots were prepared for GFP cementation and divided based on dentin pretreatment: (1) CONTROL: no pretreatment, (2) EDTA: 17% EDTA for 3 min, (3) EDTA-LIG: 17% EDTA and 2% lignin for 3 min, (4) EDTA-PAC: 17% EDTA and 2% lignin for 3 min. The GFPs were cemented using the self-adhesive resin cement Multilink Speed. The roots (n = 7) were sectioned into 1 mm-thick discs and subjected to PBS testing after 1 week or 6 months. Nanoleakage was analyzed by SEM. Statistical analysis was performed using two-factor ANOVA and Tukey's test (p < 0.05). Higher PBS was identified for the CONTROL group (p < 0.001). After 6 months, the EDTA-LIG maintained the bond strength with a predominance of mixed failures, while the EDTA-PAC, EDTA, and CONTROL groups showed reduction of bond strength, with a predominance of adhesive failures along with severe silver infiltration in the interface. LIG associated with EDTA as a pretreatment for intraradicular dentin shows significant potential for attaining stable bond strength and interfacial integrity of self-adhesive resin cement to intraradicular dentin.

Pathophysiological dynamics of acute myocardial infarction rats under chronic psychological stress at different time points

There is a lack of in-depth research on the impacts and changes in chronic psychological stress (CPS) on the cardiovascular system after acute myocardial infarction (AMI). This study aims to explore the comorbid mechanism and dynamic evolution of AMI exposed to CPS. 120 Wistar rats were randomly divided into Sham Operation group, Sham Operation + Chronic Unpredictable Mild Stress (CUMS) group, AMI group and AMI + CUMS group, with each group further divided into subgroups at days 7, 14, and 28. The AMI model was created by ligating the left anterior descending coronary artery, and CUMS model was used to induce CPS in rats. Behavioral changes were assessed through open field tests and sucrose preference tests. Cardiac function and structure were evaluated via echocardiography. The serum levels of TNFα, IL-6, NO, ET, CK-MB, cTNT, and ANP were measured using assay kits. Pathological changes in cardiac and brain tissues were observed under an optical microscope. Comparative analysis across different models revealed that CUMS significantly reduced behavioral activities in rats, with an interaction between CUMS and AMI affecting total distance (P < 0.05). Both CUMS and AMI significantly reduced cardiac function indicators, with their interaction effects on LVEF, LVFS, and CO (P < 0.05). AMI significantly altered cardiac structural parameters, particularly on day 28 (P < 0.05); while the impact of CUMS on cardiac structure was not significant, except for a notable reduction in LVAW/s on day 7 in AMI + CUMS group (P < 0.05). AMI caused significant changes in the serum biomarkers, while CUMS only significantly increased cTnT on day 7, ANP, TNFα, and IL-6 on day 14, and CK-MB on day 28, with their interaction effects on the three myocardial injury markers and TNFα (P < 0.05). Comparative analysis across different time points demonstrated that behavioral activity, cardiac function, CK-MB, cTnT, ANP, TNFα, and ET levels decreased significantly over time in the AMI model rats, while the left ventricular mass increased significantly (P < 0.05). Pathologically, compared with stress or AMI alone, the AMI + CUMS group exhibited more severe myocardia cellular degeneration and inflammatory infiltration, causing larger infract areas in myocardial tissue, as well as cell number decreases and morphological changes in hippocampal tissue. AMI with CPS exacerbates myocardial injury through sustained inflammation and endothelial dysfunction, leading to heart-brain pathology manifestations characterized by decreased cardiac function and hippocampal tissue damage.

Prognostic and Predictive Capabilities of Tumor-Infiltrating Lymphocytes in Breast Cancer

The aim of the study is to evaluate the prognostic and predictive capabilities of tumorinfiltrating lymphocytes in breast cancer (BC); to study the prognostic value of T-cell (CD8+, CD3+) lymphocyte infiltration of the tumor and the level of T-regulatory (CD4+) lymphocytes; to study the prognostic and predictive value of the expression of proteins Forkhead Box Protein 3, check-point PD-1, PDL1. Materials and methods. The content of tumor-infiltrating lymphocytes and their quantitative ratio and correlation with regulatory genes were assessed. Archival material was used in the work. The study included archival data from the cancer registry of 1240 patients treated at the N.N. Petrov Cancer Research Institute from 2000 to 2009. Histological preparations stained with hematoxylin and eosin were scanned. Ten-year relapse-free and overall survival were assessed. Results. About 80% of patients with BC have low levels of tumor infiltration by CD8+ cytotoxic T lymphocytes and CD3+ lymphocytes. Expression of the PD-L1 gene was detected more often (29.5%) in triple-negative breast cancer, 1.5 times less often (18.2%) in HER2+ BC and extremely rarely (1.5%) in luminal A subtype. Severe (more than 10%) lymphocytic infiltration of CD8+ and CD3+ with low expression of PD-L1 and FOXP3 improves relapse-free and overall survival of patients with BC. Conclusion. The inclusion of lymphocytic infiltration of the tumor as a prognostic biomarker represents an important step in understanding the biology of BC.

Management of anti-melanoma differentiation-associated gene 5 antibody-induced refractory dermatomyositis complicated by interstitial pneumonia using tofacitinib and its outcomes: a case report

BackgroundClinical amyopathic dermatomyositis is characterized by cutaneous symptoms but lacks muscle symptoms. Anti-melanoma differentiation-associated gene 5 antibodies are frequently found in Japanese patients with clinical amyopathic dermatomyositis. Patients with rapidly progressive interstitial lung disease with positive anti-melanoma differentiation-associated gene 5 antibodies have poor prognoses, and majority of them are treated with combination immunosuppressive therapy; however, the best treatment is yet to be determined.Case presentationA 52-year-old Asian male patient presented with a chief complaint of dyspnea on exertion. He had a typical skin rash and rapidly progressive interstitial pneumonia. Additionally, anti-melanoma differentiation-associated gene 5 antibodies were detected; therefore, he was diagnosed with dermatomyositis-associated interstitial pneumonia. Respiratory failure worsened despite administering steroid pulse therapy, tacrolimus, and cyclophosphamide. Consequently, plasma exchange was performed on day 13 of admission. After a slight improvement, the patient’s respiratory failure worsened. Thus, cyclophosphamide was replaced by tofacitinib on day 28. Although respiratory failure improved and the progression of interstitial pneumonia seemed under control, βD-glucan level increased and Aspergillus antigen was detected on day 49. Micafungin and voriconazole were administered, but the patient succumbed to worsening respiratory failure on day 61. The pathological autopsy revealed multiple nodular lesions with cavity formation in both lungs and the presence of Aspergillus with severe neutrophilic infiltration and necrosis, which supported the diagnosis of invasive pulmonary aspergillosis.ConclusionThe patient with anti-melanoma differentiation-associated gene 5 antibody-related rapidly progressive interstitial lung disease, whose disease was difficult to control after the administration of triple immunosuppressive therapy (steroids, tacrolimus, and cyclophosphamide), showed good response with tofacitinib. Unfortunately, the patient died of invasive pulmonary aspergillosis owing to severe immunosuppression; thus, the signs of complications should be promptly detected.

Intestinal pathology in goats challenged with Clostridium perfringens type D strain CN1020 wild-type and its genetically modified derivatives.

Clostridium perfringens type D is the causative agent of enterotoxemia in sheep, goats, and cattle. Although in sheep and cattle, the disease is mainly characterized by neurological clinical signs and lesions, goats with type D enterotoxemia frequently have alterations of the alimentary system. Epsilon toxin (ETX) is the main virulence factor of C. perfringens type D, although the role of ETX in intestinal lesions in goats with type D enterotoxemia has not been fully characterized. We evaluated the contribution of ETX to C. perfringens type D enteric pathogenicity using an intraduodenal challenge model in young goats, with the virulent C. perfringens type D wild-type strain CN1020; its isogenic etx null mutant; an etx-complemented strain; and sterile, non-toxic culture medium. The intestinal tract of each animal was evaluated grossly, microscopically, and immunohistochemically for activated caspase-3. Both ETX-producing strains induced extensive enterocolitis characterized by severe mucosal necrosis, apoptosis, and diffuse suppurative infiltrates. No significant gross or microscopic lesions were observed in goats inoculated with the non-ETX-containing inocula. These results confirm that ETX is essential for the production of intestinal lesions in goats with type D disease. Also, our results suggest that the intestinal pathology of type D enterotoxemia in goats is, at least in part, associated with apoptosis.

A new protocol for the induction of chronic mastitis with intramammary infusion of lipopolysaccharide in Balb/c mice

Mastitis is the inflammation of the mammary tissue and is commonly observed in farm animals worldwide. The problem causes severe financial losses in the dairy industry in terms of veterinary costs, milk disposal, and treatment expenses. Bacteria are the main actors in the etiology and cause acute and chronic inflammatory changes in the mammary tissue. Acute inflammatory changes are easily recognized clinically, and treatment is initiated immediately, but subacute inflammation progresses insidiously and leads to chronic inflammation with irreversible fibrotic changes. Standardized experimental models for the induction of acute mastitis in laboratory animals are available, and usually, infusion of bacteria or some bacterial structural components into mammary tissue is easily applied for this purpose. However, there are few studies on the induction of chronic mastitis with fibrotic changes, and the applications are relatively complex. In the submitted study, LPS was infused through the teat duct three times on days 0, 5, and 10 to induce chronic mastitis in mice. Tissues were sampled on days 1, 6, and 15 to evaluate histopathological changes. While severe neutrophil infiltrates, a component of acute inflammation, were observed on day 1, lymphocyte infiltrates increased on day 6, consistent with subacute inflammation. On day 15, lesions representing chronic mastitis, such as fibrosis and lymphocyte infiltration, were observed. A model similar to the lesions in chronic mastitis of dairy cattle was successfully and easily established by LPS infusion in mice.

Analysis of Risk Factors Associated with Proximal Junctional Kyphosis Following Long Instrumented Fusion from L1 to Sacrum: Age Itself Does Not Independently Increase the Risk.

Background and Objectives: This study is a retrospective analysis aimed at understanding the incidence and risk factors of proximal junctional kyphosis (PJK) following long-instrumented spinal fusion from L1 to the sacrum in patients with mild to moderate sagittal imbalance. Materials and Methods: It recruited consecutive patients undergoing instrumented fusion from L1 to the sacrum for degenerative lumbar disease between June 2006 and November 2019 in a single institution. The patients' preoperative clinical data, muscle status at T12-L1 on magnetic resonance images, and sagittal spinopelvic parameters were analyzed. Univariate analysis was used to compare clinical and radiographic data between PJK and non-PJK patients. Logistic regression analysis was used to investigate the independent risk factors for PJK. Results: A total of 56 patients were included in this study. The mean age at surgery was 67.3 years and mean follow-up period was 37.3 months. In total, 10 were male and 46 were female. PJK developed in 23 (41.1%) out of 56; of these patients, 20 (87.0%) developed PJK within 1 year postoperatively. In the univariate analysis between PJK and non-PJK patients, the PJK group showed more frequent osteoporosis, lower body mass index, smaller cross-sectional area (CSA) and more fat infiltration (FI) in erector spinae muscle at T12-L1 and larger preoperative TLK and PT with statistical significance (p < 0.05). In the logistic regression analysis, severe (>50%) FI in erector spinae muscle (OR = 43.60, CI 4.10-463.06, R2N = 0.730, p = 0.002) and osteoporosis (OR = 20.49, CI 1.58-264.99, R2N = 0.730, p = 0.021) were statistically significant. Conclusions: Preexisting severe (>50%) fat infiltration in the erector spinae muscle and osteoporosis were independent risk factors associated with PJK following instrumented fusion from L1 to the sacrum, but age was not a risk factor.

Comparative study of non-suppurative meningoencephalitis in cattle in Southern Brazil.

Viral neurologic diseases are common in cattle, although most non-suppurative meningoencephalitis (NSM) remains etiologically unknown. We compared the epidemiological, clinical, and pathological data among 79 cases of rabies, 12 cases of NSM of unknown etiology (NSM-UE), and 8 cases of herpetic meningoencephalitis previously diagnosed in cattle in Southern Brazil. Neurological clinical signs were similar among rabies and NSM-UE and different in cattle with herpetic meningoencephalitis. Only two herpetic meningoencephalitis cases had gross lesions in the central nervous system, characterized by malacia and hemorrhage. Histologically, all three groups had mild to severe multifocal infiltrates of lymphocytes, plasma cells, and macrophages/microglial cells in the Virchow-Robin space, neuropil, and leptomeninges, and gliosis. Other findings included malacia and eosinophilic intracytoplasmic inclusion in rabies, and malacia and intranuclear amphophilic inclusion in herpetic meningoencephalitis. By immunohistochemistry, the predominant inflammatory cells in all cases were T lymphocytes, followed by macrophages/microglial cells, B lymphocytes, and astrocytes. The T lymphocyte count showed statistically significant differences between the diseases. Our results revealed few differences between the groups. Although the etiological agent involved has not been identified in cases of NSM-UE, the characteristics observed in this study showed similarity with viral diseases.

SHC1 serves as a prognostic and immunological biomarker in clear cell renal cell carcinoma: a comprehensive bioinformatics and experimental analysis

SHC1 plays a crucial regulatory role in various tumors, but its significance in predicting prognosis and immune response in clear cell renal cell carcinoma (ccRCC) is yet to be determined. In this study, we conducted a bioinformatics analysis of SHC1 expression, prognosis, and immunological functions in ccRCC using multiple databases. The association between SHC1 and immune infiltration, immune escape, and immunotherapy in ccRCC was systematically established. In addition, we validated our results by western blot of tumor and adjacent-tumor samples from nine ccRCC patients, as well as three renal carcinoma cell lines compared to a normal renal cell line. Our analysis revealed that the mRNA expression level of SHC1 in ccRCC tissues is significantly higher than that in normal tissues. Consistently, western blot experiment showed ccRCC tissues and cell lines exhibit higher protein levels that normal tissues and cell lines. Importantly, patients with low expression of SHC1 demonstrated a higher survival rate, indicating that SHC1 could serve as an independent prognostic factor for predicting survival in ccRCC. Additionally, high expression of SHC1 was associated with increased severe immune cell infiltration, enhanced immune escape, and higher immunotherapy scores. Hence, SHC1 emerges as a novel and easily detectable biomarker for predicting clinical outcomes, immune escape, and immunotherapy response in patients with ccRCC.

The Role of Paraspinal Muscle in Postoperative Coronal Balance Transition in Degenerative Lumbar Scoliosis: A 2-year Follow-up Study.

Retrospective observational study. The purpose of this study was to determine whether paraspinal muscle could influence postoperative coronal balance and its transition in degenerative lumbar scoliosis (DLS). Although the importance of the paraspinal muscles (PSM) in sagittal alignment is well recognized, there is no information about its role in coronal balance. The study included 102 DLS patients. Evaluation of the PSM on magnetic resonance imaging were conducted at baseline. Coronal measurements included coronal balance distance (CBD), major Cobb angle, L4 coronal tilt, and L5 coronal tilt. The cohort was divided based on postoperative parameters into persistent coronal balance (PCB), worsened coronal imbalance (WCIB), recurrent coronal balance (RCB), and persistent coronal imbalance (PCIB) according to immediate postoperative and follow-up coronal balance. Multivariate logistic regression models for postoperative CIB, follow-up WCIB and follow-up RCB were utilized to identify statistically significant associations while accounting for confounders. The cohort was divided into 57 with PCB, 13 with WCIB, 10 with RCB, and 22 with PCIB. The follow-up groups with CIB exhibited more severe fatty infiltration in the extensor muscle compared to the balanced groups. Specifically, the WCIB group demonstrated the most severe extensor muscle degeneration, particularly on the concave sides, and the most prominent asymmetrical degeneration of the PSM among the four groups. Furthermore, patients with CIB had worse sagittal malalignment compared to those with CB at the last follow up. Patients exhibiting stronger extensor muscle mass were prone to immediate postoperative CB and more likely to experience spontaneous improvement or recurrence of coronal balance during follow-up. Severe extensor muscle degeneration and prominent asymmetrical bilateral PSM degeneration represent potential risk factors for persistent CIB and recurrent CIB. It is crucial to assess the dynamic change during the follow-up period as long-term prognosis may be impacted if CB deteriorates, or otherwise develops during follow-up. 3.

Structure-dependent destructive adsorption of organophosphate flame retardants on lipid membranes

The widespread use of organophosphate flame retardants (OPFRs), a serious type of pervasive environmental contaminants, has led to a global concern regarding their diverse toxicities to living beings. Using a combination of experimental and theoretical approaches, we systematically studied the adsorption, accumulation, and influence of a series of OPFRs on the lipid membranes of bacteria and cells. Our results revealed that OPFRs can aggregate in lipid membranes, leading to the destruction of membrane integrity. During this process, the molecular structure of the OPFRs is a dominant factor that significantly influences the strength of their interaction with the lipid membrane, resulting in varying degrees of biotoxicity. Triphenyl phosphate (TPHP), owing to its large molecular size and strong hydrophobicity, causes severe membrane disruption through the formation of nanoclusters. The corresponding severe toxicity originates from the phase transitions of the lipid membranes. In contrast, smaller OPFRs such as triethyl phosphate (TEP) and tris(2-chloroethyl) phosphate (TCEP) have weaker hydrophobicity and induce minimal membrane disturbance and ineffective damage. In vivo, gavage of TPHP induced more severe barrier damage and inflammatory infiltration in mice than TEP or TCEP, confirming the higher toxicity of TPHP. Overall, our study elucidates the structure-dependent adsorption of OPFRs onto lipid membranes, highlighting their destructive interactions with membranes as the origin of OPFR toxicity.

More severe parotid gland histopathology in paediatric-onset than in adult-onset Sjögren’s disease

ObjectivesThe aim of this study was to assess the histopathological features of the parotid glands in patients with paediatric-onset Sjögren’s disease (pedSjD) in comparison to patients with adult-onset Sjögren’s disease...

Estrogen Deficiency Exacerbates Traumatic Heterotopic Ossification in Mice.

Traumatic heterotopic ossification (HO) is a devastating sequela of orthopedic surgeries and traumatic injuries; however, few studies have explored the effects of the estrogen-deficient state on HO formation. In the present study, we investigated the impact of estrogen deficiency on ectopic cartilage and bone formation in tendon after Achilles tenotomy in an ovariectomized mouse model. A total of 45 female C57BL/6 mice were randomly divided into three groups: sham-operated (control), estrogen depletion by ovariectomy (OVX) and OVX with 17β-estradiol supplementation (OVX + E2), with 15 animals in each group. Three weeks after OVX, all mice were subjected to an Achilles tenotomy using a posterior midpoint approach to induce HO. At 1, 3 and 9 weeks after tenotomy, the left hind limbs were harvested for histology, immunohistochemistry and immunofluorescence evaluations. The volume of ectopic bone was assessed by micro-CT. Mice in the OVX group formed more ectopic cartilage 3 weeks after tenotomy, as well as ectopic bone 9 weeks after tenotomy, compared to the control group. Estrogen deficiency resulted in more severe inflammatory infiltration at the injury sites 1 week after tenotomy, involving the recruitment of more macrophages and mast cells, as well as increasing the expressions of pro-inflammatory mediators, including IL-1β, IL-6, and TNF-α. Moreover, the local TGF-β/SMAD signaling pathway was dysregulated after OVX, which manifested as upregulated expressions of TGF-β and pSMAD2/3. E2 supplementation protected against OVX-induced HO deterioration, inhibited inflammatory infiltration, and downregulated the TGF-β/SMAD signaling pathway. Estrogen deficiency exacerbated HO formation in the Achilles tenotomy model. These findings might be attributable to the disturbance of the inflammatory response and the activation of TGF-β/SMAD signaling at the injury sites during the early stages of HO development.

Effects of Aeromonas infection on the immune system, physical barriers and microflora structure in the intestine of juvenile grass carp (Ctenopharyngodon idella)

Grass carp (Ctenopharyngodon idella) is an intensively cultured and economically important herbivorous fish species in China, but its culture is often impacted by Aeromonas pathogens such as Aeromonas hydrophila and Aeromonas veronii. In this study, healthy grass carp were separately infected with A. hydrophila or A. veronii for 12, 24, 48 or 72 h. The results showed that the mRNA expression levels of intestinal inflammatory factors (tnf-α, il-1β and il-8), complement factors (c3 and c4), antimicrobial peptides (hepcidin, nk-lysin and β-defensin-1), immunoglobulins (igm and igt), and immune pathway-related signaling molecules (tlr1, tlr2, tlr4, myd88, irak4, irak1, traf6, nf-κb p65 and ap-1) were differentially upregulated in response to A. hydrophila and A. veronii challenge. Additionally, the expression levels of the intestinal pro-apoptotic genes tnfr1, tnfr2, tradd, caspase-8, caspase-3 and bax were significantly increased, whereas the expression of the inhibitory factor bcl-2 was significantly downregulated, indicating that Aeromonas infection significantly induced apoptosis in the intestine of grass carp. Moreover, the expression of intestinal tight junction proteins (occludin, zo-1, claudin b and claudin c) was significantly decreased after infection with Aeromonas. Histopathological analysis indicated the Aeromonas challenge caused severe damage to the intestinal villi with adhesions and detachment of intestinal villi accompanied by severe inflammatory cell infiltration at 12 h and 72 h. The 16S rRNA sequencing results showed that Aeromonas infection significantly altered the structure of the intestinal microflora of the grass carp at the phylum (Proteobacteria, Fusobacteria, Bacteroidetes and Firmicutes) and genus (Proteus, Cetobacterium, Bacteroides, and Aeromonas) levels. Take together, the findings of this study revealed that Aeromonas infection induces an intestinal immune response, triggers cell apoptosis, destroys physical barriers and alters microflora structure in the intestine of juvenile grass carp; the results will help to reveal the pathogenesis of intestinal bacterial diseases in grass carp.

Clinical and pathological characteristics of OPDM4 patients in advanced disease.

Oculopharyngodistal myopathy type 4 (OPDM4) arises from a CGG repeat expansion in the 5' UTR of the RILPL1 gene. Reported cases of OPDM4 have been limited. The aim of this study was to investigate the clinical and myopathological characteristics of OPDM4 patients with advanced disease. We assessed a total of 8 affected and 12 unaffected individuals in an OPDM4 family with autosomal dominant inheritance. Muscle biopsy tissue from the proband underwent histological, enzyme histochemical, and immunohistochemical stains, and electron microscopy analysis. Whole exome sequencing and repeat primer PCR (RP-PCR) were conducted to investigate underlying variants. OPDM4 patients displayed a progressive disease course. Most experienced lower limb weakness and diminished walking ability in their 20s and 30s, followed by ptosis, ophthalmoplegia, swallowing difficulties, and dysarthria in their 30s to 50s, By their 50s to 70s, they became non-ambulatory. Muscle magnetic resonance imaging (MRI) of the proband in advanced disease revealed severe fatty infiltration of pelvic girdle and lower limb muscles. Biopsied muscle tissue exhibited advanced changes typified by adipose connective tissue replacement and the presence of multiple eosinophilic and p62-positive intranuclear inclusions. Immunopositivity for the intranuclear inclusions was observed with anti-glycine antibody and laboratory-made polyA-R1 antibody. RP-PCR unveiled an abnormal CGG repeat expansion in the 5' UTR of the RILPL1 gene. The clinical and radiological features in this family broaden the phenotypic spectrum of OPDM4. The presence of intranuclear inclusions in the proliferative adipose connective tissues of muscle biopsy specimens holds diagnostic significance for OPDM4 in advanced disease.

The Impact of Pentraxin 3 Serum Levels and Angiotensin-Converting Enzyme Polymorphism on Pulmonary Infiltrates and Mortality in COVID-19 Patients.

The aim of this study was to examine the impact of the pentraxin 3 (PTX3) serum level and angiotensin-converting enzyme (ACE) gene insertion/deletion (I/D) polymorphism on the severity of radiographic pulmonary infiltrates and the clinical outcomes of COVID-19. The severity of COVID-19 pulmonary infiltrates was evaluated within a week of admission by analyzing chest X-rays (CXR) using the modified Brixia (MBrixa) scoring system. The insertion (I)/deletion (D) polymorphism of the ACE gene and the serum levels of PTX3 were determined for all patients included in the study. This study included 80 patients. Using a cut-off serum level of PTX3 ≥ 2.765 ng/mL, the ROC analysis (AUC 0.871, 95% CI 0.787-0.954, p < 0.001) showed a sensitivity of 85.7% and specificity of 78.8% in predicting severe MBrixa scores. Compared to ACE I/I polymorphism, D/D polymorphism significantly increased the risk of severe CXR infiltrates, OR 7.7 (95% CI: 1.9-30.1), and p = 0.002. Significant independent predictors of severe CXR infiltrates include hypertension (OR 7.71), PTX3 (OR 1.20), and ACE D/D polymorphism (OR 18.72). Hypertension (OR 6.91), PTX3 (OR 1.47), and ACE I/I polymorphism (OR 0.09) are significant predictors of poor outcomes. PTX3 and ACE D/D polymorphism are significant predictors of the severity of COVID-19 pneumonia. PTX3 is a significant predictor of death.