IR Group Research Articles

Rhabdomyosarcoma of the Middle Ear Case Report

Background: Rhabdomyosarcoma (RMS) is a highly malignant soft tissue tumor derived from primitive embryonal mesenchymal tissue that differentiates into striated skeletal muscle. Despite the improved outcome based on the EFS and OS using the three different treatment modalities-chemotherapy, radiotherapy and surgical treatment, the survival of patients depends on their IRS groups—pathological and surgical. On the other hand in the last thirty years a great improvement of the five-year overall survival (OS) of children with RMS have been observed based on the results of large multinational collaborative trials and successive studies dedicated to children, though prognosis is variable and dependent on several factors including histologic variant, primary sites of the tumor, extent of disease (disease resectability), and molecular-level characteristics. Case presentation: We present a clinical case of a five-year-old male with initial complains of left side peripheral facial nerve palsy and secondary cervical and retroauricular lymphadenomegaly. After an exam, surgery of the temporal bone, CT and MRI embryonal type of rhabdomyosarcoma was diagnosed, and adjuvant chemotherapy was initiated in combination with concomitant local radiotherapy. Results: The results show that in these areas surgery itself is insufficient for RMS treatment(usually it is limited to taking a biopsy only). The combination of chemotherapy and local control with radiotherapy achieved a good result in our patient. Conclusions: Middle ear Embryonal Rhabdomyosarcoma is a common solid tumor, which could mimic middle ear inflammation or mastoid inflammation in patients. The multimodal approach seemed to be the ideal management of RMS. It involves a combination of chemotherapy and local control with surgery and/or radiotherapy.

Huaxian formula prevents the progression of radiation-induced pulmonary fibrosis by inhibiting the pro-fibrotic effects of macrophages

Ethnopharmacological relevanceThe Huaxian formula (HXF), a traditional Chinese medicine (TCM) remedy, specifically targets the pathological factors of “heat toxicity” and “phlegm stasis” induced by radiation in radiation-induced pulmonary fibrosis (RIPF). It works by clearing heat and invigorating the blood, addressing these key factors in the development of RIPF. Aim of the studyThe HXF has demonstrated potential in preventing RIPF, although its underlying mechanisms remain unclear. This study aims to investigate the efficacy, molecular targets, and mechanisms of action of HXF. Materials and methodsThe major constituents of the HXF were identified by ultra performance liquid chromatography and tandem mass spectrometry (UPLC-MS). C57BL/6j mice were divided into four groups: control (Ctrl), HXF alone (HXF), 17Gy-irradiation alone (IR), and irradiation plus HXF (IR + HXF). Lung damage and fibrosis were assessed by histopathological staining, and the flow cytometry and immunohistochemistry (IHC) were used to detect the macrophages phenotype of lung tissues in vivo at 16 weeks post-irradiation. Transcriptomic sequencing and bioinformatics analyses identified key genes modulated by HXF. In vitro assays included flow cytometry, western bolt, and quantitative PCR (qPCR) explored the impact of HXF on macrophage polarization and fibrotic activity, while co-culture experiments of the macrophage conditional medium and mouse embryo fibroblast NIH/3T3 investigated macrophage-fibroblast interactions. Results20 major constituents of HXF were identified. And the in vivo results revealed significant lung damage and fibrosis in the IR group, which were notably mitigated in the IR + HXF group. And HXF has been shown to significantly inhibit the infiltration of M2-type macrophages in lung tissues. Transcriptomic analysis identified differentially expressed genes (DEGs) such as Arg1, Mmp10, and Fgf23. Bioinformatics enrichment analysis indicated that these DEGs are involved in pathways related to the inhibition of extracellular matrix formation and inflammation. In vitro, HXF-containing serum reduced M2-type macrophage polarization and decreased the secretion of Arginase1 and TGFβ1. Conditioned medium from HXF-treated macrophages suppressed fibroblast activation. ConclusionHXF's preventive effects on RIPF involve multiple targets and mechanisms, including the modulation of Arg1, Mmp10, and Fgf23 expression. By inhibiting the pro-fibrotic capacity of macrophages, HXF suppresses fibroblast activation and collagen production, thereby alleviating lung fibrosis. These findings underscore the potential of HXF as a preventive strategy in managing RIPF.

NaHS protects brain, heart, and lungs as remote organs from renal ischemia/reperfusion-induced oxidative stress in male and female rats

Acute Kidney Injury (AKI) is frequently observed in hospitalized patients in intensive care units, often caused by renal ischemia-reperfusion injury (IRI). IRI disrupts the function of various ‘remote organs’ such as the lungs, pancreas, intestine, liver, heart, and brain through inflammation, oxidative stress, apoptosis, leukocyte infiltration, and increased urea and creatinine levels. Gender differences in renal IRI-induced injury are noted. H2S, an endogenous gaseous modulator, shows potential in vasodilation, bronchodilation, and hypotension and can regulate apoptosis, inflammation, angiogenesis, metabolism, and oxidative stress. This study aims to investigate the protective effects of NaHS on brain, heart, and lung injuries following renal IR and to assess the oxidative system status as a potential mechanism in male and female rats.Forty-eight Wistar rats were randomly divided into eight groups (n = 6): Control/Saline, Sham/Saline, IR/Saline, and IR/NaHS in both sexes. Forty-five minutes of bilateral renal ischemia followed by 24-hour reperfusion was induced in the IR groups. NaHS (100µM/Kg, IP) was administered 10 min before clamp release in treated groups. BUN, SCr, BUN/SCr, albuminuria, histopathology, and oxidative stress biomarkers of the brain, heart, and lung were assessed as remote organs. IR increased serum markers of renal function, albuminuria, malondialdehyde levels, and tissue injury scores while reducing nitrite levels and superoxide dismutase and glutathione peroxidase activities. NaHS treatment reversed the adverse effects of IR in remote organs in both sexes, although it showed limited improvement in renal function. Our findings demonstrate that NaHS has a beneficial effect on remote organ injury following renal IR by mitigating oxidative stress, with noted tissue-specific and gender-specific differences in response. These findings suggest NaHS as a potential therapeutic agent for mitigating multi-organ injury after renal IR, with effects varying by tissue and gender.

Circulating extracellular vesicle-carried PTP1B and PP2A phosphatases as regulators of insulin resistance.

Metabolic disorders associated with abdominal obesity, dyslipidaemia, arterial hypertension and hyperglycaemia are risk factors for the development of insulin resistance. Extracellular vesicles (EVs) may play an important role in the regulation of metabolic signalling pathways in insulin resistance and associated complications. Circulating large EVs (lEVs) and small EVs (sEVs) from individuals with (IR group) and without insulin resistance (n-IR group) were isolated and characterised. lEVs and sEVs were administered by i.v. injection to mice and systemic, adipose tissue and liver insulin signalling were analysed. The role of phosphatases was analysed in target tissues and cells. Injection of lEVs and sEVs from IR participants impaired systemic, adipose tissue and liver insulin signalling in mice, while EVs from n-IR participants had no effect. Moreover, lEVs and sEVs from IR participants brought about a twofold increase in adipocyte size and adipogenic gene expression. EVs from IR participants expressed two types of phosphatases, phosphotyrosine 1 phosphatase (PTP1B) and protein phosphatase 2 (PP2A), IR lEVs being enriched with the active form of PTP1B while IR sEVs mainly carried active PP2A. Blockade of PTP1B activity in IR lEVs fully restored IRS1 and Akt phosphorylation in adipocytes and blunted insulin-induced Akt phosphorylation by inhibition of the macrophage secretome in hepatocytes. Conversely, blockade of PP2A activity in IR sEVs completely prevented insulin resistance in adipocytes and hepatocytes. These data demonstrate that inhibition of phosphatases carried by EVs from IR participants rescues insulin signalling in adipocytes and hepatocytes and point towards PTP1B and PP2A carried by IR EVs as being novel potential therapeutic targets against insulin resistance in adipose tissue and liver and the development of obesity.

The association of plasma asprosin with anthropometric and metabolic parameters in Korean children and adolescents.

This study aimed to determine the correlation of plasma asprosin with anthropometric and metabolic parameters in Korean children and adolescents. This single-center study included 109 Korean children and adolescents: 62 (56.9%) obese participants with a body mass index (BMI) ≥95th percentile and 47 (43.1%) healthy controls with BMI between the 15th and 85th percentile. Metabolic parameters were measured, including fasting blood glucose, fasting insulin, homeostasis model assessment of insulin resistance (HOMA-IR), triglyceride and glucose (TyG) index, and lipid profiles. Plasma asprosin levels were higher in the obese group than in the control group (mean 87.0 vs. 69.3 ng/mL; p = 0.001) and in the IR group than in the non-IR group (mean 98.6 vs. 70.2 ng/mL; p < 0.001). Plasma asprosin levels were not associated with sex or pubertal stage. Plasma asprosin levels were positively correlated with BMI SDS (r = 0.34; p = 0.002), glycated hemoglobin (HbA1c) (r = 0.25; p = 0.02), glucose (r = 0.33; p = 0.002), insulin (r = 0.44; p < 0.001), HOMA-IR (r = 0.47; p < 0.001), triglyceride (TG) (r = 0.33; p = 0.003), high-density lipoprotein (HDL) cholesterol (r = -0.29; p = 0.008), and TyG index (r = 0.38; p < 0.001). Multiple linear regression analysis indicated that plasma asprosin levels were independently associated with HOMA-IR (p < 0.001) and TG/HDL cholesterol ratio (p < 0.001). This study demonstrated an association between plasma asprosin levels and obesity and insulin resistance in Korean children and adolescents.

La reparación aislada presenta mayor altura patelar postoperatoria según el índice de Insall-Salvati en comparación con la aumentación biológica en roturas agudas del tendón rotuliano

Background and objectivePatellar tendon ruptures (PTR) are rare but highly disabling injuries. Surgical treatment is the standard approach. Postoperative patellar height (PH) is used to determine the success of the intervention, with various indices described in the literature. The objective of the study was to determine if there are differences in PH according to the Insall-Salvati index in the immediate postoperative period and at 4 months, between two groups of patients operated on for acute PTR: The first group underwent isolated repair techniques (IR) and the second group had biological augmentation with autograft or allograft added to the repair (BAR). Our hypothesis was that the BAR group would have lower PH in the immediate postoperative period and at 4 months. Materials and methodsAll patients with acute PTR treated at our center between 2016 and 2022 were reviewed retrospectively. Patients with re-rupture of the operated tendon and those with non-biological augmentation were excluded. PH according to the Insall-Salvati index was measured in radiographs taken the day after surgery and at 4 months postoperatively by three orthopedic surgeons with a subspecialty in knee surgery, independently. A fourth blind evaluator analyzed the measurement results, determining the average PH and the intraclass correlation coefficient for absolute agreement among the three evaluators. The primary outcome was the difference in PH between both groups the day after surgery and at 4 months. The secondary outcome was the difference in PH between the immediate postoperative measurement and at 4 months in both groups. ResultsThis was a retrospective observational study. Thirty-one operated knees were included. IR was performed on 15 knees (48.38%), and biological augmentation was added in 16. The intraclass correlation coefficient between observer 1 and 2 was 0.88; between 2 and 3 it was 0.89; and between observers 1 and 3 it was 0.92, showing at least substantial agreement among observers. The average Insall-Salvati index the day after surgery and at 4 months was 1.15 and 1.21 for the IR group, and 0.97 and 1.07 for the BAR group, respectively. A statistically significant difference in the Insall-Salvati index was found between the two groups, both in the measurement taken the day after surgery (p<0.0001) and at 4 months (p=0.0002). Additionally, significant differences were observed indicating an increase in average PH when comparing the immediate postoperative measurement and at 4 months in each study group (for IR p=0.0016 and for BAR p<0.0001). ConclusionsThe Insall-Salvati index was significantly lower in BAR cases both the day after surgery and at 4 months postoperatively. Additionally, a significant increase in PH was observed at 4 months, regardless of the surgical technique used. Substantial inter-observer agreement was achieved for PH measurement in this cohort.

Oleuropein Relieves Pancreatic Ischemia Reperfusion Injury in Rats by Suppressing Inflammation and Oxidative Stress through HMGB1/NF-κB Pathway.

Oleuropein (OLP) is a naturally occurring phenolic compound in olive plant with antioxidant and anti-inflammatory potential and can possibly be used in treating pancreatic injuries. This investigation aimed to follow the molecular mechanism behind the potential therapeutic effect of OLP against pancreatic injury persuaded by ischemia-reperfusion (I/R). Pancreatic I/R injury was induced by splenic artery occlusion for 60 min followed by reperfusion. Oral administration of OLP (10 and 20 mg/kg) for 2 days significantly alleviated I/R-persuaded oxidative damage and inflammatory responses in pancreatic tissue as indicated by the decreased malondialdehyde (MDA) content and increased glutathione peroxidase (GPx) activity, accompanied by the suppression of myeloperoxidase (MPO) activity and reduced levels of interleukin-1beta (IL-1β), nuclear factor kappa B (NF-κB), and tumor necrosis factor alpha (TNF-α) in pancreatic tissues. Furthermore, OLP treatment markedly restored the serum levels of amylase, trypsinogen-activated peptide (TAP), and lipase, with concurrent improvement in pancreatic histopathological alterations. Moreover, treatment with OLP regulated the pancreatic expression of inducible nitric oxide synthase (iNOS) and high-mobility group box 1 (HMGB1) relative to rats of the pancreatic IR group. Thus, OLP treatment significantly alleviates the I/R-induced pancreatic injury by inhibiting oxidative stress and inflammation in rats through downregulation of HMGB1 and its downstream NF-κB signaling pathway.

21-gene recurrence score predictive of the benefit of postoperative radiotherapy after breast-conserving surgery for elderly patients with T1N0 and luminal breast cancer.

To assess the predictive value of the 21-gene recurrence score (RS) on the survival outcomes of postoperative radiotherapy (PORT) in elderly patients with T1N0 luminal breast cancer after breast-conserving surgery. We retrospectively included patients aged ≥ 70years and diagnosed with T1N0 luminal BC between 2004 and 2015 using the data from the Surveillance, Epidemiology, and End Results. The RS groups were categorized using the TAILORx criteria as follows: low risk (RS < 11) (LR), intermediate risk (RS 11-25) (IR), and high risk (RS > 25) (HR). Kaplan-Meier analysis, propensity score matching (PSM), and Cox proportional hazards analysis were used for statistical analysis. We included 5901 patients in the analysis. Of the patients, 4492 (76.1%) underwent PORT, while 1409 (23.9%) did not receive PORT. There were 1588 (26.9%), 3613 (61.2%), and 700 (12.0%) patients classified as LR, IR, and HR, respectively. There were 1182 (74.4%), 2773 (76.8%), and 537 (76.7%) patients in the LR, IR, and HR groups receiving PORT, respectively (P = 0.182). A total of 1353 pairs of patients were completely matched using PSM. PORT was independently associated with better overall survival (OS) (P < 0.001) and breast cancer-specific survival (BCSS) (P = 0.015) in the entire cohort. The sensitivity analyses showed that the receipt of PORT was not associated with OS (P = 0.887) and BCSS (P = 0.861) in the LR group. However, the receipt of PORT was associated with OS (P < 0.001) and BCSS in the IRHR group (P = 0.026). Our study highlights the possible role of the 21-gene RS in predicting the survival outcomes of PORT following BCS in elderly patients with T1N0 luminal breast cancer.

Outcomes for patients with perineal and perianal rhabdomyosarcoma: A report from the Children's Oncology Group Soft Tissue Sarcoma Committee.

To describe clinical features, risk factors, and outcomes of patients with perineal and perianal rhabdomyosarcoma. The records of 51 patients (38 perineal and 13 perianal) enrolled on Children's Oncology Group clinical trials between 1997 and 2012 were reviewed. At presentation, 53% were female, 65% were older than 10years of age, 76% were alveolar histology, 76% were more than 5cm, 84% were invasive, 65% were regional node positive by imaging, 49% were metastatic, only 16% were grossly resected upfront, and 25% of patients had a delayed excision. At a median follow-up of 6.13years, estimated 5-year event-free survival (EFS) was 38% [22.17%-53.38%], and overall survival (OS) was 42% [26.66%-58.21%]. The rates of local, regional, and distant failure were 15.6%, 13.7%, 43.1%, respectively; all failures ultimately died. By univariate analysis, only age more than 10years negatively impacted 5-year EFS (p=.023) and OS (p=.09), and IRS Group also impacted OS (p=.043). In Cox proportional hazards model, neither of these variables were significant after adjusting for other factors. Patients with perineal and perianal rhabdomyosarcoma have a poor overall prognosis, probably related to poor patient and disease characteristics at presentation.

G-tube placement in patients with robin sequence undergoing mandibular distraction osteogenesis: A multi-institutional review

The goal of this study was to investigate the relationship between mandibular distraction osteogenesis (MDO) and rates of postoperative gastrostomy tube (G-tube) placement among patients with isolated and syndromic Robin sequence (RS). This study was a multi-institutional retrospective chart review of patients with RS who underwent MDO at one of three different pediatric tertiary medical centers. The primary aim of the study was to compare rates of G-tube placement following MDO among the three institutions. The primary outcome was analyzed using Fischer's exact test. The secondary aim of the study was to assess for other contributing factors to G-tube placement such as demographic differences, length of hospital stay, and age at MDO. Analysis of secondary outcomes was assessed using multiple logistic regression models. A total of 125 patients met the inclusion criteria, which required RS diagnosis, completion of MDO between 2004 and 2019, and adequate medical record availability. Sixty percent (n = 75) of subjects were categorized as isolated RS (iRS) and forty percent (n = 50) as syndromic RS (sRS). After MDO, 20% (n = 25) of all patients had G-tubes placed. Of the iRS group, 14.7% (n = 11) required a G-tube, while 28% (n = 14) of the sRS group required a G-tube. The post-operative G-tube rate was similar between institutions when considering all patients. When considering only those patients with iRS, the post-MDO G-tube rate at one center was significantly higher than the other two. Overall, most patients with RS did not require a G-tube after MDO, regardless of diagnosis. However, the significant differences in rates of G-tube placement among patients with iRS may indicate differing practice philosophies, surgical protocols, thresholds for G-tube placement, or regional influences between institutions.

Imaging the Area of Internal Limiting Membrane Peeling after Macular Hole Surgery.

Background: The aim of this study was to compare en-face optical coherence tomography (OCT) imaging and confocal scanning laser ophthalmoscopy (cSLO) imaging at different wavelengths to identify the internal limiting membrane (ILM) peeling area after primary surgery with vitrectomy and ILM peeling for macular hole (MH). Methods: In total, 50 eyes of 50 consecutive patients who underwent primary surgery with vitrectomy and ILM peeling for MH were studied. The true ILM rhexis based on intraoperative color fundus photography was compared to the presumed ILM rhexis identified by a blinded examiner using en-face OCT imaging and cSLO images at various wavelengths. To calculate the fraction of overlap (FoO), the common intersecting area and the total of both areas were measured. Results: The FoO for the measured areas was 0.93 ± 0.03 for en-face OCT, 0.76 ± 0.06 for blue reflectance (BR; 488 nm), 0.71 ± 0.09 for green reflectance (GR; 514 nm), 0.56 ± 0.07 for infrared reflectance (IR; 815 nm) and 0.73 ± 0.06 for multispectral (MS). The FoO in the en-face OCT group was significantly higher than in all other groups, whereas the FoO in the IR group was significantly lower compared to all other groups. No significant differences were observed in FoO among the MS, BR, and GR groups. In en-face OCT, there was no significant change in the ILM peeled area measured intraoperatively and postoperatively (8.37 ± 3.01 vs. 8.24 ± 2.81 mm2; p = 0.8145). Nasal-inferior foveal displacement was observed in 38 eyes (76%). Conclusions: En-face OCT imaging demonstrates reliable postoperative visualization of the ILM peeled area. Although the size of the ILM peeling remains stable after one month, our findings indicate a notable inferior-nasal shift of the overall ILM peeling area towards the optic disc.

Outcome of children and young adults with localized extremity rhabdomyosarcoma treated on Children's Oncology Group trials.

10006 Background: Extremity rhabdomyosarcoma (RMS) accounts for 15-20% of patients with localized RMS. The long-term survival and associated prognostic factors of extremity RMS patients treated on the contemporary Children’s Oncology Group (COG) trials have not been reported. Methods: Patients with localized extremity RMS enrolled on the D9602, D9803, ARST0331 and ARST0531 COG trials were analyzed. The Kaplan-Meyer method estimated event-free (EFS) and overall survival (OS). The Log-rank test and Cox regression were used to estimate the prognostic significance of age at diagnosis, sex, race, tumor site &amp; size, IRS group, stage, histology, FOXO1 fusion status, anaplasia, lymph node involvement, delayed primary excision (DPE) and radiation on survival. Results: We identified 159 patients with localized extremity RMS; 108 (68%) were &lt; 10 years at diagnosis, 84(53%) had primary tumor &gt; 5 cm, and 35 (22%) and 98 (62%) had IRS group II and III disease. Histology was alveolar in 106 (67%), and 43 (27%) had regional lymph node involvement on imaging or pathology. Seventeen were treated on low-risk (LR) studies D9602 (N = 10) and ARST0331 (N = 7), whereas 142 were treated on intermediate-risk (IR) trials D9803 (N = 85) and ARST0531 (N = 57). Eighty patients (50%) underwent primary surgical excision and 121 (76%) received radiotherapy. DPE was attempted in 60 patients (38%), resulting in an R0 (microscopically negative) margin in 48 (80%). Relapse/progression occurred in 52 patients: metastatic 26 (50%), local 22 (42%), and combined 3(6%). The 5-year EFS and OS of the whole cohort were 62.6% (95% CI: 54%-71.2%) and 78.7%% (95%CI: 71.4%-85.9%). LR patients treated on D9602 and ARST0331 trials had comparable 5-year EFS (70% vs. 71.4%, p-0.90) and OS (90% vs. 85.7%, p-0.54), respectively, while IR patients had significantly better 5-year EFS (68.9% vs. 51%, p-0.023) and OS (86.2% vs. 64.4%, p-0.007) on D9803 trial than ARST0531 trial. The 5-year EFS and OS of IRS group III patients with DPE with R0 margins were comparable to IRS group II patients (70.4% vs. 66.6%) &amp; (86.5% vs. 84.6%), respectively. On univariable analysis, age at diagnosis and sex were associated with EFS, while age at diagnosis, sex, tumor size, IRS group, stage and fusion status were associated with OS (p &lt; 0.05). None of the analyzed factors demonstrated independent statistical association with EFS, whereas female sex was the only factor independently associated with OS [Hazard ratio (HR): 0.4, 95% CI 0.20-0.85)]. Conclusions: IR localized extremity RMS patients treated on ARST0531 had suboptimal outcome. Survival of IRS group III patients with R0 DPE was comparable to IRS group II patients. Females with localized extremity RMS had better OS than males. Clinical trial information: NCT00354835 ; NCT00003958 ; NCT00075582 ; NCT00002995 .

Effects of fingolimod on heart injury induced by renal ischemia-reperfusion

Background. Renal ischemia-reperfusion injury (IRI) is one of the inevitable complications of surgery. The evidence shows that fingolimod, with its anti-inflammatory effects, can play a protective role in renal IRI. The main aim of the present study was to investigate the role of fingolimod against renal IRI in the heart tissue. Methods. Twenty-four male Wistar rats (220±20g) were treated with a single dose of fingolimod (1mg/kg) by intraperitoneal injection before the induction of kidney IRI. At the end of the reperfusion period, the oxidative stress biomarker (malondialdehyde) and antioxidant biomarkers (catalase, superoxide dismutase, glutathione, glutathione peroxidase, and total antioxidant capacity) were evaluated in the heart tissue. Results. Fingolimod pretreatment could increase cardiac glutathione enzyme activity in the fingolimod+IR group compared to the IR group, which was not statistically significant (P&gt;0.05). The level of total antioxidant capacity in the heart tissue was also significantly increased in the fingolimod+IR group in comparison to the IR group (P&lt;0.05). Conclusion. Fingolimod was able to prevent oxidative stress damage in the heart caused by kidney IR induction by increasing the level of total antioxidant capacity of the heart tissue. It is suggested that future studies evaluate the effects of this drug in clinical trials. Practical Implications. The total antioxidant capacity of the heart, as a distant organ affected by kidney IRI, is increased following fingolimod pre-treatment.

Correlation of Oral Microbiota With Different Immune Responses to Antiretroviral Therapy in People Living With HIV

Abstract Both HIV infection and antiretroviral therapy (ART) affect the oral microbiome. Whether successful treatment with ART in people living with HIV (PLWH), which leads to a significant decline in viral loads and immune reconstitution, is associated with changes in or recovery of the oral microbiome remains unknown. Therefore, we performed a cross-sectional study of 118 PLWH receiving regular ART and 40 healthy controls (HCs). Among the 118 PLWH, 18 immunological nonresponders (INRs; &lt;200 CD4+ T cells/μL) and 30 immunological responders (IRs; ≥500 CD4+ T cells/μL) were identified. The oral microbiota composition of all participants was analyzed using 16S rRNA gene sequencing of throat swab samples. Relative abundance of bacterial genera was compared between IRs and INRs, and Pearson correlations between bacterial abundance and peripheral blood immune cell counts were evaluated. The INR group showed lower alpha diversity than the IR and HC groups, which displayed similar alpha diversity. The genera Alloprevotella, Prevotella and Neisseria were more abundant in PLWH than in HCs, whereas the genera Rothia, Streptococcus and Fusobacterium were more abundant in HCs than in PLWH. The genus Rothia was more abundant in the INR group, whereas Prevotella, Alloprevotella, Porphyromonas and Haemophilus were more abundant in the IR group. The genera Rothia and Alloprevotella were negatively and positively associated with CD4+ T cell counts, respectively. Thus, an increased abundance of Rothia in the oral microbiome is associated with unfavorable outcomes regarding immune reconstitution in PLWH receiving regular ART, whereas Prevotella, Alloprevotella, Porphyromonas and Haemophilus are associated with favorable outcomes.

Discriminatory performance of the pulpal inflammatory biomarkers; Interleukin-8 and TNF-α in patients with symptoms indicative of reversible and irreversible pulpitis: A diagnostic accuracy study.

The success of vital pulp treatment (VPT) procedures is dependent on an accurate diagnosis of the pulpal inflammatory condition. Compared with current subjective pulpal diagnostic tests, inflammatory molecular biomarkers involved in the pathogenesis of pulpitis represent potential objective indicators of the degree of pulpal inflammation. Therefore, the aim of this study was to quantify level of inflammatory biomarkers - Interleukin 8 (IL-8) and TNF-α in patients diagnosed with reversible pulpitis (RP), irreversible pulpitis (IR) and normal pulp (NP) and investigate their diagnostic accuracy in differentiating between healthy and inflamed conditions. This prospective, cross-sectional study enrolled 72 patients aged 14-53 years with extremely deep carious lesions after establishing a clinical diagnosis of RP (n = 42), symptomatic IR (n = 22) and NP (n = 8). 50 μL of pulpal blood sample was collected from all the patients using a micropipette after pulpal exposure. The level of IL-8 and TNF-α was assessed in pg/mL using enzyme-linked immunosorbent assays. Mann-Whitney U test was applied to establish the association between IL-8/TNF-α level and degree of pulp inflammation. Receiver operating curve (ROC) analysis was carried out to calculate area under the curve (AUC) for RP versus IR. Cut-off values were established using Youden's index. IL-8 and TNF-α levels differed significantly between RP and IR groups (p ≤ .001). The median value of IL-8 in RP and IP groups was 259.8 pg/mL [187.5-310.0] and 1357.8 pg/mL [1036.7-2177.6] respectively. The AUC-ROC curve for RP versus IR was 0.997 with 95.5% sensitivity and 99.76% specificity. The median value of TNF-α in RP and IR groups was 75.4 pg/mL [62.7-95.8] and 157.6 pg/mL [94.1-347.3]. The AUC-ROC curve for TNF-α was 0.812 with a sensitivity and specificity of 59.1% and 92.1%, respectively. IL-8 and TNF-α levels were below detection levels for all NP samples. This study showed that pulpal blood could provide an excellent medium for establishing pulpal diagnosis under extremely deep carious lesions. The selected cytokines, IL-8 and TNF-α, demonstrated excellent discriminatory performance for reversible and irreversible pulpitis. Future studies should correlate the IL-8/TNF-α levels with VPT treatment outcomes.

L. plantarum and L. casei synergistically mitigate Gastro-Intestinal Acute Radiation Syndrome (GI-ARS)

Introduction: Incidental or accidental acute exposure to ionizing radiation causes radiation toxicity, for which there are no known medical countermeasures (MCMs). Ionizing radiation alters host gut epithelium as well as the gut microbiome. We sought to investigate if radiation induced gut dysbiosis occurred as a result of depletion of gut-protective bacteria, and if probiotic supplementation of such bacteria could help mitigate GI-ARS post ionizing radiation exposure. Methods: Adult C57BL/6 mice were subjected to total body irradiation, and a defined concentration of probiotics (either individually or combined) were given 24 h after radiation exposure. Mice were sacrificed 24 hours post treatment (i.e., 48 h post irradiation). Vascular-to-luminal flux of FITC-inulin was measured to evaluate intestinal mucosal permeability and endotoxemia by measuring plasma lipopolysaccharide. Gut microbiota was analyzed by 16S rRNA sequencing. Intestinal epithelial junctions were analyzed by immunofluorescence confocal microscopy and mucosal inflammatory response by RT-PCR for cytokine expression. Systemic inflammation was evaluated by ELISA for plasma cytokines. Results and conclusion: Our data show that radiation altered the taxonomic composition of the microbiota compared to age-matched sham treated mice. Shannon index analysis showed a significant decrease in α-diversity in irradiated mice. Bray-Curtis analysis indicated significant differences in microbiota composition between Sham and IR groups. E. coli abundance was increased whereas Lactobacillus abundance was decreased, particularly L. plantarum and L. casei. Probiotic supplementation with individual Lactobacillus species each mitigated radiation induced intestinal barrier dysfunction, gut specific as well as systemic inflammatory cytokine expression and endotoxemia. Our preliminary data further indicates that L. plantarum affects Epidermal Growth Factor Receptor (EGFR)-mediated signaling whereas L. casei was EGFR-independent. Interestingly, combined supplementation in a 1:1 ratio further enhanced recovery compared to adding the individual effects. Individual supplementation of L. plantarum and L. casei to irradiated mice reduced intestinal (ileal) permeability by 2-fold and 3-fold respectively, whereas combined supplementation reduced ileal permeability almost 8-fold compared to irradiated but untreated mice. This data indicates that L. plantarum and L. casei mitigate GI-ARS by distinct cellular mechanisms. Furthermore, data show that a mixture of these probiotic species is more effective in radio-mitigation than individually. Further mechanistic analysis will be required to identify cross-talking targets between mechanisms of these probiotics and synergistic radio-mitigation. In conclusion, Lactobacillus species are potential MCMs for the treatment of GI-ARS. Funding acknowledgements: NIH/NIAID, UO1-AI170019, UO1-AI172991. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

DECIPHERING THE NOVEL ROLE OF ERDOSTEINE IN PREVENTION OF MYOCARDIAL ISCHEMIA-REPERFUSION INJURY IN RAT

Objective: Myocardial ischemia-reperfusion injury (MIRI) is caused by thrombotic obstruction of the coronary artery subsequently leading to heart failure. Despite the availability of improved prognosis, MIRI is a foremost cause of morbidity and mortality worldwide. In this study, we investigated the therapeutic potential of Erdosteine (a mucoactive antioxidant molecule, with multifaceted pharmacological properties), to attenuate myocardial infarction in rat model of MIRI. Design and method: This study was conducted in adult male Wistar rats (n=40), randomized into five groups: Sham, IR-C (disease control), Erdosteine (80 mg/kg; s.c.) + IR, Telmisartan (10 mg/kg; orally) +IR and Erdosteine (80 mg/kg) perse groups. Erdosteine and telmisartan were administered for 28 days as a pre-treatment. On the 29th day, animals were subjected to LAD coronary artery occlusion for 1 h followed by reperfusion for 45 minutes. The cardioprotective potential was assessed through measurement of hemodynamic parameters, biochemical markers (MDA, GSH, SOD), cardiac injury markers (CK-MB and LDH), inflammatory markers (IL-6, TNF), histopathological evaluation, TUNEL assay and immunohistochemistry analysis (Bax, caspase 3, Bcl2). Further, the underlying mechanism was evaluated using immune-blotting of MAPK, Nrf-2/HO-1, and Akt/p-Akt. The statistics analysis was performed by one-way ANOVA followed by Turkey-Kramer post hoc tests. Results: Erdosteine and telmisartan pretreatment improves hemodynamic parameters by preserving MAP, ±LVdP/dt, and HR as compared to the IR-C group. Pretreatment markedly restored the levels of antioxidants (GSH and SOD) and decreased peroxidation marker (MDA) compared to the IR-C group. Also, the pretreatment significantly prevented the damage following IR injury as compared to the IR-C group when investigated through cardiac injury markers and histopathological evaluation. Erdosteine profoundly reversed the expressions of Bcl2/BAX ratio, in comparison to the IR-C group indicating its anti-apoptotic effect when reconfirmed through TUNEL assay. Erdosteine pretreatment preserved the levels of NF-kB, TNF, and IL-6 following IR challenge. Erdosteine pretreatment restores FABP, Nrf-2, and Hsp27. Conclusions: Erdosteine protects the myocardial cell membrane from peroxidation and inhibits oxidative burst and inflammation through NF-kB pathway activation. Erdosteine attenuates MIRI injury in rats preserving the MAPK and Nrf-2 signaling cascade.

Adipose-derived stem cells ameliorate radiation-induced lung injury by activating the DDAH1/ADMA/eNOS signaling pathway

BackgroundIonizing radiation-induced lung injury is caused by the initial inflammatory reaction and leads to advanced fibrosis of lung tissue. Adipose-derived stem cells (ASCs) are a type of mesenchymal stem cell that can differentiate into various functional cell types with broad application prospects in the treatment of tissue damage. The purpose of this study was to explore the protective effect of ASCs against radiation-induced lung injury and to provide a novel basis for prevention and treatment of radiation-induced lung injury. Materials and methodsFifty mice were randomly divided into a control group (Ctrl), radiation exposure group (IR), radiation exposure plus ASC treatment group (IR + ASC), radiation exposure plus L-257 group (IR + L-257), and radiation exposure plus ASC treatment and L-257 group (IR + ASC + L-257). Mice in IR, IR + ASC, and IR + ASC + L-257 groups were exposed to a single whole-body dose of 5 Gy X-rays (160 kV/25 mA, 1.25 Gy/min). Within 2 h after irradiation, mice in IR + ASC and IR + ASC + L-257 groups were injected with 5 × 106 ASCs via the tail vein. Mice in IR + L-257 and IR + ASC + L-257 groups were intraperitoneally injected with 30 mg/kg L-257 in 0.5 mL saline. ResultsThe mice in the IR group exhibited lung hemorrhage, edema, pulmonary fibrosis, and inflammatory cell infiltration, increased release of proinflammatory cytokines, elevation of oxidative stress and apoptosis, and inhibition of the dimethylarginine dimethylamino hydratase 1 (DDAH1)/ADMA/eNOS signaling pathway. ASC treatment alleviated radiation-induced oxidative stress, apoptosis, and inflammation, and restored the DDAH1/ADMA/eNOS signaling pathway. However, L-257 pretreatment offset the protective effect of ASCs against lung inflammation, oxidative stress, and apoptosis. ConclusionsThese data suggest that ASCs ameliorate radiation-induced lung injury, and the mechanism may be mediated through the DDAH1/ADMA/eNOS signaling pathway.

Effects of sevoflurane and fullerenol C60 on lower limb ischemia-reperfusion injury in streptozocin-induced diabetic mice.

Ischemia-reperfusion injury (IRI) poses a significant challenge for physicians, necessitating the management of cell damage and the preservation of organ functions. Various surgical procedures, such as vascular surgery on extremities, temporary cross-clamping of the abdominal aorta in aortic surgery, and the use of a tourniquet in extremity surgeries, may induce lower limb IRI. The susceptibility to IRI is heightened in individuals with diabetes. This study aimed to investigate the effects of fullerenol C60 and sevoflurane on mouse muscle tissue in a lower limb IRI model and to assess their potential in preventing complications arising from ischemia-reperfusion in mice with streptozocin-induced diabetes. A total of 36 adult Swiss albino mice were randomly divided into six groups, each consisting of six mice: control group (group C), diabetes group (group D), diabetes-ischemia/reperfusion group (group DIR), diabetes-ischemia/reperfusion-fullerenol C60 group (group DIR-FC60), diabetes-ischemia/reperfusion-sevoflurane group (group DIR-S), and diabetes-ischemia/reperfusion-sevoflurane-fullerenol C60 group (DIR-S-FC60). Streptozocin (55 mg/kg) was intraperitoneally administered to induce diabetes in the relevant groups, with mice displaying blood glucose levels of 250 mg/dL or higher at 72 h were considered diabetic. After 4 weeks, all groups underwent laparotomy under anesthesia. In DIR-FC60 and DIR-S-FC60 groups, fullerenol C60 (100 mg/kg) was intraperitoneally administrated 30 min before the ischemia period. Sevoflurane, delivered in 100% oxygen at a rate of 2.3% and 4 L/min, was administered during the ischemia period in DIR-S and DIR-S-FC60 groups. In the IR groups, a microvascular clamp was placed on the infrarenal abdominal aorta for 120 min during the ischemia period, followed by the removal of the clamp and a 120-min reperfusion period. At the end of the reperfusion, gastrocnemius muscle tissues were removed for histopathological and biochemical parameter examinations. Histopathological examination revealed a significant reduction in the disorganization and degeneration of muscle cells in the DIR-S-FC60 group compared to the DIR group (p = 0.041). Inflammatory cell infiltration was notably lower in the DIR-S, DIR-FC60, and DIR-S-FC60 groups than in the DIR group (p = 0.031, p = 0.011, and p = 0.013, respectively). The total damage scores in the DIR-FC60 and DIR-S-FC60 groups were significantly lower than in the DIR group (p = 0.018 and p = 0.008, respectively). Furthermore, the levels of malondialdehyde (MDA) in the DIR-S, DIR-FC60, and DIR-S-FC60 groups were significantly lower than in the DIR group (p < 0.001, p < 0.001, and p < 0.001, respectively). Catalase (CAT) enzyme activity in the DIR-S, DIR-FC60, and DIR-S-FC60 groups was higher than in the DIR group (p = 0.001, p = 0.014, and p < 0.001, respectively). Superoxide dismutase (SOD) enzyme activity in the DIR-FC60 and DIR-S-FC60 groups was also higher than in the DIR group (p < 0.001 and p = 0.001, respectively). Our findings indicate that administering fullerenol C60 30 min prior to ischemia in diabetic mice, in combination with sevoflurane, led to a reduction in oxidative stress and the correction of IR-related damage in muscle tissue histopathology. We believe that the administration of fullerenol C60 before IR, coupled with sevoflurane administration during IR, exerts a protective effect in mice.

Protective Role of Rapamycin in Fibrotic Liver Ischemia/Reperfusion Injury (C57bl/6 Mouse)

Liver ischemia/reperfusion injury (IRI) is a well-documented phenomenon that occurs after liver resection and transplantation, posing a significant clinical challenge. We aim to contribute valuable insights into potential therapeutic interventions for fibrotic liver IRI, ultimately advancing our understanding of liver transplantation and resection outcomes. Twenty-four mice were divided randomly into 4 equal groups: [1] the normal group, n = 6; [2] the liver fibrosis (LF) group, n = 6; [3] the LF and IR group, n = 6; and [4] the LF with treatment of rapamycin and IR group; n = 6. Key biomarkers assessing liver function, alanine aminotransferase and aspartate aminotransferase, significantly decreased with Rapamycin administration. There is a substantial decrease observed in inflammatory cytokines such as interleukin (IL) 6, IL-1B, tumor necrosis factor alpha, Transforming growth factor-beta (TGF-beta), and Inducible nitric oxide synthase (iNOS) with rapamycin treatment. Furthermore, NOX levels, caspase-3, and caspase-9 were reduced after rapamycin administration. The application of rapamycin demonstrates appropriate effects in anti-inflammation, antioxidation, and anti-apoptosis, indicating significant therapeutic potential for fibrotic liver IRI.