Target Organ Research Articles

Impact of joint peer review with a specialist radiologist in head and neck cancer radiotherapy planning.

e18059 Background: Head and neck oncology is heavily dependent on interpretation of radiology. This raises the question whether input from radiology should be formally scheduled as part of the radiotherapy contouring processes. Our aim was to assess the impact of this practice through qualitative and quantitative analysis. Methods: H & N patients treated with radiotherapy between May-2022 and August-2023 were reviewed by a specialist HN radiologist. Incidence of changes in T, N, M and TNM-stage, change as defined by The RCR guidance: 'major' (change in gross tumour volume and/or high-dose clinical target volume, dose/fractionation) or 'minor' (change in intermediate or elective dose clinical target volumes or organs at risk), was recorded. We evaluated whether any of these changes were time dependent (Time interval between MDT to RT Peer Review). Other clinical variables: age, gender, primary diagnosis, tumour site, significant incidental findings, post-operative pTNM, post-operative disease status in adjuvant setting on planning imaging (NIRADS), need for additional imaging. We also recorded number of scans reviewed per patient, types of imaging studies reviewed, time taken for review and number of radiologist PA’s required. Retrospective analysis of a prospective database was performed. Results: 410 patients (252-definitive, 137-adjuvant, 21-palliative) were reviewed of which 51.5%-oropharynx followed by 26.8%-Larynx & Hypopharynx. Mean age was 66.14 yrs (SD 11.46) and M:F was 4:1. Baseline and Therapy planning imaging was reviewed for 100% patients of which baseline included: MR (91.7%), CT (96.1%), US (34.1%), PETCT (31%) and therapy planning (62%-CECT, 19%-MR&CT, 15%-CT, 4%-PETCT). In all patients (n=410), T/N/M/TNM-stage percentage upstage was 27/36.8/8.3/42.2% and in definitive cohort (n=252), it was 41.7/56.7/7.1/59.5%. Reviewing RT-planning Imaging in adjuvant cohort (n=137), 76.6% were NIRADS-1, 11.7% NIRADS-3/4/loco-regional disease and 11.7% distant metastasis. T/N/M/TNM-stage upstage was time dependent and the mean time interval between MDT to RT was higher in upstaged patients (p <0.001). Overall percentage change in treatment plan: no/minor/major/palliative was 40.5/33.9/17.3/8.3% and in definitive cohort 30.2/42.1/20.6/7.1%. Change in plan was not dependent on mean time interval between MDT to RT. Average 10 patients (4 imaging studies/patient) were reviewed by radiologist in 1PA (time to review and peer review meeting time). Conclusions: Routine head and neck radiologist input in radiotherapy peer review is feasible and resulted in a number of major and minor changes to treatment. Inputs not only eased the contouring for oncologists but also lead to updating of current disease stage just prior to treatment. Overall, our results suggest that this is a practice changing step in the radiotherapy workflow which has a direct impact on patient management.

Effect of triptolide on reproductive toxicity in female rats with Ⅱ type collagen induced arthritis

In order to study the toxic effect and mechanism of triptolide(TP) on the reproductive system of female rats with Ⅱ type collagen induced arthritis(CIA), 50 SD rats were randomly divided into normal control group, CIA model group, and three groups receiving TP tablets at clinically equivalent doses of 0. 5, 1, and 2 times, respectively(with TP dosages of 3. 75, 7. 5, and 15 μg·kg~(-1)·d~(-1)), each comprising 10 rats. Intragastric administration was started on the day after the first immunization, once a day, for 42 days.The results were taken on the 21st and 42nd days to calculate the uterine and ovarian organ indexes; pathological and morphological changes in uterus and ovaries were observed under a light microscope; and the levels of estradiol(E_2) and cytochrome P450A1(aromatase,CYP19A1) in ovarian homogenate were detected by ELISA. Furthermore, immunohistochemistry was employed to detect the expression levels of transforming growth factor β3( TGFβ3) pathway-related proteins, mothers against decapentaplegic homolog 3(Smad3) and steroidogenic factor-1(SF-1) in ovarian tissues. In vitro, the mouse Chinese hamster ovary(CHO) cell line was established, and after 24 hours of TP administration(30, 60, 120 nmol·L~(-1)), cell proliferation was detected by the thiazolyl blue tetrazolium bromide(MTT) method, apoptosis by the flow cytometry, and TGFβ3, Smad3 and SF-1 protein expression in cells by the Western blot method, and the nuclear entry of SF-1 was detected by immunofluorescence. The results showed that compared with the CIA model group, all TP administration groups showed decreased number of uterine glands, total follicles, mature follicles, and corpus luteum on days 21 and 42 of administration, but there was no statistical difference, and only the administration of 2 times the clinically equivalent dose of TP could significantly increase the number of atretic follicles at 42 days of administration. TP at 3. 75 μg·kg-1·d-1significantly reduced the level of E_2 at 21 days of administration and the expression of TGFβ3 and Smad3 factors in ovarian tissues,but had no significant effect on the rate-limiting enzyme in estrogen synthesis CYP19A1. TP at 7. 5 and 15 μg·kg~(-1)·d~(-1) significantly reduced the expression of SF-1 regardless of administration for 21 days or 42 days. TP can significantly promote ovarian cell apoptosis in vitro, with apoptosis mainly concentrated in the late stage of apoptosis after 24 hours of administration. In addition, 60 nmol·L~(-1) TP significantly reduced the protein expression of TGFβ3, Smad3 and SF-1 in a dose-dependent manner. In summary, intragastric administration of TP at less than 2 times the clinically equivalent dose for 21 days and 42 days did not cause obvious reproductive damage to the uterus and ovarian tissues of CIA rats, and the number of atretic follicles changed significantly only when the 2 times the clinically equivalent dose was administered for 42 days. TP exerted reproductive toxicity in vivo on reproductive target organs and in vitro on ovarian cells by inhibiting the expression of TGFβ3/Smad3/SF-1 pathway.

Clinical, target organ damage and 24-hour ambulatory blood pressure monitoring characteristics in older adults with high-risk hypertension

Objectives: To investigate the clinical characteristics, target organ damage subclinical, and 24-hour ambulatory blood pressure monitoring (ABPM) in older adults with high-risk hypertension. Material and methods: A cross-sectional study was conducted on 96 older adults (≥ 60 years old) with treated high-risk HTN. The patients were performed with 24-hour ABPM. Results: The mean age was 70.88 ± 7.86 years, and 64.6% females. The prevalence of cardiovascular risk factors included dyslipidemia - 80.2%, physical inactivity - 69.8%, family history of hypertension - 54.2%, diabetes mellitus - 51.0%, obesity - 29.2%, and smoking - 24%. The target organ damage had occurred in 63.5% of the patients, with 51% having left ventricular hypertrophy on echocardiography, 11.5% showing serum creatinine levels ≥ 115 µmol/l, and 26.1% with proteinuria. On ABPM, the prevalence was as follows: uncontrolled masked hypertension - 12.5%, morning blood pressure (BP) - 40.6%, nocturnal dipping - 13.5%, non-dipping - 53.1%, nocturnal hypertension - 33.3%, isolated systolic hypertension - 37.5%. Daytime BP overload - 60.4%, nighttime BP overload - 90.6%, 50% daytime BP overload - 41.7%, 50% BP overload - 75%. Conclusions: Older adults with high-risk hypertension exhibit a high prevalence of traditional cardiovascular risk factors. The target organ damage observed in approximately 2/3 of cases. Morning BP, isolated systolic hypertension, and nocturnal hypertension were presented in over 1/3 of cases. Non-dipping observed in over 1/2 of cases. Daytime BP overload observed in in nearly 2/3 of cases, with most patients experiencing nighttime BP overload. Key words: 24-hour ambulatory blood pressure, clinical characteristics, high-risk hypertension, older adults.

Safety and efficacy of Symani robotic-assisted microsurgery: Assessment of vascular anastomosis patency, thrombus, and stenosis in a randomized preclinical study

IntroductionThe Symani Surgical System undergoes scrutiny in this study as part of a series of preclinical investigations. Previous studies compared the precision of robotic-assisted anastomoses with manual techniques. This study aims to evaluate critical, histological and gross parameters at site of anastomosis and at distance in a rat model to provide insights into the safety and efficacy of robotic-assisted microsurgery, enhancing its potential for clinical adoption. Materials and MethodsExperienced microsurgeons performed arterial and venous anastomoses in sixteen Wistar rats, randomized in four treatment groups: robotic artery, robotic vein, manual artery, and manual vein. Various parameters were evaluated at two time-points (T0 and T30) on the day of the procedure and at Day 28(T28d). In the second phase of the study animals underwent necropsy, histopathologic analysis and micro-CT scans angiography imaging of anastomosis sites, major organs and distant target organs by a blinded assessor. ResultsPatency rates were 100% at T0 and T30 for all anastomoses, remaining so at T28d for the robotic subgroups, while decreasing to 87.5% for manual arterial anastomoses due to an obstructive thrombus. No evidence of clot migration. Blood flow parameters and procedure times did not significantly differ. The blinded semi-quantitative histological analysis revealed no significant disparities between robotic and manual anastomoses across various pathological indicators. No gross abnormalities were detected in musculoskeletal examinations. ConclusionThis preclinical study demonstrates the safety of the Symani Surgical System. Results suggest equivalence between robotic and manual techniques regarding thrombus formation at the anastomotic site and distal organs.

Validation of a deep-learning modular prototype to guide novices to acquire diagnostic ultrasound images from urinary system

ImportanceArtificial intelligence (AI) application in guiding the acquisition of ultrasonography images represents a pioneering field of research. A new developed hybrid deep-learning (DL) algorithm, trained on more than high quality 60.000 curated and labelled reference images (distilled from a set of more than 600.000 abdominal ultrasound images) from La Paz Hospital, can provide real-time prescriptive guidance for novice operators to obtain standard planes images of the target organs. ObjectiveThis study aims to evaluate the capability of novice users to acquire diagnostic-quality abdominal ultrasound images of the urinary system using the deep-learning (DL)-based guiding research prototype provided by GMV. DesignSetting, and Participants: This prospective diagnostic study was conducted within the facilities of an academic hospital. A cohort of 24 technically-oriented volunteers, lacking prior knowledge in anatomy or medicine and without experience in conducting ultrasound examinations, was recruited. After a brief training session focused on various organs, each pair of volunteers performed scans of each other's urinary system, exclusively guided by AI support. These scans were subsequently repeated by experienced sonographers using identical ultrasound equipment but without AI assistance. Four radiologists, each with decades of experience, independently and blindly assessed the quality of each acquisition. ResultsOver 90.6 % of the images scanned by volunteers were identified as valuable clinical picture, using only an AI-based guidance system. This is nearly comparable to the results achieved by experienced radiologists, who attained a 98.6 % success rate. ConclusionsThis deep-learning (DL) prototype enables novices lacking experience in ultrasonography to acquire diagnostic ultrasound images suitable for subsequent expert evaluation. The modular prototype works on a large range of ultrasound device models and vendors. This advancement has the potential to extend the application of ultrasound beyond traditional clinical environments, particularly in situations requiring immediate anatomical and functional interrogation, as well as in resource-limited settings.

Ultrastructural Biomarkers in Target Organs of Fish from Algeria Coastline to Access Water Quality

Ultrastructural Biomarkers in Target Organs of Fish from Algeria Coastline to Access Water Quality

Systematic Review of the First 40 Cases of 177Lu-PSMA Therapy in the Treatment of Non-prostatic Cancer.

The current systematic review aimed to collect and analyze all available published and unpublished cases in which prostate-specific membrane antigen (PSMA)-targeted radioligand therapy (177Lu-PSMA) was used to treat non-prostatic cancer. Literature search and evidence acquisition through contacts with organizations that use 177Lu-PSMA were employed. PubMed/Medline, SCOPUS, and ScienceDirect searches were performed following PRISMA recommendations. The search strategy was to screen all articles describing 177Lu-PSMA radioligand therapy published to date with the key word "177Lu-PSMA". These articles were collected and screened for non-prostatic cancer cases. Quality assessment was performed using the GRADE criteria. A total of 713 articles were screened, and the search revealed 15 eligible records. Forty patients with a mean age of 51.2±18.5 years were treated with 177Lu-PSMA for non-prostatic cancer. Of them, 30 cases were published, and 10 were found in medical institution records. Cancers of the salivary glands were most often targeted (13/40), followed by various brain cancer types (8/40), and osteosarcoma (6/40). The authors used previously established protocols for castration-resistant prostate cancer with the dose per cycle as 6.0-7.4 GBq and the number of cycles between one and four. Toxicity was estimated as low, and 21 out of 28 patients with reported outcomes survived to the time of the publication. PSMA-targeted radioligand therapy was infrequently used to treat different non-prostatic cancer types in various target organs. These pioneering efforts indicate that 177Lu-PSMA can be used to treat non-prostatic cancer with PSMA expression. The toxicity of such treatment was low, and the outcome was relatively good.

Translocator Protein 18 kDa Tracer 18F-FDPA PET/CTA Imaging for the Evaluation of Inflammation in Vulnerable Plaques.

Inflammation induced by activated macrophages within vulnerable atherosclerotic plaques (VAPs) constitutes a significant risk factor for plaque rupture. Translocator protein (TSPO) is highly expressed in activated macrophages. This study investigated the effectiveness of TSPO radiotracers, 18F-FDPA, in detecting VAPs and quantifying plaque inflammation in rabbits. 18 New Zealand rabbits were divided into 3 groups: sham group A, VAP model group B, and evolocumab treatment group C. 18F-FDPA PET/CTA imaging was performed at 12, 16, and 24 weeks in all groups. Optical coherence tomography (OCT) was performed on the abdominal aorta at 24 weeks. The VAP was defined through OCT images, and ex vivo aorta PET imaging was also performed at 24 weeks. The SUVmax and SUVmean of 18F-FDPA were measured on the target organ, and the target-to-background ratio (TBRmax) was calculated as SUVmax/SUVbloodpool. The arterial sections of the isolated abdominal aorta were analyzed by HE staining, CD68 and TSPO immunofluorescence staining, and TSPO Western blot. The results showed that at 24 weeks, the plaque TBRmax of 18F-FDPA in group B was significantly higher than in groups A and C. Immunofluorescence staining of CD68 and TSPO, as well as Western blot, confirmed the increased expression of macrophages and TSPO in the corresponding regions of group B. HE staining revealed an increased presence of the lipid core, multiple foam cells, and inflammatory cell infiltration in the area with high 18F-FDPA uptake. This indicates a correlation between 18F-FDPA uptake, inflammation severity, and VAPs. The TSPO-targeted tracer 18F-FDPA shows specific uptake in macrophage-rich regions of atherosclerotic plaques, making it a valuable tool for assessing inflammation in VAPs.

Elucidating the mechanism of hepatotoxicity in Euodia rutaecarpa: insights from QSAR toxicity prediction and metabolomics

Objective: Euodia rutaecarpa, (Wu Zhu Yu) a Chinese medicine clinically used to treat gastrointestinal disorders, has been widely employed. However, Euodia rutaecarpa is regarded as a small toxic traditional Chinese medicine in the Chinese Pharmacopoeia and other herbal works. Using toxicity predictions combined with in vitro and in vivo studies, this study aimed to identify the toxic components and toxic target organs of Euodia rutaecarpa, and explore its toxic mechanism from a metabolic perspective. Methods: The toxic target organs of Euodia rutaecarpa were identified through in vitro and in vivo studies. In vitro toxicity screening was performed by alkaloid enrichment and isolation. The potential toxicity of compounds was predicted by Absorption, Distribution, Metabolism, Excretion, and Toxicity Predictor (ADMET Predictor) based on Quantitative Structure–Activity Relationship (QSAR) construction. In addition, the study integrated the serum metabolomic analysis after the administration of potentially toxic components to clarify the effect of potentially toxic substances on metabolism in mice. Results: Comparing the acute toxicity in mice of different extraction methods and before and after processing, it was evident that Euodia rutaecarpa alcoholic extract had the highest toxicity, and the target organ of Euodia rutaecarpa toxicity was the liver. The alkaloid fraction of alcoholic extract of Euodia showed strong cytotoxicity. The potential toxicity of Euodia rutaecarpa was calculated and predicted by ADMET Predictor, and alkaloids are suspected to be responsible for the toxicity of Euodia rutaecarpa. Evodiamine significantly reduced the number of cells and increased the mitochondrial membrane potential in vitro. Different metabolites were significantly identified by serum metabolomics, of which bile acid metabolism and steroid hormone biosynthesis are the key pathways of hepatotoxicity. Conclusions: Clarify the scientific significance of clinical use of processed products by comparing the acute toxicity of different extraction methods before and after processing. Combining the toxicity prediction based on QSAR with the toxicity screening in vitro and in vivo, the potential toxic target organs and toxic components of Euodia rutaecarpa can be identified. Through metabolomics, we preliminarily revealed that the hepatotoxicity of Euodia rutaecarpa may be related to bile acid metabolism and steroid hormone biosynthesis. This study lays the foundation for elucidating the mechanism of Euodia rutaecarpa and evaluating its safety and quality.

An overview of mycotoxicoses in rabbits.

Mycotoxicoses are usually a consideration in large animal species but can affect companion animals as well. Due to increasing interest and the ease of using rabbits as laboratory models, a growing number of published experimental studies discuss the effects of various mycotoxins on this species. However, the available evidence is fragmented and heterogeneous, and has not recently been collated in a review, to my knowledge. Although mycotoxicoses in rabbits are typically subclinical, clinical signs can include weight loss, anorexia, gastrointestinal disorders, stunted growth, reproductive abnormalities, and susceptibility to infections. An antemortem diagnosis typically relies on a comprehensive clinical history, and assessment of clinical signs and relevant laboratory findings, with confirmation of exposure achieved through the measurement of mycotoxin concentrations in feed or target organs. My review focuses on the clinicopathologic and histopathologic effects of the mycotoxins most important in rabbits, including fumonisins, ochratoxins, aflatoxins, trichothecenes, and zearalenone. This review offers a thorough overview of the effects of mycotoxins in rabbits, serving as a one-stop resource for veterinary practitioners, diagnosticians, and researchers.

Temporal dynamics of RAS mutations in circulating tumor DNA in metastatic colorectal cancer: clinical significance of mutation loss during treatment

PurposeIn metastatic colorectal cancer (mCRC), RAS mutation loss may occur during the standard-of-care regimen. In this study, we aimed to investigate the temporal dynamics of the RAS gene and its clinical significance.MethodsThis was a retrospective, single-center study that included 82 patients with tissue RAS-mutant (RAS-MT) mCRC who underwent circulating tumor DNA (ctDNA) RAS monitoring between January, 2013–April, 2023. Patients were analyzed for the rate of change over time to acquired RAS mutation loss (aRAS-ML) and clinicopathological factors. The prognostic relevance of mutation loss was assessed.ResultsaRAS-ML was detected in 33 (40.2%) patients, 32 of whom had a mutation loss in the first ctDNA RAS assay. Patients with a RAS mutation detected in the first assay had a median time of 8 months until the second ctDNA RAS assay, with 4.5% cases newly converted to aRAS-ML; no new conversions were detected at the third assay. The aRAS-ML group exhibited more single-organ metastases in the target organ during ctDNA measurement (aRAS-ML: 84.8% vs. RAS-MT: 59.2%, p = 0.02). Of the 33 patients with aRAS-ML, seven (21.2%) received anti-epidermal growth factor receptor (EGFR) therapy, with a median progression-free survival of 8 months. Multivariate analysis revealed that persistent ctDNA RAS mutation was an independent prognostic factor for overall survival (hazard ratio: 2.7, 95% confidence interval: 1.1–6.3, p = 0.02).ConclusionThe rate of ctDNA mutation loss in patients with RAS-MT mCRC decreases over time. Therefore, using a ctDNA RAS assay early in treatment will assist in challenging the use of EGFR regimens.

Comparative effects of polyvinyl chloride microplastics on the brittle star Ophiactis virens and the amphibian Xenopus laevis

In this study, we investigated the effects of PVC microplastics (PVC-MPs) using two different animal models: the brittle star Ophiactis virens, and the African clawed frog Xenopus laevis. This is the first study using an environmental relevant sample of PVC-MPs obtained through mechanical fragmentation of a common PVC plumbing pipe. Exposure experiments on brittle star were performed on the adult stage for a duration of 14 days, while those on African clawed frog were performed on the embryogenic developmental stage according to the standardized FETAX protocol (Frog Embryo Teratogenesis Assay-Xenopus). For both models, different endpoints were analysed: mortality, developmental parameters, behavioural assays and histological analyses on target organs by optical and electronic microscopy. Results showed that the concentration of 0.1 μg mL−1 PVC do not cause any adverse effects in both models (common NOEC concentration), while exposure to 1 μg mL−1 PVC adversely affected at least one species (common LOEC concentration). In particular arm regeneration efficiency was the most affected parameters in O. virens leading to a significantly lower differentiation pattern at 1 μg mL−1 PVC. On the contrary, in X. laevis larvae histopathological analyses and behavioural tests were the most susceptible endpoints, exhibiting several abnormal figures and different swimming speed at 10 μg mL−1 PVC. Histopathological analyses revealed a higher abundance of degenerating cells, pyknotic nuclei and cellular debris in the gut of exposed larvae in respect to control. The comparative analyses performed in this work allowed to characterize the specificity of action of the PVC-MPs on the two species, underlining the importance of exploring a large spectrum of endpoints to offer adequate protection in the emerging fields of microplastic research.

Human amniotic epithelial stem cell is a cell therapy candidate for preventing acute graft-versus-host disease.

Graft-versus-host disease (GVHD), an immunological disorder that arises from donor T cell activation through recognition of host alloantigens, is the major limitation in the application of allogeneic hematopoietic stem cell transplantation (allo-HSCT). Traditional immunosuppressive agents can relieve GVHD, but they induce serious side effects. It is highly required to explore alternative therapeutic strategy. Human amniotic epithelial stem cells (hAESCs) were recently considered as an ideal source for cell therapy with special immune regulatory property. In this study, we evaluated the therapeutic role of hAESCs in the treatment of GVHD, based on our previous developed cGMP-grade hAESCs product. Humanized mouse model of acute GVHD (aGVHD) was established by injection of huPBMCs via the tail vein. For prevention or treatment of aGVHD, hAESCs were injected to the mice on day -1 or on day 7 post-PBMC infusion, respectively. We showed that hAESCs infusion significantly alleviated the disease phenotype, increased the survival rate of aGVHD mice, and ameliorated pathological injuries in aGVHD target organs. We demonstrated that hAESCs directly induced CD4+ T cell polarization, in which Th1 and Th17 subsets were downregulated, and Treg subset was elevated. Correspondingly, the levels of a series of pro-inflammatory cytokines were reduced while the levels of the anti-inflammatory cytokines were upregulated in the presence of hAESCs. We found that hAESCs regulated CD4+ subset polarization in a paracrine mode, in which TGFβ and PGE2 were selectively secreted to mediate Treg elevation and Th1/Th17 inhibition, respectively. In addition, transplanted hAESCs preserved the graft-versus-leukemia (GVL) effect by inhibiting leukemia cell growth. More intriguingly, hAESCs infusion in HSCT patients displayed potential anti-GVHD effect with no safety concerns and confirmed the immunoregulatory mechanisms in the preclinical study. We conclude that hAESCs infusion is a promising therapeutic strategy for post-HSCT GVHD without compromising the GVL effect. The clinical trial was registered at www.clinicaltrials.gov as #NCT03764228.

Evaluation of the InterDosi code in estimating S-values with the digimouse voxelized mouse phantom

This study delves into the realm of internal dosimetry in small animals, focusing on the crucial role of S-values (absorbed dose rate per unit activity). The primary objective is to enhance our understanding of ionizing radiation distribution and optimize treatment strategies involving multidisciplinary approach which integrates dosimetry data, computational modeling, clinical expertise, and patient-specific factors to deliver safe, effective, and personalized cancer treatments, paving the way for more effective and tailored radiotherapeutic interventions and diagnostic cancer research. The research specifically targets the assessment of radiation doses absorbed in target organs and adjacent tissues during targeted radionuclide therapy with iodine-131 and yttrium-90. Executed on the ERSN (Radiations and Nuclear Systems Laboratory) computing grid, the study employed optimized calculations using 20 CPU (Central Processing Unit) cores for each of the 103 SAF (Specific Absorbed Fractions) grid sizes of photons and electrons. The InterDosi code, a user-friendly tool, streamlined S-value estimation within the Digimouse phantom. Nine source organs and the entire murine anatomy were comprehensively evaluated. The G4EMLiverMorePhysics package ensured high-confidence dosimetric coefficient simulations. S-values revealed insights into absorbed dose influences, including organ mass and inter-organ distance. Comparative analyses were conducted with reference data, which involved a simplified S-value calculation. The results aligned well with the reference data, especially with similar SAF grid sizes for electrons and photons. Accuracy improved with larger grid sizes. This study confirms InterDosi's effectiveness, highlighting its adaptability and precision in estimating S-values for various radionuclides, making it a valuable tool for evaluating radiation doses in murine anatomy.

Principle of contrast-enhanced ultrasonography.

Sonazoid, an ultrasound contrast agent, has been covered by insurance in Japan since January 2007 for the diagnosis of hepatic mass lesions and is widely used for diagnosing not only primary liver cancer but also liver metastases such as those from breast cancer and colorectal cancer. Contrast-enhanced ultrasound for breast mass lesions has been covered by insurance since August 2012 after phase II and phase III clinical trials showed that the diagnostic performance was significantly superior to that of B-mode and contrast-enhanced magnetic resonance imaging. This paper describes the principles of imaging techniques in contrast-enhanced ultrasonography including the filter, pulse inversion, amplitude modulation, and amplitude-modulated pulse inversion methods. The pulse inversion method, which visualizes the second-harmonic component using the nonlinear scattering characteristics of the contrast agent, is widely used regardless of the contrast agent and target organ because of its high resolution. Sonazoid has a stiffer shell and requires a higher acoustic amplitude than Sonovue to generate nonlinear vibrations. The higher transmitted sound pressure generates more tissue harmonic components. Since pulse inversion allows visualization of the tissue harmonic components, amplitude modulation and amplitude-modulated pulse inversion, which include few tissue harmonic components, are primarily used. Amplitude modulation methods detect nonlinear signals from the contrast agent in the fundamental band. The mechanism of the amplitude modulation is considered to be changes in the echo signal's phase depending on the sound pressure. Since the tissue-derived component is minor in amplitude modulation methods, good contrast sensitivity can be obtained.

A novel subunit vaccine based on Fiber1/2 knob domain provides full protection against fowl adenovirus serotype 4 and induces stronger immune responses than a Fiber2 subunit vaccine

Outbreaks of hepatitis-hydropericardium syndrome (HHS) caused by fowl adenovirus serotype 4 (FAdV-4) have resulted in huge economic losses to the poultry industry in China since 2015. However, commercially available vaccines against the FAdV-4 infection remain scarce. In our study, subunit vaccine candidates derived from the bacterially expressed recombinant Fiber1 knob domain and Fiber2 knob domain fusion protein (termed as Fiber1/2 knob subunit vaccine) and Fiber2 protein (termed as Fiber2 subunit vaccine) of the FAdV-4 SDSX strain were developed. Immunogenicity evaluation showed that the Fiber1/2 knob subunit vaccine induced the production of antibodies at 7 d postvaccination (dpv), earlier than the Fiber2 subunit vaccine. Moreover, the neutralizing antibody level of the Fiber1/2 subunit vaccine group was higher than the Fiber2 subunit vaccine group, showing significant differences at 14, 21, and 28 dpv. Immune protection test results revealed that both Fiber1/2 knob subunit and Fiber2 subunit vaccines could protect chickens from death against FAdV-4 challenge, although the weight of chickens in the Fiber1/2 knob subunit vaccine group decreased less. Furthermore, analysis of plasma Glutamic oxaloacetic transaminase (AST) and blood glutamic pyruvic transaminase (ALT) levels suggested that the Fiber1/2 subunit vaccine can significantly inhibit liver damage caused by FAdV-4 infection and is more effective in blocking the pathogenicity of FAdV-4 in target organs. In addition, the Fiber1/2 knob subunit vaccine further reduced the viral load in different tissues and virus shedding in chickens than the Fiber2 subunit vaccine. Overall, the Fiber1/2 knob subunit vaccine was more effective than the Fiber2 subunit vaccine. These findings lay the foundation for the development of more effective FAdV-4 subunit vaccines.

#2940 Expression and modulation of SGLT2 in human podocytes. Not all in tubule

Abstract Background and Aims Inhibitors of the sodium-glucose cotransporter type 2 (iSGLT2) modify important non-glycemic pathways that lead to the protection of target organs. Its renoprotective pleiotropic effects in CKD, associated or not with diabetes, justifies further studies to elucidate the mechanisms of action (MoA) of these drugs. Although the co-transporter SGLT2 is mostly expressed in the proximal tubule, its location and function in other cells, that can be potential therapeutic targets for inhibitor drugs, has not been well studied. We explored the presence of SGLT2 in human podocyte cells and whether it is modified in different situations of cellular stress. Method Differentiated immortalized human podocytes were cultured under different experimental conditions for 24 or 48 h: Normoxia (Control); Normoxia + High concentration of glucose (HG); Hypoxia(1%)+2 h Reoxygenation(H) and the combination H + HG. Cells were also incubated with 0.1 uM Dihydrotestosterone (DHT). SGLT2 gene expression was determined by quantitative PCR and SGLT2 protein was detected by Western Blot (WB) and immunofluorescence (IF). Results Sglt2 gene expression was detected in human podocytes in culture, with an increase of expression after 24 h of exposure to HG. Protein expression by WB showed a trend to increase after 48h of exposure to HG and hypoxia, which was significant with co-incubation with DHT. HG plus DHT also increased the protein expression of SGLT2 under normoxic conditions. The IF measured by intensity of area did not show an increase in the protein expression of the transporter under HG incubation. However, protein expression increased under all hypoxic conditions: Control 50.05 ± 3, Hypoxia 140 ± 16.3 (p < 0.05 vs control) and HG + Hypoxia 253 ± 21.29 (p < 0.001 vs control). Conclusion Our data suggest that podocyte may be one of the therapeutic targets on which the iSGLT2 is acting. The experimental conditions described did not show differences in the expression of the transporter before the single stimulus of glucose, but they did under hypoxia and the combination Hypoxia-high glucose. In addition, co-incubation with testosterone seems to enhance the expression. Studying the MoA of iSGLT2 will open better comprehension of the kidney pathophysiology, particularly in the podocytopathies, regardless of diabetes.

Blockade of CCR5 and CXCR3 attenuates murine acute graft-versus-host disease through modulating donor-derived T cell distribution and function.

Lymphocyte trafficking via chemokine receptors such as CCR5 and CXCR3 plays a critical role in the pathogenesis of aGVHD. Our previous studies showed that addition of CCR5 or CXCR3 antagonist could only slightly alleviate the development of aGVHD. Given the specificity of T lymphocytes bearing CXCR3 and CCR5, we investigated whether combined CCR5 and CXCR3 blockade could further attenuate murine aGVHD. A mouse model of aGVHD was established to assess the efficacy of CCR5 or/and CXCR3 blockade on the development of aGVHD. The distribution of lymphocytes was calculated by quantification of immunostaining cells. The immunomodulatory effect on T cells were assessed by evaluating T- cell proliferation, viability, and differentiation. Using murine allo-HSCT model, we demonstrated that blockade of both CCR5 and CXCR3 could efficiently alleviate the development of aGVHD. Further investigation on the immune mechanisms for this prophylactic effect showed that more T cells were detained into secondary lymphoid organs (SLOs), which may lead to reduced infiltration of T cells into GVHD target organs. Our study also showed that T cells detained into SLOs dampened the activation, suppressed the polarization toward Th1 and Tc1, and induced the production of Treg cells. These data suggest that concurrent blockade of CCR5 and CXCR3 attenuates murine aGVHD through modulating donor-derived T cell distribution and function, and this might be applicable for aGVHD prophylaxis in clinical settings.

#2199 Kidney involvement aspects in women with Fabry disease from the Romanian cohort

Abstract Background and Aims Fabry disease (FD) is an X-linked rare lysosomal storage disease causing progressive nervous system, kidney and heart disorders. Progressive kidney impairment is common, and an important cause of morbidity and mortality. In FD females, treatment guidelines recommend therapy initiation when there is evidence of organ involvement, but determining the proper moment to start FD specific treatment could be a real challenge in the cases. The aim of our retrospective study is to investigate clinical and histological aspects of kidney involvement in female patients with genetically confirmed FD evaluated in our Expert Center for Rare Diseases of the Reno-Urinary System between 2015–2023, with focus on the clinical phenotype and initiation of FD specific treatment. Method All patients had comprehensive assessment for target organ manifestations of Fabry disease. Renal function was assessed using serum creatinine, albumin creatinine ratio and proteinuria. Also, 14 (40%) FD females underwent kidney biopsy (KB) in order to reveal the FD specific lesions and/or to establish the indication for FD pathogenic therapy. Kidney biopsy specimens were analyzed by light and electron microscopy. Results Our study included 35 heterozygous adult females with FD from 25 unrelated families. Most females (91.4%) were diagnosed through family screening, while 3 females (8.6%) were index cases. The mean age at diagnosis was 43.8 ± 15.3 years (range 17-73 years), although mean age of symptoms onset was 34.3 ± 13.5 years (range 14-61 years). Six women from our cohort were considered healthy carriers, three females presented only neurological manifestations, nine patients had combined renal and neurological involvement, one patient had cardiac and renal involvement and the others presented multiple organ involvement. Significant co-morbidities were: arterial hypertension in 13 patients, diabetes mellitus in 2 patients, and obesity in 9 patients. Overall baseline mean eGFR was 79.9 ± 28.2 (range 25-135) ml/min/1.73 m2. Eleven patients had no obvious kidney involvement, 10 patients presented chronic kidney disease (CKD) stage 2, 11 patients CKD stage 3, one CKD stage 4 and 1 was hemodialyzed. Seventeen patients presented normal UACR, thirteen patients showed microalbuminuria, and four patients presented macroalbuminuria. Overt proteinuria (> 0.3 g/24 h) was noticed in 8 patients. Thirteen females performed KB before the initiation of FD specific treatment, and one patient was already treated. At the moment of KB, five patients had normal biologic kidney assessment, three patients presented CKD stage 2 (microalbuminuria in 2 cases and proteinuria in 1 case), and five patients presented CKD stage 3 (normoalbuminuria in 1 case, microalbuminuria in 1 case and overt proteinuria in 3 cases). All KBs showed lysosomal accumulation in the podocytes, 92.8% showed inclusions in the parietal cells of the Bowman capsule, 85.7% in the tubules, while vascular inclusions were found in 78.5% of cases. Also, we observed segmental glomerular sclerosis in 5 cases, interstitial fibrosis and tubular atrophy in 3 cases, each. Only 2 patients were treated with pathogenic therapy before the first evaluation in our center. After multidisciplinary evaluation, 22 patients (62.8%) from our cohort received recommendation to initiate FD specific treatment, and 11 patients entered in our follow-up program. Evidence of specific FD lesions in KB was a strong reason to support the recommendation to initiate FD specific treatment. Conclusion Our results showed that kidney involvement in FD females can vary from asymptomatic or mildly symptomatic, to severely symptomatic, with end stage kidney disease. Thus, females should be carefully evaluated and monitored in a multidisciplinary team, in order to diagnosed FD affected organs. Also, we underline that a normal standard assessment of the kidney cannot rule out kidney involvement in female patients with FD, and strengthen the importance of KB for detection of early kidney involvement and for additional support for early initiation of FD specific therapy.

Relationship between morning blood pressure variability and cerebral microbleed burden in patients with hypertension.

Cerebral microbleeds (CMBs) can be understood as a type of target organ damage caused by hypertension. We aimed to explore the association of the CMB burden with morning blood pressure (BP) variability in patients with hypertension. We divided patients with hypertension into two groups: a group with 1-10 CMBs and a group with more than 10 CMBs. The duration, grade, medication, and control of hypertension were recorded in all patients. Morning home BP measurements were performed every 3 days for a month. A total of 791 patients were recruited. Full factor model analysis showed that higher morning home diastolic BP variability (standard deviation [SD], OR=1.080, 95% CI: 1.024-1.140, P=.005; coefficient of variation [CV], OR=1.076, 95% CI: 1.028-1.128, P=.002) was associated with more than 10 CMBs. Morning home systolic and diastolic blood pressure variability (SD, CV, average real variability) in more than 10 non-lobar CMBs group was significantly higher than that in 1-10 non-lobar CMBs group (P<.05).The multivariate analysis showed higher morning home diastolic blood pressure variability (SD, OR=1.124, 95% CI: 1.031-1.224, P=.008; CV, OR=1.099, 95% CI: 1.019-1.186, P=.015; average real variability, OR=1.055, 95% CI: 0.995-1.120, P=.075) was associated with more than 10 non-lobar CMBs. There was no significant relationship between morning home systolic blood pressure variability and more than 10 non-lobar CMBs (P>.05). Higher morning home diastolic blood pressure variability was associated with more than 10 CMBs and more than 10 non-lobar CMBs.