CD34 Antibodies Research Articles

Macrophage Membrane-Biomimetic Multi-Layered Nanoparticles Targeting Synovial Angiogenesis for Osteoarthritis Therapy.

Osteoarthritis (OA) is an inflammatory and progressive joint disease characterized by angiogenesis-mediated sustained, chronic, and low-grade synovitis. Anti-angiogenesis is emerging as a strategy for attenuating OA progression, but is often compromised by poor targeted drug delivery and immune clearance. Recent studies have identified macrophages formed a "protective barrier" in the lining layer (LL) of synovium, which blocked the communication of joint cavity and sublining layer (SL) of synovium. Inspired by natural mimicry, macrophage membrane-camouflaged drug delivery is explored to avoid immune clearance. Based on the single cell RNA sequencing, the CD34+ synovial cells are identified as "sentinel cells" for synovium angiogenesis. Consequently, CD34 antibody-modified macrophage membraneis constructed to target new angiogenesis. Hence, a biomimetic multi-layered nanoparticle (NP) is developed that incorporates axitinib-loaded poly(lactic-co-glycolic) acid (PLGA) with CD34 antibody modified macrophage membrane (Atb@NP@Raw@CD34) to specifically deliver axitinib (Atb) to the SL and sustain inhibiting angiogenesis without immune elimination. It is found that the Atb@NP@Raw@CD34 can pass through macrophage "barrier", specifically targeting CD34+ cells, continuously releasing Atb and anti-angiogenesis in OA synovitis. Furthermore, in vivo data demonstrated that Atb@NP@Raw@CD34 can attenuate joint degeneration by inhibiting synovium angiogenesis-mediated synovitis. In conclusion, local injection of Atb@NP@Raw@CD34 presents a promising approach for clinically impeding OA progression.

Perfusion Patterns of Peripheral Pulmonary Metastasis Using Contrast-Enhanced Ultrasound (CEUS) and Their Correlation with Immunohistochemically Detected Vascularization Pattern.

Description of the perfusion of pulmonary metastasis by contrast-enhanced ultrasound (CEUS) and their correlation with vascularization patterns represented by immunohistochemical CD34 endothelial staining. The data of 54 patients with histologic proven peripheral pulmonary metastasis, investigated between 2004 and 2023 by CEUS. These CEUS parameters were evaluated: time to enhancement (TE), categorized as early pulmonary-arterial (PA) or delayed bronchial-arterial (BA) patterns; extent of enhancement (EE), either marked or reduced; homogeneity of enhancement (HE), homogeneous or inhomogeneous; and decrease of enhancement (DE), rapid washout (<120 s) or late washout (≥120 s). Additionally, tissue samples in 45 cases (83.3%) were stained with CD34 antibody for immunohistochemical analysis. In total, 4 lesions (7.4 %) exhibited PA enhancement, and 50 lesions (92.6%) demonstrated BA enhancement. Furthermore, 37 lesions (68.5%) showed marked enhancement, while 17 lesions (31.5%) exhibited reduced enhancement. The enhancement was homogeneous in 28 lesions (51.86%) and inhomogeneous in 26 lesions (48.14%). Additionally, 53 lesions (98.1%) displayed a rapid washout. A chaotic vascular pattern indicative of a bronchial arterial blood supply was identified in all cases (45/45, 100%), including all 4 lesions with PA enhancement. Pulmonary metastases in CEUS predominantly reveal bronchial arterial enhancement and a rapid washout. Regarding EE and HE, pulmonary metastases show heterogeneous perfusion patterns. A PA enhancement in CEUS does not exclude BA neoangiogenesis.

Role of Interstitial Cells of Cajal in Congenital Ureteropelvic Junction Obstruction.

Although congenital ureteropelvic junction (UPJ) obstruction is the most common cause of neonatal hydronephrosis, aetiopathogenesis is still inconclusive. Recently, the paucity of interstitial cells of Cajal (ICC) at the narrow adynamic part of UPJ has been implicated as a causative factor. This prospective study was conducted between October 2019 and March 2022 to find out the density of ICC by the immunohistochemical method using CD117 (c-kit) antibody, in resected segments of UPJ in obstruction patients and in renal tumour patients as control. ICC/high power field (hpf) was also studied from the margins of the resected segment in the obstruction group. The pre-operative and post-operative sonographic and renal scintigraphic features were compared. The median age of patients in the study group (n = 25) was 36 months and in the control group was 39 months. The mean ICC/hpf at the stenotic part of UPJ in the study group was 3.56 ± 1.26 and in the control group was 12.56 ± 1.89 (P = 0.0001). ICC density from the proximal and distal margins of the resected segment was 11.12 ± 2.12 and 11.68 ± 1.62, respectively (P < 0.001). The post-operative antero-posterior diameter of the renal pelvis and differential renal function showed significant improvement in comparison to the pre-operative value (P = 0.0045 and 0.0005, respectively). The significant decrease in the density of ICC at the stenotic part of UPJ compared to controls suggests a pacemaker role of these cells in ureteral peristalsis and the aetiopathogenesis of UPJ obstruction. Histopathological analysis of ICC should not only be limited to the stenotic part of UPJ but also should focus on the anastomosed ends of the ureter, which reflects post-pyeloplasty outcome.

A histology and immunochemistry study comparing open versus close and open-method endoscopic saphenous vein harvesting.

The introduction of endoscopic saphenous vein harvesting (ESVH) has been reported to reduce wound pain and infection, compared with open saphenous vein harvesting (OSVH) techniques. There are still controversies regarding this technique. The aim of our study is to investigate the endothelial preservation of saphenous vein (SV) grafts harvested by different techniques. Further observations were made for harvesting and closure time, incision length and effect of pressure distension of the veins to the vein endothelium. Prospective observational study of sixty human saphenous vein grafts was performed to evaluate endothelial preservation by haematoxylin-eosin and Cluster of Differentiation 31 (CD 31) staining. Saphenous vein was harvested endoscopically either by closed CO2 (carbon dioxide) ESVH, open CO2 ESVH or OSVH harvesting technique. Demographic data and intra-operative data were collected. Two saphenous vein samples were collected from each patient to compare differences before and after distension of the veins. Both haematoxylin-eosin and immunohistochemistry slides were imaged by a high-resolution slide scanning system. Open CO2 ESVH group showed the highest number of endothelial detachments. Mean scoring of the immunohistochemistry method using the CD31 antibody was much lower in the open CO2 ESVH group (33.25% ± 28.71, P < 0.0003). This represents a more poorly preserved endothelial cells in the Open CO2 ESVH than the closed CO2 ESVH. Closure time and incision lengths were significantly shorter in both ESVH groups compared to the OSVH group. Significant low scores of immunohistochemistry for samples were seen in distended veins (39.0% ± 30.08, p = 0.004). The OSVH in random sample B, which represents the conduit that will be used, had a far better endothelium preservation and less endothelial detachment when compared to ESVH. We observed more endothelial detachment in the open CO2 ESVH group, due to lack of subcutaneous tissue separation, poor visualization and traction stress across the wall of the saphenous vein. The closed CO2 ESVH group had more endothelial cells preserved, but the OSVH group fared the best with the least number of endothelial cell detachment and a higher score of CD31 antibody. The online version contains supplementary material available at 10.1007/s12055-024-01752-3.

Abstract P233: Amyloid Beta Plaque Deposits Co-Localize with Endothelial Cells in the Hippocampus of Female APPswe/PSEN1dE9 Mice During the Onset of Amyloid Pathology

Increasing evidence shows that cardiovascular diseases are associated with an increased risk of cognitive impairment and Alzheimer’s diseases (AD). It is unknown whether systemic vascular dysfunction occurs prior to the development of AD, if this occurs in a sex-dependent manner, or whether endothelial cells play a role in the deposition of amyloid beta (Aβ) peptides. We hypothesized that female mice present with an exacerbated formation of Aβ peptides, and vascular dysfunction would occur during the onset of amyloid pathology, thus escalating its progression. To test this hypothesis, we used a double transgenic mouse model of early onset AD (APPswe/PSEN1dE9) prior to the onset of amyloid pathology (3 months of age) and during the onset of amyloid pathology (7 months of age). Immunofluorescence was performed in the hippocampus stained with Aβ 1-42 and CD31 antibodies. High resolution images were acquired on the confocal microscope. Acetylcholine-induced relaxation (1pM-30µM) was evaluated in mesenteric resistance arteries (MRA), and the data analyzed as non-linear curve regression and maximum effect (Emax). The statistical analysis was performed using Student’s t test, or two-way ANOVA followed by Tukey post-hoc ( p &lt;0.05*). We observed that during the onset of amyloid pathology (7-months-old), there was elevated Aβ deposition which co-localized with endothelial cells in the hippocampus from female but not male APP/PS1 mice [females colocalization rate (%): control 12.9 ± 2.0 vs . APP/PS1 25.1 ± 7.8, n=4; males colocalization rate (%): control 23.2 ± 6.5 vs. APP/PS1 15.3 ± 3.3, n=4]. In addition, the 7-month-old female APP/PS1 mice showed an exacerbated endothelium-dependent relaxation compared to the 3-month-old group (female APP/PS1 7-month-old ACh Emax: 97.1 ± 1.6*%, n=4 vs. 3-month-old ACh Emax: 70.1 ± 8.3%, n=6). Furthermore, endothelial dysfunction occurred prior to the onset of amyloid pathology irrespective of sex. Overall, these data showed that the endothelial dysfunction prior to the onset of amyloid pathology may be an early marker of AD. Further, a greater burden of Aβ formation in hippocampal endothelial cells from female but not male APP/PS1 mice may be associated with vascular abnormalities and dysfunction.

Diagnostic Values of Ki67, Cox2, CD31, E-cadherin, VEGF, MMP2, and MMP9 Protein Expression in Human Melanoma

Background: Cutaneous melanoma, the most severe form of skin cancer, has an unpredictable behavior and a high mortality rate. Recent research has identified new biomarkers and their associations with certain histopathological features, offering essential insights into diagnosis, prognosis, and therapeutic strategies. Material and method: In this meticulously designed and executed study, we analyzed 85 formalin-fixed, paraffin-embedded (FFPE) tissue samples using the gold standard technique of immunohistochemistry (IHC) to characterize the protein expression profiles. We used a comprehensive panel of Ki67, Cox2, CD31, VEGF, MMP2, MMP9, and E-cadherin antibodies. Clinical stages were meticulously determined according to TNM classification, the thickness of the Breslow depth, and Clark levels. We employed robust statistical methods such as one-way ANOVA and curve estimation regression to analyze the data. The STRING database, a trusted resource, was used to identify protein-protein interactions and related pathways. Results: The results of our study unveiled novel pairwise positive correlations between CD31 and Ki67 (r = 0.72), and between VEGF and Ki67 protein expression (r = 0.63). We also found a significant inverse relationship between the expression of E-cadherin and some biomarkers, such as Ki67, CD31, and VEGF (p &lt; 0.001 for all). Additionally, our findings revealed a strong positive association between the extent of tumor invasion and the expression levels of Ki67, CD31, and VEGF. The overexpression of these proteins was significantly correlated with advanced clinical stages (p &lt; 0.001 for all). Conclusion: Our study suggests that the biomarkers we examined could be reliable potential diagnostic and prognostic biomarkers for cutaneous melanoma. These findings have the potential to significantly impact the diagnosis, prognosis, and treatment of this deadly disease, offering new avenues for improved patient care.

Inhibition of JAK/STAT3 Expression by Acute Myeloid Leukemia-Targeted Nanoliposome for Chemotherapy Enhancement.

Acute myeloid leukemia (AML) is a relatively common malignant hematological disease whose development is mostly associated with abnormal activation of the JAK/STAT3 signaling pathway. Our previous study revealed that SAR317461, a novel JAK2/STAT3 inhibitor, can effectively inhibit the activation of the JAK2/STAT3 signaling pathway and has significant damaging and pro-apoptotic effects on AML cell lines. This project aims to build upon our prior research to enhance the application of SAR317461 in AML. The surface modification of liposomes with the CD34 antibody, along with the inclusion of the SAR317461 and cytarabine (a common AML chemotherapeutic agent), is observed. Due to the high expression of CD34 on the surface of AML cells, the nanoliposome could target AML cells specifically, further achieving an effective treatment for AML through the synergistic effect of JAK2/STAT3 inhibitors and chemotherapeutic agents. The implementation of this project will provide more theoretical support and ideas for the clinical application of JAK/STAT3 inhibitors in malignant tumors and for overcoming chemotherapy resistance.

Evaluation of circulating microvesicles and their procoagulant activity in patients with COVID-19

ObjectiveSeveral pathological conditions trigger the formation of microvesicles (MVs), including infectious diseases such as COVID-19. The shedding of MVs increases the levels of inflammatory factors (e.g., interleukin-6; IL-6) and ultimately leads to an inflammatory cascade response, while also increasing the procoagulant response. The current study aimed to evaluate the level of circulating MVs and their procoagulant activity as well as the serum level of IL-6 in patients with COVID-19 and healthy controls. In this case-control study, 65 patients with COVID-19 and 30 healthy individuals were sampled after obtaining written informed consent. MVs counting was measured using conjugated CD61, CD45, CD235a, and Annexin-V antibodies. Additionally, the procoagulant activity of MVs and the IL-6 level were estimated using enzyme-linked immunosorbent assay (ELISA).ResultsThe majority of MVs were platelet-derived MVs (PMVs). Patients with COVID-19 had significantly higher levels of MVs, procoagulant MVs, and IL-6 compared to healthy controls (p < 0.001). MVs were significantly correlated with procoagulant MVs, D-Dimer levels, fibrinogen, and IL-6, but not with platelet, lymphocyte, and neutrophil counts.ConclusionElevated levels of procoagulant MVs and their association with inflammatory and coagulation markers in patients with COVID-19 are suggested as a novel circulatory biomarker to evaluate and predict the procoagulant activity and severity of COVID-19.

Decalcification-free technique on the analysis of dental pulp tissue: histological and immunohistochemical analysis.

Histological techniques are essential for the microscopic study and investigation of the human dental pulp. The aim of this study was to evaluate the impact of decalcification-free technique by examining dental pulp morphology by histological staining with haematoxylin and eosin and immunohistochemistry. The sample consisted of 30 healthy third molars extracted for orthodontic indication, the pulp tissue was obtained by removing the mineralized tissues, separating the enamel and dentine and by marking with a flexible diamond disc on the coronal surface and longitudinal axis of the root. These guides made it possible to separate the fragments and obtain the pulp tissue for fixation and staining with H&E and subsequent immunohistochemistry with CD34 and S-100 antibodies. The technique showed preservation of pulp morphology with adequate preservation of microscopic structures. No alterations in tissue viability were observed. The staining allowed an accurate assessment of vascular and nervous components by means of CD34 and S-100 markers, respectively. This technique allows preservation of pulp tissue, maintaining viable tissue for histological analysis and immunohistochemistry tests, as well as reducing sample processing time.

Peculiarities of regeneration of an experimental femur fracture in rats with intramedullary fixation with carbon-carbon composite material implants

the issue of finding modern bioinert materials for use in surgical orthopedics and traumatology, as well as other medical fields related to the surgical treatment of diseases of the musculoskeletal system, is the most urgent today. The aim of the study was to establish the features of the course of regeneration of a femur fracture in rats with intramedullary fixation of fragments with a carbon-carbon composite material. The femur of 12 Wistar rats was studied in the work. Six rats - with simulated transverse diaphyseal fracture and intramedullary fixation with carbon-carbon composite material and six rats - with simulated transverse diaphyseal fracture and intramedullary fixation with medical steel. The last one were used as a comparison group. Histological sections prepared according to standard methods for inspection microscopy were stained with hematoxylin and eosin. To study revascularization processes, an immunohistochemical reaction with CD-34 antibodies was performed. As a result of the study, it was established that in animals with a simulated complete transverse diaphyseal fracture of the femur and intramedullary fixation with implants made of carbon-carbon composite material in the bone regenerate, the relative area of ​​CD-34+ vascular endothelial cells probably increases compared to the group of animals in which fixation was performed using medical stainless steel. Thus, the use of carbon-carbon composite material reveals an angiogenic effect in bone regeneration, which accelerates the rate of repair and remodeling of fractures.

Positive Effect of Peptides Obtained from Nile Tilapia (Oreochromis niloticus) on Inflammation Regulation and Wound Healing

This study investigated the effect of natural compounds from Nile tilapia (Oreochromis niloticus) skin on wound healing in IL-10 knockout mice. The healing fraction, Fraction T19, was obtained through hydrolysis with trypsin. In vitro, T19 was not cytotoxic to RAW 264.7 macrophage cells, promoting increased cell proliferation and migration. In vivo, mice (n = 30) were divided into three groups with 12 mm wounds in the dorsal region: control (distilled water), T1 (T19 at 125 μg/mL), and T2 (T19 at 250 μg/mL). Daily applications were performed, with tissue removal after nine days. The results showed that T19 increased the production of nitric oxide (NO) and hydrogen peroxide (H2O2), preventing wound contamination. There was an increase in pro-inflammatory (IL-2 and IFNγ) and anti-inflammatory (IL-4) cytokines, as well as cell proliferation markers (PCNA and KI67). Antibodies CD31, CD163, and COX-2 indicated an increase in the formation of new vessels and a reduction in inflammation. Both groups treated with T19 showed better healing results, with better effects observed at higher doses. It was concluded that T19 can effectively modulate the skin repair process and represent an alternative therapeutic for improving the quality of wound skin, especially in the clinical context. Formulations using tilapia skin are safe and effective for accelerating wound healing.

Морфофункциональное состояние органов иммунной системы у поросят-гипотрофиков в ранний неонатальный период

Abstract. The purpose of the study was to study the state of the immune system organs in hypotrophic piglets in the early neonatal period in an industrial pig breeding complex. Methods. The experiment was conducted in 2023 in a large industrial pig farm in the Voronezh region on piglets of the wounded neonatal period received from sows 3-4 farrowing. The sows were kept at optimal microclimate parameters, taking into account their physiological state, and fed with SK-1 nutritionally balanced feed. At the initial stage of the experiment, the piglets obtained during farrowing underwent clinical examination and weighing. Animals under 800 g are counted as hypotrophic piglets (n = 30), animals over 800 g are normotrophic, respectively (n =30). After the groups were formed, animals were forced to be slaughtered before taking colostrum (n =5) and biological material was taken from each group (thymus, lymph nodes (inguinal), spleen for immunohistochemical studies. Results. The level of mitotic activity in the thymus of normotrophic piglets was 9.4 % higher (p &lt; 0.05) than in piglets with body weight deficiency. In the spleen, the level of mitotic activity had no significant differences and was approximately the same in all animals participating in the experiment. In lymph nodes, the mitotic activity of cells in normotrophics was 12.7 % higher (p &lt; 0.05). A study of positively expressed CD-3 cells in the spleen revealed significant differences, so, in normotrophic piglets, the number of cells positively stained with this marker was significantly higher than in piglets with a body weight deficiency by 10.2 % (p &lt; 0.05). The number of CD-3 cells in the thymus differed by 5.6 %, and in the lymph nodes by 2.4 % between the groups, but there were no significant differences. The cellular expression of “immature” forms of B lymphocytes (PAX-5) in the lymph nodes of normotrophics was significantly higher by 12.9 % (p &lt; 0.05) compared with hypotrophic piglets. The scientific novelty lies in the fact that for the first time a comprehensive immunohistochemical study of the organs of the immune system of piglets in the early neonatal period was carried out using monoclanal antibodies CD-3, Ki-67 and PAX-5. As a result of the experiment, it was revealed that piglets with body weight deficiency have a “depression” of the immune system, which manifests itself in hypoplasia of T-lymphocytes in the spleen, “maturing” B-lymphocytes in the lymph nodes and low mitotic potential in the thymus and lymph nodes.

Intratumoral T-cell composition predicts epcoritamab-based treatment efficacy in B-cell non-Hodgkin lymphomas.

Leveraging endogenous tumor-resident T-cells for immunotherapy using bispecific antibodies (BsAb) targeting CD20 and CD3 has emerged as a promising therapeutic strategy for patients with B-cell non-Hodgkin lymphomas. However, features associated with treatment response or resistance are unknown. To this end, we analyzed data from patients treated with epcoritamab-containing regimens in the EPCORE NHL-2 trial (NCT04663347). We observed downregulation of CD20 expression on B-cells following treatment initiation both in progressing patients and in patients achieving durable complete responses (CR), suggesting that CD20 downregulation does not universally predict resistance to BsAb-based therapy. Single-cell immune profiling of tumor biopsies obtained following one cycle of therapy revealed substantial clonal expansion of cytotoxic CD4+ and CD8+ T-cells in patients achieving CR, and an expansion of follicular helper and regulatory CD4+ T-cells in patients whose disease progressed. These results identify distinct tumor-resident T-cell profiles associated with response or resistance to BsAb therapy.

P-327 A multicenter retrospective study of the investigation of detailed immunological inflammatory grading for chronic endometritis and their correlation with the endometrial microbiome

Abstract Study question There exists refractory chronic endometritis which does not respond to antibiotic treatment. This study aims to elucidate its pathogenesis and develop detection methods. Summary answer Through treatment intervention for chronic endometritis (CE), the inflammatory grade decreased overall and the bacteria most associated with endometritis were Lactobacillus iners. What is known already CE and endometrial dysbiosis have been reported to be associated with recurrent pregnancy loss (RPL) and recurrent implantation failure (RIF), but there is insufficient data confirming the interrelation between them. Moreover, the diagnosis of CE typically involves single immunohistochemical staining with CD138, but the diagnostic criteria vary across reports (e.g., McQueen’s method, Liu’s method, etc.) and moreover, use of this classical inflammatory assessment methods raises concerns about their sufficiency in evaluating the pathogenesis of endometritis in cases of RPL and RIF. Study design, size, duration A retrospective analysis was conducted on 206 samples of endometrial specimens collected from infertile patients at seven participating assisted reproductive healthcare facilities from 2021 to 2023. Among these, 73 samples (from 48 cases) had undergone both histological examination and simultaneous endometrial microbiome testing, and data on pregnancy outcomes (including implantation success and ongoing pregnancy rate) were collected until December 2023 for these cases. Participants/materials, setting, methods The endometrial specimens from cases with RIF or RPL were obtained through intrauterine aspiration in luteal phase. Immunohistochemical staining using CD138, CD56, and CD8 antibodies was performed. A composite grading for endometritis, based on histological findings and immunohistochemistry intensity, was established (INF grading; grade 0 to 5). Furthermore, a portion of the specimens underwent microbiome analysis using next-generation sequencing targeting the 16SrRNA, exploring correlations between bacterial taxa, abundance, histological findings, and pregnancy outcomes. Main results and the role of chance Among analyzed 48 cases, 11 cases underwent repeated endometrial microbiome analysis post-treatment again, while 7 cases had additional testing without improvement in chronic endometritis (CE) or endometrial dysbiosis. The McQueen’s method diagnosed 27 cases with CE, while the Liu method identified 4 cases. According to our INF grading, 11 cases were classified as endometritis grade 3 or above. Successful embryo implantation was achieved in 18 cases (37.5%), with ongoing pregnancy in 13 cases (27.1%). CE treatment involved broad-spectrum oral antibiotics, mainly doxycycline and metronidazole, resulting in improvement to grade 2 or below in 9 out of 11 cases with INF grade 3 or above. However, clear associations between pregnancy outcomes and INF grades were not apparent, with the most significant correlation found in CE diagnosis using Liu’s method. Analyzing biomarkers for CE through LEfSe (Linear discriminant analysis effect size), both McQueen’s method and INF grading 2 or above were associated with the intrauterine presence of Lactobacillus iners. Estimating endometrial inflammation based on intrauterine Lactobacillus iners presence yielded an AUC of 0.733 for McQueen’s CE and 0.723 for INF grade 2 or above. Limitations, reasons for caution This study is a retrospective analysis, and the treatment protocols and intervention criteria for chronic endometritis vary among different facilities. The number of cases is also limited, and it is desirable to conduct a unified prospective multicenter cohort study with consistent protocols. Wider implications of the findings The combination of the endometrial microbiome testing and detailed inflammatory grading suggests the potential for establishing appropriate treatment intervention strategies and predicting the success rate of infertile treatment. Since Lactobacillus iners tends to colonize extensively in the vaginal environment of East Asian women, it is necessary to explore region-specific measures. Trial registration number not appicable

Evidence of Endothelial Bone Marrow Microenvironment Affection in Poor Graft Function Post Allogenic Stem Cell Transplanration

Abstract Allogeneic hematopoietic stem cell transplantation (HSCT) is considered a curative option for several hematological disorders. The prognosis of patients undergoing allogenic (HSCT) is dependable on various factors that affect bone marrow microenvironment and poor prognosis can results from poor graft function. On of these factors is the numbers of CD31+ endothelial cells that support the hematopoietic stem cells. Aim of the Study The aim of this study was to assess how the bone marrow microenvironment is being affected in patients with poor graft function post allogenic stem cell transplantation targeting the endothelial cells using multiparameter flow cytometric analysis of CD31 antibody. Patients and Methods Forty patients were enrolled in this study with both benign and malignant hematological diseases who underwent allogenic HSCT from matched related donors. Patients were divided into 2 equal groups: poor graft function (PGF) group and good graft function group (GGF). Patients were subjected to full history recording, clinical examination, full laboratory and radiological work-up, tissue typing and chemotherapeutic conditioning before transplantation. After transplant, bone marrow aspirate was obtained and analyzed for CD31+ endothelial cells using Quantitative tri-color Flow cytometric assay. Results PGF group showed significantly lower counts of WBCs, platelets and hemoglobin concentration compared to GGF group. In addition, CD34 dose showed significantly lower concentration in PGF group compared to GGF group. PGF group had statistically significant lower percentage of stem cells total count compared to GGF group in addition, PGF Group had a significantly higher mean fluorescence intensity (MFI) of CD31 on non-stem cells compared to GGF group. Conclusion PGF had significant increase in MFI of CD31+ve on non-stem cells compared to GGF group. In addition, PGF group had statistically significant lower percentage of CD31+ stem cells total count compared to GGF group.

P-302 Oral ofloxacin combined with vaginal metronidazole for chronic endometritis: a prospective open-label cohort study

Abstract Study question Does oral ofloxacin 2x200 mg for 10 days combined with vaginal metronidazole 1x500 mg for 10 days reduce the activity of chronic endometritis? Summary answer The investigated antibiotics may be beneficial in some cases of chronic endometritis, except in cases of concomitant endometrial polyp, where they adversely affect treatment outcome. What is known already Chronic endometritis (CE) is characterized by the presence of atypical plasma cell infiltrates (CD138 positive) in the endometrial stroma. CE may adversely affects fertility by reducing endometrial receptivity, impairing decidualization and uterine contractility, thus increasing the risk of recurrent pregnancy loss and implantation failure. Some idiopathic infertility cases are attributed to CE. The diagnostics of CE is challenging because the clinical evaluation is nonspecific and the immunohistochemical criteria are not consistent between research centers. Literature data suggested that successful treatment of CE with antibiotics could improve live birth rates, but a preferred treatment regimen has not been identified. Study design, size, duration A prospective cohort study included women undergoing office hysteroscopy (OH) at a tertiary referral center in 2021-2023. Women diagnosed with CE were assigned, based on their preference, to the intervention arm of ofloxacin 2x200 mg orally for 10 days + metronidazole 500 mg vaginally for 10 days or to the control arm (no antibiotics). 100 women underwent repeated hysteroscopy after 3 cycles, of which 77 received antibiotics and 23 were controls. Participants/materials, setting, methods Women aged 18-45 who met eligibility criteria for OH were included. Karl Storz 5.0 mm Bettocchi® operating sheath with a 2.9 mm 30° telescope and a 5Fr working channel was used. Endometrial lesions or biopsy specimens were collected with grasping forceps, blunt scissors or bipolar needle electrode. Saline was used as the dilating medium. Tissue samples were subjected to histopathological examination. Plasma cells were detected by immunohistochemical staining with Monoclonal Mouse Anti-Human CD138 antibodies. Main results and the role of chance The most common findings identified at hysteroscopy were endometrial polyps (43%), polypoid endometrium (38%), micropolyps (23%), focal (27%) and diffuse hyperemia (18%) and cesarean scar defect (14%). CE was defined as the presence of &amp;gt; 2 plasma cells/mm2. The Mann-Whitney U test showed a significantly higher plasma cell count (PCC) in the presence of a cesarean scar defect than in other cases (23 vs. 18/mm2, p = 0.031) and a tendency towards a lower PCC in micropolyps (9 vs. 21/mm2, p = 0.063). The correlation of hysteroscopic findings with CE activity was weaker than expected. Using repeated measures ANOVA, no difference in PCC between primary and follow-up hysteroscopy was found in either arm (p = 0.8), while post-hoc tests showed that the mean PCC decreased from 12 to 5/mm2 (p = 0.02) in the intervention arm, and from 14 to 6 (p = 0.23) in the control arm. On follow-up hysteroscopy, polyp remission was observed significantly more often in the control arm than in the intervention arm (61% vs. 27%, pChi2=0.02), in which de novo polyp was observed in 4%. In polypoid endometrium, the remission rate was similar in both arms (19% vs. 22%), and the rate of de novo lesions was significantly lower in interventional arm (16% vs. 39%, pChi2=0.035). Limitations, reasons for caution The limitations include the small sample size, the CE definition based on the number of plasma cells calculated using the Receiver Operating Characteristic curve for the purposes of the study, and the empirical selection of antibiotic therapy. Wider implications of the findings Due to the weak correlation of hysteroscopic findings with CE activity, the diagnostics and treatment of CE based on clinical evaluation of the uterine cavity is not justified. Poorly selected antibiotic therapy may adversely affect the outcomes of treatment. Trial registration number 1072.6120.322.2020

Long-Term Cost-Effectiveness of Case Finding and Mass Screening for Celiac Disease in Children

Background and aimsCeliac disease (CD) is a common yet underdiagnosed autoimmune disease with substantial long-term consequences. High-accuracy point-of-care tests (POCTs) for CD antibodies conducted at youth primary healthcare centers (YHCCs) may enable earlier identification of CD, but evidence about the cost-effectiveness of such strategies is lacking. We estimated the long-term cost-effectiveness of active case-finding and mass-screening compared to clinical detection in the Netherlands. MethodsA decision tree and Markov model were used to simulate a cohort of three-year-old children with CD according to each strategy, taking into account their impact on long-term costs (from a societal perspective) and quality-adjusted life-years (QALYs). Model parameters incorporated data from the GLUTENSCREEN project, the Dutch Celiac Society, the Dutch Pediatric Surveillance Unit, and published sources. The primary outcome was the incremental cost-effectiveness ratio (ICER) between strategies. ResultsMass-screening produced 7.46 more QALYs and was € 28,635 more costly compared to current care (ICER: € 3841/QALY), and case-finding produced 4.33 more QALYs and was € 15,585 more costly compared to current care (ICER: € 3603/QALY). At a willingness to pay of € 20,000 per QALY, both strategies were highly cost-effective compared to current care. Scenario analyses indicated that mass-screening is likely the optimal strategy, unless no benefit in detecting asymptomatic cases is assumed. ConclusionAn earlier identification of CD through screening or case-finding in children using a POCT leads to improved health outcomes and is cost-effective in the long-term compared to current care. If the feasibility and acceptability of the proposed strategies are successful, implementation in Dutch regular care is needed.

Evaluation of Microvascular Density in Glioblastomas in Relation to p53 and Ki67 Immunoexpression.

Glioblastoma is the most aggressive tumor in the central nervous system, with a survival rate of less than 15 months despite multimodal therapy. Tumor recurrence frequently occurs after removal. Tumoral angiogenesis, the formation of neovessels, has a positive impact on tumor progression and invasion, although there are controversial results in the specialized literature regarding its impact on survival. This study aims to correlate the immunoexpression of angiogenesis markers (CD34, CD105) with the proliferation index Ki67 and p53 in primary and secondary glioblastomas. This retrospective study included 54 patients diagnosed with glioblastoma at the Pathology Department of County Emergency Clinical Hospital Târgu Mureș. Microvascular density was determined using CD34 and CD105 antibodies, and the results were correlated with the immunoexpression of p53, IDH1, ATRX and Ki67. The number of neoformed blood vessels varied among cases, characterized by different shapes and calibers, with endothelial cells showing modified morphology and moderate to marked pleomorphism. Neovessels with a glomeruloid aspect, associated with intense positivity for CD34 or CD105 in endothelial cells, were observed, characteristic of glioblastomas. Mean microvascular density values were higher for the CD34 marker in all cases, though there were no statistically significant differences compared to CD105. Mutant IDH1 and ATRX glioblastomas, wild-type p53 glioblastomas, and those with a Ki67 index above 20% showed a more abundant microvascular density, with statistical correlations not reaching significance. This study highlighted a variety of percentage intervals of microvascular density in primary and secondary glioblastomas using immunohistochemical markers CD34 and CD105, respectively, with no statistically significant correlation between evaluated microvascular density and p53 or Ki67.

[123I]CLINDE SPECT as a neuroinflammation imaging approach in a rat model of stroke

Poststroke neuroinflammation exacerbates disease progression. [11C]PK11195-positron emission tomography (PET) imaging has been used to visualize neuroinflammation; however, its short half-life of 20 min limits its clinical use. [123I]CLINDE has a longer half-life (13h); therefore, [123I]CLINDE-single-photon emission computed tomography (SPECT) imaging is potentially more practical than [11C]PK11195-PET imaging in clinical settings. The objectives of this study were to 1) validate neuroinflammation imaging using [123I]CLINDE and 2) investigate the mechanisms underlying stroke in association with neuroinflammation using multimodal techniques, including magnetic resonance imaging (MRI), gas-PET, and histological analysis, in a rat model of ischemic stroke, that is, permanent middle cerebral artery occlusion (pMCAo). At 6 days post-pMCAo, [123I]CLINDE-SPECT considerably corresponded to the immunohistochemical images stained with the CD68 antibody (a marker for microglia/microphages), comparable to the level observed in [11C]PK11195-PET images. In addition, the [123I]CLINDE-SPECT images corresponded well with autoradiography images. Rats with severe infarcts, as defined by MRI, exhibited marked neuroinflammation in the peri-infarct area and less neuroinflammation in the ischemic core, accompanied by a substantial reduction in the cerebral metabolic rate of oxygen (CMRO2) in 15O-gas-PET. Rats with moderate-to-mild infarcts exhibited neuroinflammation in the ischemic core, where CMRO2 levels were mildly reduced. This study demonstrates that [123I]CLINDE-SPECT imaging is suitable for neuroinflammation imaging and that the distribution of neuroinflammation varies depending on the severity of infarction.

Isolation and characterization of stem cells from human exfoliated deciduous teeth.

SHEDs have been shown to have a higher rate of proliferation and raise in cell population doublings when compared to stem cells from permanent teeth. Hence, using them in tissue engineering may be advantageous over stem cells from adult human teeth. Stem cells were removed from pulpal tissues of thirty primary teeth undergoing extraction under six to fourteen year of age. The tissues were incubated after centrifuging and adding DMEM-KO following the addition of a 2 mg/ml collagenase blend for examination of plates in search of cell attachment and growth. Flow cytometric analysis showed successful isolation of SHEDs using fluoresce inisothiocyanate (FITC)-conjugated CD-34, CD-105, and PE (R-phycoerythrin)-conjugated CD-45, CD-90, CD-73, and HLA-DR antibodies. The surface antigens CD-73, CD-90 and CD-105 which are known to be present in mesenchymal lineages were positively expressed in SHEDs according to flow cytometry analysis, whereas CD-34, CD-45, and HLA-DR were not.