Immunological Reactions Research Articles

Effect of bisphenol A on formation and process of alergic reaction - review

One of the possible immunomodulatory effects of BPA is alterating the allergy process and asthma. The growth in the prevalence of asthma around the world roughly corresponds with the rise in the widespread usage of certain chemicals. Despite the fact that asthma is a complicated and varied condition brought on by exposure to a variety of environmental factors and genetic predispositions, many classes of chemicals that are frequently present in the surrounding environment and are linked to endocrine disruption have recently been linked to the disease's pathogenesis. BPA is endocrine disrupting chemical compound made up of two phenol rings joined by a methyl bridge and two methyl functional groups adhered to the bridge. BPA is ingested by humans, inhaled via home dust, and exposed through the skin. An allergy is an aberrant immunological reaction to an antigen, such as a protein or allergen. An ingested or breathed allergen encounters the epithelium as its initial barrier. The inflammatory condition and increased permeability of the epithelial barrier are associated with allergic inflammatory disorders, and this suggests that the epithelium barrier is more sensitive to allergen sensitization. The existing literature and scientific publications do not adequately address the direct influence of bisphenols on the development and influence of the allergic process. Multiple authors discuss how bisphenol affects specific allergic response components. We may conclude that exposure to bisphenol A alters the allergic response based on the findings of the studies and the analysis of the literature.

Incidental Bystander or Essential Culprit: A Systematic Review of Bacterial Significance in the Pathogenesis of Breast Implant-Associated Anaplastic Large Cell Lymphoma.

Breast implant-associated anaplastic large cell lymphoma (BIA-ALCL) is a distinct subtype of T-cell non-Hodgkin lymphoma that arises in the context of prolonged exposure to textured breast implants. The intent of this manuscript is to explore whether the bacterial presence in biofilms on these implants is a mere incidental finding or plays a pivotal role in the pathogenesis of BIA-ALCL. Our goal is to delineate the extent of bacterial involvement, offering insights into potential underlying mechanisms, and establishing future research priorities aimed at resolving the remaining uncertainties surrounding this complex association. A comprehensive systematic review of several databases was performed. The search strategy was designed and conducted by an experienced librarian using controlled vocabulary with keywords. The electronic search identified 442 publications. After evaluation, six studies from 2015 to 2021 were included, encompassing 201 female patients aged 23 to 75. The diagnosis span post-implantation ranged from 53 to 135.6 months. Studies consistently found bacteria near breast implants in both BIA-ALCL cases and controls, with varied microbial findings. Both BIA-ALCL cases and controls exhibited the presence of specific bacteria, including Pseudomonas aeruginosa, Klebsiella oxytoca, Staphylococcus aureus, and Ralstonia spp., without any statistically significant differences between groups. The use of antiseptic and antimicrobial agents during implant insertion did not demonstrate any impact on reducing or altering the risk of developing BIA-ALCL. Our systematic review reveals that the current evidence is inadequate to link bacterial etiology as a central factor in the development of BIA-ALCL. The limitations in the existing data prevent a complete dismissal of the role of biofilms in its pathogenesis. The observed gap in knowledge underscores the need for more focused and comprehensive research, which should be structured in a multi-faceted approach. Initially, this involves the utilization of sophisticated genomic and proteomic methods. Following this, it is crucial to delve into the study of immunological reactions specifically induced by biofilms. Finally, this research should incorporate extended observational studies, meticulously tracking the evolution of biofilm development and its correlation with the emergence of BIA-ALCL. In light of the inconclusive nature of current findings, further investigation is not only justified but urgently needed to clarify these unresolved issues.

The Prevalence of Serum Anti Nuclear Antibodies in Children Treated With Anti-Epileptics

To evaluate the prevalence of positive serum antinuclear antibody (ANA) in children with epilepsy using three major antiepileptic drugs (phenytoin, carbamazepine and ethosuximide), 60 children under 18 years with epilepsy who were referred to pediatric neurology clinic or had admitted to neurology ward in Children Hospital in Tehran, Iran, were entered our study. They had been treated with one of the three antiepileptic drugs (carbamazepin, phenytoin, ethosuximide) with suitable dose for at least one month. The patients were divided into two groups according to the classification of the International League Against Epilepsy (ILAE): drug-resistant and drug-responsive. We studied the epidemiological and clinical characteristics and also serum ANA of the patients in both groups. In this research, we studied ANA in 60 epileptic children. 30 patients were diagnosed with drug resistant epilepsy and the other 30 were drug responsive. None of them showed the clinical manifestations of lupus erythematosus. As a whole, 7 patients (11.7%) were ANA-positive, 6.7% of drug resistant and 16.7% of drug responsive group showed this finding. There was no relationship between drug resistancy and ANA according to statistical studies (P=0.21). Although in our study, epidemiological and clinical data of the patients was reported in two separate groups of resistant or responsive to antiepileptic drugs, and no meaningful statistical difference was found between these two groups. Overally in our study, the prevalence of positive ANA in patients receiving antiepileptic drugs was less in comparison with previous studies and was more common in males. Finally, we suggest a more comprehensive and extensive study with more cases and further follow-up period in order to find the cause of immunological reactions to antiepileptic drugs in children with epilepsy.

Advances and application of common biosensors

Biosensors are devices that can detect and measure chemical components in organisms through specialized biological properties or reactions. This paper provides an overview of the three main types of biosensors: enzymatic, immunosensors, and microbial sensors. Enzymatic biosensors utilize enzymes to catalyze specific reactions between target molecules and other components of the biosensor, creating measurable signals. Immunosensors rely on immunological reactions between antigens and antibodies to detect and quantify substances in a solution. Microbial sensors use microbial cells to produce signal outputs being tested. Each type of biosensor has unique applications and mechanisms that make them useful in various scientific studies, including clinical medicine, laboratory research, environmental monitoring, and food engineering. Ongoing advancements in technology will undoubtedly continue to expand the scope of biosensors uses, making them increasingly important in many fields. This paper aims to provide researchers with an understanding of the mechanisms behind biosensors and strategies to improve their performance for better measurement, detection, and monitoring capabilities.

Prevalence and Risk Factors for IgE and IgG Cow's Milk Protein Allergies in Saudi Arabia.

Cow's milk protein allergies (CMPAs) particularly occur in infancy and early childhood due to an immunological allergic reaction to milk proteins. This issue is increasing in Saudi Arabia and requires research to improve health status and provide alternatives. Our study aims to investigate the important aspects of immunoglobulin E(IgE) and IgG CMPA in Saudi Arabia regarding its prevalence and association with demographic and health-related factors in both adult and pediatric populations. A descriptive retrospective cross-sectional study was conducted on 376 patients, comprising 314 adults aged between 19 and 86 years, and 62 pediatric patients aged between one and 12 years. The study focused on individuals who attended a private medical center in Jeddah city over the past five years. Laboratory results for food allergy of IgE and IgG tests, including cow's milk proteins (CMPs), serum 25-hydroxyvitamin D (25OHD), specific IgE inhalant allergy results, and other health-related factors were collected from an electronic record system. Results have shown that casein was the most common sensitizing allergen of CMPs in adults, whereas α-lactalbumin was the most common sensitizing allergen in pediatrics. The most frequent sensitizing allergen in IgG CMP was cow's milk in 54/92 (58.7%) adults, followed by cow's sour milk products 41 (44.6%). Cow's milk was the most common sensitizing allergen in 20/20 (100%) children. The rate of CMPA was significantly higher in children younger than five years old (P =0.003), while children who interacted with pets had a marginal significantly reduced rate (P = 0.054). Thus, cow's milk is the most sensitizing allergen in IgG CMPs in adults and pediatrics.

FEATURES OF USING ORTHOPEDIC IMPLANTS ON AN EXPERIMENTAL MODEL OF SENSITIZATION TO NICKEL (NI)

Introduction. The theme of metal hypersensitivity (MHS) reactions is among the most controversial and complex issues in orthopedic and trauma practice. MHS diagnoses are diagnostically challenging because they are exclusionary diagnoses. It is currently uncertain which biomarkers can reliably predict a potential pathological response to implants.
 The aim of this research is to investigate hypersensitivity reactions to orthopedic implants containing nickel (Ni).
 Materials and methods. This research was conducted on female Wistar rats in accordance with the standards of bioethical principles. To obtain conclusive results, the animals were divided into two experimental groups: with prior sensitization to Freund's adjuvant containing Ni and without it. Nickel plates with a total surface area of 24 mm2 were implanted in the rats under general anesthesia. Five months after the intervention, the animals were removed from the experiment, and histological examination of the obtained samples was conducted. Extracted implants were examined using electron microscopy to assess the implant surface. The local elemental composition of the implants was analyzed using an energy-dispersive spectrometer.
 Results. Under the experimental conditions, it was demonstrated that after 5 months following the operation, a dense connective tissue capsule with an inflammatory infiltrate in the capsule lumen had formed around the implants in the rats. This suggests a possible manifestation of hypersensitivity reactions to implants containing Ni. Electron microscopy of the surface of the extracted implants revealed corrosion phenomena in all the samples. The degree of corrosion was more pronounced in the group of animals with prior sensitization to Ni, and distant particles of Ni were detected, which can be characterized as the beginning of implant degradation.
 Conclusions. A connective tissue capsule forms around the implants, and it was found to be 34.8% denser in animals sensitized to Ni prior to implantation, which may indicate tissue reactions with signs of hypersensitivity. Further research will provide a deeper understanding of the fundamental inflammatory and immunological reactions to metals present in implants. This, in turn, will facilitate the identification of clinically useful applications necessary for the development of diagnostic or prognostic tests for patients with metal implants.

Microbial mavericks: unleashing bacteria’s tumor-seeking superpowers in the fight against cancerMicrobial mavericks: unleashing bacteria’s tumor-seeking superpowers in the fight against cancer

Capitalizing on bacteria's natural capacity to infiltrate tumor microenvironments, bacterial cancer therapy offers a fresh paradigm for targeting solid tumors. Bacterial cancer therapies have a unique mechanism to cure cancer that is unachievable with current methods. This article addresses the possibility for "Microbial Mavericks," or bacteria, to accumulate specifically in cancerous tissues by taking advantage of the special characteristics of tumors’ micro-environment including hypoxia and acidity. Bacteria can be simply genetically modified to develop and deliver anticancer agents according to clinical needs. Researchers want to improve therapeutic efficacy while reducing systemic negative effects by utilizing bacteria's tumor tropism. The need to carefully weigh the potential advantages of bacterial therapeutics with the risks of infections, immunological reactions, and bacterial resistance is emphasized, along with safety issues and challenges. Over the last decades, Salmonella, Clostridium, Listeria, and many other genera have been researched to inhibit cancer growth and increase survival in animal models. This manuscript provides insights into the potential, challenges, and ethical ramifications of the developing field of bacterial cancer therapy. The symbiotic interaction between bacteria and tumors may usher in a new era of cancer treatment as clinical research advances, leveraging the strength of microbial allies in the battle against cancer.

Microbial mavericks: unleashing bacteria’s tumor-seeking superpowers in the fight against cancer

Capitalizing on bacteria's natural capacity to infiltrate tumor microenvironments, bacterial cancer therapy offers a fresh paradigm for targeting solid tumors. Bacterial cancer therapies have a unique mechanism to cure cancer that is unachievable with current methods. This article addresses the possibility for "Microbial Mavericks," or bacteria, to accumulate specifically in cancerous tissues by taking advantage of the special characteristics of tumors’ micro-environment including hypoxia and acidity. Bacteria can be simply genetically modified to develop and deliver anticancer agents according to clinical needs. Researchers want to improve therapeutic efficacy while reducing systemic negative effects by utilizing bacteria's tumor tropism. The need to carefully weigh the potential advantages of bacterial therapeutics with the risks of infections, immunological reactions, and bacterial resistance is emphasized, along with safety issues and challenges. Over the last decades, Salmonella, Clostridium, Listeria, and many other genera have been researched to inhibit cancer growth and increase survival in animal models. This manuscript provides insights into the potential, challenges, and ethical ramifications of the developing field of bacterial cancer therapy. The symbiotic interaction between bacteria and tumors may usher in a new era of cancer treatment as clinical research advances, leveraging the strength of microbial allies in the battle against cancer.

Paradoxical increase of neurofilaments in SMA patients treated with onasemnogene abeparvovec-xioi.

Neurofilament light chain (NfL) has been proposed as a biomarker reflecting disease severity and therapy response in children with spinal muscular atrophy type 1 and 2 (SMA1 and 2). The objective of this study was to examine how serum NfL changes after gene replacement therapy (GRT) with onasemnogene abeparvovec-xioi. We measured NfL in serum probes from 19 patients (10 SMA 1 and 6 SMA 2; 15 previously treated with nusinersen or risdiplam; 12 male) before and at variable time points after GRT. These values were related to motor scores (CHOP-Intend, HFMSE and RULM). Median age at GRT was 19 months (range 2-46 months). Median NfL of all patients before GRT was 39 pg/ml (range 0-663 pg/ml; normal values <25 pg/ml), increased significantly to 297 pg/ml (range 61-1,696 pg/ml; p<0,002) 1 month after GRT, and decreased to 49 pg/ml (range 24-151 pg/ml) after 6 months. Subjects pre-treated with nusinersen or risdiplam had lower baseline NfL levels than naïve patients (p<0,005), but absolute increases of NfL were similar in both groups. While motor scores were improved in 14 out of 18 SMA patients (78%) 6 months after GRT NfL values differed not significantly from those measured at baseline (p = 0,959). Serum NfL showed a paradoxical transient increase after GRT in both, pre-treated and naïve patients, which may reflect an immunological reaction in the CNS related to transfection of neuronal cells by AAV9. The clinical meaning of this increase should be assessed in future studies. Our findings encourage regular monitoring of NfL in OA treated patients.

Diagnosis and Treatment of Leprosy in Taiwan during the COVID-19 Pandemic: A Retrospective Study in a Tertiaty Center.

Currently, over 200,000 new cases of leprosy are reported annually worldwide. Although leprosy was thought to have been eradicated in Taiwan, a few new cases still occur annually. Protean clinical manifestations of leprosy and immunological reactions result in delayed diagnoses. In addition, drug-resistant leprosy is emerging and poses treatment challenges. In this retrospective study, we collected and analyzed the clinicopathological features, leprosy type, treatment response, and relapse rate of patients with leprosy in our hospital between January 2009 and November 2022. We found that 54% of patients were Indonesian, and borderline lepromatous leprosy was predominant (39%); moreover, histoid leprosy and the Lucio phenomenon were also reported. Polymerase chain reaction analysis identified four positive cases, including a dapsone-resistant (4%) case. Our findings indicated good control of leprosy and a lower rate of dapsone resistance than that reported by the World Health Organization (4% vs. 13%) from 2009 to 2015. We found that the patient profile in terms of the treatment duration, recurrence rate, systemic symptoms, and neurological symptoms did not differ between before and during the pandemic. We report the recent advances in leprosy diagnosis, drug-resistant gene mutations, post-exposure prophylaxis, vaccination, and the effect of coronavirus disease 2019 on leprosy to facilitate updated leprosy diagnosis and management.

CRISPR/Cas9 Technology: Challenges and drawbacks

Genome engineering has been transformed in recent years by the introduction of the CRISPR technology for a variety of cancer research projects spanning from fundamental science to translational medicine and precision cancer treatment. Although there have been tremendous advancements in this area, a number of technical issues still need to be resolved, including off- target activity, inadequate indel or poor homology-directed repair (HDR) efficiency, in vivo distribution of the Cas system components, and immunological reactions. Chromosome rearrangements brought on by off-target effects might unintentionally affect some poorly matched genomic locations and restrict the use of CRISPR-Cas editing technologies for therapeutic reasons. Studies have shown that CRISPR-Cas tools may be more susceptible to off-target effects than some of the other common gene-editing techniques because a Cas protein is a monomer that might accidentally enhance the identification of shorter target sequences, whereas the TALEN and ZFN assemblies are dimeric. Off-target effects often come from Cas enzymes that cleave bystanders (not intended targets) and guide RNA to recognize mismatches. CRISPR systems delivered in vivo can trigger immune responses against foreign substances by significantly increasing people's innate immunity and/or adaptive immunity. Guide RNAs may be used to initiate innate immune responses. This article provides an overview of CRISPR-Cas applications from the lab to the clinic, identifies current barriers that may restrict the use of CRISPR-Cas systems as gene-editing toolkits in precision medicine, and offers some perspectives on how to address these barriers and speed up technical advancement

Evaluation of a decellularized bronchial patch transplant in a porcine model

Biological scaffolds for airway reconstruction are an important clinical need and have been extensively investigated experimentally and clinically, but without uniform success. In this study, we evaluated the use of a decellularized bronchus graft for airway reconstruction. Decellularized left bronchi were procured from decellularized porcine lungs and utilized as grafts for airway patch transplantation. A tracheal window was created and the decellularized bronchus was transplanted into the defect in a porcine model. Animals were euthanized at 7 days, 1 month, and 2 months post-operatively. Histological analysis, immunohistochemistry, scanning electron microscopy, and strength tests were conducted in order to evaluate epithelialization, inflammation, and physical strength of the graft. All pigs recovered from general anesthesia and survived without airway obstruction until the planned euthanasia timepoint. Histological and electron microscopy analyses revealed that the decellularized bronchus graft was well integrated with native tissue and covered by an epithelial layer at 1 month. Immunostaining of the decellularized bronchus graft was positive for CD31 and no difference was observed with immune markers (CD3, CD11b, myeloperoxidase) at two months. Although not significant, tensile strength was decreased after one month, but recovered by two months. Decellularized bronchial grafts show promising results for airway patch reconstruction in a porcine model. Revascularization and re-epithelialization were observed and the immunological reaction was comparable with the autografts. This approach is clinically relevant and could potentially be utilized for future applications for tracheal replacement.

Post COVID - 19 Infection Diffuse Patch Granuloma Annulare in a Female Patient with Rheumatoid Arthritis: A Case Report

Introduction: Granuloma Annulare (GA) is a benign, usually self-limited, inflammatory granulomatous skin disorder of unknown etiology and with a diversity of clinical variants. Genetic factors, associations with predisposing diseases along with several triggers such as certain infections and vaccines, have been described, SARS-CoV-2 virus being one of them. Case Presentation: Localized, generalized and subcutaneous forms of GA have been reported following either SARS-CoV-2 infection or vaccination against the virus. Herein we present a case of diffuse patch GA in a middle-aged female patient with medical history of rheumatoid arthritis, one month after catching COVID -19 despite being fully vaccinated against the virus. Discussion: Overall, taking into account the relevant literature, the occurrence of GA after infection with SARS-CoV-2 as well as after vaccination seems to be attributed to the activation of an immunologic reaction rather than to the direct effect of the virus on the skin lesions, with the latest also appearing in the form of diffuse patch rashes. Conclusion: Dermatologists should be aware of such lesions appearing after COVID-19 infection or even vaccination especially in patients with autoimmune diseases.

Investigating Spine and Spinal Cord Complications of COVID-19

Background and Aim: SARS-COV-2 can present with pulmonary, renal, gastrointestinal, hematological, and neurological manifestations. Neurological manifestations may occur after or before COVID-19 symptoms and signs. Spine and spinal cord complications are documented as neurological complications of COVID-19. Spinal cord pathology following COVID-19 showed inflammatory myelopathy and suspected cord ischemia. The most frequent presentation of COVID-19 myelitis is non-enhancing central expansile cord T2 signal changes, but it can present with lateral and dorsal column-specific disease and in some cases with negative magnetic resonance imaging (MRI). There is no known documented mechanism for spinal cord involvement in COVID-19 infection, but it seems as a post-infectious immunological and post-inflammatory disorder and reaction. Viral infection of SARS-CoV-2 can cause demyelination of the brain and spinal cord and also can exacerbate the known primary demyelinating disorders. Methods and Materials/Patients: This is a narrative study about the spinal cord complications of COVID-19. To provide up-to-date information, we precisely reviewed COVID-19 articles on spine and spinal cord complications. Based on the keywords COVID-19, SARS-CoV-2, spine, and spinal cord, all the related articles were taken from Google Scholar, PubMed, and Medline and were precisely studied. Results: There are reported cases of COVID-19 spine and spinal cord involvement. There is no documented mechanism for these involvements but the possible mechanisms are direct invasion, cytokine storm, coagulopathy, and an autoimmune response. The routine therapy of such complications is the treatment of these complications with other primary causes with a poor and unsatisfactory response of myelopathy to treatment; however, early diagnosis and vigilance of such involvement improve outcome. Conclusion: COVID-19 can cause spine and spinal cord complications in some patients without a known incidence rate of such complications. The pathogenesis is not completely known; therefore, more conclusive studies are obligatory to improve our information on COVID-19 spinal cord and spine complications.

Immune response status and development of streptococcus pneumoniae carriage as health risk factors for workers engaged in coke production and basic oxygen steelmaking

Occupational hazards affect immunological reactivity and increase the risk of infection with respiratory pathogens. The purpose of our work was to study the mechanisms and causes of developing carriage of streptococci possessing genetic determinants of resistance to antibiotics as health risk factors for workers engaged in coke production and basic oxygen steelmaking. We examined 136 workers of a ferrous metals industry and compared the prevalence of the carriage of Streptococcus pneumoniae, Streptococcus spp. and macrolide resistance genes (Mef, ErmB) in those engaged in coke production and basic oxygen steelmaking. We also collated the results of immunological examination of the carriers and non-carriers of S. pneumoniae. The control cohort was formed of engineering and technical personnel. We established that, compared with the controls, Streptococcus pneumoniae was a more frequent finding in the workers engaged in coke production and basic oxygen steelmaking (p ˂ 0.05). The cycle threshold value of Streptococcus spp. in them was also statistically higher. The Mef gene was more abundant in the steelmakers, including those carrying Streptococcus pneumoniae (p ˂ 0.05), and a change in immunological reactivity was detected in the cases carrying Streptococcus pneumoniae. In those engaged in basic oxygen steelmaking, significant changes were detected at the cellular level expressed by a reduced absolute number of mature T-lymphocytes (CD3+), T-helpers (CD4+), and T-suppressor/cytotoxic cells (CD8+) (p ≤ 0.05). At the humoral level, both the relative and absolute number of B-lymphocytes (CD19+) and the level of IgM were reduced, while that of IgG was, on the opposite, increased (p ≤ 0.05). In the coke plant workers carrying Streptococcus pneumoniae, the level of IgG was significantly higher than the control and reference values. We conclude that Streptococcus pneumoniae carriage in the steelmakers as a health risk factor was accompanied by a change in immunological reactivity and higher abundance of the Mef gene compared to the controls while in the coke production workers the related differences were detected only in the IgG level.

Состояние иммунного ответа и формирование носительства streptococcus pneumoniae как факторы риска здоровью работников коксохимического и конвертерного производства

Occupational hazards affect immunological reactivity and increase the risk of infection with respiratory pathogens. The purpose of our work was to study the mechanisms and causes of developing carriage of streptococci possessing genetic determinants of resistance to antibiotics as health risk factors for workers engaged in coke production and basic oxygen steelmaking. We examined 136 workers of a ferrous metals industry and compared the prevalence of the carriage of Streptococcus pneumoniae, Streptococcus spp. and macrolide resistance genes (Mef, ErmB) in those engaged in coke production and basic oxygen steelmaking. We also collated the results of immunological examination of the carriers and non-carriers of S. pneumoniae. The control cohort was formed of engineering and technical personnel. We established that, compared with the controls, Streptococcus pneumoniae was a more frequent finding in the workers engaged in coke production and basic oxygen steelmaking (p ˂ 0.05). The cycle threshold value of Streptococcus spp. in them was also statistically higher. The Mef gene was more abundant in the steelmakers, including those carrying Streptococcus pneumoniae (p ˂ 0.05), and a change in immunological reactivity was detected in the cases carrying Streptococcus pneumoniae. In those engaged in basic oxygen steelmaking, significant changes were detected at the cellular level expressed by a reduced absolute number of mature T-lymphocytes (CD3+), T-helpers (CD4+), and T-suppressor/cytotoxic cells (CD8+) (p ≤ 0.05). At the humoral level, both the relative and absolute number of B-lymphocytes (CD19+) and the level of IgM were reduced, while that of IgG was, on the opposite, increased (p ≤ 0.05). In the coke plant workers carrying Streptococcus pneumoniae, the level of IgG was significantly higher than the control and reference values. We conclude that Streptococcus pneumoniae carriage in the steelmakers as a health risk factor was accompanied by a change in immunological reactivity and higher abundance of the Mef gene compared to the controls while in the coke production workers the related differences were detected only in the IgG level.

Abstract B033: Multiplex analysis of the tumor immune microenvironment during treatment with atezolizumab/pelareorep/letrozole reveals novel immune-tumor interactions

Abstract Background: Pelareorep (pela) is a non-modified intravenously administered oncolytic reovirus exhibiting effective tumor suppression through both innate and adaptive immune responses, as well as direct tumor lysis. Findings from the AWARE-1 window of opportunity study previously demonstrated the synergistic potential of combining pela with atezolizumab (atezo), showcasing promising immunological reactions within tumors of early breast cancer (eBC) patients. To gain deeper insights into the complex tumor immune microenvironment (TiME) pre- and post-treatment, we employed imaging mass cytometry (IMC) to conduct high-dimensional, single-cell analysis of tissue samples. Methods: Patients (n=10) received pela (days 1, 2 and 8, 9), atezolizumab (day 3), and letrozole (day 1 to 21). Tumor biopsies (FFPE samples) were collected pre-treatment on day 3 (prior to atezolizumab administration) and on day ~21 (surgical excision). Samples from 8 out of 10 patients were subjected to IMC with a comprehensive panel of 37 antibodies for single cell investigation of TiME changes. Tumor, proliferation, T cell, macrophage, NK, and immunoregulatory markers were included in the panel. Results: Our first pass analysis of IMC images, showed an increase in proliferating (Ki67+) cytotoxic T cells adjacent to apoptotic (caspase 3+) tumor cells post-treatment (both D3 and D21) samples indicating immunogenic cell death. Conversely, we observed a decrease in Ki67+ and ER-positive tumor cells after treatment indicating decreased tumor cell proliferation and hormone expression. In 3 out of 8 patients, treatment was associated with a shift in monocyte/macrophages from an M1 (CD68+/CD163-) to M2 (CD68+/CD163+) phenotype associated with significant tumor infiltration. Surprisingly, in these patients we also noted a significant increase in apoptotic M2 macrophages on D21. Consistent with the known immune priming effects of pela, both PD-1 and PD-L1 increased on D3. Subsequently, on day 21, post atezo, tumor PD-L1 decreased on tumor cells while PD-1 expression persisted. Pela induced IDO expression on both tumor and monocyte/macrophage cells. Unlike its effect on PD-L1, atezo did not appear to attenuate IDO expression on day 21. We were also able to detect a treatment related increase in immune cell perivascular localization on D3 providing further evidence of TiME priming. Conclusions: The immune system plays a crucial role in regulating cancer progression; however, our understanding of immune interactions with tumors remains limited. Our study utilizing IMC revealed complex immune-tumor interactions changes during treatment with atezo/pela/letrozole. These findings provide valuable insights into the intricate dynamics of the TiME and may inform new therapeutic combination approaches in breast cancer treatment. Citation Format: Homa Dadrastoussi, Julian Olea, Eduardo Fernandez Hernandez, Hugo Lara Martinez, Kaijin Wu, Houra Loghmani, Thomas Heineman, Richard Trauger, Matt Coffey, Akil Merchant, Kevin Kelly. Multiplex analysis of the tumor immune microenvironment during treatment with atezolizumab/pelareorep/letrozole reveals novel immune-tumor interactions [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Tumor Immunology and Immunotherapy; 2023 Oct 1-4; Toronto, Ontario, Canada. Philadelphia (PA): AACR; Cancer Immunol Res 2023;11(12 Suppl):Abstract nr B033.

The use of aromatherapy in pediatric practice

Aromatherapy is a method of treatment using natural essential oils injected into the body through the respiratory tract, skin and (or) mucous membranes. Currently, about 3,000 essential oil plants have been studied, from which over 500 organic compounds have been isolated and identified, the number of which in the composition of one essential oil can vary from 120 to 500, and no essential oil has yet been fully investigated.&#x0D; Essential oils are multicomponent organic compounds of terpenes, alcohols, aldehydes, ketones and other hydrocarbons produced by essential oil plants. Biologically active substances of essential oils have a low molecular weight and therefore easily penetrate the skin through the bloodstream. The effect exerted on a person by essential oils is determined in two ways ― neuro-reflex and humoral. Each essential oil is multifunctional. The composition of essential oils can include up to 300 components, so they can have several medicinal properties at the same time, but there is a general trend in the action of essential oils. All of them have immunomodulatory, pronounced bactericidal and antiseptic effects, antiviral and anti-inflammatory properties.&#x0D; In medical practice, the method of hardware aromatherapy is used, the active factor of which is a therapeutic respiratory environment that simulates the natural air background over plants. The study of the effect of essential oils in children's institutions showed their positive effect on the general condition of children, emotional tone, immunological reactivity and nonspecific protection factors, the frequency and severity of respiratory pathology, the course of the adaptation period. The beneficial effect of the complex effect of aromatherapy in the sensory room is shown.&#x0D; Thus, the high efficiency of hardware aromatherapy, good tolerability, the availability of domestic equipment allows us to recommend the method for wide use in therapeutic and preventive and sanatorium-resort institutions.

Poly(N-methyl-N-vinylacetamide): A Strong Alternative to PEG for Lipid-Based Nanocarriers Delivering siRNA.

Lipid-based nanocarriers have demonstrated high interest in delivering genetic material, exemplified by the success of Onpattro and COVID-19 vaccines. While PEGylation imparts stealth properties, it hampers cellular uptake and endosomal escape, and may trigger adverse reactions like accelerated blood clearance (ABC) and hypersensitivity reactions (HSR). This work highlights the great potential of amphiphilic poly(N-methyl-N-vinylacetamide) (PNMVA) derivatives as alternatives to lipid-PEG for siRNA delivery. PNMVA compounds with different degrees of polymerization and hydrophobic segments, are synthesized. Among them, DSPE (1,2-distearoyl-sn-glycero-3-phosphoethanolamine)-PNMVA efficiently integrates into lipoplexes and LNP membranes and prevents protein corona formation around these lipid carriers, exhibiting stealth properties comparable to DSPE-PEG. However, unlike DSPE-PEG, DSPE-PNMVA24 shows no adverse impact on lipoplexes cell uptake and endosomal escape. In in vivo study with mice, DSPE-PNMVA24 lipoplexes demonstrate no liver accumulation, indicating good stealth properties, extended circulation time after a second dose, reduced immunological reaction, and no systemic pro-inflammatory response. Safety of DSPE-PNMVA24 is confirmed at the cellular level and in animal models of zebrafish and mice. Overall, DSPE-PNMVA is an advantageous substitute to DSPE-PEG for siRNA delivery, offering comparable stealth and toxicity properties while improving efficacy of the lipid-based carriers by minimizing the dilemma effect and reducing immunological reactions, meaning no ABC or HSR effects.

Interaction between JAK2-Mutated Neutrophils and Platelets Initiates Thrombosis Via Neutrophil Extracellular Traps

Thrombosis is one of the most common complications of myeloproliferative neoplasms with JAK2-V617F mutation. Recently, individuals with clonal hematopoiesis (CH) harboring JAK2-V617F mutation were also shown to have 12-fold higher risk of thrombosis compared to JAK2 non-mutated individuals (N Engl J Med. 2017;377:111-121). The finding that individuals with JAK2-V617F CH do not have altered blood counts suggests the mechanisms other than increases in the hematopoietic cells which induce thrombosis associated with JAK2-V617F hematopoiesis. JAK2-V617F neutrophils are primed to neutrophil extracellular traps (NETs), which can promote thrombosis (Sci Transl Med. 2018;10: eaan8292). NETs were initially reported as an immunological reaction against infection. However, NETs also play various roles in non-infectious (sterile) settings, and the mechanism how sterile NETs are initiated in vivo remains largely unknown. Here, we studied the mechanisms how NETs are initiated and promote thrombosis in the presence of JAK2-V617F hematopoiesis. First, we isolated neutrophils and platelets from Jak2-V617F transgenic ( Jak2VF) and wild-type (WT) mice, and co-cultured in each cell/genotype combination (see Figure1) at a 1:50 ratio. Of note, Jak2VF neutrophils formed NETs with Jak2VF platelets, but not with WT platelets. Also, Jak2VF platelets slightly increased formation of NETs by WT neutrophils. However, culture supernatant of Jak2VF platelets did not induce formation of NETs by Jak2VF neutrophils. These findings indicated that direct interaction, between Jak2VF neutrophils and Jak2VF platelets, initiates sterile NETs in the presence of JAK2-V617F hematopoiesis. Surprisingly, platelet activation of Jak2VF platelets measured by P-selectin release was inferior to WT platelets. Thus, it was unlikely that overactivation of Jak2VF platelets fully initiate NETs. Subsequently, we sought an initiator on Jak2VF platelets to form NETs in the presence of JAK2-V617F hematopoiesis. Unbiased proteomics analysis of WT and Jak2VF platelets identified several proteins significantly enriched in Jak2VF platelets, including a membrane protein PODXL, which is expressed on podocytes in kidney as well as platelets and other hematopoietic cells. Interestingly, PODXL on platelet is associated with aggregation with other hematopoietic cells depending on CD41, SLC44A2 and P-selectin. Therefore, we hypothesized that PODXL enhances aggregation of Jak2VF platelets with neutrophils and initiates formation of NETs (Figure 2). We confirmed overexpression of PODXL in Jak2VF platelets and in platelets of another murine model, hematopoietic cell-specific Jak2V617F/+ mice by FACS and immunofluorescence imaging. The formations of NETs measured by intensity of citrullinated histone H3 (citH3) correlated with aggregation of PODXL-expressing Jak2VF platelets on cell surface. Presently, we are conducting to confirm the findings utilizing patients' samples with JAK2-V617F. To test if a JAK inhibitor prevents PODXL-associated formation of NETs, we collected platelets from ruxolitinib-treated Jak2VF mice. Platelets from ruxolitinib-treated mice similarly expressed PODXL and induced formation of NETs with neutrophils from untreated Jak2VF mice, compared to platelets from untreated Jak2VF mice, suggesting that ruxolitinib does not alter platelets' ability to initiate formation of NETs. Next, we tested if these mechanisms induce venous and arterial thrombosis in vivo, utilizing models of half ligation of inferior vena cava (IVC) and photochemically induced thrombosis (PIT) in femoral artery. In both models, Jak2VF or Jak2V617F/+ mice formed thrombosis more quickly than WT controls. Jak2-mutated mice formed white thrombus presenting citH3-positive neutrophils aggregated with PODXL-expressing platelets at the core of the thrombus, while WT mice mainly formed red thrombus without any nucleated cells in these models. Therefore, we are generating platelet-specific knockout mice of PODXL to directly clarify if PODXL on Jak2VF-platelet is the initiator of NETs and thrombosis. In conclusion, PODXL on JAK2-V617F platelets may be a key player in the pathogenesis of thrombosis associated with formation of NETs with JAK2-V617F neutrophils in myeloproliferative neoplasms.