Higher Levels Of TNF-alpha Research Articles

DUMB CANE (DIEFFENBACHIA SEGUINE (JACQ.) SCHOTT) EXTRACT NANOEMULSION: PREPARATION, CHARACTERIZATION AND ITS ACTIVITY AS AN INFLAMMATION-INDUCING AGENT IN RATS

Objective: This study aimed to characterize Dumb cane extract nanoemulsion for injection preparation and test its activity as an inflammation-inducing agent in rat intraplantar with IL-6 and TNF-alpha parameters in blood serum as well as the swelling response in paw. Methods: Nanoemulsion was made from an emulsion base of olive oil, coconut oil, tween 80 propylene glycol, and water for injection, as well as varying concentrations of Dumb cane extract (1%, 2%, and 4%) using vortex mixing and sonication methods. The characterization included particle size, zeta potential, and polydispersity index using the Zetasizer tool. Induction was carried out intraplantar in male rats. The parameters observed were the volume of swelling in paw and IL-6 and TNF-alpha in blood serum. Results: Dumb cane extract nanoemulsion concentrations of 1%, 2%, and 4% have particle size characteristics in the range of 20.3±0.17–30.1±0.68 nm, and zeta potential -31.4±1.59–33.1±1.33 mV. Dumb cane nanoemulsion can induce intraplantar inflammation with high IL-6 and TNF-alpha levels, significantly different from normal controls. Volume swelling occurred 4 h after intraplantar induction at a nanoemulsion concentration of 4%. Conclusion: Dumb cane extract nanoemulsion can be an alternative agent for inducing intraplantar inflammation in rat models.

Predicting Retinopathy of Prematurity Risk Using Plasma Levels of Insulin-like Growth Factor 1 (IGF1), Tumor Necrosis Factor-Alpha (TNF-Alpha), and Neonatal Parameters.

Retinopathy of Prematurity (ROP) remains a leading cause of vision impairment in premature infants, especially those with Respiratory Distress Syndrome (RDS) necessitating respiratory support. This study aimed to identify correlations between plasma levels of Insulin-like Growth Factor 1 (IGF1) and Tumor Necrosis Factor-alpha (TNF-alpha), and the risk of developing ROP. Additionally, it explored the association of ROP severity grades with plasma levels of glucose, lactate dehydrogenase (LDH), creatin phosphokinase (CPK), and other biomarkers, aiming to uncover predictive markers for ROP risk and severity in this population. This prospective study included premature neonates admitted with RDS requiring respiratory support, conducted over 18 months at the Neonatal Intensive Care Unit of the Louis Turcanu Emergency Clinical Hospital for Children, Timisoara. Plasma levels of IGF1 and TNF-alpha were measured on days 1 and 14 post-birth, alongside the initial assessment of glucose, LDH, and CPK levels. Significant correlations were observed between lower gestational age and elevated LDH levels on day 7-10 (rho = -0.341, p = 0.0123) and between TNF-alpha levels at 2 weeks and ROP severity (rho = 0.512, p = 0.0004). Elevated IGF1 levels were protective against ROP, with Beta coefficients of 0.37 (p = 0.0032) for the first collection and 0.32 (p = 0.0028) for the second, suggesting their potential as biomarkers for ROP risk assessment. Higher levels of TNF-alpha at 2 weeks were associated with an increased risk of ROP (Beta = -0.45, p = 0.0014), whereas higher IGF1 levels offered protective effects against ROP, with Beta coefficients of 0.37 (p = 0.0032) for the first collection and 0.32 (p = 0.0028) for the second. Elevated LDH levels on day 7-10 post-birth were linked to an increased risk of ROP (Beta = 0.29, p = 0.0214). These findings highlight the potential of IGF1 and TNF-alpha as predictive biomarkers for ROP, offering avenues for early intervention and improved management strategies in this high-risk group.

Abstract 3615: Targeting high-risk neuroblastoma by TGFβ imprinted natural killer cells

Abstract Background: Neuroblastoma (NB) is a common malignant extracranial solid tumor in children, responsible for 11% of cancer deaths in pediatric patients under 15 (Smith et al, Cancer, 2014). High-risk neuroblastoma is a metastatic disease in children aged 18 months or older with L2, M, or MS disease with amplification of the MYCN oncogene (Chuang Pediatr Blood Cancer 2021). Recent analyses by COG show that HR Neuroblastoma still has a 5-year event free survival of 51% despite an extensive treatment regimen (Steven et al, ASCO, 2022). To address improved tumor targeting and overcome antigen escape, chimeric antigen receptor (CAR) technology to Natural Killer (NK) cell therapy has been adapted (Chu/Cairo, et al, Frontiers in Oncology, 2019). TGFβ is a potent mechanism of the NK cell anti-tumor resistance and is present at elevated levels in the NB tumor microenvironment (TME) (Foltz, Cancers, 2018). Studies demonstrated that blockade of TGF-β signaling in NK cells caused the accumulation of NK cells that produce IFN-γ and neutralization of TGF-β prevented NKG2D downregulation and also restored NK cell anti-tumor reactivity (Nayyar/Chu/Cairo, et al Frontierls in Oncology, 2018). ROR1, a transmembrane protein is highly expressed on the majority of NB (Rebagay/Yan Front Oncol, 2012). Our group has successfully expanded functional and active peripheral blood NK cells (exPBNK) with irradiated feeder cells and electroporated CAR mRNA to exPBNK (Chu/Cairo, et al Cancer Immunol Res, 2015). Objective: To determine in vitro cytotoxicity against neuroblastoma cells by ex vivo expanded TGFβ imprinting (TGFβi) NK cells. Methods: TGFβi NK cells are generated by culturing expanded NK cells with irradiated K562-41BBL-mbIL21 in 50 IU/mL IL-2 plus 10 ng/mL TGFβ every 2-3 days for two weeks. NK and TGFβi NK cells specific in-vitro cytotoxicity were compared via luciferase reporter assay against ROR1+ NB cell lines as we previously described (Chu/Cairo, et al, JITC, 2022). Cytokine/chemokine levels secreted by NK and TGFβi NK cells were compared using ELISA assays (Chu/Cairo, et al, JITC, 2022). Results: We have successfully expanded TGFβi NK cells. These TGFβi NK cells showed significantly enhanced in vitro cytotoxicity against NB cells compared to NK cells under TGFβ environment (20+2.4% vs 0%; p<0.001) at E:T=3:1 and consistently, TGFβ-imprinting NK secreted significantly high level of TNF-alpha (393.7pg/ml+16.77pg/ml) against NB cells (p<0.001) compared to NK cells (79.7pg/ml+14.7pg/ml) under TGFβ environment. Conclusion: Our data indicate TGFβi NK cells may be a promising novel therapy to overcome the immunosuppression of TGFβ in the tumor microenvironment for pediatric NB tumors. We will further engineer these TGFβi NK cells to express anti-ROR1 CAR to enhance the targeting specificity against NB tumors and investigate the anti-tumor effects of anti-ROR1 CAR TGFβi NK cells against NB tumors in-vitro and in-vivo (Funded by U54 CA232561). Citation Format: Jessica A. Ayala-Cuesta, Yaya Chu, Keira Foley, Meijuan Tian, Kayleigh Klose, Alyssa S. Mendelowitz, Timothy P. Cripe, Dean A. Lee, Mitchell S. Cairo. Targeting high-risk neuroblastoma by TGFβ imprinted natural killer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 3615.

INFLAMMATORY CYTOKINE, SOCIAL SUPPORT, AND HEALTH DISPARITIES IN PERSONS WITH COGNITIVE IMPAIRMENT AND DEMENTIA

Abstract The protective role of social support on cognitive functioning is extensively documented. A pro-inflammatory cytokine, Tumor necrosis factor-alpha (Tnf-alpha), might have a role in the connection between social support and dementia as a marker of systemic inflammation. This study describes the influence of lack of friends’ support and Tnf-alpha on cognitive functioning. We conducted a cross-sectional study on Health and Retirement Study participants aged 65 and above, using datasets from the 2016 interviews. Of the 9,262 participants, 72.4% had normal cognition. 21.3% had cognitive impairment, and 6.3% had dementia. Our regression model controlled with depression scores revealed that participants with a lack of friends’ support were 51% more likely to have cognitive impairment and 54% more likely to have dementia. The effect of Tnf-alpha was comparably higher on participants with dementia and cognitive impairment than on participants with normal cognition. Compared to quartile-1 (lowest Tnf-alpha levels), Quartile-4 (highest Tnf-alpha levels) was significantly associated with cognitive impairment (OR=1.60) and dementia (OR=2.37). Quartile-3 was only a significant predictor of dementia (OR=2.08). The effect of Tnf-alpha was insignificant in quartile-2 for both groups. Hispanics were 65% more, and African Americans were 67% more likely to have cognitive impairment than non-Hispanic whites. The likelihood of having dementia was even higher in racial/ethnic minorities: Hispanics were 3.3 times more, and African Americans were 2.3 times more likely to have dementia than non-Hispanic whites. Future research can focus on racial and ethnic variations in pro-inflammatory cytokines and social support among people with dementia.

Molecular Characterisation of Blood Microbiome in Patients with Ankylosing Spondylitis and Healthy Controls.

In human and animal studies, ankylosing spondylitis (AS) has been increasingly linked to changes in the microbial inhabitants in the human body (microbiome). These studies have primarily now concentrated on the microbial communities that live in the gastrointestinal tract. However, evidence suggests that various molecular techniques can be used to detect microbial DNA in blood circulation. This DNA might be an unknown reservoir of biomarkers with the potential to track alterations in the microbiomes of remote locations, such as the gut. To this end, we compared the presence and identity of microbial DNA in blood samples taken from ankylosing spondylitis patients to healthy control subjects by amplifying and sequencing the bacterial 16S rRNA variable region four. The study's design is a case study based on the presence and identity of bacterial DNA in the blood of Ankylosing spondylitis (AS) patients (n = 10) and healthy control subjects (n = 10) was investigated by amplifying and sequencing the bacterial 16S rRNA gene. Blood concentrations of the cytokines TNF alpha, IL-17A, and IL-23 were determined by the Human Magnetic Luminex Screening, and data were analysed using an Unpaired T-test. Using PCR amplification, 8 of 10 AS patients (80%) and 8 of 10 healthy control samples (80%) had microbial 16S rRNA in their blood. At the phylum level, Proteobacteria (Control = 48.5%, AS = 52%), Firmicutes (Control = 27.8%, AS = 26.1%), Actinobacteria (Control = 15.4%, AS = 10.7%), and Bacteroidetes (Control = 6.5%, AS = 10%) dominated the blood microbiome. A two-tailed Mann-Whitney test found that Ankylosing Spondylitis was associated with significantly elevated Bacteroides (P < 0.05), Prevotella (P < 0.001), and Micrococcus (P < 0.01), and significantly reduced levels of Corynebacterium 1 (P < 0.001), Gemella (P < 0.01), and Alloprevotella (P < 0.05), compared to healthy controls. Additionally, it was shown that the presence of the Prevotella genus was highly positively correlated with higher levels of TNF-alpha (P < 0.05; r = 0.8) in AS patients' blood. This article reveals that a blood microbiome exists in healthy individuals and identifies particular taxa modulated in disease. These blood-derived signatures indicate that this field needs more research and may be helpful as disease biomarkers.

Inflammatory CSF profiles and longitudinal development of cognitive decline in sporadic and GBA-associated PD

Inflammation modifies the incidence and progression of Parkinson’s disease (PD). By using 30 inflammatory markers in CSF in 498 people with PD and 67 people with dementia with Lewy bodies (DLB) we show that: (1) levels of ICAM-1, Interleukin-8, MCP-1, MIP-1 beta, SCF and VEGF were associated with clinical scores and neurodegenerative CSF biomarkers (Aβ1-42, t-Tau, p181-Tau, NFL and α-synuclein). (2) PD patients with GBA mutations show similar levels of inflammatory markers compared to PD patients without GBA mutations, even when stratified by mutation severity. (3) PD patients who longitudinally developed cognitive impairment during the study had higher levels of TNF-alpha at baseline compared to patients without the development of cognitive impairment. (4) Higher levels of VEGF and MIP-1 beta were associated with a longer duration until the development of cognitive impairment. We conclude that the majority of inflammatory markers is limited in robustly predicting longitudinal trajectories of developing cognitive impairment.

Effect of Melatonin on Feeding Intolerance and Incidence of Necrotizing Enterocolitis in Preterm Infants

Abstract Background Necrotizing enterocolitis (NEC) is one of the most dangerous gastrointestinal diseases that affects neonates especially preterm neonates. It is considered one of the oxygen radical diseases of neonatology as it is attributed to accumulation of reactive oxygen species (ROS) and free radicals leading to cellular oxidative damage. Melatonin proved to be an effective antioxidant and free radical scavenger with no documented side effects. Objective We aimed to evaluate the efficacy and safety of melatonin as adjuvant therapy in feeding intolerance and prophylaxis against NEC. Methods This was an interventional trial conducted on 90 preterm neonates with feeding intolerance. They were simply randomized to Group I that received melatonin treatment (n = 45) and Group II that received traditional treatment only (n = 45). Melatonin was administered at a total dose of 20 mg enterally in two divided doses of 10 mg each, 1 h apart. Tumor necrosis factor alpha (TNFα) level was assessed in patients 72 hours after starting treatment and both groups were observed for incidence of NEC, clinical improvement of feeding intolerance and time needed to reach full intake. Results Group I reported significantly less incidence of NEC in Group I treated with melatonin. Improved tolerance to feeds was observed in Group I resulting in earlier achievement of full enteral intake. Significantly higher levels of TNF alpha were found in Group II that received traditional treatment, indicative of higher level of oxidative stress and free radicals. Conclusion Melatonin in an effective and safe antioxidant and free radical scavenger associated with improvement of clinical and laboratory data in preterm infants with feeding intolerance and significantly less incidence of NEC.

Role of TNF-a and Interleukin 6 Serum against Ovarian Reserve in Endometriosis Cysts

Objective: To investigate the correlation between proinfl ammatory factors (TNF-alpha and IL-6) with ovarian reserve in patients with endometriosis cysts. Methods: This is a cross sectional study. The number of samples is 25 people. Sampling technique consecutive sampling. The study began in June 2019 until the number of samples was fulfi lled at the Obstetrics and Gynecology Polyclinic of the Dr. General Central Hospital M. Djamil Padang and at the Biomedical Laboratory of the Faculty of Medicine, Andalas University, Padang. To determine the correlation of TNF-alpha, IL-6, and AMH in patients with endometriosis cysts. Examination of TNF-alpha, IL-6, and AMH levels was carried out in a quantitative manner, using the ELISA method.Result: The mean AMH level was 2 ± 0.40 pmol / L andTNF-alpha level was 11.16 ± 4.79 pg / ml. Correlation of AMH level with TNF-alpha indicates the value of r = -0.049 which has weak strength and negative pattern means that the higher the TNF-alpha level, the lower the AMH level. The analysis showed that there was no correlation between TNF-alpha levels and AMH levels in patients with endometriosis cysts (p&gt;0.05). Correlation of AMH levels with IL-6 shows the value of r = 0.35 which has moderate strength and positive pattern means that the higher the IL-6 level, the higher the AMH level. The analysis showed that there was no correlation between IL-6 levels and AMH levels in patients with endometriosis cysts (p&gt; 0.05). Conclusions: There is a negative correlation of TNF-alpha levels with AMH levels in patients with endometriosis cysts, namely the higher TNF-alpha levels, the lower AMH levels and there is a positive correlation of IL-6 levels with AMH levels in patients with endometriosis cysts. Where the higher levels of IL-6, the higher levels of AMH. Keywords: AMH, endometriosis, TNF-alpha, IL-6.

Abstract 106: Assessing the impact of staggered light exposure on breast cancer risk and metabolism

Abstract Background: Numerous epidemiological studies, including the Nurses' Health Study II, found that shift workers have a higher risk for breast cancer. These studies hypothesize that suppressed melatonin and disrupted clock gene expression likely lead to an immune compromised, proliferative phenotype. However, the majority of studies to date rely on correlative evidence. With the onset of the COVID-19 pandemic, shift work has become increasingly prevalent, making the need to understand the mechanisms promoting disease, including breast cancer, even more important. Objective: The objective of this study is to analyze the impact of staggered light exposure on the immune function and breast tissue gene expression in order to identify how breast cancer risk is mediated. Methods: An 18-week study with 10 C57BL/6 female mice was conducted whereby half of the mice were exposed to a staggered light/dark (LD) cycle, as would be experienced by shift workers, and half were exposed to a regular LD cycle. Body weight was monitored weekly and sera and mammary tissues were collected at termination. The sera from each group was pooled and cytokine levels assessed using an antibody-pair-based assay. Results: Intriguingly, although the mice from the two study groups did not significantly differ in average body weight, the staggered LD exposure group had higher levels of inflammatory cytokines indicative of chronic, low-grade inflammation. Notably, the staggered LD group had 75% higher IL-6 levels, 157% higher IL-10 levels, and 49% higher TNF-alpha levels compared to the control group. To determine the impact of staggered light exposure on localized breast tissue gene expression, qPCR arrays of breast cancer risk and inflammatory genes will be done comparing expression levels between the control and staggered LD groups. IHC techniques will be used to determine Ki67 and macrophage subtype infiltration in the breast tissue of the two groups. Conclusion: Our data show that staggered LD exposure induces chronic, low-grade inflammation independent from body mass, which can be indicative of increased breast cancer risk. These data help elucidate the metabolic and immune responses induced by staggered light exposure, helping to define the mechanisms promoting health risks imposed by shift work, with the ultimate goal to develop preventive measures to limit breast cancer development. Citation Format: Lauren Palluth, Brittany Harlow, Bryan McCellan, Christopher Jolly, Molly Bray, Linda deGraffenried. Assessing the impact of staggered light exposure on breast cancer risk and metabolism [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 106.

Cytokine Level and Symptoms of Schizophrenia

BACKGROUND: The study was intended to find one of the cytokines, that is, tumor necrosis factor (TNF)-alpha correlation with PANSS scores of people with schizophrenia (PwS) and their differences compared to the healthy people of Malayan- Mongoloid Race. AIM: The aim of this study was providing a profile of TNF-alpha level in PwS of Malayan-Mongoloid race and its correlation with positive and negative symptoms in schizophrenia. METHODS: It was conducted using cross-sectional and following sampling methods, with inclusion and exclusion criteria set for the PwS and control groups. RESULTS: The result was no correlation between TNF-alpha levels with PANSS scores on PwS (p &gt; 0.005), with very weak correlation and positive correlation direction, and there were differences in TNF-alpha levels.between PwS and control groups. CONCLUSION: It can be concluded that the higher levels of TNF-alpha would impact on the severity symptoms experienced by PwS.

FEATURES OF THE BLOOD PRESSURE DYNAMICS AND PRO-INFLAMMATORY CYTOKINES IN PATIENTS WITH RESISTANT HYPERTENSION AND CORONARY ATHEROSCLEROSIS AFTER RENAL DENERVATION

Objective: Inhibition of sympathetic activity via renal denervation (RDN) is associated with a reduction of blood pressure and variety of inflammatory markers in patients with resistant hypertension (RH). The purpose of this study was to investigate the effects of RDN on BP, TNF-alpha and hs-CRP in patients with confirmed atherosclerosis, to compare the studied parameters in revascularized and conservatively treated patients groups. Design and method: In 35 patients with RH associated with CAD and confirmed by coronary angiography atherosclerosis, levels of 24-h BP, TNF-alpha and hs-CRP analyzed by ELISA were identified before and 12 months after RDN. Baseline 24-h systolic (SBP), diastolic (DBP) and pulse (PBP) BP were 154 ± 12 mmHg, 82 ± 13 mmHg and 73 ± 13 mmHg respectively. Then patients retrospectively where divided into two groups according to the severity of coronary artery disease (CAD): the first group underwent coronary stenting or coronary artery bypass grafting, the second had no need for revascularization and was treated conservatively. Results: 12 month after RDN all the patients had significant (p &lt; 0,05) 24-h SBP (144 ± 15 mmHg), DBP (77 ± 11 mmHg) and PBP (67 ± 14 mmHg) and TNF-alpha from 7,0 ± 3,4 ng/ml to 6,6 ± 3,0 ng/ml, p = 0,03 reduction and downward trend in hs-CRP from 4,0 ± 3,5 ng/ml to 2,8 ± 2,4 ng/ml, p = 0,07, that correlated with PBP reduction (r = 0,47, p &lt; 0,05). It was no difference between SBP and hs-CRP levels in the two groups. In the first group baseline levels of PBP and TNF-alpha where higher than in the second: 78,9 ± 12,8 mmHg vs 69,0 ± 12,1 mmHg, p = 0,03; 9,0 ± 1,2 ng/ml vs 5,7 ± 3,7 ng/ml, p = 0,01. Downward trend in TNF-alpha and hs-CRP persisted in both groups. In conservatively treated group there was significant 24-h SBP/DBP reduction 12 months after RDN (p &lt; 0,05) on 15 (3; 23) / 7 (4; 16) mmHg, whereas in the revascularized group SBP, DBP and PBP did not change (p &gt; 0,05). Conclusions: Patients with RH associated with CAD have significant TNF-alpha reduction, downward trend in hs-CRP 12 months after RDN. Patients with severe coronary atherosclerosis have higher levels of TNF-alpha and are non-responders for RDN.

Improved Spatial Memory And Neuroinflammatory Profile Changes in Aged Rats Submitted to Photobiomodulation Therapy.

Recent evidences have shown the therapeutic potential of transcranial photobiomodulation on traumatic brain injury and Alzheimer's disease. Despite the promising benefits in the brain, little is known about the laser's effects in the absence of pathological conditions. We submitted young (4months old) and aged (20months old) rats to transcranial low-level laser and evaluated their exploratory activity and habituation in open field, anxiety in elevated plus maze, spatial memory in Barnes maze, and aversive memory in a step-down inhibitory avoidance task. Additionally, the levels of a panel of inflammatory cytokines and chemokines were quantified in two different brain regions: the cerebral cortex and the hippocampus. Young and aged rats submitted to transcranial laser exhibited better cognitive performance in Barnes maze than did control rats. Transcranial laser therapy decreased cortical levels of GM-CSF, IL-10, MCP-1, LIX, and TNFα in young rats and IL-5 in aged rats. High levels of IL-6, IL-10, and TNF-alpha were found in the cerebral cortex of aged rats submitted to transcranial laser. In the hippocampus, a decrease in IP-10 and fractalkine levels was observed in the aged rats from the laser group when compared to the aged rats from the control group. Our data indicate that transcranial photobiomodulation improves spatial learning and memory and alters the neuroinflammatory profile of young and aged rats' brains.

The Association Between Shift Work and Immunological Biomarkers in Nurses.

Objectives: Shift work is associated with several negative health effects. The underlying pathophysiological mechanisms are unclear, but low-grade inflammation has been suggested to play a role. This project aimed to determine whether levels of immunological biomarkers differ depending on work schedule, self-reported sleep duration, self-reported sleep quality, and presence of shift work disorder (study 1). Furthermore, we aimed to determine whether these biomarkers differ after a night of sleep vs. at the end of a night or a day shift (study 2).Methods: In study 1, 390 nurses provided blood samples after a night of sleep with the dried blood spot method. In study 2, a subset of 55 nurses also provided blood samples after a day shift and after a night shift. The following biomarkers were measured: interleukin-1alpha, interleukin-1beta, interleukin-4, interleukin-6, interleukin-8, interleukin-10, interleukin-13, monocyte chemoattractant protein-1, interferon-gamma, and tumor necrosis factor-alpha. Multiple linear regressions with adjustment for age, sex and body mass index (study 1) and ANOVAs with repeated measures (study 2) were conducted.Results: In study 1, neither work schedule, number of night shifts, number of quick returns (<11 h between consecutive shifts), sleep duration, poor sleep quality, nor shift work disorder were systematically associated with most of these biomarkers. Compared with day only work, day-evening work was associated with higher levels of IL-1alpha and IL-13, quick returns were associated with higher levels of IL-1beta and MCP-1, short sleep duration (<6 h) was associated with lower levels of IL-1beta and higher levels of TNF-alpha, and long sleep duration (8+ h) was associated with higher levels of IL-13. In study 2, IL-1beta levels were higher (large effect size) both after a day shift (14% increase) and a night shift (75% increase) compared with levels after a night of sleep. Similarly, TNF-alpha levels were higher (moderate-large effect size) after a day shift (50% increase) compared to after a night of sleep. In contrast, MCP-1 levels were lower (large effect size) both after a day shift (22% decrease) and a night shift (12% decrease) compared with after a night of sleep.Conclusions: We found some indications that shift work influenced immunological biomarkers. The results should be interpreted with caution due to limitations, e.g., related to the sampling procedure and to low levels of biomarkers in the blood samples.

236. Ectopic bone formation by submicron structured calcium phosphates: role of the innate immune system

BACKGROUND CONTEXT Ectopic implantation of calcium phosphates (CaP) with an advanced submicron surface topography can lead to de novo bone formation. Since implantation initiates a wound healing response, the innate immune system may play a role in this process. PURPOSE Tis study is to investigate the role of immune cells (ie, macrophages) and osteoclasts in material-driven bone formation. STUDY DESIGN/SETTING TCP-B with micron-scaled surface (1-2mm, 1cc/implant) and BCP with submicron-scaled surface (MagnetOs, 1-2mm, 1cc/implant) were implanted in the dorsal muscles of nine beagles for 3 days, 1, 3, 6 and 12 weeks. Histology, immune-histochemistry and bioassays were performed to detect alkaline phosphatase (ALP), tartrate-resistant acid phosphatase (TRAP), Cathepsin K (CTSK), Tumor necrosis factor-alpha (TNF-alpha), Interleukin-10 (IL-10) and bone formation. PATIENT SAMPLE N/A OUTCOME MEASURES Bone formation was identified by histology and ALP expression. M1/M2 macrophages were characterized by IL-10 level (M2) and TNF-alpha level (M1) and the presence of osteoclasts was determined by TRAP level, TRAP staining and CTSK staining. Histomorphometry was also performed to quantify bone formation. One-way ANOVA with Tukey pos-hoc test was applied to distinguish differences between groups (p METHODS At harvesting, samples (n=4 per condition) were cut into halves for bioassays, histology and immune-histochemistry (IHC). Bioassays (Elisa for IL-10 and TNF-alpha; pNPP method for ALP and TRAP) were performed with lysates and the data were normalized to the mass of the samples. Histology (HE, Masson-Golder, ALP and TRAP) and IHC stain (CTSK) were applied on undecalcified sections. HE-stained sections were used to quantify bone (area percentage of bone in available space). RESULTS Bone was absent in TCP-B implants at all time-point, while bone was formed in BCP at week 6 (3/4, CONCLUSIONS Higher IL-10 level in BCP than TCP-B up to week 3, indicates that BCP favors pro-healing M2 macrophage polarization. The higher TNF-alpha level in BCP than TCP-B up to week 1, showed that initially more pro-inflammatory M1 macrophages are present in BCP compared to TCP-B. TRAP/CTSK-positive multinucleated cells were present at week 3, before bone formation (at week 6), which suggests that osteoclastogenesis occurs prior to osteogenesis. These data imply that calcium phosphates with a submicron topography facilitate macrophage recruitment/proliferation, enhance pro-healing M2 macrophage polarization and osteoclast-like cell formation, ultimately leading to ectopic bone formation. FDA DEVICE/DRUG STATUS MagnetOs Granules (Not approved for this indication)

DNA Vaccine Treatment in Dogs Experimentally Infected with Trypanosoma cruzi.

Chagas disease is a chronic and potentially lethal disorder caused by the parasite Trypanosoma cruzi, and an effective treatment has not been developed for chronic Chagas disease. The objective of this study was to determine the effectiveness of a therapeutic DNA vaccine containing T. cruzi genes in dogs with experimentally induced Chagas disease through clinical, pathological, and immunological analyses. Infection of Beagle dogs with the H8 T. cruzi strain was performed intraperitoneally with 3500 metacyclic trypomastigotes/kg body weight. Two weeks after infection, plasmid DNA immunotherapy was administered thrice at 15-day intervals. The clinical (physical and cabinet studies), immunological (antibody and cytokine profiles and lymphoproliferation), and macro- and microscopic pathological findings were described. A significant increase in IgG and cell proliferation was recorded after immunotherapy, and the highest stimulation index (3.02) was observed in dogs treated with the pBCSSP4 plasmid. The second treatment with both plasmids induced an increase in IL-1, and the third treatment with the pBCSSP4 plasmid induced an increase in IL-6. The pBCSP plasmid had a good Th1 response regulated by high levels of IFN-gamma and TNF-alpha, whereas the combination of the two plasmids did not have a synergistic effect. Electrocardiographic studies registered lower abnormalities and the lowest number of individuals with abnormalities in each group treated with the therapeutic vaccine. Echocardiograms showed that the pBCSSP4 plasmid immunotherapy preserved cardiac structure and function to a greater extent and prevented cardiomegaly. The two plasmids alone controlled the infection moderately by a reduction in the inflammatory infiltrates in heart tissue. The immunotherapy was able to reduce the magnitude of cardiac lesions and modulate the cellular immune response; the pBCSP treatment showed a clear Th1 response; and pBCSSP4 induced a balanced Th1/Th2 immune response that prevented severe cardiac involvement. The pBCSSP4 plasmid had a better effect on most of the parameters evaluated in this study; therefore, this plasmid can be considered an optional treatment against Chagas disease in naturally infected dogs.

Abstract A39: Inflammatory macrophage derived TNF alpha induces downregulation of estrogen receptor alpha in breast cancer cells by inactivation of Foxo3A

Abstract Breast cancers are divided into different subtypes based on receptor expression status: estrogen receptor (ER), progesterone receptor (PR) and Her2. The luminal A subtype (ER+PR+Her2-) is most often associated with a good prognosis while the triple-negative (TN) cancers (ER-PR-Her2-) have a poor prognosis. ER+ cancer cells depend on estrogen for their growth and can be treated with drugs to block its effects. While most human breast cancers express ER alpha, some tumors are hormone independent despite the presence of ER and others become ER negative over time. Myeloid cells are known to affect tumor development, and infiltration of tumor-associated macrophages is associated with a worse prognosis. Theoretically, however, macrophages can be both beneficial and bad for the patient. Proinflammatory M1 macrophages (CD68+ CD163-) can eliminate cancer cells and promote antitumor immune responses while anti-inflammatory M2 macrophages (CD163+CD68+) or anti-inflammatory myeloid cells (CD68-CD163+) can promote tumor progression by inhibiting proinflammatory immune responses and induce wound-healing processes. This includes angiogenesis and matrix degradation, both of which promote metastatic spread and possibly even EMT. Macrophages have also been proposed to downregulate ER in breast cancer cells through an unknown mechanism. Due to the recent reports suggesting that macrophages can induce EMT and downregulate ER, we evaluated the effect of human primary macrophages on ER expression in breast cancer cells. Immunohistochemistry analyses of breast cancer tumor xenografts, established with co-transplantation of primary human myeloid cells together with ER+ breast cancer cells in NSG mice, showed association of monocytes with downregulation of ER-receptor expression. Using primary human monocyte-derived macrophage cultures In vitro, we show that the different subtypes of macrophages have unique cytokine secretion profiles with M1 macrophages secreting significantly higher levels of TNF alpha compared to M2 macrophages. ER alpha positive MCF-7 and ER alpha negative MDA-MB-231 breast cancer cells were cultured in macrophage condition media, and both Western blot and qPCR analysis showed downregulation of ER was caused by M1 macrophage-derived TNF alpha that inhibits FoxO3A transcription of ER alpha. We propose that proinflammatory macrophages (M1), despite being tumoricidal, may have direct effects on endocrine resistance in breast cancer patients. Our findings suggest that different macrophage subtypes have various and unique impacts on breast cancer progression, and that proinflammatory macrophages, despite being tumoricidal, may have unwanted and direct effects on endocrine resistance mechanisms in breast cancer patients. Citation Format: Frida Björk Gunnarsdottir, Catharina Hagerling, Caroline Bergenfelz, Camilla M. Rydberg, Meliha Mehmeti, Roni Allaoui, Sofie Mohlin, Sven Påhlman, Daniel Bexell, Karin Leandersson. Inflammatory macrophage derived TNF alpha induces downregulation of estrogen receptor alpha in breast cancer cells by inactivation of Foxo3A [abstract]. In: Proceedings of the AACR Special Conference on Tumor Immunology and Immunotherapy; 2018 Nov 27-30; Miami Beach, FL. Philadelphia (PA): AACR; Cancer Immunol Res 2020;8(4 Suppl):Abstract nr A39.

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An Experimental Model for Peri-conceptual COVID-19 Pregnancy Loss and Proposed Interventions to Optimize Outcomes.

Reports appear to give reassurance that vertical transmission near term is unlikely, but risks of incidental SARS-CoV-2 infection during fertility treatments, at embryo implantation, or in the first trimester remain unknown. If early pregnancy sequela in the current COVID-19 pandemic are modeled from the 2004 Coronavirus outbreak data, then SARS-CoV-2 infection proximate to blastocyst nidation is likely to cause implantation failure or spontaneous abortion. Our model explains why this outcome is less attributable to virus-associated maternal pulmonary distress and instead derives from systemic inflammation and interference with trophectoderm-endometrium molecular signaling required for implantation. COVID-19 is often accompanied by high levels of IL-6, IL-8, TNF-alpha and other cytokines, a process implicated in pulmonary collapse and systemic organ failure. Yet when regarded in an early reproductive context, this “cytokine storm” of COVID-19 triggers a pro-coagulative state hostile to normal in utero blastocyst/fetal development. Evidence from obstetrics is accumulating to show that mothers with SARS-CoV-2 deliver placentas with abnormal interstitial villi fibrin deposits, diffuse infarcts, and hemangiomatous changes. This model classifies such lesions as permissive at term but catastrophic near embryo implantation or early first trimester pregnancy. Clinical experience with recurrent pregnancy loss offers workable interventions to address this challenge, but success will depend on prompt and accurate SARS-CoV-2 diagnosis. Although no professional guidelines currently exist for SARS-CoV-2 in early pregnancy, this model would warrant a high-risk designation for such cases; these patients should receive priority access to screening and treatment resources.

Correlation of tumor necrosis factor alpha, interleukin 6 and interleukin 10 with blood pressure, risk of preeclampsia and low birth weight in gestational diabetes.

Several pathophysiological mechanisms have been proposed in the development of pregnancy complications, including endothelial dysfunction, an inflammatory pathway and oxidative stress. The aim of the present study was to evaluate the correlation between proinflammatory cytokines TNF-alpha, IL-6 and dual cytokine IL-10 in the mother's peripheral blood and systolic blood pressure, risk of preeclampsia and low birth weight in gestational diabetes (GDM). We observed 40 women with GDM divided into a gestational hypertension group (n=20) and comparison group (n=20) with normal blood pressure. We found a significant positive correlation between TNF-alpha; IL-6; IL-10 levels and systolic blood pressure (SBP) in the second trimester (p<0.001; p<0.001; p<0.001); the third trimester (p<0.001; p<0.001; p<0.05). We also proved correlations for diastolic blood pressure (DBP) during the second; third trimester (p<0.001; p<0.001; p<0.001); (p<0.001; p<0.001; p<0.0015). We demonstrated a statistically significant positive association between high TNF-alpha group and preeclampsia risk in the third trimester (p=0.04). We also determined the negative correlation in the second trimester between birth weight and TNF-alpha; IL-6, IL-10 levels (p<0.05; p<0.001; p<0.001). To conclude, our data highlight the importance of cytokines TNF-alpha, IL-6 and IL-10 in blood pressure regulation. In addition, high levels of TNF-alpha have been associated with increased risk of preeclampsia. We found a significant negative correlation between levels of TNF-alpha, IL-6, IL-10 and birth weight.

Level of Vitamin D, TNF-Alpha and Calcium in Patients with Dengue Fever, Dengue Hemorrhagic Fever and Dengue Shock Syndrome

Background: Vitamin D has been widely studied for its beneficial role in treating different diseases. Dengue epidemic has affected many house hold since last decade. Vitamin D seems a gate through in its treatment as it inhibits Th1 cytokine activities, responsible for the development of dengue complications.Objective: To determine the levels of vitamin D, TNF- ? and calcium in patients with variation of dengue fever stages.Methods: A cross sectional analytical study conducted on dengue confirmed patients (fever, hemorrhagic, shock) admitted in tertiary care hospitals of Lahore (IRBNo.1165). After obtaining informed consent the study objectives were explained to the patients and data collected. Patients were classified into three groups (a) dengue fever (DF) (b) dengue hemorrhagic fever(DHF) and (c) dengue shock syndrome (DSS) (40+30+10 cases). Serum from 5cc blood was stored at –20? C until analysis. The data was analyzed using SPSS version 21.Results: The study was conducted in 4 hospitals of Lahore on 80 NS1confirmed cases (age: 5-70 years). The age difference, vitamin D and calcium levels were significant (p-values 0.030, 0.047 and 0.003 respectively). Calcium levels were lowest in DSS and significantly lower than DHF patients. Vitamin D levels were higher among those with higher severity of disease with p-value 0.031. The patients did not have significantly different TNF ? levels. There were 90.0% of DSS cases that had TNF ? levels &gt; 100, (p-value 0.109).Conclusion: High levels of TNF-alpha are associated with dengue complications regardless of vitamin D levels and severity of disease.