CTRL Group Research Articles

Dietary supplementation of archaeal carotenoids improved antioxidative capacity and regulated immune‐related gene expression of golden trout Oncorhynchus mykiss against challenge

Archaeal carotenoids, as novel bioactive compound, are stronger free radical scavenger. The effects of archaea Halorubrum carotenoids on skin coloration, antioxidative status and immune-related genes expression of golden trout Oncorhynchus mykiss were investigated. In total, 420 fish (10.90 ± 0.06 g) were divided into three groups with four replicates, and fed with commercial diets containing 0 (Ctrl), 25 mg/kg (P25) and 50 mg archaeal carotenoids/kg feed (P50) for 8 weeks. Although no significant difference was observed on growth, the fish fed archaeal carotenoids had significantly higher yellowness and carotenoid content (p < 0.05) in skin. Compared with the Ctrl group, the total antioxidant capacity (T-AOC) in serum was significantly enhanced in the P25 and P50 groups (p < 0.05), but no significant difference on lysozyme activity was obtained (p > 0.05). In liver, the catalase activity significantly decreased in the P50 group (p < 0.05), but T-AOC and superoxide dismutase activity was not significantly regulated by archaeal carotenoids (p > 0.05). The significant improvement of liver glutathione peroxidase in P50 group was observed, while malondialdehyde content significantly decreased (p < 0.05). After ammonia exposure and Aeromonas hydrophilia challenge, higher survival was observed in the P25 and P50 groups, associating with reducing the expression of pro-inflammation cytokines il-1β, but enhancing the expression of anti-inflammation cytokines tgf-β and hsp70. However, ammonia stress and pathogenic challenge caused opposite trends in the expression of tnf-α. Our study confirmed the beneficial role of archaeal carotenoids on skin coloration, antioxidative and immune systems of golden trout, and provided evidence for the potential application of this novel carotenoid in health management of aquaculture.

Long-term effects of multiple concussions on prefrontal cortex oxygenation during repeated squat-stands in retired contact sport athletes

ABSTRACT Background This study investigated the long-term effects of multiple concussions on prefrontal cortex oxygenation using near-infrared spectroscopy (NIRS) during a squat-stand maneuver that activated dynamic cerebral autoregulation. Methods Active male retired contact sport athletes with a history of 3+ concussions (mTBI; n = 55), and active retired athletes with no concussion history (CTRL; n = 29) were recruited. Participants completed a 5-min squat-stand maneuve (10-s squat, 10-s stand, 0.05 Hz; 15 times). Oxygenated (O2Hb), deoxygenated (HHb), total (tHb) hemoglobin, and hemoglobin difference (HbDiff) were analyzed through the change in maximal and minimal values during the test (∆MAX), Z-scores, and standard deviations. Results mTBI group showed left prefrontal cortex O2Hb ∆MAX (p = 0.046) and HbDiff ∆MAX (p = 0.018) were significantly higher. Within-group analyses showed significantly higher left HHb ∆MAX (p = 0.003) and lower left HbDiff Z-scores (p = 0.010) only in the mTBI group. The CTRL group demonstrated significantly lower left HbDiff SD (p = 0.039), tHb Z-scores (p = 0.030), and HbDiff ∆MAX (p = 0.037) compared to right prefrontal cortex response. Conclusion These preliminary results suggest changes in prefrontal cortex oxygenation potentially affecting the brain’s ability to adapt to changing cerebral perfusion pressure after multiple previous concussions.

Severe COVID-19 correlates with lower mitochondrial cristae density in PBMCs and greater sitting time in humans.

An interaction between mitochondrial dynamics, physical activity levels, and COVID‐19 severity has been previously hypothesized. However, this has not been tested. We aimed to compare mitochondrial morphology and cristae density of PBMCs between subjects with non‐severe COVID‐19, subjects with severe COVID‐19, and healthy controls. Additionally, we compared the level of moderate‐vigorous physical activity (MVPA) and sitting time between groups. Blood samples were taken to obtain PBMCs. Mitochondrial dynamics were assessed by electron microscopy images and western blot of protein that regulate mitochondrial dynamics. The International Physical Activity Questionnaire (IPAQ; short version) was used to estimate the level of MVPA and the sitting time The patients who develop severe COVID‐19 (COVID‐19++) not present alterations of mitochondrial size neither mitochondrial density in comparison to non‐severe patients COVID‐19 (COVID‐19) and control subjects (CTRL). However, compared to CTRL, COVID‐19 and COVID‐19++ groups have lower mitochondrial cristae length, a higher proportion of abnormal mitochondrial cristae. The COVID‐19++ group has lower number (trend) and length of mitochondrial cristae in comparison to COVID‐19 group. COVID‐19, but not COVID‐19++ group had lower Opa 1, Mfn 2 and SDHB (Complex II) proteins than CTRL group. Besides, COVID‐19++ group has a higher time sitting. Our results show that low mitochondrial cristae density, potentially due to physical inactivity, is associated with COVID‐19 severity.

P-478 Deleterious influence of cyclophosphamide on primordial follicles derives from increased DNA damage and reduced proliferation in xenotransplanted human ovarian tissue

Abstract Study question What are the acute effects of cyclophosphamide (Cp) on primordial follicles (PrFs) in human ovarian tissue? Summary answer Administration of Cp damages PrFs via DNA fragmentation and results in reduced proliferation with no increase in PrF activation markers. What is known already Alkylating agents are highly gonadotoxic, and in cases where freezing oocytes, embryos, or ovarian tissue is impractical, a better understanding of the underlying damage mechanism could enable development of fertoprotective approaches (1). Studies from different groups suggest conflicting mechanisms of ovarian damage, with either PrF activation (2) or cell damage and apoptosis (3-4) proposed as drivers of PrF depletion. Few studies have examined this question using human tissue. We performed xenografts using ovarian tissue from a 17-month-old girl to ensure a graft with plentiful PrFs to test the acute effect of Cp. Study design, size, duration Cross-sectional study. We utilized a xenotransplantation model in which human ovarian tissue is co-transplanted with endothelial cells (ECs) into immunocompromised mice (NSG) (5). Three weeks after xenotransplantation, time 0, intraperitoneal (IP) injection of (saline/Cp) was followed by an IP injection of ethynyl-deoxyuridine (Edu), at 0 and 24 hours, followed by 5-chloro-2′-deoxyuridine (CldU), at 48 and 72 hours. Grafts were harvested at 96 hours. Participants/materials, setting, methods We co-xenotransplanted human ovarian cortical tissue from a 17-month-old girl, cryopreserved for fertility preservation, into immunocompromised mice. After 3 weeks, Cp (75mg/Kg)/saline was administered IP. Mice were then sequentially injected with EdU, followed by CldU (both at 100mg/kg for 2 days) 24 hours apart. Twenty-four hours later, mice were sacrificed and xenografts were harvested and sectioned. Slides were stained for EdU, CldU, VASA, γ-H2AX, FOXO3a, and 4',6-Diamidino-2-Phenylindole-Dilactate (DAPI) and imaged using a confocal microscope. Main results and the role of chance We used anti-VASA staining to evaluate oocyte morphology, confirming that chemotherapy was not sterilizing; we counted 76.5% of morphologically normal PrF in the control group (Ctrl) (130/170) and 35.1% (13/37) in the Cp group. In the Ctrl group, 11% stained positively for DNA fragmentation using anti-γ-H2AX, whereas 43% were positive in the Cp group (p = 0.0073). To evaluate activation, we stained for FOXO3a in the oocyte nucleus. No difference was found between the groups: 52.4% (74/144) versus 44.6% (37/83) in Ctrl versus Cp, respectively. Interestingly, when comparing EdU incorporation, the Ctrl group had higher incorporation at 72%, versus the Cp group with 40% of incorporation (p = 0.0485), and no difference was found with CldU incorporation: Ctrl 22% versus 27% in the Cp group, respectively (P = 0.6960). Limitations, reasons for caution We measured acute effects, DNA fragmentation, and proliferation, at five days post-Cp administration; however, this falls short of evaluating processes that appear later (fibrosis and neovascularization). Also, the results may partly stem from the patient’s young age. Repeating the experiment using adult-derived tissue might yield different results. Wider implications of the findings A better understanding of both the timeline and underlying mechanisms that contribute to chemotherapy-related gonadotoxicity could diminish the damage and depletion of the ovarian reserve (PrF). These results provide evidence that direct damage to PrFs, and not increased activation, contributes to gonadotoxicity in the acute phase following administration of Cp. Trial registration number NA

Nutritional Properties of Milk from Dairy Ewes Fed with a Diet Containing Grape Pomace.

The aim of the present study was to evaluate the effects of a diet containing a 10% of grape pomace (GP) on the milk yield, chemical-nutritional characteristics, total phenolic compounds (TPCs), antioxidant activity (AOA), fatty acids and proteins profile of dairy ewe’s milk. Forty-six ewes were dived into two groups: a control group (Ctrl), fed a standard diet, and an experimental group (GP+), whose diet was supplemented with 10% of GP on dry matter. The trial lasted 60 days and milk samples were collected and analyzed at the beginning (T0) and after 60 (T60) days. Dietary enrichment with GP did not affect the yield and the chemical composition of the milk. TPCs and AOA were not affected by the diet. After 60 days, the diet induced an increase in monounsaturated fatty acids (MUFA) and a decrease in medium chain saturated fatty acids (MCSFA), but the total saturated fatty acids (SFA), polyunsaturated fatty acids (PUFA), short chain saturated fatty acids (SCSFA) and long chain saturated fatty acids (LCSFA) were not modified. A decrease in the C14 desaturation index and an increase in the C18 index were also detected. Total caseins and whey protein were not affected by GP, even if a lower content of k-casein in GP+ milk compared to Ctrl milk was observed on the 60th day. The results of the present study suggest that 10% of GP can be included in the diet of lactating ewes without modifying milk gross composition but inducing significantly changes the fatty acid profile.

The injuries of spleen and intestinal immune system induced by 2-Gy 60Co γ-ray whole-body irradiation

Purpose The aim of the present study was to investigate the injuries of spleen and intestinal immune system induced by 2 Gy 60Co γ ray in mice. Materials and methods A total of 120 Balb/c mice were randomly divided into two groups: blank control (Ctrl) and model (IR). The IR mice were exposed to a single dose of total body irradiation (2 Gy, dose rate: 1 Gy/min) and sacrificed on 1st, 3rd, 7th, 14th and 21st day after irradiation. The indicators including general observations and body weight, the changes in peripheral hemogram, spleen index, histopathology examination and lymphocyte subsets of spleen. As well as the count and subsets of lymphocyte in gut-associated lymphoid tissue. Results Compared with the Ctrl group, the body weight, spleen index, peripheral blood cell and splenocyte amounts, intraepithelial lymphocytes number decreased significantly after exposure, accompanied by a notable decreased count of lymphocytes in Peyer’s patch and mesenteric lymph nodes. Moreover, ionizing radiation also broke the balance of CD4+/CD8+ and increased the Treg proportion in spleen, which then triggered immune imbalance and immunosuppression. In general, the spleen injuries occurred on 1st day after exposure, worse on 3rd day, and were relieved on 7th day. The intestinal immune injuries were observed on 1st day, and attenuated on 3rd day. On 21st day after exposure, the spleen volume and index have returned to normal, except for the distribution of lymphocyte subpopulations. Furthermore, all indicators of gut-associated lymphoid tissue, except for mesenteric lymph nodes lymphocyte count, had returned to normal levels on 21st day. Conclusion In conclusion, our data showed the injuries of spleen and intestinal immune system induced by 2 Gy 60Co γ ray whole-body irradiation. These findings may provide the bases for further radiation protection in the immunity.

P833: CATEGORIZING HEMATOLOGICAL RESPONSE TO PEGCETACOPLAN IN PATIENTS WITH PAROXYSMAL NOCTURNAL HEMOGLOBINURIA: A POST HOC ANALYSIS OF THE PHASE 3 PRINCE STUDY DATA

Background: Paroxysmal nocturnal hemoglobinuria (PNH) is a rare complement-mediated hematological disorder characterized by anemia, hemolysis, and bone marrow failure. Pegcetacoplan (PEG), an FDA/EMA-approved targeted C3 therapy for the treatment of PNH, can control intravascular and prevent extravascular hemolysis. A prior post hoc analysis of PEGASUS reported a higher proportion of patients presenting good or better hematologic responses to PEG versus eculizumab (Risitano A, et al., Blood, 2021;138 [Supplement 1]: 1104). The PRINCE study demonstrated efficacy/safety of PEG compared to control treatment (CTRL; supportive treatment of PNH without complement inhibitors) in complement-inhibitor naïve patients with PNH. Aims: This post hoc analysis categorized the hematologic response to PEG and CTRL at baseline and Week 26 in the PRINCE Trial (phase 3, multicenter, randomized, open-label, controlled trial [NCT04085601]). Methods: Adult complement-inhibitor naïve (no complement-inhibitor treatment within 3 months prior to screening) patients with PNH who had low hemoglobin (Hb) levels and high lactate dehydrogenase levels were included. Patients were randomized 2:1 to receive PEG or CTRL through Week 26. Hematologic response to PEG and CTRL was categorized as complete, good, partial, or minor at baseline and at Week 26 using the number of packed red blood cells transfusions needed in the last 6 months and Hb level at the specified timepoints (revised categorization criteria per Debureaux PE, et al. Bone Marrow Transplant. 2021; 56(10):2600-2602). Hematologic response categorization was performed according to the following criteria: Complete response: no transfusions, no anemia (≥12 g/dL); Good response: no transfusions, but with chronic mild anemia (10-12 g/dL); Partial response: no/occasional transfusions (≤2 units/6 months) and chronic moderate anemia (8-10 g/dL); Minor response: regular transfusions (≥3 units/6 months) and chronic moderate/severe anemia (<10 g/dL), or no/occasional transfusions (≤2 units/6 months) and chronic severe anemia (<8 g/dL). Any patients with missing data at Week 26 were evaluated using the most recent value within the 6 weeks prior. Statistical tests assessed significance for responses of ‘Good’ or ‘Complete’, collectively (Fisher’s Exact Test between PEG and CTRL groups at baseline; McNemar’s test within PEG group at baseline vs. Week 26). Results: In this study, 53 patients were randomized to PEG (n=35) or CTRL (n=18). At baseline, most patients had a partial or minor response to supportive therapy prior to randomization, with only 5.7% of patients in the PEG group and 0.0% in the CTRL group presenting a Good or Complete hematologic response (p= 0.5428). Hematologic response categorization for patients in the PEG group are shown for baseline and Week 26 (Figure). At Week 26, 80.0% of patients in the PEG group exhibited a Good or Complete response. The proportion of patients in the PEG group who demonstrated a Good or Complete response was significantly higher at Week 26 as compared to baseline (p<0.0001). One patient treated with PEG had missing data at Week 26 due to being lost to follow-up and was not evaluated. Image:Summary/Conclusion: This post hoc analysis of PRINCE trial data revealed that a substantial proportion of patients achieved good or complete hematologic response to PEG after 26 weeks. These findings further support the positive efficacy of PEG on hematologic parameters and provide evidence for the role of proximal complement inhibition on hematological and clinical improvements for patients with PNH.

Two-Dimensional Ultrasound and Triplane Tissue Doppler Ultrasound of Patients with Severe Preeclampsia.

This study was to investigate the cardiac function characteristics under two-dimensional ultrasound and triplane tissue Doppler imaging (TDI) of patients with severe preeclampsia (SPE). 28 SPE patients with singleton pregnancy from January 2018 to December 2020 were included in the SPE group. 25 healthy nonpregnant women of reproductive age were taken as the control group (Ctrl group), and 26 normal pregnant women with singleton pregnancy were selected as the normal group (Norm group); all the research objects underwent ultrasonography. The morphological and functional indexes of left and right ventricles were compared among the cases in different groups. The results showed that the left ventricular end-diastolic period diameter (LVEDd), left ventricular relative wall thickness (LV-RWT), left ventricular mass index (LVMi), left anterior descending (LAd), left ventricular E/e and e/a values, right ventricular diameter (RV-D), right ventricular anterior wall thickness (RVAW), a value, right atrial septum (RA-S), pulmonary artery systolic pressure (PASP), left ventricular end-systolic period diameter (LVEds), interventricular septal thickness (IVSd), posterior wall thickness (PWd), end-diastolic period volume (EVD), end-systolic period volume (ESV), relative wall thickness (RWT), sphericity index (SpI), left atrium volume index (LAVi), and E/e value of patients in the SPE group were higher than those in the Ctrl group and the Norm group (P < 0.05). The mitral annular plane systolic excursion (MAPSE), s value, tricuspid annual plane systolic excursion (TAPSE), ratio of early diastolic blood flow velocity to late diastolic blood flow velocity (E/A), ratio of peak early diastolic velocity to peak late diastolic velocity (e/a), peak early diastolic velocity (e), and ejection fraction (EF) of the SPE group were lower than those of the Ctrl group and the Norm group (P < 0.05). The ratio of mitral valve early diastolic blood flow velocity to peak early diastolic velocity (E/e) of the Norm group was higher than that of the Ctrl group (P < 0.05). In two-dimensional ultrasound of the SPE group, the maximum difference in time from the start to the peak of systole (Ts) of the right ventricle between the basal and middle segments of the lateral wall and that of interventricular septum (RV-Ts-max) was 31.56 ± 0.39%. The maximum difference in time to peak of early diastole (Te) under the same condition (RV-Te-max) was 47.16 ± 0.19%. Left ventricular LV-Ts-max and LV-Te-max were 9.83 ± 0.80% and 8.37 ± 0.68%, respectively, in triplane TDI, which were considerably higher than those in the Ctrl and Norm groups (P < 0.05). It suggested that two-dimensional ultrasound and triplane TDI could reflect the ventricular morphology as well as diastolic and systolic function injury in patients, which offered a reference basis for the diagnosis of SPE.

Complementarity of surgical therapy, photobiomodulation, A-PRF and L-PRF for management of medication-related osteonecrosis of the jaw (MRONJ): an animal study

BackgroundThis study aimed to evaluate the complementarity of surgical therapy, photobiomodulation (PBM), advanced platelet-rich fibrin (A-PRF), and Leukocyte and platelet-rich fibrin (L-PRF) for the management of medication-related osteonecrosis of the jaw (MRONJ).MethodsSixty rats underwent injection of zoledronate followed by left mandibular first and second molar extractions to induce MRONJ lesions. All rats were examined for the signs of MRONJ 8 weeks post-dental extraction. Forty-nine rats with positive signs of MRONJ were appointed to seven different groups as follows: control (Ctrl); surgery alone (Surg); surgery and PBM (Surg + PBM); surgery and A-PRF insertion (Surg + APRF); surgery and L-PRF insertion (Surg + LPRF); surgery, A-PRF insertion, and PBM (Surg + APRF + PBM); surgery, L-PRF insertion, and PBM (Surg + LPRF + PBM). Euthanasia was carried out 30 days after the last treatment session. The lesions' healing was evaluated clinically, histologically, and radiographically. Data were analyzed using STATA software version 14, and the statistical significance level was set at 5% for all cases.ResultsAccording to the present study, A-PRF and L-PRF treatment resulted in significant improvements in clinical, histological, and radiographical parameters compared to the Ctrl group (P < 0.05). The PBM also decreased wound dimensions and the number of empty lacunae compared to the Ctrl group (P < 0.05). Surg + APRF + PBM and Surg + LPRF + PBM were the only groups that presented a significantly higher mean number of osteocytes (P < 0.05). No significant differences were observed between A-PRF and L-PRF treatment groups (P > 0.05).ConclusionsSurgical resection followed by applying A-PRF or L-PRF reinforced by PBM showed optimal wound healing and bone regeneration in MRONJ lesions.

A Single Session of SMR-Neurofeedback Training Improves Selective Attention Emerging from a Dynamic Structuring of Brain-Heart Interplay.

Research on sensorimotor rhythms (SMR) based on neurofeedback (NFb) emphasizes improvements in selective attention associated with SMR amplification. However, the long-term training proposed in most studies posed the question of acceptability, which led to the evaluation of the potential of a single NFb session. Based on cognitive and autonomic controls interfering with attention processes, we hypothesized changes in selective attention after a single SMR-NFb session, along with changes in brain–heart interplay, which are reflected in the multifractality of heartbeat dynamics. Here, young healthy participants (n = 35, 20 females, 21 ± 3 years) were randomly assigned either to a control group (Ctrl) watching a movie or to a neurofeedback (NFb) group performing a single session of SMR-NFb. A headset with EEG electrodes (positioned on C3 and C4) connected to a smartphone app served to guide and to evaluate NFb training efficacy. A Stroop task was performed for 8 min by each group before and after the intervention (movie vs. SMR-NFb) while collecting heart rate variability and C4-EEG for 20 min. When compared to Ctrl, the NFb group exhibited better Stroop performance, especially when facing incongruent trials. The multifractality and NFb training efficacy were identified as strong predictors of the gain in global Stroop performance, while multifractality was the only predictor regarding incongruent trials. We conclude that a single session of SMR-NFb improves selective attention in healthy individuals through the specific reorganization of brain–heart interplay, which is reflected in multifractal heartbeat dynamics.

The Kisspeptin analogue C6 induces ovulation in jennies

Kisspeptins (KPs) are the most potent stimulating neurotransmitters of GnRH release, and consequently KP administration triggers LH and/or FSH release. In small ruminants, KP or its analogs induced an LH surge followed by ovulation in both cyclic and acyclic animals, while in the mare KP only increased LH plasma levels but failed to induce ovulation. This study in jennies compares the endocrinological effects, ovulatory and pregnancy rates of the KP analog C6 and the GnRH analog buserelin acetate. The ovarian activity of nine Amiata jennies was monitored daily by transrectal ultrasound for three complete estrous cycles. Jennies in estrus were assigned, to one of three treatment groups: 50 nmol of the KP analog C6 (injected twice, 24 h apart, C6 group); 0.4 mg buserelin acetate (injected once, Bu group); and 2 mL of saline (injected once, CTRL group). Blood samples were collected at Day-1 (−24 h) Day0 (h0, before treatment), h2, h4, h6, h8, h10, h24 (before second treatment with C6), h26, h28, h30, h32, h34, h48 and every 24 h until ovulation. Jennies were inseminated once at h24 with fresh extended semen from a donkey stallion. Pregnancy diagnoses were performed 14 days after ovulation. On days 5, 10, and 14 after ovulation, for every CL the cross-sectional area (CSA) and the vascularized area (VA) were recorded by color doppler ultrasound and measured. Significantly higher plasma LH levels were found after induction between the Bu and CTRL groups at h6 and h8 (P < 0.05), while tendentially higher differences were found between the Bu/C6 groups and CTRL at h10. Five/9, 4/9, and 2/9 jennies ovulated between 24 and 48 h after induction from the Bu, C6, and CTRL groups respectively, (P > 0.05). Correlations between corpora lutea CSA and VA with serum progesterone concentration were r = 0.31, P = 0.01, r = 0.38, P = 0.01, respectively. Pregnancy rates after artificial insemination did not differ among groups (CTRL: 6/9, 66.7%; C6: 7/9, 77.8%; Bu: 6/9, 66.7%; P > 0.05). Ovulation rates after C6 treatment were comparable to that of Bu, although not different from the CTRL. Pregnancy rates were comparable to the literature in terms of fresh extended donkey semen in every group. This study suggests that stimulation of the Kp system in jennies, in contrast to findings observed in mares, induces ovulation. Further studies using higher doses and/or more animals are needed to better characterize the efficacy of C6 in jennies.

Abstract LB057: Bevacizumab leaves a photoacoustic fingerprint in breast cancer mouse models

Abstract Background: Photoacoustic tomography (PAT) provides a direct readout of tumor haemoglobin (Hb) concentration and oxygenation. This readout could be used to provide an early indication of response to anti-angiogenic drugs, thus optimizing the management of these therapies. Experimental Procedures: Two cohorts of nude Balb/c were inoculated with either estrogen-dependent MCF-7 (n=7) or estrogen-independent MDA-MB-231 (n=19) cells. Mice were randomly split into Control (Ctrl) and Bevacizumab (Bev,10 mg/Kg, IP, weekly) groups. PAT was performed weekly, starting just before enrolment. Oxy- and deoxy-Hb were quantified in the tumour and reference areas. Total Hb (THb) and O2 saturation (SO2) were calculated. Blood samples and tissues were collected after imaging. Hypoxia (Hypoxyprobe and CA-IX) and vascular density/maturity (CD31 and ASMA) were analyzed by immunohistochemistry. Circulating levels of human and mouse vascular endothelial growth factor (hVEGF and mVEGF) and Hb were measured. Summary of New Data: Reflecting clinical observations, the MCF-7 Bev group and most of the mice from the MDA-MB-231 Bev group (8/11) showed no improvement in survival with Bev treatment. All resistant tumours (Bev MCF-7 and Bev MDA-MB-231 non-responders (Bev-NR)) showed an increase in hVEGF production (Ctrl vs Bev; 73.0513.06 vs. 497103.9 for MCF-7; 247.984.81 vs. 330.4 85.97 for Bev NR). MDA-MB-231 responders (Bev-R) showed a significant decrease in hVEGF (247.984.81 vs. 80.53 26.13). These results indicate that VEGF was only successfully sequestered in a small subset of animals (3/11) and that VEGF overload might be a resistance mechanism. At the final time point, PAT showed no difference between Ctrl and Bev-NR in THb for either group (THbMDA-MB-231 7.2±1.3 vs. 5.5±1.2). THb was found significantly increased in the Bev-R group by PAT (13.9±3.3 p-valuevs.Crtl=0.0339; p-valuevs.Bev=0.0072) and biochemical measurements. SO2 showed a slight but significant decrease between Ctrl and Bev-NR (0.58±0.03 vs. 0.48±0.01). A dramatic decrease was seen in the Bev-R group (0.31±0.07) and significantly different from Ctrl and Bev-NR. SO2 was sensitive to early changes, SO2 values increased in Ctrl and Bev-NR 48h after enrolment (p-values paired t-test pCtrl= 0.0134; pNR=0.0268) while Bev-R shows a trend (p=0.0649). Analysis of SO2MSOT over time shows a significant (p&amp;lt;0.0001) change in slope at 3 weeks after enrolment in the Bev-R group compared to either of the Ctrl or Bev-NR groups. Our results indicate that PAT SO2 can differentiate responders at very early stages of the treatment. Histologically, we observed fewer blood vessels but better structured in Bev-NR than in Bev-R, which showed higher vascular density and also higher levels of hypoxia makers. Statement of the Conclusions: Our results postulates PAT as a low-cost, label-free and non-invasive candidate to monitor response to Bevacizumab over time. Our results also indicate that tumours with functional vasculature may be resistant to Bevacizumab treatment. Citation Format: Isabel Quiros-Gonzalez, Michal R. Tomaszewski, Monika A. Golinska, Emma Brown, Laura Ansel-Bollepalli, Lina Hacker, Dominique-Laurent Couturier, Rosa M. Sainz, Sarah E. Bohndiek. Bevacizumab leaves a photoacoustic fingerprint in breast cancer mouse models [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr LB057.

A 12-Week Cycling Workstation Intervention Improves Cardiometabolic Risk Factors in Healthy Inactive Office Workers.

The aim of this study was to evaluate the effects of a portable pedal machine intervention (60 minutes per working day) for 12 weeks on healthy tertiary employees' cardiometabolic risk factors. Anthropometric parameters, body composition, cardiometabolic/inflammatory markers, physical fitness, physical activity, and sedentary time measured before and after the intervention were compared between office healthy workers who used a portable pedal machine (INT, n = 17) and those who did not (CTRL, n = 15). The INT group improved Δultrasensitive C-reactive protein ( P = 0.008), Δtotal cholesterol ( P = 0.028), and Δlight-density lipoprotein cholesterol ( P = 0.048) compared with the CTRL group (Δ: T1-T0). The intervention reduced daily sitting time ( P ≤ 0.01) and increased time spent at light intensity ( P ≤ 0.01) and moderate-to-vigorous ( P ≤ 0.01) physical activity compared with baseline values. These findings suggest that promoting physical activity during workdays can reduce the negative health effects of spending too much time sitting and inactive.

Exaggerated Ventilator-Induced Lung Injury in an Animal Model of Type 2 Diabetes Mellitus: A Randomized Experimental Study.

Although ventilator-induced lung injury (VILI) often develops after prolonged mechanical ventilation in normal lungs, pulmonary disorders may aggravate the development of adverse symptoms. VILI exaggeration can be anticipated in type 2 diabetes mellitus (T2DM) due to its adverse pulmonary consequences. Therefore, we determined whether T2DM modulates VILI and evaluated how T2DM therapy affects adverse pulmonary changes. Rats were randomly assigned into the untreated T2DM group receiving low-dose streptozotocin with high-fat diet (T2DM, n = 8), T2DM group supplemented with metformin therapy (MET, n = 8), and control group (CTRL, n = 8). In each animal, VILI was induced by mechanical ventilation for 4 h with high tidal volume (23 ml/kg) and low positive end-expiratory pressure (0 cmH2O). Arterial and venous blood samples were analyzed to measure the arterial partial pressure of oxygen (PaO2), oxygen saturation (SaO2), and the intrapulmonary shunt fraction (Qs/Qt). Airway and respiratory tissue mechanics were evaluated by forced oscillations. Lung histology samples were analyzed to determine injury level. Significant worsening of VILI, in terms of PaO2, SaO2, and Qs/Qt, was observed in the T2DM group, without differences in the respiratory mechanics. These functional changes were also reflected in lung injury score. The MET group showed no difference compared with the CTRL group. Gas exchange impairment without significant mechanical changes suggests that untreated diabetes exaggerates VILI by augmenting the damage of the alveolar–capillary barrier. Controlled hyperglycemia with metformin may reduce the manifestations of respiratory defects during prolonged mechanical ventilation.

Effects of Outdoor Walking on Positive and Negative Affect: Nature Contact Makes a Big Difference.

It has been consistently demonstrated that physical exercise is a cost-effective way to promote emotional well-being. However, the environment in which it takes place might amplify or mitigate this beneficial effect. The present study aimed at comparing the effects of walking in a natural or urban field setting on positive and negative affect. For this purpose, 150 students (46 female, 104 male; mean age: 20.2 years) were randomized into one of three groups: Green Walking (GW, n = 50), Urban Walking (UW, n = 50), or no-exercise (control; CTRL, n = 50). Positive and negative affect ratings were collected for each participant before and after walking (or before and after attending a class in the CTRL group). Exercise parameters (duration, intensity, weather conditions, group size) were identical in the GW and UW groups. The walking routes differed in terms of vegetation density, proximity of water, presence of traffic, and amount of asphalted surfaces. Participants in the GW and UW groups reported significant reductions in negative affect pre- to post walking. However, positive affect was increased only for participants in the GW group. This finding may have meaningful implications for mental health professionals who treat patients with significant emotional distress or mood instability. Several explanations are discussed as potential mechanisms for the more beneficial effect of Green walking, and presented as an important avenue for future research.

Adhesion to a new CAD/CAM resin composite: Effects of the machining roughness simulation, surface treatments, and long-term aging

ObjectivesTo evaluate the effect of surface treatments, finishing condition (polished vs. ground) and aging on the microtensile bond strength of a CAD/CAM resin composite to resin cement. Materials and methodsCAD/CAM resin composite blocks (Tetric CAD HT) were sectioned (7 × 12 × 4 mm), randomly divided according to finishing condition (polished or ground) and surface treatment groups (Ctrl: no treatment; AA: air-abrasion with 110 μm Al2O3 powder; TBS: tribochemical silica coating). A coupling agent corresponding to each surface treatment was applied for cementation (Ctrl and AA groups: ExciTE F DSC; TBS group: RelyX Ceramic Primer) and each block was adhesively cemented (Variolink N) with another one from the same conditioning. Next, 1 mm2 cross-section beams were produced after sectioning, with half being tested under microtensile test after 24 h, while the other half was aged (25,000 cycles of thermocycling + 90 days of storage) (n = 50). Contact angle and Scanning Electron Microscopy (SEM) analysis were performed. Bond strength data (MPa) were statistically analyzed using Three-way ANOVA and Tukey tests (α = 0.05). The failure types were examined and classified as predominantly adhesive, cohesive or mixed under stereomicroscopy. ResultsThe surface treatments (AA and TBS) promoted higher bond strength than the Ctrl group. Grinding finishing improved the bond strength only when no treatment was performed (Ctrl group), having a negative effect for the TBS treatment. The long-term aging damaged the resin bond, independent of the condition. The failure analysis showed that 92% of the failures were classified as adhesive in the baseline conditions, and 94.7% in aged specimens. TBS showed the lowest contact angle among the tested groups for both conditions (baseline and aged). ConclusionSurface conditioning with alumina particle air-abrasion or tribochemical silica coating improves the bond strength. CAD/CAM milling roughness induces better surface condition for adhesion, in particular when no treatment is performed. Bond degradation occurs after long-term aging for all the test conditions. Clinical relevanceWhen luting CAD/CAM resin composite restorations, the surface treatment and roughness due to the grinding by burs in the manufacturing process influence the adhesion of restoration to resin cement.

REDUCTION OF HEART RATE VIA P2X4 RECEPTOR ACTIVATION AS A PROMISING TREATMENT STRATEGY IN PULMONARY ARTERIAL HYPERTENSION

Objective: Mounting evidence suggest that the ATP-sensitive ionotropic P2X4 receptor (P2X4R) improves cardiac performance and optimizes myocardial energy demand by reducing chronotropy. Here, we set to investigate whether the digitalis-like effect of the P2X4R in the sinus node was preserved in rats with right heart failure due to pulmonary arterial hypertension (PAH). Design and method: Male Wistar rats received a single subcutaneous injection of either monocrotaline (MCT; PAH group) or vehicle (CTRL group). Right heart failure associated to PAH normally occurs in Wistar rats 21 to 25 days after MCT administration. Myographic recordings were performed in spontaneously beating atria (chronotropic effect) isolated from both animal groups. Results: Baseline spontaneous atrial rate was 250 ± 5 (n = 26) bpm in CTRL and 237 ± 7 bpm (n = 23) in PAH group (p &gt; 0.05). ATP (0.01–1 mM) concentration-dependently reduced the spontaneous atrial rate of both animal groups. The negative chronotropic effect of ATP (100 M) was smaller in PAH rats (-9 ± 2%; n = 14 vs. -22 ± 4%; n = 25, p &lt; 0.05), but this divergence was attenuated upon increasing the concentration of ATP to 1 mM (CTRL: -54 ± 5%; n = 15 vs. MCT: -50 ± 7%; n = 8; p &gt; 0.05). The P2X4R antagonist, 5-BDBD (10 M), attenuated the negative chronotropic action of ATP (100 M) in CTRL (from -19 ± 5% to -13 ± 3%; n = 5, p &gt; 0.05) and PAH (from -8 ± 3% to 1 ± 8%; n = 4, p &gt; 0.05) rats. The positive allosteric modulator of P2X4R, ivermectin (30 M), potentiated the negative chronotropic effect of ATP (100 M) in CTRL (from -31 ± 5% to -56 ± 14%; n = 5, p &gt; 0.05) and PAH (from -12 ± 6% to -24 ± 9%; n = 4, p &lt; 0.05). The non-selective P2XR agonist, CTP (1 mM), which contrary to ATP does not breakdown into adenosine, also decreased atrial chronotropy in CTRL (-24.31 ± 6.73%; n = 6) and PAH (-11.82 ± 1.98%; n = 6) rats. Conclusions: Data suggest that the negative chronotropic effect of P2X4R activation is preserved in atria of rats with heart failure associated to PAH, thus strengthening the hypothesis that heart rate reductions using this ATP-sensitive receptor may increase cardiac performance in patients with PAH-induced heart failure.

THERAPEUTIC EFFECTS OF NEBULIZATION- BASED DELIVERY OF ANTI-MIR-146A IN EXPERIMENTAL PULMONARY ARTERIAL HYPERTENSION

Objective: Pulmonary arterial hypertension (PAH), is a chronic disorder characterized by excessive pulmonary vascular remodelling, resulting in elevated pulmonary vascular resistance and right ventricle (RV) overload and failure. PAH remains incurable, and new therapeutic approaches are required. The microRNA-146a (miR-146a) promotes vascular smooth muscle cell proliferation and vascular neointimal hyperplasia, both important hallmarks of PAH. This study aimed to investigate the role of miR-146a in the pathophysiology of PAH and in the progression to RV hypertrophy and failure. Design and method: Sprague Dawley rats received a subcutaneous injection of monocrotaline (MCT group) or vehicle (CTRL group) and, after fourteen days they were treated, by intratracheal nebulization, with an anti-miR-146a, forming four distinct groups: CTRL-treated; CTRL-untreated, MCT-treated, and MCT-untreated. Twenty-seven days after MCT injection, echocardiographic and invasive hemodynamic evaluations were performed in all animal groups. Also, wild-type (WT) and miR-146a knock-out (KO) (miR-146a-/-) mice were submitted to either 3 weeks of chronic hypoxia-induced PAH with weekly Sugen 5416 administration (SuHx). Results: Compared to MCT-untreated rats, the MCT-treated rats showed decreased RV hypertrophy, (as measured by the Fulton index), decreased RV end-diastolic diameter/left ventricle end-diastolic diameter (RVEDD/LVEDD) ratio, and decreased in RV diastolic pressures. KO-SuHx group, when compared to WT-SuHx group, showed decreased RV hypertrophy (as measured by the Fulton index), and decreased RV systolic pressures and dilation, as well as lower RV end end-diastolic diameter (RVEDDi) and right atria area (RAAi). Conclusions: Our findings show that miR-146a pharmacological or genetic inhibition reduces RV remodelling and improves cardiac function. Thus, miR-146a represents a promising therapeutic target in PAH.

CHANGES OF VASCULAR REACTIVITY TO ADENOSINE A1 AND A2A RECEPTOR AGONISTS AFTER HYPERBARIC OXYGENATION

Objective: Hyperbaric oxygenation (HBO) affects vascular reactivitiy, which plays an important role in the pathophysiology of hypertension and other cardiovascular diseases. New data suggest important effects of HBO on oxidative stress and alteration of production of vasoactive metabolites in blood vessels; however, the role of adenosine A1 and A2a receptors is still underinvestigated.This study aimed to determine the functional role of adenosine A1 and A2a receptors in changes of vascular reactivity in isolated middle cerebral arteries (MCA) of Spague-Dawley rats, after acute and intermittent HBO. Design and method: Male and female healthy Sprague-Dawley rats aged 8–10 weeks were divided into 3 groups: CTRL (control non HBO), A-HBO (single animals exposed to HBO for 2 hours) and 4D-HBO (animals exposed to HBO at a pressure od 2 bars for 2 hours daily for 4 days, with vascular experiments conducted on the fifth day). The rats were anesthetized with ketamine-chloride (3 ml/kg) and midazolam (0,5 ml/kg) and decapitated. The MCA were isolated, cannulated and pressurized for 60’at 80 mmHg to assess basal diameter using pressure myograph Danish Myo Technology. A1 adenosin receptor selective agonist CCPA or A2a adenosin receptor agonist CGS in concentrations from 10–5 to 10–10 M were added to the vessel chamber, and blood vessel diameter was measured after incubation of the drug for 15 minutes. Statistical analyses were performed with Two-way ANOVA tests; p &lt; 0.05 was considered significant. Results: The vasoconstriction in response to A1 agonist administration was similar between the CTRL and 4D-HBO group, while it was significantly reduced in A-HBO compared to the CTRL and 4D-HBO groups. The vasodilation in response to A2a stimulation was significantly reduced in rats exposed to A-HBO and I-HBO compared to CTRL at higher drug concentrations (10–7 to 10–5 M). Conclusions: Acute exposure of rats to HBO leads to decrease in the vasoconstrictive response to adenosine A1 receptor stimulation, while acute and intermittent exposure of rats to HBO is associated with a decrease in the vasodilatory response of MCA to adenosine A2a receptor activation, suggesting that HBO alters sensitivity to or expression of receptors.

Effect of Oral Supplementation of Gordonibacter urolithinfaciens on Gut Microbiota and Bioavailability of Ellagic Acid and Urolithins in Mice

ObjectivesGordonibacter urolithinfaciens (GUro) is one of the three human bacteria that can convert ellagic acid (EA) to urolithins (Uros). Individuals deficient in Uro-producing bacteria may not benefit from EA-rich diets due to poor bioavailability of EA. Thus, GUro supplementation could be a solution. This work aims to examine 1) the viability of naked and protected GUro after gastrointestinal (GI) digestion; 2) colonization of GUro in vivo and the impact of GUro on gut microbiota; 3) the effect of GUro supplementation on the bioavailability of EA and Uros. MethodsIn vitro studies: Simulated GI digestion was conducted with GUro in naked (NG), alginate-chitosan encapsulated (ACG) or hydrogel protected (HGG) forms. Bacteria enumeration was performed by plate count. In vivo studies: Male C57BL/6J mice deficient in GUro were divided into 2 groups: Control (Ctrl, vehicle) and Experiment (Exp, GUro at 107 CFU/mouse). Mice received vehicle or GUro on days 1, 2, 3 and 7 by oral gavage, and then 20 mg/kg EA 12 h after the final dose of GUro or vehicle on D8. Feces were collected for gut microbiota analysis. Blood and tissues were collected at 0.5, 12, 24 and 36 h after EA intake. 36 h excretion of urine and feces were collected. The change in the main genera of gut microbiota was measured by qPCR. Colon tissue histology was examined by H&E staining. EA and Uros were extracted from biosamples and analyzed by UHPLC with triple quadrupole mass spectrometry. ResultsNG showed comparable viability as ACG and HGG after in vitro GI digestion indicating GUro’s suitability to be ingested without protection. For in vivo study, the level of GUro increased 8 folds 12 h after the last dose of GUro but returned to baseline in 24–36 h. For Ctrl group, Akkermansia (Akk) increased substantially at all time points. Similarly, Akk bloomed post-gavage of EA but most strikingly at 0.5 h in Exp group. GUro administration significantly enhanced the bioavailability of EA and Uros. Substantial amounts of Uro C, dimethyl Uro C, Uro A and methyl Uro A were excreted in the urine of Exp group, but barely detected in the Ctrl. ConclusionsSupplementation of GUro improved the bioavailability of EA and bioactive metabolites. Moreover, the high survival rate of naked GUro after GI digestion indicated its potential to serve as a probiotic. Funding SourcesNational Natural Science Foundation of China; PolyU Research Institute for Future Food.