Impacts of different intensities of exercise on inflammation and hypoxia markers in low altitude

Background This study aims to determine and compare the effects of exercise modalities with different intensities on the secretion of key inflammation and hypoxia markers in amateur athletes. Methods Twenty-three athletes with a mean age of 20.1 years, living at low altitude (1850 m) participated in this study. The participants' maximal oxygen consumption values (VO2 max) were determined with an incremental cycle exercise test as 54.15 ± 6.14 mL kg min−1. Athletes performed four protocols: at rest, 50% VO2 max, 75% VO2 max and 100% VO2 max (until exhaustion) with one-week intervals. 50% VO2 max, 75% VO2 max sessions were performed continuously for 30 min on a bicycle ergometer and 100% VO2 max session was performed by cycling until exhaustion. Blood samples were obtained at rest and immediately after each exercise session. Serum tumor necrosis factor alpha (TNF-α), C-reactive protein (CRP), interleukin-10 (IL-10), and hypoxia inducible factor-1 alpha (HIF-1α) levels were measured. Results There were significant differences in serum TNF-α levels in 75% VO2 max and 100% VO2 max sessions (489.03 ± 368.37 and 472.70 ± 365.21 ng/L, respectively) compared to rest conditions (331.65 ± 293.52 ng/L). Serum CRP levels of 50% VO2 max and 75% VO2 max sessions (1.19 ± 0.50; 1.07 ± 0.52 mg/L) were significantly higher than the rest condition (0.74 ± 0.35 mg/L). There were significant differences in serum IL-10 levels of rest condition and 50% VO2 max; 50% VO2 max, and 100% VO2 max sessions (328.09 ± 128.87; 446.36 ± 142.84; 347.44 ± 135.69; 324.88 ± 168.06 pg/mL). Serum HIF-1α levels were significantly higher in 75% VO2 max session compared to rest (1.26 ± 0.16; 1.08 ± 0.19 ng/mL) (P < 0.05 for all comparisons). Conclusions Both inflammatory and anti-inflammatory pathway is induced on different exercise intensities. Exercise protocols performed until exhaustion may lead to activation of inflammatory pathways and hypoxia-induced damage.

Derivation of a Novel CIHI in Patients with Lung Adenocarcinoma for Estimating Tumor Microenvironment and Clinical Prognosis

An interaction between hypoxia and immunity has been confirmed in tumor tissue. However, there is no combined biomarker for diagnosis on this basis. Therefore, we developed a scoring formula based on markers of hypoxia and immunity. Firstly, the hypoxia-immune formula of lung adenocarcinoma (LUAD) was derived using LASSO-Cox regression in three cohorts from public database, and the corresponding score was calculated for each patient. The formula is as follows: combined hypoxia and immune index CIHI = LDHA expression × 0.2252 + GAPDH expression × 0.0727 + ANGPTL 4 expression × 0.0724 + VEGFC expression × 0.1911 + DKK 1 expression × 0.1355 + ADM expression × 0.0588 + BTK expression × − 0.1659 . Meanwhile, patients were divided into groups according to high and low CIHI, and expression profiles of hypoxia markers and immune markers were analyzed in different groups. CIHI was used to confirm that patients with high CIHI represented a state of hypoxiahigh-immunitylow, which had worse overall survival. We also discussed the evaluation value in the immune microenvironment and clinical application of CIHI. In conclusion, this study developed and validated a hypoxia-immune formula that can guide hypoxia modifier treatment and immunotherapy in LUAD.

Expression of Myoglobin in Normal and Cancer Brain Tissues: Correlation With Hypoxia Markers

BackgroundMyoglobin (MB) is increasingly recognized as a key player in cancer growth and metastasis. Low oxygen tensions, commonly associated with highly aggressive and recurrent cancers, have been shown to regulate its expression in several cancers such as lung, neck, prostate and breast cancer. However, it is not yet known whether it contributes to the growth and spread of brain cancers especially Glioblastoma multiforme (GBM).MethodsHere we investigate the expression of MB, and its correlation with the hypoxia markers carbonic anhydrase IX (CAIX) and lactate dehydrogenase A (LDHA), in human tissue microarrays of multiple organ tumors, brain tumors, and GBM tumors, and their respective cancer-adjacent normal tissues. Correlation between MB protein expression and tumor grade was also assessed.ResultsWe show that MB protein is expressed in a wide variety of cancers, benign tumors, cancer-adjacent normal tissues, hyperplastic tissue samples and normal brain tissue, and low oxygen tensions modulate MB protein expression in different brain cancers, including GBM. Enhanced nuclear LDHA immune-reactivity in GBM was also observed. Finally, we report for the first time a positive correlation between MB expression and brain tumor grade.ConclusionOur data suggest that hypoxia regulate MB expression in different brain cancers (including GBM) and that its expression is associated with a more aggressive phenotype as indicated by the positive correlation with the brain tumor grade. Additionally, a role for nuclear LDHA in promoting aggressive tumor phenotype is also suggested based on enhanced nuclear expression which was observed only in GBM.

Synthesis and Characterisation of a Boron-Rich Symmetric Triazine Bearing a Hypoxia-Targeting Nitroimidazole Moiety

Boron Neutron Capture Therapy (BNCT) is a binary therapy that promises to be suitable in treating many non-curable cancers. To that, the discovery of new boron compounds able to accumulate selectively in the tumour tissue is still required. Hypoxia, a deficiency of oxygen in tumor tissue, is a great challenge in the conventional treatment of cancer, because hypoxic areas are resistant to conventional anticancer treatments. 2-Nitroimidazole derivatives are known to be hypoxia markers due to their enrichment by bioreduction in hypoxic cells. In the present work, 2-nitroimidazole was chosen as the starting point for the synthesis of a new boron-containing compound based on a 1,3,5-triazine skeleton. Two o-carborane moieties were inserted to achieve a high ratio of boron on the molecular weight, exploiting a short PEG spacer to enhance the polarity of the compound and outdistance the active part from the core. The compound showed no toxicity on normal human primary fibroblasts, while it showed noteworthy toxicity in multiple myeloma cells together with a consistent intracellular boron accumulation.

Perioperative neorectal ischemia. Experimental model

Objective. To experimentally determine and analyze the role of perioperative tissue tension in the intestinal area brought down to the proper topographic and anatomical position (neorectum) and its vascularization impairments during anorectoplasty. Materials and methods. This experiment performed on laboratory rats consisted of 3 parts: clinical (creation of an operation model and assessment of clinical parameters in the early postoperative period); morphological (histological examination of biopsy specimens taken from the surgical area); and laboratory (measurement of tissue hypoxia markers (including vascular endothelial growth factor-C (VEGF-C) and hypoxia-induced factor 1-alpha (HIF-1-alpha)) in serum and intestinal homogenate using enzyme-linked immunosorbent assay (ELISA) (specimens were collected 3 days postoperatively). Forty laboratory rats weighing from 370 to 550 g (mean weight 482 g) were divided into four equal groups: group I – ‘impaired neorectal vascularization;’ group II – ‘neorectal tension;’ group III – ‘rectal replantation;’ and the control group. During the first stage of the experiment (3 days), we assessed overall condition of the animals, number of stools, type of defecation, and pain severity. The animals received no treatment in the postoperative period. On day 3, all rats were euthanized. Tissue specimens were collected from the neorectum to evaluate its morphological characteristics and to produce intestinal homogenate for subsequent ELISA. Blood specimens were obtained by puncturing the left ventricle and then centrifuged at 3000g for 15 minutes. Blood supernatants and neorectum tissues were stored at -80°C for maximum 1 month before testing. Intestinal tissue samples for histological examination were stored in a 10% formalin solution. The levels of hypoxia biomarkers were measured in serum and intestinal homogenate using the Stat Fax 2100. Data analysis was conducted using the SPSS software (with no preliminary calculation of the sample size). We used the Mann–Whitney U-test [H.B.Mann, D.R.Whitney] and Kruskal–Wallis test [W.Kruskall, W.A.Walles]. The differences were considered significant at р < 0.05. Results. The most common morphological changes were caused by impaired vascularization of the rectum. We have found a significant increase (p < 0.001) in the levels of hypoxia markers in intestinal homogenates in all three experimental compared to controls. However, there was no significant difference in the levels of these markers between the experimental groups. The defecation process was abnormal in rats from groups I and II. Conclusion. In anorectoplasty, excessive mobilization of the rectum with its improper vascularization and formation of the anus under conditions of intestinal tissue tension lead to perioperative neorectal ischemia. This experiment should be continued to evaluate long-term results. The proposed models of perioperative neorectal ischemia can be used to analyze the role of many factors that can potentially affect the outcome of anorectal malformation correction. In the future, this problem may be investigated in clinical studies. Key words: anorectal malformations, perioperative ischemia, neorectum, tension, devascularization

Mixing and delivery of multiple controlled oxygen environments to a single multiwell culture plate

Precise oxygen control is critical to evaluating cell growth, molecular content, and stress response in cultured cells. We have designed, fabricated, and characterized a 96-well plate-based device that is capable of delivering eight static or dynamically changing oxygen environments to different rows on a single plate. The device incorporates a gas-mixing tree that combines two input gases to generate the eight gas mixtures that supply each row of the plate with a different gas atmosphere via a removable manifold. Using air and nitrogen as feed gases, a single 96-well plate can culture cells in applied gas atmospheres with Po2 levels ranging from 1 to 135 mmHg. Human cancer cell lines MCF-7, PANC-1, and Caco-2 were grown on a single plate under this range of oxygen levels. Only cells grown in wells exposed to Po2 ≤37 mmHg express the endogenous hypoxia markers hypoxia-inducible factor-1α and carbonic anhydrase IX. This design is amenable to multiwell plate-based molecular assays or drug dose-response studies in static or cycling hypoxia conditions.