Breath Fingerprint of Colorectal Cancer Patients Based on the Gas Chromatography-Mass Spectrometry Analysis.

The human body emits a multitude of volatile organic compounds (VOCs) via tissues and various bodily fluids or exhaled breath. These compounds collectively create a distinctive chemical profile, which can potentially be employed to identify changes in human metabolism associated with colorectal cancer (CRC) and, consequently, facilitate the diagnosis of this disease. The main goal of this study was to investigate and characterize the VOCs' chemical patterns associated with the breath of CRC patients and controls and identify potential expiratory markers of this disease. For this purpose, gas chromatography-mass spectrometry was applied. Collectively, 1656 distinct compounds were identified in the breath samples provided by 152 subjects. Twenty-two statistically significant VOCs (p-xylene; hexanal; 2-methyl-1,3-dioxolane; 2,2,4-trimethyl-1,3-pentanediol diisobutyrate; hexadecane; nonane; ethylbenzene; cyclohexanone; diethyl phthalate; 6-methyl-5-hepten-2-one; tetrahydro-2H-pyran-2-one; 2-butanone; benzaldehyde; dodecanal; benzothiazole; tetradecane; 1-dodecanol; 1-benzene; 3-methylcyclopentyl acetate; 1-nonene; toluene) were observed at higher concentrations in the exhaled breath of the CRC group. The elevated levels of these VOCs in CRC patients' breath suggest the potential for these compounds to serve as biomarkers for CRC.

The Detection of Colorectal Cancer through Machine Learning-Based Breath Sensor Analysis.

Colorectal cancer (CRC) is the third most common malignancy and the second most common cause of cancer-related deaths worldwide. While CRC screening is already part of organized programs in many countries, there remains a need for improved screening tools. In recent years, a potential approach for cancer diagnosis has emerged via the analysis of volatile organic compounds (VOCs) using sensor technologies. The main goal of this study was to demonstrate and evaluate the diagnostic potential of a table-top breath analyzer for detecting CRC. Breath sampling was conducted and CRC vs. non-cancer groups (105 patients with CRC, 186 non-cancer subjects) were included in analysis. The obtained data were analyzed using supervised machine learning methods (i.e., Random Forest, C4.5, Artificial Neural Network, and Naïve Bayes). Superior accuracy was achieved using Random Forest and Evolutionary Search for Features (79.3%, sensitivity 53.3%, specificity 93.0%, AUC ROC 0.734), and Artificial Neural Networks and Greedy Search for Features (78.2%, sensitivity 43.3%, specificity 96.5%, AUC ROC 0.735). Our results confirm the potential of the developed breath analyzer as a promising tool for identifying and categorizing CRC within a point-of-care clinical context. The combination of MOX sensors provided promising results in distinguishing healthy vs. diseased breath samples. Its capacity for rapid, non-invasive, and targeted CRC detection suggests encouraging prospects for future clinical screening applications.

Identification of Volatile Markers of Colorectal Cancer from Tumor Tissues Using Volatilomic Approach.

The human body releases numerous volatile organic compounds (VOCs) through tissues and various body fluids, including breath. These compounds form a specific chemical profile that may be used to detect the colorectal cancer CRC-related changes in human metabolism and thereby diagnose this type of cancer. The main goal of this study was to investigate the volatile signatures formed by VOCs released from the CRC tissue. For this purpose, headspace solid-phase microextraction gas chromatography-mass spectrometry was applied. In total, 163 compounds were detected. Both cancerous and non-cancerous tissues emitted 138 common VOCs. Ten volatiles (2-butanone; dodecane; benzaldehyde; pyridine; octane; 2-pentanone; toluene; p-xylene; n-pentane; 2-methyl-2-propanol) occurred in at least 90% of both types of samples; 1-propanol in cancer tissue (86% in normal one), acetone in normal tissue (82% in cancer one). Four compounds (1-propanol, pyridine, isoprene, methyl thiolacetate) were found to have increased emissions from cancer tissue, whereas eleven showed reduced release from this type of tissue (2-butanone; 2-pentanone; 2-methyl-2-propanol; ethyl acetate; 3-methyl-1-butanol; d-limonene; tetradecane; dodecanal; tridecane; 2-ethyl-1-hexanol; cyclohexanone). The outcomes of this study provide evidence that the VOCs signature of the CRC tissue is altered by the CRC. The volatile constituents of this distinct signature can be emitted through exhalation and serve as potential biomarkers for identifying the presence of CRC. Reliable identification of the VOCs associated with CRC is essential to guide and tune the development of advanced sensor technologies that can effectively and sensitively detect and quantify these markers.

The Association of Circulating L-Carnitine, γ-Butyrobetaine and Trimethylamine N-Oxide Levels with Gastric Cancer.

Our study aimed to evaluate the association between gastric cancer (GC) and higher concentrations of the metabolites L-carnitine, γ-butyrobetaine (GBB) and gut microbiota-mediated trimethylamine N-oxide (TMAO) in the circulation. There is evidence suggesting that higher levels of TMAO and its precursors in blood can be indicative of either a higher risk of malignancy or indeed its presence; however, GC has not been studied in this regard until now. Our study included 83 controls without high-risk stomach lesions and 105 GC cases. Blood serum L-carnitine, GBB and TMAO levels were measured by ultra-high-performance liquid chromatography-mass spectrometry (UPLC/MS/MS). Although there were no significant differences between female control and GC groups, we found a significant difference in circulating levels of metabolites between the male control group and the male GC group, with median levels of L-carnitine reaching 30.22 (25.78-37.57) nmol/mL vs. 37.38 (32.73-42.61) nmol/mL (p < 0.001), GBB-0.79 (0.73-0.97) nmol/mL vs. 0.97 (0.78-1.16) nmol/mL (p < 0.05) and TMAO-2.49 (2.00-2.97) nmol/mL vs. 3.12 (2.08-5.83) nmol/mL (p < 0.05). Thus, our study demonstrated the association between higher blood levels of L-carnitine, GBB, TMAO and GC in males, but not in females. Furthermore, correlations of any two investigated metabolites were stronger in the GC groups of both genders in comparison to the control groups. Our findings reveal the potential role of L-carnitine, GBB and TMAO in GC and suggest metabolic differences between genders. In addition, the logistic regression analysis revealed that the only significant factor in terms of predicting whether the patient belonged to the control or to the GC group was the blood level of L-carnitine in males only. Hence, carnitine might be important as a biomarker or a risk factor for GC, especially in males.

Randomised clinical trial: comparison of efficacy and adverse effects of a standard triple clarithromycin-containing regimen with high-dose amoxicillin and bismuth therapy in Helicobacter pylori eradication.

BACKGROUND: The clarithromycin-based triple therapy is the most prescribed Helicobacter pylori eradication regimen in Europe; it causes adverse effects in a significant proportion of subjects, leading to discontinuation. Alternative therapies are required because of increasing clarithromycin resistance or to decrease the adverse effects. AIMS: We compared the efficacy and spectrum of adverse effects of clarithromycin-based triple therapy with the high-dose amoxicillin/bismuth regimen. METHODS: A randomised clinical trial enrolled healthy individuals aged 40-64 years. H. pylori was assessed with a 13C-urea breath test. In total 579 H. pylori-positive subjects were randomly allocated in two groups: group 1: clarithromycin 500 mg, amoxicillin 1000 mg, esomeprazole 40 mg, all twice daily; group 2: bismuth subcitrate 240 mg twice daily, amoxicillin 1000 mg three times daily, esomeprazole 40 mg twice daily. Regimens were administered for 14 days.Information on treatment completion and adverse effects were collected via a telephone interview at 21-28 days after medication delivery. The efficacy was assessed by UBT 6 months after the treatment. RESULTS: We analysed 483 subjects for adverse effects (248 vs. 235 respectively). Furthermore, 316 subjects were analysed for efficacy. In per-protocol analysis, a higher efficacy was seen in group 1 (88.4 vs. 77.0%; P < 0.001); no difference was observed in compliance (90.3 and 91.2%). Therapy-related adverse effects were more common in group 1 (56.9 vs. 40.0%; P < 0.01). In intention-to-treat analysis no statistical difference in efficacy was revealed. CONCLUSIONS: Bismuth-based high-dose amoxicillin therapy showed a lower efficacy but was less frequently associated with adverse effects. Further research is required to examine the high-dose amoxicillin and bismuth-containing regimens in various populations to maximise eradication efficacy.

Breath Sensor Technology for the Use in Mechanical Lung Ventilation Equipment for Monitoring Critically Ill Patients.

BACKGROUND: The need for mechanical lung ventilation is common in critically ill patients, either with COVID-19 infection or due to other causes. Monitoring of patients being ventilated is essential for timely and improved management. We here propose the use of a novel breath volatile organic compound sensor technology to be used in a mechanical lung ventilation machine for this purpose; the technology was evaluated in critically ill COVID-19 patients on mechanical lung ventilation. METHODS: Based on the consistency results of our study data, the breath sensor device with metal oxide gas sensors and environment-controlling sensors was mounted on the ventilation exhaust port of the ventilation machine; this allowed to ensure additional safety since the device was placed outside the contour between the patient and equipment. RESULTS: The sensors allowed stable registration of the signals for up to several weeks for 10 patients in total, depending on the storage amount; a proportion of patients were intubated or received tracheostoma during the evaluation period. Future studies are on the way to correlate sensor readings to other parameters characterizing the severity of the patient condition and outcome. CONCLUSIONS: We suppose that such technology will allow patient monitoring in real-time for timely identification of deterioration, potentially requiring some change of management. The obtained results are preliminary and further studies are needed to examine their clinical significance.

The activity of selective sigma-1 receptor ligands in seizure models in vivo.

Sigma-1 receptor (Sig1R) is a ligand-regulated protein which, since its discovery, has been widely studied as a novel target to treat neurological disorders, including seizures. However, the roles and mechanisms of Sig1R in the regulation of seizures are not fully understood. The aim of the present study was to test and compare effects of often used selective Sig1R ligands in models of experimentally induced seizures. The anti-seizure activities and interactions of selective Sig1R agonist PRE-084, selective Sig1R antagonist NE-100 and novel positive allosteric Sig1R modulator E1R were evaluated in pentylenetetrazol (PTZ) and (+)-bicuculline (BIC)-induced seizure models in mice. Sig1R antagonist NE-100 at a dose of 25mg/kg demonstrated pro-convulsive activity on PTZ-induced seizures. Agonist PRE-084 did not change the thresholds of chemoconvulsant-induced seizures. Positive allosteric modulator E1R at a dose of 50mg/kg showed anti-convulsive effects on PTZ- and BIC-induced clonic and tonic seizures. The anti-seizure activity of E1R was blocked by NE-100. Surprisingly, NE-100 at a dose of 50mg/kg induced convulsions, but E1R significantly alleviated the convulsive behaviour induced by NE-100. In conclusion, the selective Sig1R antagonist NE-100 induced seizures that could be partially attenuated by positive allosteric Sig1R modulator. Our results confirm that Sig1R could be a novel molecular target for new anti-convulsive drugs.

Selective inhibition of OCTN2 is more effective than inhibition of gamma-butyrobetaine dioxygenase to decrease the availability of l-carnitine and to reduce myocardial infarct size.

l-Carnitine is a cofactor in the energy metabolism pathways where it drives the uptake and oxidation of long chain fatty acids (LCFA) by mitochondria. LCFA lipotoxicity causes mitochondrial damage and results in an insufficient energy supply and a decrease in l-carnitine content limits LCFA flux and protects mitochondria. Here, we tested whether the inhibition of GBB dioxygenase (BBOX) or organic cation transporter 2 (OCTN2) is the most effective strategy to decrease l-carnitine content. The activity of 51 compounds was tested and we identified selective inhibitors of OCTN2. In contrast to selective inhibitors of BBOX, OCTN2 inhibitors induced a 10-fold decrease in l-carnitine content in the heart tissues and a significant 35% reduction of myocardial infarct size. In addition, OCTN2 inhibition correlated with the inhibitor content in the heart tissues, and OCTN2 could potentially be an efficient target to increase drug transport into tissues and to reduce drug elimination by urine. In conclusion, the results of this study confirm that selective inhibition of OCTN2, compared to selective inhibition of BBOX, is a far more effective approach to decrease l-carnitine content and to induce cardioprotective effects. OCTN2 could potentially be an efficient tool to increase drug transport in tissues and to reduce drug elimination via urine.

Effects of meldonium on sexual performance, sperm motility, testes morphology and blood biochemical markers in boars.

The objective of this study was to evaluate effects of long term (90-day) administration of meldonium [3-(2,2,2-trimethylhydrazinium) propionate] (mildronate, quaterin, MET-88) on sexual performance, sperm motility, testes morphology and biochemical blood markers in boars. Boars were treated with 2.0g of meldonium daily for 90 days. Administration of meldonium improved sexual performance and sperm motility. Thus, the reaction time (time from exposure to the dummy to the start of ejaculation) was reduced and the progressive motility of spermatozoa was significantly increased in the meldonium-treated boars compared to that of the boars of control group. In addition, the spermatogenic epithelium was thicker and proliferation of interstitial endocrine cells (Leydig cells) was observed in meldonium-treated boars. The concentration of blood serum testosterone was higher in the meldonium-treatment group than in the control group. Meldonium did not affect the concentration of creatinine, total bilirubin, total cholesterol, glucose and aspartate aminotransferase/AST, alanine aminotransferase/ALT activity in blood plasma. In conclusion, 90-day administration of meldonium improved sexual performance and sperm motility of boars and it also increased concentration of testosterone in blood serum. Further studies are necessary to substantiate the potential use of meldonium as a sperm motility and/or sperm quality-enhancing agent in livestock.

Expression of bioactive bone morphogenetic proteins in the subacromial bursa of patients with chronic degeneration of the rotator cuff.

Degeneration of the rotator cuff is often associated with inflammation of the subacromial bursa and focal mineralization of the supraspinatus tendon. Portions of the supraspinatus tendon distant from the insertion site could transform into fibrous cartilage, causing rotator-cuff tears owing to mechanical instability. Indirect evidence is presented to link this pathology to ectopic production and secretion of bioactive bone morphogenetic proteins (BMPs) from sites within the subacromial bursa. Surgically removed specimens of subacromial bursa tissue from patients with chronic tears of the rotator cuff were analyzed by immunohistochemistry and reverse transcription-PCR. Bioactive BMP was detected in bursa extracts by a bioassay based on induction of alkaline phosphatase in the osteogenic/myogenic cell line C2C12. Topical and differential expression of BMP-2/4 and BMP-7 mRNA and protein was found in bursa tissue. The bioassay of C2C12 cells revealed amounts of active BMP high enough to induce osteogenic cell types, and blocking BMP with specific antibodies or soluble BMP receptors Alk-3 and Alk-6 abolished the inductive properties of the extract. Sufficient information was gathered to explain how ectopic expression of BMP might induce tissue transformation into ectopic bone/cartilage and, therefore, promote structural degeneration of the rotator cuff. Early surgical removal of the subacromial bursa might present an option to interrupt disease progression.

Direct influence of mild hypothermia on cytokine expression and release in cultures of human peripheral blood mononuclear cells.

Hypothermia is associated with elevated frequency of infectious complications. Dysfunction of the immune response caused by hypothermia has been demonstrated in both clinical and animal studies, but it still remains unclear to what extent immunocompetent cells are directly influenced by hypothermia. To estimate the direct influence of mild hypothermia on cytokine expression and release by human peripheral blood mononuclear cells (PBMC), primary cultures of PBMC were incubated at 34 degrees C or 32 degrees C activated by lipopolysaccharide (LPS), phytohemagglutinin (PHA), or tumor necrosis factor-alpha (TNF-alpha). The cytokine gene expression was evaluated by RT-PCR. Release of interleukin-2 (IL-2), IL-6, IL-10, and TNF-alpha was measured by ELISA. Mild hyperthermia significantly impaired IL-2 gene expression in PHA-stimulated cultures of PBMC and decreased IL-2 release in all variants of cultures. Secretion of IL-6, IL-10, and TNF-alpha was decreased in hypothermic cultures of PBMC stimulated with the T lymphocyte activator PHA. Slight suppression of IL-10 secretion was observed also in TNF-alpha-stimulated hypothermic cultures of PBMC. TNF-alpha release increased slightly in mild hypothermia control cultures. Our data demonstrate that the direct influence of hypothermia on cytokine expression and release from PBMC is not uniform. Reduction of IL-2 production might play a crucial role in the impairment of immune response in hypothermia.