Serum Interleukins 8, 17, and 33 as Potential Biomarkers of Colon Cancer.

This research investigated the serum levels of three interleukins (IL8, IL17A, and IL33) and the possible relationships between them in healthy people and colon cancer patients at different stages. This study involved 82 participants, 42 of whom had colon cancer and 40 were healthy individuals. The cancer patients were classified into four groups according to the TNM staging classification of colon and rectal cancer. Serum levels of the interleukins were measured by the ELISA test. The data were analyzed statistically to compare the demographic characteristics, the interleukin levels across cancer stages, and the correlation between interleukins in both groups. The results showed that women had more early-stage colon cancer diagnoses, while men had more advanced-stage cancer diagnoses. Stage two colon cancer was more common in older people. Younger people, men, and those with early-stage colon cancer had higher levels of interleukins. The levels of IL8 and IL17A were higher in the cancer group, while the level of IL33 was higher in the healthy group. There was a strong correlation between IL8 and IL17A levels in both groups (p = 0.001). IL17A influenced the level of IL33 in the cancer group (p = 0.007). This study suggested that cytokine variation profiles could be useful for detecting colon cancer and predicting its outcome.

A Group of Tumor-Suppressive micro-RNAs Changes Expression Coordinately in Colon Cancer.

MicroRNAs (miRNAs) are molecules with a role in the post-transcriptional regulation of messenger RNA, being involved in a wide range of biological and pathological processes. In the present study, we aim to characterize the behavior of a few miRNAs with roles in the cell cycle and differentiation of colon cancer (CC) cells. The present work considers miRNAs as reflections of the complex cellular processes in which they are generated, their observed variations being used to characterize the molecular networks in which they are part and through which cell proliferation is achieved. Tumoral and adjacent normal tissue samples were obtained from 40 CC patients, and the expression of miR-29a, miR-146a, miR-215 and miR-449 were determined by qRT-PCR analysis. Subsequent bioinformatic analysis was performed to highlight the transcription factors (TFs) network that regulate the miRNAs and functionally characterizes this network. There was a significant decrease in the expression of all miRNAs in tumor tissue. All miRNAs were positively correlated with each other. The analysis of the TF network showed tightly connected functional modules related to the cell cycle and associated processes. The four miRNAs are downregulated in CC; they are strongly correlated, showing coherence within the cellular network that regulates them and highlighting possible approach strategies.

A role for serum cytokines and cell adhesion molecules in the non-invasive diagnosis of colorectal cancer.

Colorectal cancer (CRC) remains a major cause of cancer-related mortality. Consequently, new diagnostic and therapeutic approaches are being investigated including the serum levels of cytokines and other molecules, although the results are often inconclusive. Thus, the present study aimed to determine whether serum level of cytokines, cell adhesion molecules or matrix metalloproteinases (MMPs), alone or in combination, may contribute to the non-invasive diagnosis of CRC. The serum levels of nine cytokines [ILs; IL-1ß, IL-4, IL-6, IL-8, IL-10, IL-17, IL-22 and IL-33, and interferon (IFN)-γ], two cell adhesion molecules (intercellular adhesion molecule-1 and P-selectin) and an MMP-7 were measured by ELISA in 33 patients with CRC and 35 healthy controls. Combined capacity of all molecules to detect the presence of CRC was assessed by logistic regression. Molecules and molecule combinations were tested for all stages and tumor grades. A significant increase was identified for IL-8 in patients compared with healthy controls; IL-10 was found to be significantly decreased. The biomarker potential of each significantly modified molecule was tested: IL-8 had a sensitivity of 0.865, a specificity of 0.600 and an area under the curve (AUC) of 0.777; for IL-10, sensitivity was 0.65, specificity was 0.69, with an AUC of 0.689. Logistic regression determined the best discriminative potential between patients and control groups for the combination IL-4 + IL-6 + IL-8 + IFN-γ, with 0.97 sensitivity and 0.58 specificity. For the early stages of CRC, the combination IL-6 + IL-8 + IL-22 showed good performance. It was concluded that increased IL-8 had potential as single biomarker in CRC. Cytokine combinations are superior to single cytokine analysis in showing the presence of CRC.

COVID-19 and Pneumocystis jirovecii Pulmonary Coinfection-The First Case Confirmed through Autopsy.

Background: Establishing the diagnosis of COVID-19 and Pneumocystisjirovecii pulmonary coinfection is difficult due to clinical and radiological similarities that exist between the two disorders. For the moment, fungal coinfections are underestimated in COVID-19 patients. Case presentation: We report the case of a 52-year-old male patient, who presented to the emergency department for severe dyspnea and died 17 h later. The RT-PCR test performed at his admission was negative for SARS-CoV-2. Retesting of lung fragments collected during autopsy revealed a positive result for SARS-CoV-2. Histopathological examination showed preexisting lesions, due to comorbidities, as well as recent lesions: massive lung thromboses, alveolar exudate rich in foam cells, suprapleural and intra-alveolar Pneumocystisjirovecii cystic forms, and bilateral adrenal hemorrhage. Conclusion: COVID-19 and P.jirovecii coinfection should be considered, particularly in critically ill patients, and we recommend the systematic search for P. jirovecii in respiratory samples.

An overview of the tumor microenvironment, from cells to complex networks (Review).

For a long period, cancer has been believed to be a gene disease, in which oncogenic and suppressor mutations accumulate gradually, finally leading to the malignant transformation of cells. This vision has changed in the last few years, the involvement of the tumor microenvironment, the non-malignant part of the tumors, as an important contributor to the malignant growth being now largely recognized. There is a consensus according to which the understanding of the tumor microenvironment is important as a means to develop new approaches in the therapy of cancer. In this context, the present study is a review of the different types of non-malignant cells that can be found in tumors, with their pro or antitumoral actions, presence in tumors and therapeutic targeting. These cells establish complex relations between them, through cytokines, exosomes, cell adhesion, co-stimulation and co-inhibition; these relations will also be examined in the present work.

A Machine Learning Approach in Analyzing Bioaccumulation of Heavy Metals in Turbot Tissues.

Metals are considered to be one of the most hazardous substances due to their potential for accumulation, magnification, persistence, and wide distribution in water, sediments, and aquatic organisms. Demersal fish species, such as turbot (Psetta maxima maeotica), are accepted by the scientific communities as suitable bioindicators of heavy metal pollution in the aquatic environment. The present study uses a machine learning approach, which is based on multiple linear and non-linear models, in order to effectively estimate the concentrations of heavy metals in both turbot muscle and liver tissues. For multiple linear regression (MLR) models, the stepwise method was used, while non-linear models were developed by applying random forest (RF) algorithm. The models were based on data that were provided from scientific literature, attributed to 11 heavy metals (As, Ca, Cd, Cu, Fe, K, Mg, Mn, Na, Ni, Zn) from both muscle and liver tissues of turbot exemplars. Significant MLR models were recorded for Ca, Fe, Mg, and Na in muscle tissue and K, Cu, Zn, and Na in turbot liver tissue. The non-linear tree-based RF prediction models (over 70% prediction accuracy) were identified for As, Cd, Cu, K, Mg, and Zn in muscle tissue and As, Ca, Cd, Mg, and Fe in turbot liver tissue. Both machine learning MLR and non-linear tree-based RF prediction models were identified to be suitable for predicting the heavy metal concentration from both turbot muscle and liver tissues. The models can be used for improving the knowledge and economic efficiency of linked heavy metals food safety and environment pollution studies.

Combining cytogenetic and epigenetic approaches in chronic lymphocytic leukemia improves prognosis prediction for patients with isolated 13q deletion.

Background: Both defective DNA methylation and active DNA demethylation processes are emerging as important risk factors in chronic lymphocytic leukemia (CLL). However, associations between 5-cytosine epigenetic markers and the most frequent chromosomal abnormalities detected in CLL remain to be established. Methods: CLL patients were retrospectively classified into a cytogenetic low-risk group (isolated 13q deletion), an intermediate-risk group (normal karyotype or trisomy 12), and a high-risk group (11q deletion, 17p deletion, or complex karyotype [≥ 3 breakpoints]). The two 5-cytosine derivatives, 5-methylcytosine (5-mCyt) and 5-hydroxymethylcytosine (5-hmCyt), were tested by ELISA (n = 60), while real-time quantitative PCR was used for determining transcriptional expression levels of DNMT and TET (n = 24). Results: By using global DNA methylation/demethylation levels, in the low-risk disease group, two subgroups with significantly different clinical outcomes have been identified (median treatment-free survival [TFS] 45 versus > 120 months for 5-mCyt, p = 0.0008, and 63 versus > 120 months for 5-hmCyt, p = 0.04). A defective 5-mCyt status was further associated with a higher percentage of 13q deleted nuclei (> 80%), thus suggesting an acquired process. When considering the cytogenetic intermediate/high-risk disease groups, an association of 5-mCyt status with lymphocytosis (p = 0.0008) and the lymphocyte doubling time (p = 0.04) but not with TFS was observed, as well as a reduction of DNMT3A, TET1, and TET2 transcripts. Conclusions: Combining cytogenetic studies with 5-mCyt assessment adds accuracy to CLL patients' prognoses and particularly for those with 13q deletion as a sole cytogenetic abnormality.

Alterations in DNA methylation/demethylation intermediates predict clinical outcome in chronic lymphocytic leukemia.

Cytosine derivative dysregulations represent important epigenetic modifications whose impact on the clinical outcome in chronic lymphocytic leukemia (CLL) is incompletely understood. Hence, global levels of 5-methylcytosine (5-mCyt), 5-hydroxymethylcytosine (5-hmCyt), 5-carboxylcytosine (5-CaCyt) and 5-hydroxymethyluracil were tested in purified B cells from CLL patients (n = 55) and controls (n = 17). The DNA methylation 'writers' (DNA methyltransferases [DNMT1/3A/3B]), 'readers' (methyl-CpG-binding domain [MBD2/4]), 'editors' (ten-eleven translocation [TET1/2/3]) and 'modulators' (SAT1) were also evaluated. Accordingly, patients were stratified into three subgroups. First, a subgroup with a global deficit in cytosine derivatives characterized by hyperlymphocytosis, reduced median progression free survival (PFS = 52 months) and shorter treatment free survival (TFS = 112 months) was identified. In this subgroup, major epigenetic modifications were highlighted including a reduction of 5-mCyt, 5-hmCyt, 5-CaCyt associated with DNMT3A, MBD2/4 and TET1/2 downregulation. Second, the cytosine derivative analysis revealed a subgroup with a partial deficit (PFS = 84, TFS = 120 months), mainly affecting DNA demethylation (5-hmCyt reduction, SAT1 induction). Third, a subgroup epigenetically similar to controls was identified (PFS and TFS > 120 months). The prognostic impact of stratifying CLL patients within three epigenetic subgroups was confirmed in a validation cohort. In conclusion, our results suggest that dysregulations of cytosine derivative regulators represent major events acquired during CLL progression and are independent from IGHV mutational status.

Anti-CD20 monoclonal antibodies in chronic lymphocytic leukemia: from uncertainties to promises.

Over the last two decades, anti-CD20 monoclonal antibody (mAb) therapy has improved patient outcome in B-cell malignancies, and confirmed CD20 as an important target in chronic lymphocytic leukemia (CLL). Until recently, the gold standard was based on the utilization of rituximab combined with chemotherapy (fludarabine and cyclophosphamide), but patients often relapse. Next, with our better understanding of mAb engineering, anti-CD20 mAb therapy has evolved with the development of new mAb permitting significant clinical responses by improving pharmacokinetics, safety, activity and immunogenicity. Last but not least, the development of key tumoral tyrosine kinase inhibitors and their association with anti-CD20 mAb is a work in progress with promising results.

Total Intravenous Versus Inhalation Anesthesia in Patients Undergoing Laparoscopic Cholecystectomies. Effects on Two Proinflammatory Cytokines Serum Levels: Il-32 and TNF-Alfa.

INTRODUCTION: It has been reported that as compared with total intravenous anesthesia (TIVA), inhalation anesthesia is increasing the postoperative level of proinflammatory interleukins.The aim of the study is to investigate if there is an in-vivo relationship between proinflammatory cytokines, Interleukin-32 (IL-32) and Tumour necrosis factor - α (TNF-α), in patients undergoing laparoscopic cholecystectomies with two different anesthetic techniques, TIVA or inhalation anesthesia. MATERIAL AND METHODS: Twenty two consecutive patients undergoing laparoscopic cholecystectomies were prospectively randomized into two groups: Group 1: TIVA with target-controlled infusion (TIVA-TCI) (n=11) and Group 2: isoflurane anesthesia (ISO) (n=11). IL-32 and TNF-α were determined before the induction of anesthesia (T1), before incision (T2) and at 2h (T3) and 24h (T4) postoperatively. Our primary outcome was to compare plasma levels of IL-32 and TNF-α concentrations (expressed as area-under-the-curve) over 24 hours between study groups. Our secondary outcome was to establish whether there is a correlation between plasma levels of IL-32 and of TNF-α at each time point between the two groups. RESULTS: Area-under-the-curve (AUC) of IL-32 plasma concentration was 7.53 in Group 1 (TIVA) versus 3.80 in Group 2 (ISO), p= 1. For TNF-α, AUC of plasma concentration was 733.9 in Group 1 (TIVA) and 668.7 in Group 2 (ISO), p=0.066. There were no significant differences in plasma concentrations of both IL-32 and TNF-α between the groups. CONCLUSIONS: IL-32 expression in response to minor surgery is very low. There were no significant difference between plasma levels of TNF-α and IL-32 after TIVA versus inhalation anesthesia during the first 24 hours postoperatively. Further studies are needed on larger groups to investigate whether there can be a correlation between these interleukins after 2 different anesthetic techniques and the impact of this correlation on postoperative outcome.

Phytochemical Screening: Antioxidant and Antibacterial Properties of Potamogeton Species in Order to Obtain Valuable Feed Additives.

The alcoholic extracts from three submerged perennial plants Potamogeton crispus L., P. pusillus L. and P. pectinatus L. were analyzed by gas chromatography-mass spectrometry coupled with solid phase microextraction (SPME-GC/MS) and by High Performance Liquid Chromatography (HPLC) and their volatile fingerprint and polyphenols composition was mutually compared. Twenty-nine chemical compounds were detected and identified in ethanolic and methanolic extracts; the highest abundance (over 5%) in descending order, was detected for 9,9-dimethyl-8,10- dioxapentacyclo (5,3,0(2,5) 0(3,5,)0 (3,6) decane (21.65%), phenol 2,6 bis (1,1 dimethyletyl) 4-1-methylpropil (20.8%), pentadecanoic acid (14.3%), 2-(5-chloro-2-Methoxyphenyl) pyrrole (8.66%), propanedioic (malonic) acid 2-(4-methylphenyl) sulfonyl ethylidene (5.77%), 2 hydroxy-3 tert butyl-5-isopropyl-6 methyl phenyl ketone (5.76%). The highest total polyphenols and flavonoids content was found in the methanolic extract of P. crispus (112.5±0.5 mg tannic acid/g dry extract; 64.2±1.2 mg quercitin/g dry extract). Antioxidant activities (2,2-difenil-1-picrilhidrazil, hydrogen peroxide and reducing power assays) of obtained extracts are comparable with the standard compounds, butylated hydroxytoluene, rutin and ascorbic acid. Antibacterial efficiency of methanolic extracts was notably demonstrated against Gram negative (Escherichia coli, Enterobacter hormaechei) and Gram positive bacteria (Enterococcus casseliflavus). The data reported for the first time for Romanian Potamogeton species, provides extensive support for the chemical investigations of these plants of the aquatic anthropogene ecosystems in order to obtain valuable bioadditives for animal feed and/or pharmaceutical/food industry.

IgG antibodies against immunodominant C-terminal epitopes of BP230 do not induce skin blistering in mice.

Bullous pemphigoid, the most common autoimmune blistering disease in Western Europe and the USA is characterized by the presence of circulating and tissue-bound autoantibodies against the hemidesmosomal proteins BP230 and BP180/collagen XVII. After binding to their target antigens at the basement membrane of the dermal-epidermal junction these autoantibodies are thought to trigger an inflammatory cascade comprising complement- and granulocyte-dependent reactions that result in tissue damage. Whereas the role of anti-BP180 antibodies has been extensively characterized, few and conflicting data is available on the contribution of anti-BP230 antibodies to bullous pemphigoid pathogenesis. Therefore, we addressed in the present study the role of autoantibodies to BP230 in experimental bullous pemphigoid. Rabbit polyclonal antibodies generated against epitopes of the C-terminal fragment of murine BP230 bound to the basement membrane and activated the complement system ex vivo. Affinity-purified antibodies were subsequently subcutaneously transferred into neonatal and adult BALB/c mice. In vivo, we observed a dose-dependent binding of transferred antibodies in the murine skin; however, there was no complement activation and these mice showed no clinical or histological signs of inflammatory disease, in contrast to mice receiving anti-BP180 antibodies. We further conducted ex vivo experiments and demonstrated that rabbit IgG anti-BP230-specific antibodies, in contrast to antibodies from bullous pemphigoid patients or rabbit IgG anti-BP180 antibodies used as positive controls, did not activate human granulocytes to induce dermal-epidermal separation in skin cryosections. Our present findings demonstrate that antibodies against BP230 are non-pathogenic in experimental models of bullous pemphigoid and suggest that proper activation of the complement and granulocytes represent prerequisites for conferring bullous pemphigoid autoantibodies their tissue destructive potential.

The role of Staphylococcal enterotoxin A in achieving oral tolerance to myelin basic protein in adult mice.

BACKGROUND: Oral tolerance is the biological process explaining the non-responsiveness of gut lymphoid tissue to intestinal content. Our study tested a new approach for the enhancement of oral tolerance to a multiple sclerosis-triggering auto-antigen-myelin basic protein, by its oral administration with the Staphylococcal enterotoxin A. METHODS: Immune tolerance thus stimulated was assessed in adult BALB/c mice, by measuring different cytokines from the supernatant of mesenteric lymph nodes cells (IFN-γ, IL-4, IL-10, IL-17, and TGF-ß), and in a SJL/E mouse model of experimental autoimmune encephalomyelitis, by evaluating the development of regulatory T cells in mesenteric lymph nodes and the clinical outcome of the intervention. RESULTS: We obtained a significant rise in the levels of IL-10 and TGF-ß compared with control and a significant decrease of IFN-γ, IL-4 (p < 0.05). Regulatory T cells were increased compared with control (p < 0.05). These results were attributable both to myelin basic protein and to Staphylococcal enterotoxin A. The clinical outcome of experimental autoimmune encephalomyelitis was influenced only by the administration of myelin basic protein. CONCLUSION: In our experiment, Staphylococcal enterotoxin A enhanced the immune tolerance to myelin basic protein in the gut mucosa, but had no impact on the clinical evolution of experimental autoimmune encephalomyelitis.

Magnesium and selenium in diabetics with peripheral artery disease of the lower limbs.

BACKGROUND AND AIMS: Knowing that diabetes mellitus (DM) often leads to cardiovascular injury such as peripheral artery disease (PAD) of the lower limbs and that some microelements like magnesium (Mg) and Selenium (Se) might be involved in the alterations entailed by these complications, we aimed to investigate the behavior of those microelements in these diseases. METHODS: We studied 114 patients with type 2 DM, treated not with insulin but only by diet or oral antidiabetics, mean aged 56.6 having a mean duration of the diabetes of 14.5 years, mostly men (80), 64 of them (40 men) having also PAD stage 2 without severe complications. We had a control group of 40 similarly-aged subjects without DM or PAD. The measurements were performed on a Konelab 30I device and serum selenium was quantified by atomic absorption spectroscopy. All patients underwent clinical, paraclinical and arterial Doppler echography for establishing the diagnosis of PAD. RESULTS: The results for controls were 2.2±0.4 mg/dl for Mg, 130±6 µg/dl for Se; diabetics had 1.68±0.4 mg/dl Mg, 88±6 µg/dl Se; diabetics with PAD had significantly lower levels: 1.36±0.6 mg/dl Mg, 66±6 µg/dl Se (p<0.05). Age inversely correlated with the values of both microelements especially in women. Mixed dysipidemia, present in DM, correlated with PAD. Blood pressure also inversely correlated with Mg and Se. Coronary heart disease (ischemia) linked to lower blood values of Mg and Se. Left Ventricle Hypertrophy (as proved by echography) also correlated to lower blood values of Mg and Se, as did glycated hemoglobin (HbA1c). CONCLUSION: We may conclude that in DM there is a Mg deficit which becomes significant in DZ+PAD especially in forms with high blood pressure (HBP), severe dyslipidemia and in the presence of early microvascular complications, our paper suggesting the opportunity of a substitutive treatment with Mg and Se in selected cases.

Novel cellular and molecular approaches to stratification and treatment of colorectal cancer.

Colorectal cancer is an important disease of the modern world, generating significant mortality and morbidity rates. Its therapy, although considerably improved, continues to be unhelpful for a large percentage of patients, especially for those in advanced stages. This justifies the efforts toward producing new therapies, as well as for stratifying patients according to risk factors and prediction of therapeutic response. In this respect, the contributions of modern science are essential for defining the most intimate mechanisms and players of tumorigenesis and for proposing new biomarkers. The study of antitumor immune responses has revealed new insights into the tumor microenvironment, leading to the development of vaccines and adoptive transfer of immunocompetent cells. Circulating tumor cells are a real opportunity to detect early relapses and to define risk categories, while miRNAs, a family of small non-coding RNA implicated in regulation of gene expression, evolved as a new class of biomarkers with high potential for diagnosis, prognosis and prediction of colorectal cancers.

Gold nanoparticles conjugated with cisplatin/doxorubicin/capecitabine lower the chemoresistance of hepatocellular carcinoma-derived cancer cells.

BACKGROUND AND AIMS: The aim of the current study was to evaluate in vitro the anti-tumor efficacy of gold nanoparticles (GNPs) conjugated with conventional chemotherapy drugs for the treatment of liver cancer. This approach based on gold proposes a novel platform therapy with minimal toxicity and increased efficacy profiles for the destruction of hepatic cancer cells. METHODS: GNPs, stabilized with a monolayer of L-aspartate and additional cytostatic drugs, were successfully used as a complex tumor-targeting drug-delivery system. The drugs (doxorubicin, cisplatin, and capecitabine) were non-covalently conjugated onto the hydrophilic assemblies of GNPs-L-Aspartate nanostructure. Transmission electron microscopy was used to characterize the morphological and structural properties of these drug-metallic nanostructures. RESULTS: The cellular proliferation rates in the presence of the anti-cancer drugs delivered by the GNPs were found to be statistically lower than those of cells exposed to the cytostatic drugs alone, indicating that GNPs facilitated an increased susceptibility of cancer cells to cisplatin, doxorubicin, and capecitabine plus ribavirin. CONCLUSION: This approach could offer a new chemotherapy strategy for patients diagnosed with unresectable hepatocellular carcinoma (HCC).