Beta-3 adrenergic receptor blockade prevents alterations in feeding behavior in lymphoma-bearing mice in a sex-dependent manner

Introduction: Cancer cachexia is a metabolic syndrome defined by an ongoing loss of adipose and muscular tissue and is characterized by a reduced food intake. Local b-3 adrenergic receptor activation in adipose tissue induces lipolysis, whereas b-3 signaling in tumors has been shown to be tumorigenic in some cancer models. Therefore, b-3 signaling may be a therapeutic target in cancer cachexia. The aim of this study was to assess the role of b-3 adrenergic signaling in feeding behavior, body composition, and tumor progression in the L5178Y-R murine lymphoma model. Methods: In our study we used BALB/c mice of both sexes, which were divided in tumor-free and tumor-bearing groups. For the tumor model, L5178Y-R lymphoma cells were subcutaneously administered into animals right flank. These groups of mice intraperitoneally received L-748,337, a beta-3 antagonist, at a 50 mg/kg/day dose, starting the day after tumor implantation. Food and water intake were monitored every other day and body mass index (BMI) was calculated at the end of the experiment. Animals were euthanized for necropsy, when endpoint criteria were achieved. Transcriptional expression of Ucp-1, a molecular marker of thermogenesis, was quantified in interscapular adipose tissue. Results: We observed a 15% and 35% reduction in food intake in tumor-bearing male and female mice, respectively. This effect was not observed in male mice treated with the b-3 adrenergic receptor antagonist L-748,337. In females, such an effect persisted despite beta-3 blocking. Reduced water intake was also observed in tumor-bearing animals, which was not altered by beta-3 antagonism. We also observed that tumor-free mice of both sexes showed reduced water intake after L-748,337 treatment. Furthermore, reduced BMI was observed in tumor-bearing animals of both sexes, which was not changed by b-3 blocking. Interscapular adipose tissue loss was observed in females (51.06%) but not in males. Additionally, 1.7-fold and 4.4-fold reduction in Ucp-1 gene expression was shown in tumor-bearing males and females, respectively. Decreased final tumor weight was observed only in tumor-bearing females treated with L-748,337 (p \u3c 0.05). Conclusion: In L5178Y-R tumor-bearing BALB/c mice, selective blocking of beta-3 adrenergic signaling prevents alterations in food intake in a sex-dependent manner

Development and evaluation of a nanofiber membrane in vitro as a therapeutic alternative for the post treatment in breast cancer cell in a murine model

Worldwide female breast cancer is the most commonly diagnosed cancer, with an estimated 2.3 million new cases in 2020, reason for the need of targeted therapies that can maximize treatment success and minimize toxicity. Nanoparticles of gold (AuNps) exhibit cytotoxic properties against certain types of cancer cell lines. Nanofibers have been use in the drug delivery systems due to its degradability and high surface area. We proposed a membrane with nanometric fibers using polivinilic alcohol (PVA) and chitosan (Qts) loaded with AuNps and Doxorubicin (Doxo) with the purpose of diminish tumor regression. PVA-Qts membrane was develop with electrospinning, the injection, voltage, distance and relative humidity parameters were standardized and it were characterized by Microscopic Atomic Force. The cytotoxicity with a median lethal dose (DL50) in two cell lines, breast adenocarcinoma murine (4T1) and murine fibroblast (NiH3T3) as a healthy control were evaluated. AuNps had a size of 3 nanometers (nm) with a Z potential of 13.2 mVolts and a DL50 of 75 µM in the cell line 4T1. Doxo was decrease in 95% with a final concentration of 0.03 mg/cm2. Both doses were loaded in the PVA-Qts solution. PVA-Qts-Doxo and PVA-Qts-AuNps-Doxo decrease the viability in 4T1 in 24 hours with a 15%, 72 hours with a 28%, the first with 60% and the latter with 82%. PVA-Qts-Doxo and PVA-Qts-AuNps-Doxo in NiH3T3 diminish incrementing with the time reaching a 40% in 120 hours. Finally, The viability for 4T1 cultured on PVA-Qts-Doxo was minor than in NiH3T3. The amount of Doxo in the membrane synthetized was 95% less than the employ doses, demonstrating that the fiber improves the delivery of the chemotherapeutic in a palatine time

Secretion of biologically active interferon-gamma inducible protein-10 (IP-10) by Lactococcus lactis

<p>Abstract</p> <p>Background</p> <p>Chemokines are a large group of chemotactic cytokines that regulate and direct migration of leukocytes, activate inflammatory responses, and are involved in many other functions including regulation of tumor development. Interferon-gamma inducible-protein-10 (IP-10) is a member of the C-X-C subfamily of the chemokine family of cytokines. IP-10 specifically chemoattracts activated T lymphocytes, monocytes, and NK cells. IP-10 has been described also as a modulator of other antitumor cytokines. These properties make IP-10 a novel therapeutic molecule for the treatment of chronic and infectious diseases. Currently there are no suitable live biological systems to produce and secrete IP-10. <it>Lactococcus lactis </it>has been well-characterized over the years as a safe microorganism to produce heterologous proteins and to be used as a safe, live vaccine to deliver antigens and cytokines of interest. Here we report a recombinant strain of <it>L. lactis </it>genetically modified to produce and secrete biologically active IP-10.</p> <p>Results</p> <p>The IP-10 coding region was isolated from human cDNA and cloned into an <it>L. lactis </it>expression plasmid under the regulation of the pNis promoter. By fusion to the usp45 secretion signal, IP-10 was addressed out of the cell. Western blot analysis demonstrated that recombinant strains of <it>L. lactis </it>secrete IP-10 into the culture medium. Neither degradation nor incomplete forms of IP-10 were detected in the cell or supernatant fractions of <it>L. lactis</it>. In addition, we demonstrated that the NICE (nisin-controlled gene expression) system was able to express IP-10 "de novo" even two hours after nisin removal. This human IP-10 protein secreted by <it>L. lactis </it>was biological active as demonstrated by Chemotaxis assay over human CD3+T lymphocytes.</p> <p>Conclusion</p> <p>Expression and secretion of mature IP-10 was efficiently achieved by <it>L. lactis </it>forming an effective system to produce IP-10. This recombinant IP-10 is biologically active as demonstrated by its ability to chemoattract human CD3+ T lymphocytes. This strain of recombinant <it>L. lactis </it>represents a potentially useful tool to be used as a live vaccine <it>in vivo</it>.</p

Mortality from gastrointestinal congenital anomalies at 264 hospitals in 74 low-income, middle-income, and high-income countries: a multicentre, international, prospective cohort study

Summary Background Congenital anomalies are the fifth leading cause of mortality in children younger than 5 years globally. Many gastrointestinal congenital anomalies are fatal without timely access to neonatal surgical care, but few studies have been done on these conditions in low-income and middle-income countries (LMICs). We compared outcomes of the seven most common gastrointestinal congenital anomalies in low-income, middle-income, and high-income countries globally, and identified factors associated with mortality. Methods We did a multicentre, international prospective cohort study of patients younger than 16 years, presenting to hospital for the first time with oesophageal atresia, congenital diaphragmatic hernia, intestinal atresia, gastroschisis, exomphalos, anorectal malformation, and Hirschsprung’s disease. Recruitment was of consecutive patients for a minimum of 1 month between October, 2018, and April, 2019. We collected data on patient demographics, clinical status, interventions, and outcomes using the REDCap platform. Patients were followed up for 30 days after primary intervention, or 30 days after admission if they did not receive an intervention. The primary outcome was all-cause, in-hospital mortality for all conditions combined and each condition individually, stratified by country income status. We did a complete case analysis. Findings We included 3849 patients with 3975 study conditions (560 with oesophageal atresia, 448 with congenital diaphragmatic hernia, 681 with intestinal atresia, 453 with gastroschisis, 325 with exomphalos, 991 with anorectal malformation, and 517 with Hirschsprung’s disease) from 264 hospitals (89 in high-income countries, 166 in middleincome countries, and nine in low-income countries) in 74 countries. Of the 3849 patients, 2231 (58·0%) were male. Median gestational age at birth was 38 weeks (IQR 36–39) and median bodyweight at presentation was 2·8 kg (2·3–3·3). Mortality among all patients was 37 (39·8%) of 93 in low-income countries, 583 (20·4%) of 2860 in middle-income countries, and 50 (5·6%) of 896 in high-income countries (p<0·0001 between all country income groups). Gastroschisis had the greatest difference in mortality between country income strata (nine [90·0%] of ten in lowincome countries, 97 [31·9%] of 304 in middle-income countries, and two [1·4%] of 139 in high-income countries; p≤0·0001 between all country income groups). Factors significantly associated with higher mortality for all patients combined included country income status (low-income vs high-income countries, risk ratio 2·78 [95% CI 1·88–4·11], p<0·0001; middle-income vs high-income countries, 2·11 [1·59–2·79], p<0·0001), sepsis at presentation (1·20 [1·04–1·40], p=0·016), higher American Society of Anesthesiologists (ASA) score at primary intervention (ASA 4–5 vs ASA 1–2, 1·82 [1·40–2·35], p<0·0001; ASA 3 vs ASA 1–2, 1·58, [1·30–1·92], p<0·0001]), surgical safety checklist not used (1·39 [1·02–1·90], p=0·035), and ventilation or parenteral nutrition unavailable when needed (ventilation 1·96, [1·41–2·71], p=0·0001; parenteral nutrition 1·35, [1·05–1·74], p=0·018). Administration of parenteral nutrition (0·61, [0·47–0·79], p=0·0002) and use of a peripherally inserted central catheter (0·65 [0·50–0·86], p=0·0024) or percutaneous central line (0·69 [0·48–1·00], p=0·049) were associated with lower mortality. Interpretation Unacceptable differences in mortality exist for gastrointestinal congenital anomalies between lowincome, middle-income, and high-income countries. Improving access to quality neonatal surgical care in LMICs will be vital to achieve Sustainable Development Goal 3.2 of ending preventable deaths in neonates and children younger than 5 years by 2030

Additional file 1: Figure S1. of IMMUNEPOTENT CRP induces cell cycle arrest and caspase-independent regulated cell death in HeLa cells through reactive oxygen species production

(A) ROS levels were measured by flow cytometry through DCFDA staining in SiHa cells left alone or pretreated with NAC or QVD.oph and then treated with I-CRP (1.25 U/mL) for 24 h. (B) The effect on cell death of cells left alone or pretreated with NAC or QVD.oph and then treated with I-CRP (1.25 U/mL) for 24 h, was analyzed by flow cytometry through Annexin-V staining. The results were analyzed and graphed. (PDF 20 kb

Additional file 2: Figure S2. of IMMUNEPOTENT CRP induces cell cycle arrest and caspase-independent regulated cell death in HeLa cells through reactive oxygen species production

Left, caspase-3 activity of HeLa cells left untreated or pretreated with Nac, and then treated with I-CRP. Right, the results obtained were analyzed and graphed as the percentage of HeLa cells positive for caspase-3 activity. (PDF 37 kb

Clinical trial evaluating the effectiveness of biocompound IMMUNEPOTENT CRP in the third-molar extraction

A controlled, parallel, randomized and comparative trial was carried out to evaluate the anti-inflammatory efficacy of IMMUNEPOTENT CRP versus ibuprofen in patients after third-molar surgery over seven days. The anti-inflammatory efficacy of IMMUNEPOTENT CRP was evaluated using the method of Amin and Laskin, and the analysis of cytokine production (IL-2, IL-4, IL-6, IL-10, TNF-α, INF-γ) in saliva was done by flow cytometry. The swelling process after surgery was significant (p < 0.05) and the treatments with IMMUNEPOTENT CRP or ibuprofen controlled this process properly; no difference between the groups was found (p < 0.05). Both treatments were shown to modulate the cytokine production. These results demonstrate the anti-inflammatory activity of the natural compound IMMUNEPOTENT CRP and suggest it could be used in clinical dental practice

Increase of the antitumour efficacy of the biocompound IMMUNEPOTENT CRP by enzymatic treatment

IMMUNEPOTENT CRP is a dialyzable leukocyte extract obtained from bovine spleen with immunomodulatory and antitumour properties; therefore, when administrated as an adjuvant therapy for cancer patients, it has increased their survival and quality of life. The bioavailability of any orally administered compound can be reduced due to gastrointestinal enzymes. In this study, we evaluated if IMMUNEPOTENT CRP is resistant to the treatment with different enzymes (proteases, nucleases, polysaccharide-degrading enzymes or lipase), using as parameters for biological activity measurement its in vitro antitumour and antioxidant properties and in vivo the antitumour effect of IMMUNEPOTENT CRP treated with proteinase K. In conclusion, we consider necessary to include the antioxidant and cytotoxic activity on the MCF-7 cancer cell line as parameters for the quantitative determination of biological activity or potency tests for batch release. Additionally, the results showed that different enzymatic treatments do not affect the antitumour and antioxidant activities of IMMUNEPOTENT CRP in vitro, suggesting that this product can be administered orally without any loss of biological activity. Furthermore, IMMUNEPOTENT CRP treatment with proteinase K increases the antitumour activity in vivo