Chimeric Peptides from Californiconus californicus and Heterodontus francisci with Antigen-Binding Capacity: A Conotoxin Scaffold to Create Non-Natural Antibodies (NoNaBodies).

Research into various proteins capable of blocking metabolic pathways has improved the detection and treatment of multiple pathologies associated with the malfunction and overexpression of different metabolites. However, antigen-binding proteins have limitations. To overcome the disadvantages of the available antigen-binding proteins, the present investigation aims to provide chimeric antigen-binding peptides by binding a complementarity-determining region 3 (CDR3) of variable domains of new antigen receptors (VNARs) with a conotoxin. Six non-natural antibodies (NoNaBodies) were obtained from the complexes of conotoxin cal14.1a with six CDR3s from the VNARs of Heterodontus francisci and two NoNaBodies from the VNARs of other shark species. The peptides cal_P98Y vs. vascular endothelial growth factor 165 (VEGF165), cal_T10 vs. transforming growth factor beta (TGF-ß), and cal_CV043 vs. carcinoembryonic antigen (CEA) showed in-silico and in vitro recognition capacity. Likewise, cal_P98Y and cal_CV043 demonstrated the capacity to neutralize the antigens for which they were designed.

Ten-Week Sucralose Consumption Induces Gut Dysbiosis and Altered Glucose and Insulin Levels in Healthy Young Adults.

Sucralose consumption alters microbiome and carbohydrate metabolism in mouse models. However, there are no conclusive studies in humans. Our goals were to examine the effect of sucralose consumption on the intestinal abundance of bacterial species belonging to Actinobacteria, Bacteroidetes, and Firmicutes and explore potential associations between microbiome profiles and glucose and insulin blood levels in healthy young adults. In this open-label clinical trial, volunteers randomly drank water, as a control (n = 20), or 48 mg sucralose (n = 20), every day for ten weeks. At the beginning and the end of the study, participants were subjected to an oral glucose tolerance test (OGTT) to measure serum glucose and insulin every 15 min for 3 h and provided fecal samples to assess gut microbiota using a quantitative polymerase chain reaction. Sucralose intake altered the abundance of Firmicutes without affecting Actinobacteria or Bacteroidetes. Two-way ANOVA revealed that volunteers drinking sucralose for ten weeks showed a 3-fold increase in Blautia coccoides and a 0.66-fold decrease in Lactobacillus acidophilus compared to the controls. Sucralose consumption increased serum insulin and the area under the glucose curve compared to water. Long-term sucralose ingestion induces gut dysbiosis associated with altered insulin and glucose levels during an OGTT.

The Combined Use of Cytokine Serum Values with Laboratory Parameters Improves Mortality Prediction of COVID-19 Patients: The Interleukin-15-to-Albumin Ratio.

Laboratory parameters display limited accuracy in predicting mortality in coronavirus disease 2019 (COVID-19) patients, as with serum albumin. Emerging evidence suggests that cytokine serum values may enhance the predictive capacity of albumin, especially interleukin (IL)-15. We thus investigated whether the use of the IL-15-to-albumin ratio enables improving mortality prediction at hospital admission in a large group of COVID-19 patients. In this prospective cross-sectional study, we enrolled and followed up three hundred and seventy-eight patients with a COVID-19 diagnosis until hospital discharge or death. Two hundred and fifty-five patients survived, whereas one hundred and twenty-three died. Student's T-test revealed that non-survivors had a significant two-fold increase in the IL-15-to-albumin ratio compared to survivors (167.3 ± 63.8 versus 74.2 ± 28.5), a difference that was more evident than that found for IL-15 or albumin separately. Likewise, mortality prediction considerably improved when using the IL-15-to-albumin ratio with a cut-off point > 105.4, exhibiting an area under the receiver operating characteristic curve of 0.841 (95% Confidence Interval, 0.725-0.922, p < 0.001). As we outlined here, this is the first study showing that combining IL-15 serum values with albumin improves mortality prediction in COVID-19 patients.

Use of Nasoil® via intranasal to control the harmful effects of Covid-19.

In the absence of vaccines and antiviral drugs available to prevent and treat COVID-19, it becomes imperative to find or use all those products with the potential to fight this virus. This article is an attempt to propose ways to prevent, treat and control the COVID-19 virus, using a product based on plant extracts with the potential to reduce the symptoms caused by the SARS-CoV-2 virus. Nasoil® counts as one of its main components, Asclepias curassavica extracts, and in the present study it has been shown that it is an effective adjuvant in the treatment of Covid-19, increasing the respiratory capacity of the patients (SpO2> 90%) and reducing the symptoms from the first application, improving the patients around the fifth to the eighth application. At a preventive level, the individuals in this study who have applied it (400 individuals) only a 3.15% of these presented symptoms, disappearing when increasing the weekly applications.

Marine peptides as immunomodulators: Californiconus californicus-derived synthetic conotoxins induce IL-10 production by regulatory T cells (CD4+Foxp3+).

Context: CD4+ T lymphocytes are able to differentiate into distinct subtypes according to several immunological scenarios, including T helper (Th)1, Th2, Th17 and regulatory T (Treg) cells. CD4+ T cells are phenotypically flexible and have specific ion channels, such as the nicotinic acetylcholine receptors (nAChR) that could be modulated by peptides produced by marine snails, known as conotoxins. Their effect on T lymphocytes has not been explored and emerging evidence suggests that these peptides may have immunomodulatory activities. Objective: This study investigated the effect of two Californiconus californicus-derived synthetic conotoxins on the proliferation and differentiation of T lymphocyte subpopulations Th1, Th2, Th17 and Treg. Methods: Cells from lymph nodes of BALB/c mice were cultured in the presence of conotoxins cal14.1b and cal14.2c (5.5 µM), during 96 h. Cell proliferation and intracellular cytokine production (IFN-γ, IL-4, IL-17 and IL-10) were analyzed by flow cytometry. Results and Discussion: cal14.1b and cal14.2c increased intracellular IL-10 production in Treg (CD3+CD4+Foxp3+) cells and decreased intracellular IL-17 production (CD3+CD4+) after 72 h of culture. Conotoxins did not show any effect on T cell proliferation nor Th1/Th2 balance. Conclusion: These results suggest that synthetic conotoxins exert immunomodulatory activity, especially by regulating specific functions on T lymphocytes.

1 H-NMR Based Metabolomics Profiling of Citrus Juices Produced in Veracruz, México.

This study describes the 1 H-NMR-based metabolomics profiling of juices from citrus fruits harvested in the state of Veracruz, México. The hydrophilic profile of commercial lemons (Agrio and Persian), tangerines (Fremont and Mónica), oranges (Valencia and Washington Navel), and grapefruits (Red Ruby and Rio Red) was determined. According to our results, 35 metabolites were identified in the 1 H-NMR profiling. The statistical differences obtained by PCA and OPLS-DA revealed that specific amino acids, sugars, and organic acids were differential metabolites in the species and cultivars studied. High endogenous levels of sucrose (10-190 mM), α-glucose, ß-glucose (α- and ß-isomers, 40-205 mm), and fructose (36-170 mm) were detected in the juices of grapefruits, oranges, and tangerines, whereas citric acid (40-530 mm) was the principal organic acid in the juices of lemons. To calculate the specific amounts of metabolites from these species and their cultivars, the results were finely analyzed using the qNMR method. According to these calculations, Valencia oranges had the highest concentration of ascorbic acid (>2 mm). The described 1 H-NMR method is highly reproducible, inexpensive, and highly robust in comparison to other analytical methods used to determine the hydrophilic profile of citrus juices.

Semi-Automatic Elemental Identification of Laser-Induced Breakdown Spectra Using Wavelength Similarity Coefficient.

This work proposes a method to perform elemental identification on plasmas produced using the laser-induced breakdown spectroscopy (LIBS) technique. The method is based on the preservation of the relative relevance of the spectral line emission intensities, which is lost during the parametric correlation procedure, by the introduction of a similitude coefficient called wavelength similarity coefficient. Furthermore, it was shown that for identification purposes, a simplified plasma model is sufficient to predict adequately the relative emission intensities in LIBS plasmas. As a result, it is possible to automatically identify the species with high emission signals, while trace detection is also possible by relaxing search conditions, although manual refinement is still required.

A non-hepatotropic parasite infection increases mortality in the acetaminophen-induced acute liver failure murine model: possible roles for IL-5 and IL-6

We evaluated the effects of a non-hepatotropic parasite infection (Taenia crassiceps) on the outcome of acetaminophen-induced acute liver failure in mice. Uninfected and T. crassiceps infected mice orally received either 300 mg/kg acetaminophen or water as vehicle (n = 5 per group). Survival analysis, hepatocyte necrosis, alanine aminotransferase (ALT) levels, CYP2E1 protein, interleukin (IL-) 5, and IL-6 were assessed for all groups. All infected mice died within 16 h after exposure to acetaminophen (Tc+APAP group), whereas only one-third of uninfected animals exposed to acetaminophen (APAP group) died. Uninfected (Control group) and infected (Tc group) mice that received the vehicle showed no liver damage. Tc+APAP mice exhibited massive liver necrosis characterised by marked balloning degeneration of hepatocytes and higher serum ALT compared to Control, Tc, and APAP animals. Liver tissue from Tc+APAP mice also displayed increased expression of CYP2E1 protein and higher mRNA and protein levels of IL-5 and IL-6 compared to the other groups. These findings suggest that non-hepatotropic parasite infections may increase mortality following acute liver failure by promoting hepatocyte necrosis via IL-5 and IL-6-dependent CYP2E1 overproduction. This study identifies new potential risk factors associated with severe acute liver failure in patients.

A non-hepatotropic parasite infection increases mortality in the acetaminophen-induced acute liver failure murine model: possible roles for IL-5 and IL-6.

We evaluated the effects of a non-hepatotropic parasite infection (Taenia crassiceps) on the outcome of acetaminophen-induced acute liver failure in mice. Uninfected and T. crassiceps infected mice orally received either 300 mg/kg acetaminophen or water as vehicle (n = 5 per group). Survival analysis, hepatocyte necrosis, alanine aminotransferase (ALT) levels, CYP2E1 protein, interleukin (IL-) 5, and IL-6 were assessed for all groups. All infected mice died within 16 h after exposure to acetaminophen (Tc+APAP group), whereas only one-third of uninfected animals exposed to acetaminophen (APAP group) died. Uninfected (Control group) and infected (Tc group) mice that received the vehicle showed no liver damage. Tc+APAP mice exhibited massive liver necrosis characterised by marked balloning degeneration of hepatocytes and higher serum ALT compared to Control, Tc, and APAP animals. Liver tissue from Tc+APAP mice also displayed increased expression of CYP2E1 protein and higher mRNA and protein levels of IL-5 and IL-6 compared to the other groups. These findings suggest that non-hepatotropic parasite infections may increase mortality following acute liver failure by promoting hepatocyte necrosis via IL-5 and IL-6-dependent CYP2E1 overproduction. This study identifies new potential risk factors associated with severe acute liver failure in patients.

In Vitro Effect of the Synthetic cal14.1a Conotoxin, Derived from Conus californicus, on the Human Parasite Toxoplasma gondii.

Toxins that are secreted by cone snails are small peptides that are used to treat several diseases. However, their effects on parasites with human and veterinary significance are unknown. Toxoplasma gondii is an opportunistic parasite that affects approximately 30% of the world's population and can be lethal in immunologically compromised individuals. The conventional treatment for this parasitic infection has remained the same since the 1950s, and its efficacy is limited to the acute phase of infection. These findings have necessitated the search for new drugs that specifically target T. gondii. We examined the effects of the synthetic toxin cal14.1a (s-cal14.1a) from C. californicus on the tachyzoite form of T. gondii. Our results indicate that, at micromolar concentrations, s-cal14.1a lowers viability and inhibits host cell invasion (by 50% and 61%, respectively) on exposure to extracellular parasites. Further, intracellular replication decreased significantly while viability of the host cell was unaffected. Our study is the first report on the antiparasitic activity of a synthetic toxin of C. californicus.

Two-photon excited fluorescence of silica nanoparticles loaded with a fluorene-based monomer and its cross-conjugated polymer: their application to cell imaging.

In this work the two-photon activity of nanoparticles obtained from a fluorene monomer (M1) and its cross-conjugated polymer (P1) is reported. Aqueous suspensions of M1 and P1 nanoparticles prepared through the reprecipitation method exhibited maximum two-photon absorption (TPA) cross-sections of 84 and 9860 GM (1 GM = 10(-50) cm(4) s) at 740 nm, respectively, and a fluorescence quantum yield of ~1. Such a two-photon activity was practically equal with respect to that for molecular solutions of M1 and P1. These materials were then successfully encapsulated into silica nanoparticles to provide bio-compatibly. A lung cancer cell line (A549) and a human cervical cancer cell line (HeLa cells) were incubated with our fluorescent silica nanoparticles to carry out two-photon imaging. By means of these studies we demonstrate that optimized nonlinear optical polymers loaded in silica nanoparticles can be used as efficient probes with low cytotoxicity and good photostability for two-photon fluorescence microscopy. To the best of our knowledge, studies concerning polymer-doped silica nanoparticles exhibiting large two-photon activity have not been reported in the literature.

Neonatal exposure to estradiol induces resistance to helminth infection and changes in the expression of sex steroid hormone receptors in the brain and spleen in adult mice of both sexes.

A single injection of 17beta-estradiol administered to 4-day-old male and female mice increased the cellular immune response, and induced resistance to Taenia crassiceps cysticercosis as well as changes in the expression pattern of progesterone (PR) and estrogen receptor (ER) isoforms in the brain and splenocytes. Regardless of gender, when treated mice reached adulthood, they were highly resistant to infection. Female mice presented early vaginal opening and altered estrous cycles. In male and female mice, the expression of the PR and ER isoforms in the brain was differentially regulated after neonatal exposure to estradiol. Moreover, an increase in the expression of IL-4 and IFN-gamma was found in the serum of experimentally infected neonatally estrogenized animals, which correlated with the observed protection against T. crassiceps infection. In conclusion, early exposure to estradiol permanently modifies immune system activity and sex steroid hormone receptors in the brain, and causes profound changes in sex-associated susceptibility, leading to resistance to helminth parasite infection.

Immune sexual dimorphism: effect of gonadal steroids on the expression of cytokines, sex steroid receptors, and lymphocyte proliferation.

The aims of this study were, first, to explore the differences in the expression of Th1/Th2 cytokines and of steroid receptors in spleen of intact and gonadectomized mice of both sexes; second, to evaluate the effect of estradiol (E2), progesterone (P4) and testosterone (T) on cytokine production and lymphocyte proliferation, and third, to determine the percentage of spleen cell subpopulations in both sexes. Results indicated dimorphic expression of IFN-gamma and IL-4, which was affected by gonadectomy. CD4+ T lymphocytes were the most frequent type of cell in the spleen, followed by B lymphocytes (CD19+). Interestingly, there was no dimorphic pattern of cell subtypes, and gonadectomy had no effect. Regarding lymphocyte proliferation, E2 inhibited both cells of male (19.51%) and female (24.62%). P4 diminished lymphocyte proliferation by 22% in cells of female and had no effect on cells of male. It is very interesting to note that the sex steroid receptors mRNA was highly expressed in all splenocytes, and that this expression was dimorphic. However, flow cytometry analysis confirmed that only expression of progesterone receptor was dimorphic. This dimorphic pattern was, however, only seen in lymphocytes. Present evidence indicates that sex steroids are capable of affecting crucial immune system functions dimorphically.

[Immune sexual dimorphism: can sex steroids affect the Th1/Th2 cytokine profile?]. / Dimorfismo sexual inmunitario: pueden los esteroides sexuales polarizar el perfil de citocinas Th1/Th2?

Cytokine secretion is a crucial aspect in immune system modulation. The secretion pattern of these molecules determines the immune response type that will confront a particular antigen, and this pattern can be at least of two types. A Th1 pattern, effective to eliminate mainly intracellular pathogens and a Th2 pattern, crucial to eradicate extra cellular pathogens. There are many immunological factors that affect expression of these proteins and auto regulate the Th1/Th2 balance, but there are few evidences about effect of other protagonists of mammals physiology. This review focuses on the regulation of the Th1/Th2 cytokine secretion pattern of immune cells by sexual steroids. The evidences indicate that cytokines and steroids form a common chemical language effective to keep the balance between immune and endocrine systems. Alterations of this delicate network can explain different pathologies where gender-associated differences exist and where sexual steroids are crucial factors.

Dimorfismo sexual inmunitario: ¿pueden los esteroides sexuales polarizar el perfil de citocinas Th1/Th2? / Immune sexual dimorphism: Can sex steroids affect the Thl/Th2 cytokine profile?

Cytokine secretion is a crucial aspect in immune system modulation. The secretion pattern of these molecules determines the immune response type that will confront a particular antigen, and this pattern can be at least of two types. A Thl pattern, effective to eliminate mainly intracellular pathogens and a Th2 pattern, crucial to eradicate extra cellular pathogens. There are many immunological factors that affect expression of these proteins and auto regulate the Th1/Th2 balance, but there are few evidences about effect of other protagonists of mammals physiology. This review focuses on the regulation of the Th1/Th2 cytokine secretion pattern of immune cells by sexual steroids. The evidences indicate that cytokines and steroids form a common chemical language effective to keep the balance between immune and endocrine systems. Alterations of this delicate network can explain different pathologies where gender-associated differences exist and where sexual steroids are crucial factors.

La secreción de citocinas es uno de los aspectos más importantes en la modulación de la respuesta inmune. El patrón de secreción de estas moléculas determina el tipo de respuesta inmune que confrontará a un antígeno particular. Ésta puede ser al menos de dos tipos: la respuesta Th1 (encargada principalmente de controlar patógenos intracelulares) y la respuesta Th2 (involucrada en el control de patógenos extracelulares). Existe una autorregulación del balance del tipo de respuesta Th1/Th2 por mecanismos inmunológicos que pueden afectar la expresión de estas proteínas, pero poco se sabe con respecto al papel de otros protagonistas de la fisiología de los mamíferos. En esta revisión se discuten los trabajos referentes al efecto de los esteroides sexuales en la regulación de la secreción de citocinas Th1/Th2 por parte de células del sistema inmune. Las evidencias indican que las citocinas y los esteroides constituyen un lenguaje químico común para el funcionamiento balanceado de los sistemas inmune y endocrino. La alteración de la delicada comunicación entre estos sistemas puede explicar diversas patologías en que existe susceptibilidad asociada al sexo, y en las que los esteroides sexuales son factores clave.

Regulation of the immune response to cestode infection by progesterone is due to its metabolism to estradiol.

The aim of this work was to investigate the role of progesterone during Taenia crassiceps cysticercosis, and the immunological mechanisms involved in its effects, by relating progesterone treatment to whole parasite counts, to host humoral and cellular immune response, to the presence or absence of nuclear receptors to sex steroids in splenocytes, and to serum sex steroid levels in infected mice of both genders. Progesterone treatment increased parasite loads two-fold in females and three-fold in males compared with control mice. The expression of the Th2 cytokine profile (IL-4, IL-6 and IL-10) was markedly increased in infected mice of both genders, while progesterone treatment returned this expression to basal levels. However, the Th1 cytokine profile (IFN-gamma and TNF-alpha) was not affected by infection, whilst progesterone treatment increased the expression of both cytokines two-fold compared to uninfected, infected and placebo-treated mice. Testosterone serum levels decreased in infected male mice by 95%, and treatment with progesterone did not affect them. In females, no change in testosterone levels was observed. Progesterone levels increased three-fold only in progesterone-treated infected mice of both sexes, while estradiol levels in female and male progesterone-treated infected mice increased two-fold compared to infected control mice. The infection markedly induced the expression of progesterone receptor (PR) isoforms A and B in splenocytes of infected mice of both genders (five-fold). Metabolism of progesterone to estradiol was demonstrated by the use of the anti-estrogen tamoxifen, which reduced parasite loads 100% in infected mice of both sexes treated with progesterone. These results suggest that progesterone, possibly through its metabolism to estradiol, affects establishment, growth and reproduction of the helminth parasite T. crassiceps.