Effect of an Alliaceae Encapsulated Extract on Growth Performance, Gut Health, and Intestinal Microbiota in Broiler Chickens Challenged with Eimeria spp.

This study analyzed the effects of an Alliaceae encapsulated extract (AE-e) on daily gain (ADG), feed intake (ADFI), feed conversion ratio (FCR), oocysts per gram of feces (OPG), intestinal lesion (LS), and microbiota composition in broilers challenged with Eimeria spp. A total of 4800 one day Cobb-500 were allotted into 10 treatment groups with 12 replicates of 40 birds in a 2 × 4 + 2 factorial arrangement. The first factor was non-challenged (NC) or challenged (C), the second was four levels of AE-e added in the basal diet, 0 (AE0), 250 (AE250), 500 (AE500), and 750 mg·kg-1 (AE750), plus two ionophore controls, non-challenged (NC-Ion) and challenged (C-Ion). No interactions were observed between factors (NC0, NC250, NC500, NC750, C0, C250, C500, and C750), while C-Ion improved FCR at 21 d. The challenge affected negatively ADG and FCR and promoted enteropathogens in cecum. AE750 improved FCR in the finisher and cumulative phases, while C-Ion had fewer total OPG than C0 and C250. Likewise, at 21d, C250, C500, and C-Ion had fewer LS than C0, while at 28 d, C750 showed lower than C-Ion. In the cecum microbiota, C500 had more Ruminococcus, Firmicutes b, and Intestinimonas than C-Ion. In summary, AE-e showed beneficial results in broilers infected with Eimeria spp.

The Effect of Encapsulated Propyl Propane Thiosulfonate (PTSO) on Apparent Ileal Digestibility and Productive Performance in Broiler Chickens.

This study analyzed the effects of different dietary doses of encapsulated propyl propane thiosulfonate (Pe-PTSO) on the apparent ileal digestibility (AID) of nutrients and productive performance in broilers. A total of 100 one-day-old Cobb 500 were housed in battery cages for 20 days. At 10 days of age, the birds were assigned to one of five diets: negative control (P0), 250 mg/kg of Pe-PTSO (P250), 500 mg/kg of Pe-PTSO (P500), 750 mg/kg of Pe-PTSO (P750), and positive control, nicarbazin-narasin (ION). Titanium dioxide was the external marker, which was added to the diets from day 17 to 20. In the birds fed the P250 diet, there was a significant difference (p ≤ 0.05) in the AID values for amino acids and energy compared to those that consumed the P0 diet. Furthermore, the P250 diet significantly increased (p ≤ 0.05) the average daily weight gain compared to the P0 diet. No significant differences were observed between treatments in average daily feed intake and feed conversion ratio. In summary, the inclusion of 250 mg of encapsulated PTSO per kg in broiler chickens diet improved the digestibility of amino acids and energy, as well as weight gain.

Pharmacological tumor PDL1 depletion with chlorambucil treats ovarian cancer and melanoma: improves antitumor immunity and renders anti-PDL1-resistant tumors anti-PDL1-sensitive through NK cell effects.

BACKGROUND: Tumor intracellular programmed cell death ligand-1 (PDL1) mediates pathologic signals that regulate clinical treatment responses distinctly from surface-expressed PDL1 targeted by αPDL1 immune checkpoint blockade antibodies. METHODS: We performed a drug screen for tumor cell PDL1 depleting drugs that identified Food and Drug Administration (FDA)-approved chlorambucil and also 9-[2-(phosphonomethoxy)ethyl] guanine. We used in vitro and in vivo assays to evaluate treatment and signaling effects of pharmacological tumor PDL1 depletion focused on chlorambucil as FDA approved, alone or plus αPDL1. RESULTS: PDL1-expressing mouse and human ovarian cancer lines and mouse melanoma were more sensitive to chlorambucil-mediated proliferation inhibition in vitro versus corresponding genetically PDL1-depleted lines. Orthotopic peritoneal PDL1-expressing ID8agg ovarian cancer and subcutaneous B16 melanoma tumors were more chlorambucil-sensitive in vivo versus corresponding genetically PDL1-depleted tumors. Chlorambucil enhanced αPDL1 efficacy in tumors otherwise αPDL1-refractory, and improved antitumor immunity and treatment efficacy in a natural killer cell-dependent manner alone and plus αPDL1. Chlorambucil-mediated PDL1 depletion was relatively tumor-cell selective in vivo, and treatment efficacy was preserved in PDL1KO hosts, demonstrating tumor PDL1-specific treatment effects. Chlorambucil induced PDL1-dependent immunogenic tumor cell death which could help explain immune contributions. Chlorambucil-mediated PDL1 reduction mechanisms were tumor cell-type-specific and involved transcriptional or post-translational mechanisms, including promoting PDL1 ubiquitination through the GSK3ß/ß-TRCP pathway. Chlorambucil-mediated tumor cell PDL1 depletion also phenocopied genetic PDL1 depletion in reducing tumor cell mTORC1 activation and tumor initiating cell content, and in augmenting autophagy, suggesting additional treatment potential. CONCLUSIONS: Pharmacological tumor PDL1 depletion with chlorambucil targets tumor-intrinsic PDL1 signaling that mediates treatment resistance, especially in αPDL1-resistant tumors, generates PDL1-dependent tumor immunogenicity and inhibits tumor growth in immune-dependent and independent manners. It could improve treatment efficacy of selected agents in otherwise treatment-refractory, including αPDL1-refractory cancers, and is rapidly clinically translatable.

Pharmacologic Tumor PDL1 Depletion with Cefepime or Ceftazidime Promotes DNA Damage and Sensitivity to DNA-Damaging Agents.

The interaction between tumor surface-expressed PDL1 and immune cell PD1 for the evasion of antitumor immunity is well established and is targeted by FDA-approved anti-PDL1 and anti-PD1 antibodies. Nonetheless, recent studies highlight the immunopathogenicity of tumor-intrinsic PDL1 signals that can contribute to the resistance to targeted small molecules, cytotoxic chemotherapy, and αPD1 immunotherapy. As genetic PDL1 depletion is not currently clinically tractable, we screened FDA-approved drugs to identify those that significantly deplete tumor PDL1. Among the candidates, we identified the ß-lactam cephalosporin antibiotic cefepime as a tumor PDL1-depleting drug (PDD) that increases tumor DNA damage and sensitivity to DNA-damaging agents in vitro in distinct aggressive mouse and human cancer lines, including glioblastoma multiforme, ovarian cancer, bladder cancer, and melanoma. Cefepime reduced tumor PDL1 post-translationally through ubiquitination, improved DNA-damaging-agent treatment efficacy in vivo in immune-deficient and -proficient mice, activated immunogenic tumor STING signals, and phenocopied specific genetic PDL1 depletion effects. The ß-lactam ring and its antibiotic properties did not appear contributory to PDL1 depletion or to these treatment effects, and the related cephalosporin ceftazidime produced similar effects. Our findings highlight the rapidly translated potential for PDDs to inhibit tumor-intrinsic PDL1 signals and improve DNA-damaging agents and immunotherapy efficacy.

Immune checkpoint expression and relationships to anti-PD-L1 immune checkpoint blockade cancer immunotherapy efficacy in aged versus young mice.

Introduction: Aging is the biggest cancer risk, and immune checkpoint (IC) inhibition (ICI) is a revolutionary cancer immunotherapy approach. Nonetheless, there are limited preclinical/clinical data regarding aging effects on ICI outcomes or age effects on IC expression in different organs or tumors. Methods: Flow cytometry assessed IC on immune and non-immune cells in various organs in young and aged BL6 mice. Comparisons: aged versus young naïve WT versus interferon-γ KO mice and WT challenged with B16F10 melanoma and treated with αPD-1 or αPD-L1 ICI. We co-cultured young and aged T cells and myeloid cells in vitro and used OMIQ analyses to test cell-cell interactions. Results: αPD-1 ICI treated melanoma in young and aged hosts, whereas αPD-L1 ICI was only effective in young. We found considerable, previously undescribed age effects on expression of various IC molecules participating in the ICI treatment, including PD-1, PD-L1, PD-L2, and CD80, in distinct organs and in the tumor. These data help explain differential ICI efficacy in young and aged hosts. Host interferon-γ influenced age effects on IC expression in both directions depending on specific IC molecule and tissue. IC expression was further affected by tumor challenge on immune, non-immune, and tumor cells in tumor and other organs. In in vitro co-culture, αPD-1 versus αPD-L1 distinctly influenced polyclonal T cells in young versus aged, suggesting mechanisms for distinct age-related ICI outcomes. Conclusion: Age affects IC expression on specific immune cells in an organ- and tissue-specific manner. ICs were generally higher on aged immune cells. High immune-cell PD-1 could help explain αPD-1 efficacy in aged. High co-expression of CD80 with PD-L1 on dendritic cells could help explain lack of αPD-L1 efficacy in aged hosts. Factors other than myeloid cells and interferon-γ also affect age-related IC expression and T cell function, meriting additional studies.

CD122-directed interleukin-2 treatment mechanisms in bladder cancer differ from αPD-L1 and include tissue-selective γδ T cell activation.

BACKGROUND: Anti-programmed death-ligand 1 (αPD-L1) immunotherapy is approved to treat bladder cancer (BC) but is effective in <30% of patients. Interleukin (IL)-2/αIL-2 complexes (IL-2c) that preferentially target IL-2 receptor ß (CD122) augment CD8+ antitumor T cells known to improve αPD-L1 efficacy. We hypothesized that the tumor microenvironment, including local immune cells in primary versus metastatic BC, differentially affects immunotherapy responses and that IL-2c effects could differ from, and thus complement αPD-L1. METHODS: We studied mechanisms of IL-2c and αPD-L1 efficacy using PD-L1+ mouse BC cell lines MB49 and MBT-2 in orthotopic (bladder) and metastatic (lung) sites. RESULTS: IL-2c reduced orthotopic tumor burden and extended survival in MB49 and MBT-2 BC models, similar to αPD-L1. Using antibody-mediated cell depletions and genetically T cell-deficient mice, we unexpectedly found that CD8+ T cells were not necessary for IL-2c efficacy against tumors in bladder, whereas γδ T cells, not reported to contribute to αPD-L1 efficacy, were indispensable for IL-2c efficacy there. αPD-L1 responsiveness in bladder required conventional T cells as expected, but not γδ T cells, altogether defining distinct mechanisms for IL-2c and αPD-L1 efficacy. γδ T cells did not improve IL-2c treatment of subcutaneously challenged BC or orthotopic (peritoneal) ovarian cancer, consistent with tissue-specific and/or tumor-specific γδ T cell contributions to IL-2c efficacy. IL-2c significantly altered bladder intratumoral γδ T cell content, activation status, and specific γδ T cell subsets with antitumor or protumor effector functions. Neither IL-2c nor αPD-L1 alone treated lung metastatic MB49 or MBT-2 BC, but their combination improved survival in both models. Combination treatment efficacy in lungs required CD8+ T cells but not γδ T cells. CONCLUSIONS: Mechanistic insights into differential IL-2c and αPD-L1 treatment and tissue-dependent effects could help develop rational combination treatment strategies to improve treatment efficacy in distinct cancers. These studies also provide insights into γδ T cell contributions to immunotherapy in bladder and engagement of adaptive immunity by IL-2c plus αPD-L1 to treat refractory lung metastases.

In vitro assessment of two novel Cellulases from Trabulsiella odontotermitis for agricultural waste utilization.

BACKGROUND: The production of agricultural wastes still growing as a consequence of the population growing. However, the majority of these residues are under-utilized due their chemical composition, which is mainly composed by cellulose. Actually, the search of cellulases with high efficiency to degrade this carbohydrate remains as the challenge. In the present experiment, two genes encoding an endoglucanase (EC 3.2.1.4) and ß-glucosidase (EC 3.2.1.21) were overexpressed in Escherichia coli and their recombinant enzymes (egl-FZYE and cel-FZYE, respectively) characterized. Those genes were found in Trabulsiella odontermitis which was isolated from the gut of termite Heterotermes sp. Additionally, the capability to release sugars from agricultural wastes was evaluated in both enzymes, alone and in combination. RESULTS: The results have shown that optimal pH was 6.0 and 6.5, reaching an activity of 1051.65 ± 47.78 and 607.80 ± 10.19 U/mg at 39 °C, for egl-FZYE and cel-FZYE, respectively. The Km and Vmax for egl-FZYE using CMC as substrate were 11.25 mg/mL and 3921.57 U/mg, respectively, whereas using Avicel were 15.39 mg/mL and 2314.81 U/mg, respectively. The Km and Vmax for cel-FZYE using Avicel as substrate were 11.49 mg/mL and 2105.26 U/mg, respectively, whereas using CMC the enzyme did not had activity. Both enzymes had effect on agricultural wastes, and their effect was improved when they were combined reaching an activity of 955.1 ± 116.1, 4016.8 ± 332 and 1124.2 ± 241 U/mg on corn stover, sorghum stover and pine sawdust, respectively. CONCLUSIONS: Both enzymes were capable of degrading agricultural wastes, and their effectiveness was improved up to 60% of glucose released when combined. In summary, the results of the study demonstrate that the recombinant enzymes exhibit characteristics that indicate their value as potential feed additives and that the enzymes could be used to enhance the degradation of cellulose in the poor-quality forage generally used in ruminant feedstuffs.

Genomic and functional characterisation of two Enterococcus strains isolated from Cotija cheese and their potential role in ripening.

Enterococcus spp. are present in the native microbiota of many traditional fermented foods. Their ability to produce antibacterial compounds, mainly against Listeria monocytogenes, has raised interest recently. However, there is scarce information about their proteolytic and lipolytic potential, and their biotechnological application is currently limited because enterococcal strains have been related to nosocomial infections. In this work, next-generation sequencing and optimised bioinformatic pipelines were used to annotate the genomes of two Enterococcus strains-one E. faecium and one E. faecalis-isolated from the Mexican artisanal ripened Cotija cheese. A battery of genes involved in their proteolytic system was annotated. Genes coding for lipases, esterases and other enzymes whose final products contribute to cheese aroma and flavour were identified as well. As for the production of antibacterial compounds, several peptidoglycan hydrolase- and bacteriocin-coding genes were identified in both genomes experimentally and by bioinformatic analyses. E. faecalis showed resistance to aminoglycosides and E. faecium to aminoglycosides and macrolides, as predicted by the genome functional annotation. No pathogenicity islands were found in any of the strains, although traits such as the ability of biofilm formation and cell aggregation were observed. Finally, a comparative genomic analysis was able to discriminate between the food strains isolated and nosocomial strains. In summary, pathogenic strains are resistant to a wide range of antibiotics and contain virulence factors that cause host damage; in contrast, food strains display less antibiotic resistance, include genes that encode class II bacteriocins and express virulence factors associated with host colonisation rather than invasion.

Draft Genome Sequences for Five Strains of Trabulsiella odontotermitis, Isolated from Heterotermes sp. Termite Gut.

Trabulsiella odontotermitis represents a novel species in the genus Trabulsiella with no complete genome reported yet. Here, we describe the draft genome sequences of five isolates from termites present in the north of Mexico, which have an interesting pool of genes related to cellulose degradation with biotechnological application.

Genome Sequence of Citrobacter sp. CtB7.12, Isolated from the Gut of the Desert Subterranean Termite Heterotermes aureus.

The draft genome of Citrobacter sp. CtB7.12, isolated from termite gut, is presented here. This organism has been reported as a cellulolytic bacterium, which is biotechnologically important because it can be used as a gene donor for the ethanol and biofuel industries.

Prevalence of overweight and risk of overweight among 3- to 5-year-old Chicago children, 2002-2003.

This article reports the first estimates of overweight prevalence in Chicago children entering school (aged 3-5 years). Chicago data are compared with those from the National Health and Nutrition Examination Survey (NHANES) and the Early Childhood Longitudinal Study (ECLS). Data were from 2 separate convenience samples of children aged 3-5 years attending either 18 Chicago Public Schools or 10 Chicago Catholic School pre-K programs (n = 1517). Data were taken from students' Certificate of Child Health Examination (CCHE), completed by a health professional. Overall, the prevalence of overweight subjects (body mass index >or=95th percentile) was 24%, more than twice that of the national prevalence of 10% for 2- to 5-year olds documented by NHANES (1999-2002) and 3 times that of the 1998-1999 ECLS prevalence estimate of 8% for 5- to 7-year olds in the Midwest region. The data reported here document that nearly one quarter of children entering school in Chicago are already overweight. This clearly establishes a need for local schools to develop protocols and procedures to support the physical and mental health needs of affected and at-risk children. The findings also make it plain that ongoing weight status monitoring is needed and that current plans to implement this should go forward.

Diferencia en la composición porcentual de los poliinsaturados de cadena larga en eritrocitos materno-fetales en nacimientos de término o pretérmino en humanos / Differences in the percentual composition of the long chain polyunsatureted in maternal and fetal erythrocytes in term and preterm birth in human

Se investiga el contenido y la distribución de los ácidos grasos altamente poliinsaturados de cadena larga (APCL) en los fosfolípidos de eritrocitos materno-fetales en sagre materna venosa y de cordón umbilical obtenida al momento de partos de término o pretérmino en humanos. Se selecionaron madres del mismo nivel socioeconómico, sanas. Once madres dieron a luz niños sanos de término (40 semanas), y veintidós niños sanos de pretérmino (32-34 semanas). Se obtuvo sangre venosa materna y sangre umbilical al momento del parto. Se aislaron los fosfolípidos de los eritrocitos, los ácidos grasos de los fosfolípidos se analizaron por cromatografía gas-líquida. Los resultados informan que en los fetos de término el contenido de todos los APCL omega 6 y omega 3 particularmente 20:4w6 (ARA), y 22:6w3 (DHA), se encontraron signigicativamente más altos que en su madre, a la inversa todos los APCL fueron significativamente menores en el feto pretérmino en relación a su madre. Al comparar contenidos de ARA y DHA entre los fetos se observó que sólo el DHA se encontró significativamente disminuido en fetos pretérmino. Las mujeres que dieron a luz niños de pretérmino demostraron una alta relación ARA/DHA en su sangre en relación a las mujeres con parto de término. De este estudio y tomando en cuenta la literatura preexistente se puede sugerir que un alto contenido de ARA en eritrocitos materno y fetales podría ser una señal de riesgo de prematuridad.