Ex vivo analysis of pancreatic cancer-infiltrating T lymphocytes reveals that ENO-specific Tregs accumulate in tumor tissue and inhibit Th1/Th17 effector cell functions

Pancreatic cancer (PC) is an aggressive disease with dismal prognosis. Surgical resection is the recommended treatment for long-term survival, but patients with resectable PC are in the minority (with a 5-year survival rate of 20 %). Therefore, development of novel therapeutic strategies, such as anti-PC immunotherapy, is crucial. α-Enolase (ENO1) is an enzyme expressed on the surface of pancreatic cancer cells and is able to promote cell migration and cancer metastasis. The capacity of ENO1 to induce an immune response in PC patients renders it a true tumor-associated antigen. In this study, we characterized the effector functions of ENO1-specific T cells isolated from PC patients, and we specifically evaluated the successful role of intra-tumoral T helper 17 (Th17) cells and the inhibitory role of regulatory T (Tregs) cells in respectively promoting or reducing the cancer-specific immune response. In this ex vivo study, we have demonstrated, for the first time, that ENO1-specific Th17 cells have a specific anti-cancer effector function in PC patients, and that there are decreased levels of these cells in cancer compared to healthy mucosa. Conversely, there are elevated levels of ENO1-specific Tregs in PC patients which lead to inhibition of the antigen-specific effector T cells, thus highlighting a possible role in promoting PC progression. These results may be relevant for the design of novel immunotherapeutic strategies in pancreatic cancer.     

The complete amino acid sequence of the major Kunitz trypsin inhibitor from the seeds of Prosopsis juliflora.

The major inhibitor of trypsin in seeds of Prosopsis juliflora was purified by precipitation with ammonium sulphate, ion-exchange column chromatography on DEAE- and CM-Sepharose and preparative reverse phase HPLC on a Vydac C-18 column. The protein inhibited trypsin in the stoichiometric ratio of 1:1, but had only weak activity against chymotrypsin and did not inhibit human salivary or porcine pancreatic alpha-amylases. SDS-PAGE indicated that the inhibitor has a Mr of ca 20,000, and IEF-PAGE showed that the pI is 8.8. The complete amino acid sequence was determined by automatic degradation, and by DABITC/PITC microsequence analysis of peptides obtained from enzyme digestions of the reduced and S-carboxymethylated protein with trypsin, chymotrypsin, elastase, the Glu-specific protease from S. aureus and the Lys-specific protease from Lysobacter enzymogenes. The inhibitor consisted of two polypeptide chains, of 137 residues (alpha chain) and 38 residues (beta chain) linked together by a single disulphide bond. The amino acid sequence of the protein exhibited homology with a number of Kunitz proteinase inhibitors from other legume seeds, the bifunctional subtilisin/alpha-amylase inhibitors from cereals and the taste-modifying protein miraculin.     

Xanthones and Cancer: from Natural Sources to Mechanisms of Action

Xanthones are a class of heterocyclic natural products that have been widely studied for their pharmacological potential. In fact, they have been serving as scaffolds for the design of derivatives focusing on drug development. One of the main study targets of xanthones is their anticancer activity. Several compounds belonging to this class have already demonstrated cytotoxic and antitumor effects, making it a promising group for further exploration. This review therefore focuses on recently published studies, emphasizing their natural and synthetic sources and describing the main mechanisms of action responsible for the anticancer effect of promising xanthones.     

Rapid conversion of cyclohexenone,cyclohexanone and cyclohexanol to ε-caprolactone by whole cells of Geotrichum candidum CCT 1205

?-Caprolactone (?-CL) was obtained with excellent conversion and short reaction times from the substrates cyclohexenone, cyclohexanone and cyclohexanol using whole cells of Brazilian Geotrichum candidum (CCT 1205). The reactions were monitored over time by gas chromatography, and the intermediates of the one-pot cascade biotransformation involving reductions of C=C and C=O bonds as well as the Baeyer–Villiger oxidation were identified and quantified. The Baeyer–Villiger monooxygenase (BVMO) enzyme was predominant, and all three substrates were completely converted into ?-CL. Furthermore, the whole cells of Geotrichum candidum were recycled and reutilized in the biotransformation of cyclohexanone, producing ?-CL at least six consecutive times without a significant loss of activity, reaction yields or product purity.     

Validation of the hedonic threshold methodology in determining the hedonic rejection threshold

The hedonic thresholds methodology was recently proposed which allows for determination of two new sensorial thresholds, the compromised acceptance threshold and the hedonic rejection threshold (HRT). For a new methodology to be accepted by the scientific community, its ability to produce reliable results must be demonstrated. Thus, this study sought to validate this new methodology when used to calculate the HRT. In order to do so, the analytical performance indices of precision (repeatability, intermediate precision, and reproducibility), accuracy and robustness for the results for the HRT calculation were measured. The results of seven experiments demonstrated that the methodology possesses high repeatability, intermediate precision and robustness, and satisfactory accuracy and reproducibility. It was therefore concluded that the HRT determination methodology satisfactorily met the analytical performance criteria evaluated, therefore generating reliable results.

Practical applications

The hedonic thresholds methodology (HTM) has numerous applications, including the application of a more severe thermal treatment to obtain greater microbial or enzymatic inactivation, the determination of food shelf life based on its sensory acceptance, the reduction of specific, expensive ingredients in food formulations, reduction of ingredients that are harmful to health when consumed in excess (e.g., sucrose, sodium, and fat), determination of the maximum “defect” threshold that may be present in food and others. The HTM is a recently proposed methodology; therefore, the reliability of the values of the hedonic rejection threshold must be validated, calculated via the HTM. Reliability of the HRT results was confirmed when HTM validation was performed. Thus, the methodology can undoubtedly be applied for the intended purpose.     

Thermophysiological indicators of lambs during pre-slaughter handling

The aim of this study was to evaluate the thermophysiological indicators (TI) of the lambs during the different phases of pre-slaughter handling. Twenty animals were evaluated in two sample days. At each phase, the TI (rectal temperature, respiratory rate, and heart rate) and the environmental variables were measured. Additionally, the body surface temperature was measured by infrared thermography. During the landing phase, the air temperature was the highest (33.1 °C) and the relative humidity was the lowest (45.2%). ANOVA results showed a significant effect of the pre-slaughter phases on all the TI, except the heart rate. Beyond the pre-slaughter phases, the body region and its interaction with the pre-slaughter phases also showed significant influence on the surface temperature. We conclude that the pre-slaughter handling adopted in Northeast Brazil negatively influences the TI of small ruminants, specifically the lambs. Landing was the phase that rendered maximum physiological damage to the animals.     

High-Virulence Cat-Transmitted Ocular Sporotrichosis

Mycopathologia - The article describes an atypical case of cat-transmitted ocular sporotrichosis due Sporothrix brasiliensis in a young patient. This report contributes significantly to knowledge...     

Residual concentrations of antimicrobial growth promoters in poultry litter favour plasmid conjugation among Escherichia coli

Considering that plasmid conjugation is a major driver for the dissemination of antimicrobial resistance in bacteria, this study aimed to investigate the effects of residual concentrations of antimicrobial growth promoters (AGPs) in poultry litter on the frequencies of IncFII-FIB plasmid conjugation among Escherichia coli organisms. A 2 × 5 factorial trial was performed in vitro, using two types of litter materials (sugarcane bagasse and wood shavings) and five treatments of litter: non-treated (CON), herbal alkaloid sanguinarine (SANG), AGPs monensin (MON), lincomycin (LCM) and virginiamycin (VIR). E. coli H2332 and E. coli J62 were used as donor and recipient strains, respectively. The presence of residues of monensin, lincomycin and virginiamycin increased the frequency of plasmid conjugation among E. coli in both types of litter materials. On the contrary, sanguinarine significantly reduced the frequency of conjugation among E. coli in sugarcane bagasse litter. The conjugation frequencies were significantly higher in wood shavings compared with sugarcane bagasse only in the presence of AGPs. Considering that the presence of AGPs in the litter can increase the conjugation of IncFII-FIB plasmids carrying antimicrobial resistance genes, the real impact of this phenomenon on the dissemination of antimicrobial resistant bacteria in the poultry production chain must be investigated.     

Experimental Study of Nest-site Selection in the Biscutate Swift (Streptoprocne biscutata, Aves: Apodidae) in Southern Brazil

Biscutate swift Streptoprocne biscutata nests are usually built on protected rocky cliff walls. Birds often renest at the same location. Remains of previous nests may offer information about potential nests and quality of nest-sites. Here, we experimentally study nest-site selection to test the hypothesis that information from previous nests is used in current nest-site selection. We placed old nest material at artificial nest-sites to test whether new nest-sites are chosen based on the presence of nesting material. We also tested whether the use of natural nest-sites is influenced by nesting material by creating two types of natural sites: previously used natural nest-sites with vestiges of old nests removed and never used natural nest-sites to which vestiges of old nests were added. In the first experiment, in 139 nest-use opportunities, 16 artificial nest-sites were used, all of which included vestiges. In the second experiment, in 91 nest-use opportunities, four nests were in previously unused but natural locations to which vestiges had been added, 22 nests were in previously used sites without vestiges, and the remaining 65 nests remained unused. Two processes are apparently in action: first, prior experience and memory; second, vestiges indicate where nesting has occurred, possibly useful for first breeding, or for imperfect memory. Previous nesting information may explain why swifts use nesting locations for many years and why new nesting colonies seldom form. This transmission of information suggests that swifts tend to be conservative and nest where previous nesting has occurred.