Partial replacement of soybean meal with microalgae biomass on in vitro ruminal fermentation may reduce ruminal protein degradation.

The objective of this study was to evaluate the effects of partially replacing soybean meal (SBM) with algal sources on in vitro ruminal fermentation. Using 6 fermenters in a 3 × 3 replicated Latin square with 3 periods of 10 d each, we tested 3 treatments: a control diet (CRT) with SBM at 17.8% of the diet dry matter (DM); and 50% SBM biomass replacement with either Chlorella pyrenoidosa (CHL); or Spirulina platensis (SPI). The basal diet was formulated to meet the requirements of a 680-kg Holstein dairy cow producing 45 kg/d of milk with 3.5% fat and 3% protein. All diets had a similar nutritional composition (16.0% CP; 34.9% NDF; 31.0% starch, DM basis) and fermenters were provided with 106 g DM/d split into 2 portions. After 7 d of adaptation, samples were collected for 3 d of each period for analyses of ruminal fermentation at 0, 1, 2, 4, 6, and 8 h after morning feeding for evaluation of the ruminal fermentation kinetics. For the evaluation of the daily production of total metabolites and for the evaluation of nutrient degradability, samples from the effluent containers were collected daily. Statistical analysis was performed with the MIXED procedure of SAS with treatment, time, and their interactions considered as fixed effects; day, square, and fermenter were considered as random effects. Orthogonal contrasts (CRT vs. algae; and CHL vs. SPI) were used to depict the treatment effect, and significance was declared when P ≤ 0.05. Fermenters that received algae-based diets had a greater propionate molar concentration and molar proportion when compared with the fermenters fed CRT diets. In addition, those algae-fed fermenters had lower branched short-chain fatty acids (BSCFA) and isoacids (IA), which are biomarkers of ruminal protein degradation, along with lower ammonia (NH3-N) concentration and greater nonammonia nitrogen (NAN). When contrasting with fermenters fed SPI-diets, fermenters fed based CHL-diets had a lower molar concentration of BSCFA and IA, along with lower NH3-N concentration and flow, and greater NAN, bacterial nitrogen flow, and efficiency of nitrogen utilization. Those results indicate that CHL protein may be more resistant to ruminal degradation, which would increase efficiency of nitrogen utilization. In summary, partially replacing SBM with algae biomass, especially with CHL, is a promising strategy to improve the efficiency of nitrogen utilization, due to the fact that fermenters fed CHL-based diets resulted in a reduction in BSCFA and IA, which are markers of protein degradation, and it would improve the efficiency of nitrogen utilization. However, further validation using in vivo models are required.

Gelatin edible coatings with mint essential oil (Mentha arvensis): film characterization and antifungal properties.

In this work, mint essential oil (MEO) was added into gelatin films and antifungal activity was evaluated. Five concentrations of MEO (0, 0.06, 0.13, 0.25, 0.38, 0.50% (g/g gelatin)) were incorporated into gelatin solutions. The films were prepared by casting and characterized for their barrier properties, mechanical resistance, morphology, thermal and antifungal activity. The addition of oil into the solution slightly improved water vapor barrier, increased thickness and opacity, decreased transparency and modified thermal and mechanical properties of films. With addition of oil above 0.38%, the films were effective against the growth of Botrytis cinerea and Rhizopus stolonifer, indicating an inhibitory activity. Thus, gelatin-based edible films incorporated with MEO showed to be an effective way to inhibit microbial growth on the film surface.

Haplotype frequencies based on eight polymorphic sites at the 3' untranslated region of the HLA-G gene in individuals from two different geographical regions of Brazil.

The Brazilian population represents an admixture of native Amerindians, Portuguese settlers and Africans who were brought as slaves during the colonization period that began in the 16th century and was followed by waves of immigrations of Europeans and Asians in the 20th century. The contribution of these different ethnic groups to the constitution of Brazilian populations from different geographic regions is variable and, in addition to environmental factors, might act by determining different allele profiles among Brazilian populations from different regions. We studied polymorphic sites at the 3' untranslated region of the HLA-G gene in individuals from a Northeastern Brazilian region and compared them to our previously published data about a Southeastern Brazilian region, located at a distance of 2589 km. Our results showed that most polymorphic sites present a similar distribution in both populations, except for the lower frequency of the +3003C allele in the Northeastern population compared to the Southeastern population. Although differences in genotypic distribution were only significant for the +3003 locus (P = 0.0201), the diversity of haplotypes was distinct for each population. These results are important for case-control studies on the association of human leucocyte antigen-G polymorphism with disease and also in terms of the genetic structure of two distinct Brazilian populations.

Therapeutic impact of determination of RAS mutations in the plasma of patient with colorectal cancer.

Stage IV colorectal cancer treatment includes targeted therapy depending on RAS status. During disease progression, loss or gain of RAS mutations could happen, supporting the hypothesis of the evolutionary pressure of therapy. Circulating tumor DNA (ctDNA) are nucleic acids released to the bloodstream by the tumor during its development and may be detected by liquid biopsy. The Idylla© Biocartis, a fully automated real-time-PCR-based molecular diagnostic system, was used in a patient with metastatic colorectal cancer with a NRAS mutation in progression after several therapeutic lines. The ctDNA mutational analysis was performed and revealed the absence of mutations in the KRAS, NRAS, and BRAF genes. The patient started the third line of palliative chemotherapy with irinotecan + cetuximab and achieved a partial response for the first time. The authors describe a case in which liquid biopsy determined the higher progression-free survival achieved.

Lipid nanoparticles strategies to modify pharmacokinetics of central nervous system targeting drugs: Crossing or circumventing the blood-brain barrier (BBB) to manage neurological disorders.

The main limitation to the success of central nervous system (CNS) therapies lies in the difficulty for drugs to cross the blood-brain barrier (BBB) and reach the brain. Regarding its structure and enzymatic complexity, crossing the BBB is a challenge, although several alternatives have been identified. For instance, the use of drugs encapsulated in lipid nanoparticles has been described as one of the most efficient approaches to bypass the BBB, as they allow the passage of drugs through this barrier, improving brain bioavailability. In particular, solid lipid nanoparticles (SLN) and nanostructured lipid carriers (NLC) have been a focus of research related to drug delivery to the brain. These systems provide protection of lipophilic drugs, improved delivery and bioavailability, having a major impact on treatments outcomes. In addition, the use of lipid nanoparticles administered via routes that transport drugs directly into the brain seems a promising solution to avoid the difficulties in crossing the BBB. For instance, the nose-to-brain route has gained considerable interest, as it has shown efficacy in 3D human nasal models and in animal models. This review addresses the state of the art on the use of lipid nanoparticles to modify the pharmacokinetics of drugs employed in the management of neurological disorders. A description of the structural components of the BBB, the role of the neurovascular unit and limitations for drugs to entry into the CNS is first addressed, along with the developments to increase drug delivery to the brain, with a special focus on lipid nanoparticles. In addition, the obstacle of BBB complexity in the creation of new effective drugs for the treatment of the most prevalent neurological disorders is also addressed. Finally, the proposed strategies for lipid nanoparticles to reach the CNS, crossing or circumventing the BBB, are described. Although promising results have been reported, especially with the nose-to-brain route, they are still ongoing to assess its real efficacy in vivo in the management of neurological disorders.

[The family and the mentally ill using day-hospitals: a case study]. / A família e o doente mental usuário do hospital-dia--estudo de um caso.

The day-hospital, as a modality of psychiatric care, provides families with the possibility to experience greater contact with their mentally-ill relative during treatment and leads them to searching, together with such relative, for ways to facilitate the adaptation process of interaction in the family. This study aims at understanding how families and the users of day-hospitals interact out of their homes and at describing the experience narrated by a family. The family in this study was selected from one of the users of a day-hospital who had a previous history of hospital admissions for psychiatric treatment. Five members of the family under study were interviewed. Based on the experiences described by the subjects, two great thematic categories emerged: the family experiencing the day-hospital with its family member and the family-user relationships. The experiences narrated by the subjects in this study are rather significant for the construction of new possibilities in caring for the mentally-ill person's family.

[Work-mental patient: the family's perception]. / Trabalho x doente mental: percepção da família.

The relationship family x mental ill, much more present in the psychiatric reform, constitutes a possibility to create new ways of understanding how the family perceives the relationship work x mental ill. It is considered as a fundamental element in this research, the speech of the components of the family, due to their experience. In relationship mental ill x work, the family describes several difficulties, where the idleness is present, revealing the inability of his component for work, or appointing him a different occupation, adapted to his condition of sick, without perceiving the possible depression felt as a consequence of his pathology.

Stability of thermolabile (LT) enterotoxin produced from enterotoxigenic Escherichia coli strains maintained in vitro.

The stability of thermolabile (LT) enterotoxin in 26 strains of porcine enterotoxigenic Escherichia coli (PETEC) belonging to serogroups 08 and 0149 was assayed by the passive immune hemolysis (PIH) test, over a period of 9 months at -70 degrees C. It was found that the percentage of LT+ colonies (% LT+) and the mean value of hemoglobin release (XHb), could predict a change from LT+ to LT-.

Factors affecting detection of Yersinia enterocolitica heat-stable enterotoxin by the infant mouse test.

With regard to the assay of heat-stable enterotoxin (ST) from Yersinia enterocolitica, we made a comparative study of the conventional infant mouse test read at 4 h and a modified infant mouse test read at 2 h. The influence of several factors, such as the medium used to prepare ST, lysing of bacterial cells from the broth cultures used to prepare ST, and the temperature at which the inoculated mice were kept during the test, was also investigated. Thus, with a few exceptions, Pai-Mors medium was more suitable than Casamino Acids-yeast extract medium, for the preparation of yersinial ST. Gut/carcass weight ratios obtained with lysed supernatants or with supernatants from whole cultures of Y. enterocolitica were similar, suggesting that most of the ST produced by this microorganism in broth cultures is extracellular. The amount of ST produced by Y. enterocolitica, as well as the ambient temperature at which inoculated mice were kept during the assay, was found to influence gut/carcass weight ratios obtained with both tests. Enterotoxigenicity and the temperature at which mice were kept were interrelated, such that for weakly enterotoxigenic strains there were no significant differences among gut/carcass weight ratios for the conventional and modified infant mouse tests carried out at 18 or 25 degrees C, but at 30 degrees C the values in the modified test were higher for most ST preparations with Pai-Mors medium. The influence of ambient temperature was more pronounced at 37 degrees C, since most strains produced negative results in the conventional test at this temperature. We conclude that the conventional infant mouse test is adequate for assaying yersinial ST, provided that the temperature at which mice are kept during the assay is fixed at around 25 degrees C.

Single radial immune hemolysis test for detection of Escherichia coli thermolabile enterotoxin.

A single radial immune hemolysis test for the detection of thermolabile enterotoxin has been developed for routine purposes. Stationary cultures from enterotoxigenic Escherichia coli in Casamino Acids-yeast extract medium may be used for the detection of this enterotoxin, and under the conditions of the experiment, the single radial immune hemolysis test was as sensitive as the passive immune hemolysis test. The results obtained in the single radial immune hemolysis test agreed entirely with those obtained in the passive immune hemolysis test, and no false-positive reactions were obtained when cholera antitoxin diluted 1:80 was used. The assay is easy to perform, inexpensive, and specially designed for less-equipped laboratories.