Dairy farmers across Brazil were invited to participate in a study on silage production and utilization practices. Two hundred sixty farmers filled out a questionnaire, which was made available on a ...website. The questionnaire consisted of 14 questions, including information about the characteristics of the herd (n=3), the crop(s) used in the ensiling process, the use of additives, the harvest (n=3), the type of silo (n=1), aspects related to sealing (n=2), and management practices applied during feed-out (n=3). Farmers were also asked a final question about the main barriers they faced when producing and using silage. The main dairy-producing regions of Brazil had a strong influence on the number of participants. The profiles of farmers were heterogeneous and divided into 5 groups, which was considered a positive attribute of the study, allowing better analysis and assessment of current circumstances. Corn was the most widely grown crop for silage. Sorghum, tropical grasses, and sugarcane were the other species most cited. Additives were used by a small number of farmers (27.7%). Approximately 40% of farmers still depended on loaned equipment or outsourced services. The pull-type forage harvester was the main piece of equipment used on dairy farms (90.4%). Only 54.6% of respondents answered that they sharpen their harvester knives daily. Horizontal silos (bunker and stack) were the structures most commonly used to store silage. Most farmers sealed silos with double-sided plastic film (black-on-white) and with soil. However, almost one-fifth of all farmers still use black plastic. Manual removal of silage from the silos was practiced at most farms (i.e., the lack of equipment was also reflected in the stage of silage utilization). Disposal of spoiled silage before inclusion in the livestock feed was not a common practice on the farms. The main barriers encountered on the farms were lack of equipment, lack of manpower, and climatic variations. The results of this research may guide researchers, industries, extension workers, and governments to seek efficiency in milk production on farms using silage in the diet of livestock throughout the year or during part of the year in Brazil.
Specific folate receptors are abundantly overexpressed in chronically activated macrophages and in most cancer cells. Directed folate receptor targeting using liposomes is usually achieved using ...folate linked to a phospholipid or cholesterol anchor. This link is formed using a large spacer like polyethylene glycol. Here, we report an innovative strategy for targeted liposome delivery that uses a hydrophobic fragment of surfactant protein D linked to folate. Our proposed spacer is a small 4 amino acid residue linker. The peptide conjugate inserts deeply into the lipid bilayer without affecting liposomal integrity, with high stability and specificity. To compare the drug delivery potential of both liposomal targeting systems, we encapsulated the nuclear dye Hoechst 34580. The eventual increase in blue fluorescence would only be detectable upon liposome disruption, leading to specific binding of this dye to DNA. Our delivery system was proven to be more efficient (2-fold) in Caco-2 cells than classic systems where the folate moiety is linked to liposomes by polyethylene glycol.
Vascular complication is one of the causes of skin flap healing failure. Sildenafil and tadalafil, a type-5 phosphodiesterase inhibitor, can improve flap viability, however, the action mechanisms ...involved in this process are still unclear. To assess the effects of orally administered sildenafil and tadalafil on the healing kinetics and skin flap viability, sixty-two Wistar rats were divided into three groups: control (
n
= 22), sildenafil (
n
= 20), and tadalafil (
n
= 20). The solutions were administered orally (dose: 10 mg/kg) immediately after the surgical procedure and then every 24 h. At postoperative days 7 and 14, the skin flap samples were collected, submitted to histological processing and evaluated under optical microscopy. In experimental groups (sildenafil and tadalafil), we found an increased vascularization (
p
< 0.05) on the 7th and 14th day associated with the ulcer size decrease on the 14th day, although it was not significant. There was a higher influx of neutrophils and a decrease of mononuclear population on the 7th day (
p
< 0.05). On the 14th day, these differences were observed only in the tadalafil group (
p
< 0.05). This study suggested positive results with the use of sildenafil and tadalafil as adjuvant drugs in skin flap viability.
Leprosy is a chronic human disease caused by the yet-uncultured pathogen Mycobacterium leprae. Although readily curable with multidrug therapy (MDT), over 200,000 new cases are still reported ...annually. Here, we obtain M. leprae genome sequences from DNA extracted directly from patients' skin biopsies using a customized protocol. Comparative and phylogenetic analysis of 154 genomes from 25 countries provides insight into evolution and antimicrobial resistance, uncovering lineages and phylogeographic trends, with the most ancestral strains linked to the Far East. In addition to known MDT-resistance mutations, we detect other mutations associated with antibiotic resistance, and retrace a potential stepwise emergence of extensive drug resistance in the pre-MDT era. Some of the previously undescribed mutations occur in genes that are apparently subject to positive selection, and two of these (ribD, fadD9) are restricted to drug-resistant strains. Finally, nonsense mutations in the nth excision repair gene are associated with greater sequence diversity and drug resistance.
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•Stable BSA nanoemulsions are produced by high pressure homogenization.•Linear correlation is found between Poloxamer 407 levels and size of nanoemulsions.•Presence of BSA-FA ...conjugate results in specific FA mediated internalization in folate receptor positive cells.
Bovine serum albumin (BSA) nanoemulsions were produced by high pressure homogenization with a tri-block copolymer (Poloxamer 407), which presents a central hydrophobic chain of polyoxypropylene (PPO) and two identical lateral hydrophilic chains of polyethylene glycol (PEG). We observed a linear correlation between tri-block copolymer concentration and size – the use of 5mg/mL of Poloxamer 407 yields nanoemulsions smaller than 100nm. Molecular dynamics and fluorescent tagging of the tri-block copolymer highlight their mechanistic role on the size of emulsions. This novel method enables the fabrication of highly stable albumin emulsions in the nano-size range, highly desirable for controlled drug delivery. Folic Acid (FA)-tagged protein nanoemulsions were shown to promote specific folate receptor (FR)-mediated targeting in FR positive cells. The novel strategy presented here enables the construction of size controlled, functionalized protein-based nanoemulsions with excellent characteristics for active targeting in cancer therapy.
The use of Carbon Monoxide (CO) as a therapeutic agent has already been tested in human clinical trials. Pre-clinically, CO gas administration proved beneficial in animal models of various human ...diseases. However, the use of gaseous CO faces serious obstacles not the least being its well-known toxicity. To fully realise the promise of CO as a therapeutic agent, it is key to find novel avenues for CO delivery to diseased tissues in need of treatment, without concomitant formation of elevated, toxic blood levels of carboxyhemoglobin (COHb). CO-releasing molecules (CO-RMs) have the potential to constitute safe treatments if CO release in vivo can be controlled in a spatial and temporal manner. It has already been demonstrated in animals that CO-RMs can release CO and mimic the therapeutic effects of gaseous CO. While demonstrating the principle of treatment with CO-RMs, these first generation compounds are not suitable for human use. This tutorial review summarises the biological and chemical behaviour of CO, the current status of CO-RM development, and derives principles for the creation of the next generation of CO-RMs for clinical applications in humans.
A study on laboratory scale to evaluate the environmental compatibility of electric arc furnace dust (EAFD) is reported in this article. EAFD, a waste by-product of the steel-making process, was ...generated on a steel plant located in Brazil. Different leaching tests, NBR10005 (Brazilian), AFNORX31-210 (French), JST-13 (Japanese), DIN38414-S4 (German), TCLP (American), and NEN 7343 (Netherland) were conducted. These leaching procedures are batch tests and are columns conducted in a way that an equilibrium condition should be achieved. The pH of the medium showed a crucial parameter governing the release of metals from the solid phase into solution. As the pH of the medium varies with the leachant used, this determines the dissolution of the elements. Zn, Pb, Mn, Cd, and Cu presented high leachability at NBR10005 procedures (acid pH). Except Pb and Cr, the leachability of all others metals in leaching tests with alkaline pH decreases with the increase of the pH. NBR10005 classifies the EAFD as a hazardous waste due to high concentration of Pb and Cd in leachate. The column tests are presented in the following order of leaching: Pb
>
Cr
>
Zn
>
Mn
>
Cu
>
Cd.
The aim of this study was to elucidate the differential gene expression in the RNA sequencing transcriptome of isolated perfused udders collected from 4 slaughtered Holstein × Zebu crossbred dairy ...cows experimentally inoculated with Streptococcus agalactiae. We studied 3 different statistical tools (edgeR, baySeq, and Cuffdiff 2). In summary, 2 quarters of each udder were experimentally inoculated with Strep. agalactiae and the other 2 were used as a control. Mammary tissue biopsies were collected at times 0 and 3 h after infection. The total RNA was extracted and sequenced on an Illumina HiSeq 2000 (Illumina Inc., San Diego, CA). Transcripts were assembled from the reads aligned to the bovine UMD 3.1 reference genome, and the statistical analyses were performed using the previously mentioned tools (edgeR, baySeq, and Cuffdiff 2). Finally, the identified genes were submitted to pathway enrichment analysis. A total of 1,756, 1,161, and 3,389 genes with differential gene expression were identified when using edgeR, baySeq, and Cuffdiff 2, respectively. A total of 122 genes were identified by the overlapping of the 3 methods; however, only the platelet activation presented a significantly enriched pathway. From the results, we suggest the FCER1G, GNAI2, ORAI1, and VASP genes shared among the 3 methods in this pathway for posterior biological validation.
Alternatives need to be addressed for reducing losses in elephant grass (EG; Pennisetum purpureum) silages. Furthermore, smallholders lack information on the nutritional aspects of dairy farms. The ...aim of this study was to evaluate total mixed ration silage (TMR) combining fresh EG and concentrate ingredients, creating the following treatments: (i) EG silage (control); (ii) EG, corn and soya bean meal; (iii) EG, corn, soya bean meal and molasses; (iv) EG, citrus pulp and soya bean meal; and (v) EG, citrus pulp, soya bean meal and molasses. Five replicates of each treatment were ensiled in 15‐L plastic jars. The fermentation profile, chemical composition, microbial counts and aerobic stability were assessed. Three contrasts were tested, as follows: (i) control vs. TMR; (ii) TMR with corn vs. TMR with citrus pulp; and (iii) TMR with molasses vs. TMR without molasses. Variables were analysed through the PROC MIXED procedure of the SAS at the 5% level. The TMR had better fermentation profile, lower effluent production and longer aerobic stability compared to control. When the effect of corn was compared to citrus pulp among the TMR, silages with citrus pulp showed lower fermentation losses, non‐protein nitrogen and effluent production. The aerobic stability also improved with citrus pulp. Molasses did not affect the fermentation profile. Overall, lactic acid was the primary acid in all TMR. Nitrogen source (e.g., soya bean meal) can be used without compromising the fermentation process. TMR with citrus pulp showed better results than corn. TMR may be an alternative to optimize the use of EG on smallholdings.
Pretargeted imaging has emerged as an effective multistep strategy aiming to improve imaging contrast and reduce patient radiation exposure through decoupling of the radioactivity from the targeting ...vector. The inverse electron-demand Diels–Alder (IEDDA) reaction between a trans-cyclooctene (TCO)-conjugated antibody and a labeled tetrazine holds great promise for pretargeted imaging applications due to its bioorthogonality, rapid kinetics under mild conditions, and formation of stable products. Herein, we describe the use of functionalized carbonylacrylic reagents for site-specific incorporation of TCO onto a human epidermal growth factor receptor 2 (HER2) antibody (THIOMAB) containing an engineered unpaired cysteine residue, generating homogeneous conjugates. Precise labeling of THIOMAB–TCO with a fluorescent or radiolabeled tetrazine revealed the potential of the TCO-functionalized antibody for imaging the HER2 after pretargeting in a cellular context in a HER2 positive breast cancer cell line. Control studies with MDA–MD-231 cells, which do not express HER2, further confirmed the target specificity of the modified antibody. THIOMAB–TCO was also evaluated in vivo after pretargeting and subsequent administration of an 111In-labeled tetrazine. Biodistribution studies in breast cancer tumor-bearing mice showed a significant activity accumulation on HER2+ tumors, which was 2.6-fold higher than in HER2– tumors. Additionally, biodistribution studies with THIOMAB without the TCO handle also resulted in a decreased uptake of 111In–DOTA–Tz on HER2+ tumors. Altogether, these results clearly indicate the occurrence of the click reaction at the tumor site, i.e., pretargeting of SK-BR-3 HER2-expressing cells with THIOMAB–TCO and reaction through the TCO moiety present in the antibody. The combined advantages of site-selectivity and stability of TCO tagged-antibodies could allow application of biorthogonal chemistry strategies for pretargeting imaging with minimal side-reactions and background.
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